Dissertations / Theses on the topic 'Seedlings – Diseases and pests – Control'

South Africa is the world's second largest exporter of fresh citrus and is ranked 14th in citrus production. Fungal pathogens such as Phytophthora and Pythium cause economic losses as a result of root rot and brown rot. Mycorrhizal fungi are specialized members of the fungal community forming a mutualistic relationship with plant roots. Mycorrhizal fungal structures are known to associate with other soil microorganisms and these may contribute to improved plant growth. A diverse group of bacteria that interact with the mycorrhizal fungi are known as Mycorrhizal Helper Bacteria (MHB). The aim of this study was to investigate the role of arbuscular mycorrhiza and associated bacteria isolated from spores and determine whether they had any plant growth promoting potential. A total of 19 bacteria were isolated from arbuscular mycorrhizal spores and were molecularly identified as belonging to several Bacillus, Micrococcus, Onchrobactrum and Staphylococcus sp. All bacterial isolates were tested for plant growth promotion abilities. One Bacillus isolate was able to solubilise phosphate. Four isolates Micrococcus sp, Micrococcus leteus, Ochrobacterum sp and Ochrobacterum antropi were able to produce Indole Acetic Acid and three isolates showed potential to reduce growth of Phytophthora nicotianae, P. citrocola and P. citrophthora in in vitro plate cultures. Further tests using culture supernatants of the Bacillus sp, Micrococcus sp and Bacillus cereus confirmed their ability to inhibit or reduce growth of the three Phytophthora species in a 96 well bioassay. Bacillus sp and Bacillus cereus were able to inhibit Phytophthora spp by 95 to 100 % and Micrococcus spp was able to decrease pathogen growth by 60 to 94 %. These bacterial isolates were further evaluated for plant growth promoting abilities on citrus rough lemon seedlings alone or in combination with arbuscular mycorrhizal inoculum. Bacterial and mycorrhizal inoculants influence the increase in shoot and root biomass. Bacillus cereus in combination with mycorrhizal inoculum significantly increased seedling shoot to root ratio while root biomass was significantly increased with mycorrhizal inoculation. Due to the short duration of the trial mycorrhizal colonisation could not be assessed. It is evident that selected combinations of bacteria and mycorrhizal fungi could promote citrus seedling growth and potentially improve seedling health. Further studies under nursery conditions are recommended.

Thesis (MScAgric)--University of Stellenbosch, 2005.<br>ENGLISH ABSTRACT: Grey mould, caused by Botrytis cinerea, is the most important foliar disease of rooibos seedlings. Although the disease is primarily controlled with applications of fungicides, the improvement of cultural methods of disease management should lessen this dependence on chemical control. Such improvements would, however, not be possible without knowledge of the inoculum sources and dispersal of the pathogen. The aim of this study was to investigate the inoculum ecology of B. cinerea in rooibos nurseries in order to identify primary sources of inoculum and to improve the environmentally friendly management of the disease. The study was conducted in four nurseries over two production seasons (March to July 2003 and 2004). Levels of airborne inoculum of B. cinerea were monitored on a monthly basis inside and around the nurseries with spore traps. Samples of plant material and organic debris were taken in the corresponding areas to determine the incidence of plant material infected by the pathogen and the incidences of grey mould in the nurseries were recorded. Low numbers of B. cinerea colonies were observed on the spore traps. Similar levels of airborne inoculum were observed inside and around the nurseries. The incidence of plant material yielding B. cinerea was higher outside the nurseries than inside, indicating the importance of such materials as potential sources of inoculum. Since patterns of airborne inoculum observed in this study confirmed reports of the local dispersal of B. cinerea, the removal of possible hosts outside the nurseries could aid in the management of grey mould in rooibos nurseries. Resistance to dicarboximide fungicides is a genetically stable trait in B. cinerea, and therefore has the potential to be used as a phenotypic marker. This marker can be used to gain knowledge on the dispersal of B. cinerea inoculum inside and outside rooibos nurseries. Isolates of B. cinerea collected from the air and from plant material in and around four rooibos nurseries were assessed for resistance to iprodione at 1 and 3 μg/ml a.i. Some of the isolates showed resistance to iprodione at 1 μg/ml a.i. However, none of the isolates showed resistance at 3 μg/ml a.i. iprodione. The initial incidence of dicarboximide-resistance at the nurseries was slightly higher than expected. As the season progressed, the incidence of iprodione-resistant isolates decreased towards May, after which an increase was observed towards July. A relatively high percentage of isolates collected outside the nurseries was found to be dicarboximide-resistant. Two of the nurseries had a significant higher incidence of resistant isolates on plant material collected inside, than on plant material collected outside the nursery. However, when looking at resistance levels of airborne isolates, no significant differences were found in the incidence of resistant isolates sampled inside and outside the four nurseries. The data indicated the importance of organic debris and seed-borne infections in the survival and dispersal of dicarboximide-resistant isolates of the pathogen. With the current emphasis on organic agriculture the knowledge gained in this study presents valuable possibilities of improving the cultural management of grey mould in rooibos nurseries.<br>AFRIKAANSE OPSOMMING: Vaalvrot, veroorsaak deur Botrytis cinerea, is die belangrikste bo-grondse siekte van rooibossaailinge. Alhoewel die beheer van die siekte hoofsaaklik op die gebruik van fungisiede berus, behoort die verbetering van verbouingspraktyke hierdie afhanklikheid van chemiese beheer te verminder. Sulke verbeteringe sal egter slegs moontlik wees indien voldoende kennis van die inokulumbronne en verspreiding van die patogeen beskikbaar is. Die doel van hierdie ondersoek was om die inokulum ekologie van B. cinerea in rooibos kwekerye te ondersoek sodat primêre inokulumbronne opgespoor en omgewingsvriendelike siektebestuurspraktyke verbeter kan word. Die ondersoek is in vier kwekerye oor twee produksie seisoene (Maart tot Julie 2003 en 2004) uitgevoer. Vlakke van luggedraagde inokulum van B. cinerea is op ’n maandelikse basis met behulp van spoorvangers binne en buite die kwekerye gemonitor. Monsters van plantmateriaal en organiese materiaal is in ooreenstemmende areas geneem om die voorkoms van B. cinerea geïnfekteerde plantmateriaal vas te stel en die voorkoms van vaalvrot in die kwekerye is aangeteken. Min B. cinerea kolonies is op die spoorvangers waargeneem. Soortgelyke vlakke van luggedraagde inokulum is binne en buite die kwekerye waargeneem. Die hoër voorkoms van B. cinerea geïnfekteerde plantmateriaal buite die kwekerye as binne, dui op die belang van sulke materiaal as potensiële inokulumbronne. Aangesien die patrone van luggedraagde inokulum, soos waargeneem in hierdie ondersoek, ander berigte van B. cinerea se beperkte verspreidingsvermoë bevestig, kan die verwydering van moontlike alternatiewe gashere buite die kwekerye die bestuur van die siekte binne die kwekerye verbeter. Weerstand teen dikarboksimied fungisiede is ’n geneties-stabiele kenmerk in B. cinerea en het daarom potensiaal om as ’n fenotipiese merker gebruik te word. Hierdie merker kan gebruik word om kennis aangaande die verspreiding van B. cinerea in en om rooibos kwekerye in te samel. Botrytis cinerea isolate in lug en op plantmateriaal in en om vier rooibos kwekerye is gedurende 2003 en 2004 versamel. Die isolate is vir weerstandbiedendheid teen iprodioon by konsentrasies van 1 en 3 μg/ml aktiewe bestandeel (a.b.) getoets. Isolate met weerstand teen 1 μg/ml a.b. iprodioon is waargeneem, maar nie teen 3 μg/ml nie. Die aanvanklike voorkoms van dikarboksimiedweerstand by die kwekerye was hoër as verwag. Hierdie vlak het egter gedaal met die verloop van die seisoen tot in Mei, waarna ’n toename tot in Julie waargeneem is. Die persentasie dikarboksimied-weerstandbiedende isolate buite die kwekerye was relatief hoog. In twee van die kwekerye was die voorkoms van weerstandbiedende isolate op plantmateriaal in die kwekerye betekenisvol hoër as op plantmateriaal buite die kwekerye. Daar was egter geen betekenisvolle verskille in die voorkoms van luggedraagde weerstandbiedende isolate nie, ongeag van die kwekery of posisie. Die data dui op die belang van organiese materiaal en saadgedraagde infeksies in die oorlewing en verspreiding van dikarboksimied-weerstandbiedende isolate van die patogeen. Met die huidige klem op organiese landbou bied die inligting wat in hierdie ondersoek versamel is moontlike praktyke wat geïmplementeer kan word om die beheer van vaalvrot in kwekerye met behulp van verbouingspraktyke te verbeter.

Lettuce (Lactuca sativa L.) seedlings diseases caused by soilborne pathogens are characterised by root rot, stem rot and damping-off of the seedlings that can occur at any time during growth. Fusarium solani, Pythium ultimum and Rhizoctonia solani are known to be the important destructive pathogens of lettuce, causing severe yield losses in South Africa. The aim of this research was to evaluate the effects of three selected fungicides to control these pathogens on lettuce seedlings. In this study the fungicides metalaxyl (Apron®), fludioxonil (Celest®) and mefenoxam (Subdue®) were applied at two concentrations as single and double doses on lettuce seedlings to determine their efficacy to control the pathogens Fusarium solani, Pythium ultimum and Rhizoctonia solani after significant reduction of mycelia growth was observed in vitro. Cultures of P. ultimum (UPGH024), R. solani (UPGH122) and F. solani (UPGH122) were obtained from the culture collection of the Department of Microbiology and Plant Pathology, University of Pretoria and cultivated on PDA for 2 days at 25ºC. Pasteurised soil was artificially inoculated with these pathogens. For the first experiment lettuce seeds were planted in polystyrene seedling trays at a depth of 1.0 cm. There were four replications of 50 seeds per treatment. In Experiment 2 pots (12 cm x 7 cm) were filled with pasteurised growing medium and 3-week old seedlings were transplanted. There were three replications of six pots containing three plants each. Seedling trays and pots were drenched with fungicides and placed in a randomised block design in a controlled environment room at 20- 26°C with a 12h-light/dark regime. The seedling trays and pots were rotated daily in the room. Seedling trays and pots were watered daily to maintain field capacity. The seedlings were able to grow larger in the pots than in seedling trays. It was confirmed that the treatment with fludioxonil (Celest®) at double and single dose inhibited the growth of the three fungi F. solani, P. ultimum and R. solani on lettuce seedlings without causing phytotoxicity. All three fungicides significantly reduced the diseases caused by the three pathogens. These findings are consistent with previous reports that fludioxonil, metalaxyl and mefenoxam can control oomycete fungi. There are few registered fungicides for the control of Fusarium solani, Pythium ultimum and Rhizoctonia solani on lettuce, therefore further work will aim to confirm these results in the field.<br>Dissertation (MInstAgrar)--University of Pretoria, 2008.<br>Microbiology and Plant Pathology<br>unrestricted

Dissertation (PhD (Agric))--University of Stellenbosch, 2002.<br>ENGLISH ABSTRACT: Fungi belonging to the genus Botryosphaeria are heterotrophic micromycetes that can be pathogens on woody plants. They cause serious, and in some cases devastating losses to crops through leaf necrosis, stem cankers and plant death. The Proteaceae cut-flower industry in South Africa accounts for 70% of the national cut-flower enterprise. Botryosphaeria diseases are a major impediment to production and trade of Proteaceae and there is an urgent need to investigate the etiology, epidemiology and control of these diseases. Losses of one of the most important proteas, P. magnifica, amount to 50% or more, locally. The main aims of this study were therefore to establish the etiology and aspects of epidemiology of Botryosphaeria stem cankers on P. magnifica and other Proteaceae, and to investigate methods of disease control. Although there is a vast body of information pertaining to this fungus, which was reviewed in Chapter 1, there is relatively little information available on Botryosphaeria on Proteaceae. The taxonomy of Botryosphaeria requires thorough review, and molecular techniques need to be employed to resolve species identities. In Chapter 2, it was found that Phyllachora proteae, a leaf pathogen of proteas, produced a Fusicoccum anamorph, which is described as F. proteae. A sphaeropsis-like synanamorph was associated with F. proteae and a new combination for P. proteae is proposed in Botryosphaeria, as B. proteae. The taxonomy of Botryosphaeria is in disarray at both the generic and the specific level. In Chapter 3 the taxonomic history of Botryosphaeria is reviewed, and the genus circumscribed and distinguished from other morphologically similar genera. Although several anamorph genera have been linked to Botryosphaeria, based on morphological observations and phylogenetic analysis of lTS rDNA sequence data, two anamorph genera are now recognised, those with pigmented conidia (Diplodia), and those with hyaline conidia (Fusicoccum). Botryosphaeria proteae should thus be excluded from Botryosphaeria. Several pathogenic Botryosphaeria spp. have an endophytic phase within their hosts. They are therefore imported unwittingly into other countries where they may pose a risk to agriculture and indigenous vegetation. The current global distribution of Botryosphaeria spp. associated with Proteaceae is clarified and a key to these taxa associated with Proteaceae is provided in Chapter 4. Five Botryosphaeria spp. are associated with cut-flower Proteaceae worldwide viz. B. lute a, B. obtusa, B. protearum, B. proteae and B. rib is. B. protearum is described as a new species. A thorough understanding of disease epidemiology is essential to effect a reduction of losses. In Chapter 5, I show that on P. magnifica, lesions caused by Botryosphaeria protearum, which lead to the formation of stem cankers, are initiated in the mid-rib vein or margin of leaves. Koch's postulates were satisfied and it was found that the number of lesions that developed from artificial inoculations correlated with starch levels present in leaves at the time of inoculation. In Chapter 6 it is shown that B. protearum exists as an endophyte in leaves of P. magnifica in naturally occurring as well as cultivated plants. In natural stands of proteas stem cankers are rare, but in cultivated plantations the incidence is high. Nutritional analyses indicate that higher levels of nitrogen occur in leaves of cultivated plants in spring, which could enhance disease development. High levels of sodium in the leaves of wild plants may restrict disease development. The severe economic losses caused by B. protearum make the search for improved methods of disease control essential. Fungicide applications form an important component of an integrated approach to disease management. In Chapter 7, in vitro tests demonstrate that tebuconazole, benomyl, prochloraz me, iprodione and fenarimol reduce the mycelial growth of B. protearum effectively. In the field there was a 25-85% reduction in the occurrence of stem cankers by applying fungicides or sanitation pruning. The best control was achieved by using benomyl, bitertanol, fenarimol, iprodione, prochloraz manganese chloride alternated with mancozeb and tebuconazole prophylactically. If sanitation pruning is combined with regular applications of fungicides, disease can be combated.<br>AFRIKAANSE OPSOMMING: Mikrofungi wat tot die genus Botryosphaeria behoort, is heterotrofiese organismes, wat patogenies op houtagtige plante kan wees. Hulle veroorsaak ernstige, en in sommige gevalle, verwoestende verliese, deur blaarnekrose, stamkankers en plantafsterwing. Die Proteaceae snyblom-industrie in Suid-Afrika maak 70% van die nasionale snyblomindustrie uit. Botryosphaeria siektes is 'n belangrike struikelblok in die produksie en handeldryf van Proteaceae, en daar is 'n ernstige behoefte om die etiologie, epidemiologie en beheer van siektes te ondersoek. Verliese van een van die belangrikste proteas, P. magnifica, beloop plaaslik 50% of meer. Die hoof doelstellings van hierdie studie was dus om die etiologie en epidemiologie van Botryosphaeria stamkankers op P. magnifica en ander Proteaceae vas te stel en metodes van siektebeheer te ondersoek. Hoewel daar 'n wye hoeveelheid inligting rakende die swam bestaan, wat in Hoofstuk I hersien is, is daar relatief min inligting oor Botryosphaeria op Proteaceae beskikbaar. Die taksonomie van Botryosphaeria benodig deeglike hersiening, en molekulêre tegnieke word benodig om spesie-identiteite op te klaar. In Hoofstuk 2 is gevind dat Phyllachora proteae, 'n blaarpatogeen van proteas, 'n Fusicoccum anamorf produseer, wat as F. proteae beskryf word. 'n Sphaeropsis-agtige synanamorf is met F. proteae geassosieer en 'n nuwe kombinasie vir P. proteae is as B. proteae in Botryosphaeria voorgestel. Die taksonomie van Botryosphaeria is, beide op die genus- as die spesievlak, in wanorde. In Hoofstuk 3 word die taksonomiese geskiedenis van Botryosphaeria hersien, en die genus word omskryf en van ander morfologies soortgelyke genera onderskei. Hoewel verskeie anamorf genera al met Botryosphaeria op grond van morfologiese waarnemings en filogenetiese analise van ITS rDNA volgorde data verbind is, word twee anamorf genera nou herken, dié met gepigmenteerde konidia (Diplodia), en dié met deurskynende konidia (Fusicoccum). Botryosphaeria proteae moet dus van Botryosphaeria uitgesluit word. Verskeie patogeniese Botryosphaeria spp. het 'n endofitiese fase in hul lewenssiklus. Hulle word dus onwetend in ander lande ingevoer waar hulle 'n gevaar vir landbou en inheemse plantegroei kan inhou. Die huidige wêreldverspreiding van Botryosphaeria spp. wat met Proteaceae geassosieer word is opgeklaar, en in Hoofstuk 4 word 'n sleutel tot die taksa wat met Proteaceae geassosieer word verskaf. Vyf Botryosphaeria spp. word met snyblom Proteaceae wêreldwyd geassosieer, naamlik B. lutea, B. protearum, B. proteae, B. ribis en B. obtusa. B. protearum word as 'n nuwe spesie beskryf. 'n Deeglike kennis van siekte-epidemiologie is noodsaaklik ten einde verliese te verminder. In Hoofstuk 5 dui ek aan dat letsels wat lei tot stamkankers, veroorsaak deur Botryosphaeria protearum op P. magnifica, in die hoofnerf of rant van blare ontstaan. Koch se postulate is uitgevoer en daar is vasgestel dat die aantal letsels wat vanuit kunsmatige inokulasies ontwikkel het korreleer met die styselvlakke teenwoordig in die blare ten tye van die inokulasie. In Hoofstuk 6 word getoon dat B. protearum as 'n endofiet in die blare van P. magnifica. In natuurlike standplase van proteas is stamkankers skaars, maar in verboude plantasies is die voorkoms hoog. Voedingsanalises dui aan dat hoër vlakke van stikstof in die blare van verboude plante in die lente voorkom, wat siekte-ontwikkeling moontlik kan bevorder. Hoë vlakke van natrium in die blare van natuurlike plante mag siekteontwikkeling beperk. Die ernstige ekonomiese verliese wat deur B. protearum veroorsaak word, maak die soektog na verbeterde metodes van siektebeheer noodsaaklik. Fungisiedtoedienings maak 'n belangrike deel uit van 'n geïntegreerde benadering tot siektebeheer. In Hoofstuk 7 dui in vitro toetse aan dat tebuconazole, benomyl, prochloraz me, iprodione en fenarimol die miseliumgroei van B. protearum effektief verminder. 'n Vermindering van 25-85% is aangetoon in die voorkoms van stamkankers in die veld, deur die toediening van fungisiedes en sanitasiesnoei. Die beste beheer is verkry deur die voorkomende toediening van benomyl, bitertanol, fenarimol, iprodione en prochloraz manganese chloride, afgewissel met mancozeb en tebuconazole, op plante in die veld. Indien sanitasiesnoei met gereelde toedienings van fungisiedes gekombineer word, kan die siekte bekamp word.

Thesis (PhD)--University of Stellenbosch, 2000.<br>ENGLISH ABSTRACT: The population density of the white wax scale, Ceroplastes destructor Newstead, has increased since 1994 in certain areas of Western and parts of Eastern Cape Provinces of South Africa where citrus is grown, particularly on Citrus reticulata (Blanco). A study was conducted to investigate its morphology, biology and ecology as contributions to the development of a sound integrated management programme. Characteristics of the immature stages and adult females were described and illustrated from field-collected and slide-mounted specimens. A key to the different stages and morphometeric characteristics useful for separating them are provided. No significant differences in female fecundity were found between orchards (P > 0.05). However, fecundity varied significantly between female size classes from the same orchard (P < 0.001). Female body-size also differed significantly between orchards (P < 0.05) and was significantly positively correlated with fecundity (P < 0.01). C. destructor has one discrete generation per year in South Africa. Oviposition commenced in November and continued through to the end of December with a few females ovipositing until mid January. Population density of the second instar peaked in February while the third instar extended from March to the end of July, followed by a peak population of adults in August. Seven primary and three secondary parasitoids, as well as four predator species attacking C. destructor were identified. Aprostocetus (= Tetrastichus) ceroplastae (Girault) was the dominant species, accounting for 78.87% of the total primary parasitoids reared. Peak numbers of parasitoids and predators were synchronized with peak emergence of susceptible scale stages, indicating that the host-parasitoid/predator system contained a density-dependent regulatory mechanism. Key mortality factors varied slightly between two of the orchards. Key stage mortality determined from a cohort life table was generally in the third instar (LIlI) and preovipositional female (POF) stage. Significant density-dependent mortality factors were demonstrated for the first instar (LI) and PDF stage. Dispersal of C. destructor is by first instar nymphs and the numbers caught on a series of yellow sticky traps varied significantly between crawler densities at the source, trap distances and trap directions from the source (P < 0.001). The numbers caught were positively correlated to the initial crawler density at the source (P < 0.01), suggesting that dispersal was density dependent. Trap distance and the numbers caught were inversely correlated (P < 0.01). Evaluation of effects of different densities of C. destructor on growth, survivorship and reproduction of scales as well as on leaf bearing ability of trees and area of leaf surface covered with sooty mould fungus was carried out on naturally infested Syzygium (= Eugenia) malaccensis (L.) plants. Scale body size and fecundity were inversely related to scale density (P < 0.01), suggesting density-dependent intraspecific competition. Scale survivorship generally declined with increasing density whereas scale parasitism and predation were positively correlated with density (P < 0.05). At high scale densities production of new leaves was significantly reduced (P < 0.01), reducing the resource base for subsequent generations of scale. Scale density and leaf area covered with sooty mould fungus were significantly positively correlated (P < 0.05). The toxicity of four synthetic insecticides against the three immature stages of C. destructor and of eight insecticides against the parasitoid A. ceroplastae was evaluated. Development of the first and second instars of C. destructor was completely arrested by the chemicals. Female fecundity, fertility and body sizes of survivors of treatments applied at the LIII stage were not significantly affected by any of the chemicals (P > 0.05). All the chemicals exhibited high toxicity to A. ceroplastae and hence are not recommended for integrated management of C. destructor in citrus orchards where A. ceroplastae plays an important role.<br>AFRIKAANSE OPSOMMING: Die populasiedigtheid van die witwasdopluis, Ceroplastes destructor Newstead, het sedert 1994 toegeneem in sekere gebiede van die Weskaap en Ooskaap provinsies van Suid-Afrika waar sitrus verbou word, veralop Citrus reticulata (Blanco). 'n Studie van hierdie insek se morfologie, biologie en ekologie is onderneem as bydrae tot die ontwikkeling van 'n geïntegreerde bestuursprogram. Die karaktertrekke van die onvolwasse stadia en die volwasse wyfies is beskryf en geïllustreer vanaf eksemplare wat in die veld versamel is en op g1asplaatjies gemonteer is. 'n Sleutel vir die verskillende stadia en morfometriese kenmerke wat nuttig is om hulle te onderskei, word voorsien. Geen beduidende verskille in die vrugbaarheid van wyfies van verskillende boorde is gevind nie (P < 0.05). Vrugbaarheid het egter betekenisvol verskil by die verskillende grootteklasse van wyfies uit dieselfde boord (P < 0.001). Die liggaamsgrootte van wyfies uit verskillende boorde het betekenisvol verskil (P < 0.05) en was betekenisvol positief gekorreleer met vrugbaarheid (P < 0.01). C. destructor het een generasie per jaar in Suid-Afrika. Eierlegging het in November begin en aangehou tot aan die einde van Desember, met enkele wyfies wat nog tot in middel Januarie eiers gelê het. Die populasiedigtheid van die tweede instar het 'n hoogtepunt in Februarie bereik, terwyl die derde instar van Maart tot aan die einde van Julie geduur het, gevolg deur 'n piekbevolking van volwassenes in Augustus. Sewe primêre en drie sekondêre parasitoïde asook vier predator spesies wat C destructor aanval, is geïdentifiseer. Aprostocetus (=Tetrastichus) ceroplastae (Girault) was die dominante spesies wat 78.87% van die totale aantal primêre parasitoïde wat uitgeteel is, uitgemaak het. Die pieke in die getalle van parasitoïde en predatore was gesinchroniseer met pieke in die verskyning van die gevoelige stadia, wat dui op die aanwesigheid van 'n digtheidsafhanklike regulatoriese meganisme. Die sleutel mortaliteitsfaktore het effens gevarieer tussen twee van die boorde. Die sleutelstadium van mortaliteit, soos bepaal m.b.v. 'n kohort lewenstabel, was gewoonlik die derde instar (LIlI) en die preoviposisionele wyfie (POW). Betekenisvolle digtheidsafhanklike mortaliteitsfaktore IS aangetoon vir die eerste instar (LI) en die POW. Die verspreiding van C.destructor vind plaas deur die eerste instar nimfe en die getalle wat op 'n reeks van taai geel valle gevang is, het betekenisvol gewissel volgens kruiperdigthede by die bron, asook die afstand en rigting van die valle vanaf die bron (P < 0.001). Die getalle wat gevang is, was positief gekorreleer met die aanvanklike kruiperdigtheid by die bron (P < 0.01), wat daarop dui dat verspreiding digtheidsafhanklik was. Die afstand van die valle en die aantal wat gevang is, was omgekeerd gekorreleer (P < 0.01). 'n Evaluering van die invloed van verskillende digthede van C. destructor op die groei, oorlewing en reproduksie van dopluise, asook die vermoë van bome om blare te dra en die area van die blaaroppervlak wat met roetskimmel besmet is, is uitgevoer op plante van Syzygium (= Eugenia) malaccensis (L.) met 'n natuurlike besmetting. Die liggaamsgrootte en vrugbaarheid van die dopluise was omgekeerd gekorreleer met hulle digtheid (P < 0.01), wat dui op digtheidsafhanklike intraspesifieke kompetisie. Die oorlewing van die dopluise het oor die algemeen afgeneem met toenemende digtheid, terwyl parasitisme en predasie positief gekorreleer was met digtheid (P < 0.05). By hoë dopluisdigthede het die produksie van nuwe blare betekenisvol afgeneem (P < 0.01), wat die hulpbronbasis vir daaropvolgende generasies van dopluise verswak. Die dopluisdigtheid en blaaroppervlak wat met roetskimmel bedek was, was positief gekorreleer (P < 0.05). Die toksisiteit van vier sintetiese insektemiddels teenoor die drie onvolwasse stadia van C. destructor en van agt insektemiddels teenoor die parasitoïd A. ceroplastae is geëvalueer. Die ontwikkeling van die eerste en tweede instars van C. destructor is heeltemal stopgesit deur die middels. Die fekunditeit, fertiliteit en liggaamsgrootte van wyfies wat toedienings op die LIIl stadium oorleef het, is nie betekenisvol ge-affekteer deur enige van die middels nie (P < 0.05). Al die middels was baie toksies teenoor A. ceroplastae en word dus nie aanbeveel vir die geïntegreerde bestuur van C. destructor waar A. ceroplastae 'n belangrike rol speel nie.

Field experiments were conducted in two different growing environments to evaluate the spread and movement of Dickeya dadantii. A procedure to inoculate seed potatoes with Dickeya dadantii was developed to use during this study. Spread of Dickeya dadantii from inoculated potato seed to healthy potato seed during the handling, cutting and planting procedures was not detected at either location. Spread of Dickeya dadantii from inoculated seed to surrounding progeny tubers in the field was documented in both locations. In Florida, 33% of progeny tubers tested positive for Dickeya using PCR, and in North Dakota, 13% of the progeny tubers tested positive. Stunting was observed in plants grown from Dickeya dadantii inoculated seed tubers in North Dakota, but not in Florida. These results indicate that Dickeya dadantii may spread during the seed handling and cutting processes and can spread in the field from infected seed tubers to progeny tubers.

Thesis (MScAgric)--Stellenbosch University, 2012.<br>ENGLISH ABSTRACT: Leaf blackening, a postharvest disorder which is characterized by a dark brown to black discoloration, is found in most commercially important Protea cut flower species and cultivars. As this disorder is known to increase with storage time, it is a major concern to the South African industry as the use of sea freight is increasingly preferred due to lower transport costs and a more favourable carbon footprint. The cause of leaf blackening has been strongly linked to a carbohydrate stress exerted by the large inflorescence, thus requiring the utilization of sugar bound polyphenols in the foliage, which when removed, can oxidize enzymatically or non-enzymatically. A study where harvesting was done throughout the season as well as on selected days at 08:00, 12:00, 15:00 and 17:00, concluded that leaf blackening incidences in Protea cv. Sylvia stems varies significantly throughout the season, between years and even with the harvest time of day. Leaf blackening incidences increased from October onwards and remained high until February, before decreasing to acceptably lower levels towards March to May. Carbohydrate- and phenolic content together with water status of leaves at harvest was not able to accurately predict incidence of the associated leaf blackening. However, irrespective of the season of harvesting, leaf blackening was significantly lower when stems were harvested later in the day than compared to stems harvested in the morning. Low sucrose and high water content at these harvest times was positively correlated to high incidences of leaf blackening. In a next study where uptake dynamics of glucose pulsing was investigated, Protea cv. Sylvia was harvested at different times throughout the day, dehydrated to various levels and pulsed with an increasing range of glucose concentrations. Pulsing solution uptake per stem was found to be highly influenced by these factors, as dehydration of stems and a harvest time later during the day both decreased stem water potential, which then increased pulse-solution uptake within a certain time period. The daily harvest time influenced transpiration, whilst pulse-solution uptake decreased with an increase in glucose pulse concentration. When stems were pulsed pre-storage with an increasing range of glucose concentrations, not only did pulses of between 4.7 – 13.7% glucose significantly delayed the incidence of leaf blackening, but it also maintained a positive water balance longer in stems during vase life. Ethanol or acetaldehyde vapour did not provide a viable alternative for reducing leaf blackening incidence in Protea cv. Sylvia, although a synergistic effect was found when ethanol vapour or pulsing was used in combination with glucose. A commercial verification trial disclosed that Protea magnifica and Protea ‘Pink Ice’ reacted more beneficial to ethanol vapour than was observed in ‘Sylvia’. This study confirms that carbohydrate availability within the Protea cut stem remains a key factor in the control of leaf blackening. Factors which assist in maintaining high internal carbohydrate levels, such as enhanced glucose pulse uptake or effective vase solution utilization will contribute to providing an optimum control of leaf blackening during vase life following long-term cold storage.<br>AFRIKAANSE OPSOMMING: Loofblaarverbruining is ‘n na-oes defek wat gekarakteriseer word deur ‘n donker bruin na swart verkleuring wat voorkom in meeste kommersieël belangrike Protea snyblom spesies en kultivars. Hierdie defek is bekend daarvoor dat dit toeneem met stoortyd, dus is dit ‘n groot kommer vir die Suid-Afrikaanse industrie, met toenemende gebruik van seevrag as vervoer keuse wat laer vervoer kostes en meer gunstige ‘koolstof voetspoor’ bevoordeel. Die oorsaak van loofblaarverbruining word sterk gekoppel aan ‘n koolhidraat stres wat uitgeoefen word deur die groot bloeiwyse op die loofblare, waar suiker-gebonde polifenoliese verbindings ensiematies of nieensiematies geoksideer word met die verwydering van die suiker verbinding. 'n Studie waar geoes was regdeur die seisoen, sowel as op geselekteerde dae om 08:00, 12:00, 15:00 en 17:00, het bevind dat die voorkoms van loofblaarverbruining in stele van Protea kv. Sylvia aansienlik geskil regdeur die seisoen, tussen jare en selfs met die oes tyd gedurende die dag. Die voorkoms van loofblaarverbruining het toegeneem vanaf Oktober en het hoog gebly tot en met Februarie, voordat dit gedaal het tot aanvaarbare laer vlakke teen Maart, tot en met Mei. Koolhidraat-en fenoliese inhoud sowel as die water status van die blare by oes was onsuksesvol om die voorkoms van die gepaardgaande loofblaarverbruining akkuraat te voorspel. Loofblaarverbruining was egter aansienlik laer as stele geoes later in die dag teenoor stele geoes in die oggend, ongeag die seisoen van oes. Lae sukrose en 'n hoë water inhoud geassosieer met hierdie oes-tye was positief gekorreleerd met ‘n hoë voorkoms van loofblaarverbruining. In 'n volgende studie waar die opname dinamika van glukose pulsing ondersoek was, is Protea kv. Sylvia stele geoes op verskillende tye dwarsdeur die dag, gedehidreer tot verskillende vlakke en met 'n toenemende reeks van glukose konsentrasies gepuls. Pulsoplossing opname per steel is sterk beïnvloed deur hierdie faktore, aangesien dehidrasie van die stele asook stele geoes later gedurende die dag die afname van steel waterpotensiaal veroorsaak het, terwyl die puls-oplossing opname versnel het binne ‘n bepaalde tyd. Die tyd van oes beïnvloed ook transpirasie, terwyl vaas oplossing opname afgeneem met 'n toename in glukose puls konsentrasie. Wanneer ‘Sylvia’ stele gepuls was voor stoor met 'n reeks van toenemende glukose konsentrasies, het nie net die puls van tussen 4.7 – 13.7% glukose aansienlik die voorkoms van loofblaarverbruining vertraag nie, maar dit het ook ‘n positiewe water balans langer in stele gedurende die vaas lewe behou. Nie etanol of asetaldehied dampe is bevind as geskikte alternatief vir glukose pulsing om die voorkoms van loofblaarverbruining in Protea kv. Sylvia te verlaag nie, alhoewel ‘n sinergistiese effek waargeneem was wanneer etanol in kombinasie met glukose gebruik was. ‘n Kommersieële bevestigingstoetsing het bevind dat Protea magnifica en ‘Pink Ice’ meer voordeel uit ‘n ethanoldamp behandeling kon trek teenoor ‘Sylvia’. Hierdie studie het bevestig die belangrikheid van koolhidraat beskikbaarheid in die Protea snyblom, vir beheer van loofblaarverbruining. Faktore wat die handhawing van hoë interne koolhidrate vlakke, soos bevorderde glukose puls opname of effektiewe vaas oplossing benutting sal bydra tot ‘n optimal beheer van loofblaarverbruining tydens vaas lewe na langtermyn koueopberging.<br>National Research Fund (NRF) for their financial support in 2009; Protea Producers of South Africa (PPSA) and Productschap Tuinbouw (PT) as well as the Frank Batchelor Will Trust Grant for the financial support.

Thesis (MScAgric)--University of Stellenbosch, 2000.<br>ENGLISH ABSTRACT: Petri grapevme decline, also known as black goo, slow die-back and Phaeoacremonium grapevine decline, causes significant losses of young vines worldwide. Species of Phaeoacremonium, Phaeomoniella chlamydospora and related genera are associated with this grapevine disease. This study investigates the Phaeoacremonium-complex and Phaeomoniella chlamydospora, focussing on the species isolated from grapevines. Fungicide sensitivity of Pa. chlamydospora and the possibility of employing molecular techniques for the detection of Pa. chlamydospora in grapevines were also investigated. In an overview of the literature on Petri grapevine decline the disease history and the relatedness of Petri grapevine decline to esca is discussed. Petri grapvine decline occurs in propagation material or young vines. Infected material can appear asymptomatic and therefore the possibilities of molecular techniques for identification were also investigated in the literature. In South Africa Pa. chlamydospora is the dominant organism causing Petri grapevine decline and therefore different fungicides were evaluated to control this fungus. Six isolates of Pa. chlamydospora, from Stellenbosch, Wellington, Somerset West and Malmesbury of Western Cape province, South Africa, were screened against twelve fungicides testing their effect on mycelial inhibition in vitro. These fungicides included benomyl, chlorothalonil, fenarimol, fosetyl-Al, iprodione, kresoxim-methyl, mancozeb, metalaxyl, prochloraz manganese chloride, quintozene, tebuconazole and thiram. Results provided the base-line sensitivity of South African isolates of Pa. chlamydospora. Benomyl, fenarimol, kresoxim-methyl, prochloraz manganese chloride and tebuconazole were the most effective (with EC50 values ranging from 0.01 to 0.05 ug/ml) for inhibiting mycelial growth of Pa. chlamydospora in vitro. This in vitro test gave a good indication of which fungicides could be selected for further studies in glasshouses and nurseries. The molecular phylogeny of Phaeoacremonium and Phaeomoniella isolates from grapevines of South Africa, or isolates obtained from the Centraalbureau voor Schimmelcultures (CBS) in the Netherland, were investigated. Sequence data were created from the rONA region and partial B-tubulin gene of 33 of these isolates using the PCR technique. This sequence data were analysed with PAUP* version 4.Ob2a. An analysis of the sequence data confirmed the genus Phaeomoniella to be distinct from Phaeoacremonium (Pm.) based on DNA phylogeny. Although morphologically similar, the species status of Pm. aleophi/um and Pm. angustius was confirmed with DNA phylogeny and cultural characteristics. Pm. aleophilum has an optimum growth rate at 30°C and the ability to grow at 35°C, where as Pm. angustius has an optimum growth rate at 25°C and cannot grow at 35°C_ Pm. viticola was shown to be synonymous with Pm. angustius, and a new species, Pm. mortoniae, was newly described from grapevine occurring in California. Futhermore, Pm. aleophilum was newly reported from South Africa and grapevine isolates thought to be Pm. inflatipes were all re-identified as Pm. aleophilum. These findings therefore also shed some doubt on the possible role of Pm. inflatipes in Petri grapevine decline. It was confirmed that Pa. chlamydospora, Pm. aleophilum and Pm. angustius are the species involved in Petri grapevine decline. Pm. mortoniae was isolated from grapevines, but its pathogenicity should still be confirmed and the role of Pm. injlatipes in Petri grapevine decline remains unclear. Pa. chlamydospora has been routinely isolated from symptomless propagation and nursery material. Because the disease can take years to develop, it is crucial that healthy propagation material is used at planting. Pa. chlamydospora is a slowgrowing fungus, and positive identification from symptomless grapevine tissue can take up to 4 wks. The possibility of employing molecular techniques for the detection of Pa. chlamydospora in apparently healthy grapevines was investigated. Speciesspecific primers (PCLI and PCL2) based on the regions ITSI and ITS2 were designed for Pa. chlamydospora. These primers were highly sensitive and amplification was achieved from genomic DNA of Pa. chlamydospora from as low as 16 pg. Phaeoacremonium spp., related genera and common fungal taxa from grapevines were tested with these primers, but positive amplification was achieved for Pa. chlamydospora only. The presence of Pa. chlamydospora in symptomless grapevine tissue culture plants was confirmed by PCR within 24 hours. These primers therefore allow rapid and accurate identification of Pa. c~lamydospora. Testing on a larger scale with nursery material should be conducted to determine the feasibility of using these species-specific primers in the grapevine industry.<br>AFRIKAANSE OPSOMMING: Petri-terugsterwing van jong wingerde, ook algemeen bekend as "black goo" en Phaeoacremonium-terugsterwing, veroorsaak wêreldwyd groot geldelike verliese in die wingerdbedryf. Spesies van Phaeoacremonium, Phaeomoniella chlamydospora en verwante genera word met hierdie wingerdsiekte geassosieer. In die tesis word In oorsig gegee van die geskiedenis van hierdie siekte, die verwantskap tussen Petriterugsterwing en esca, en moontlike maniere van siektebestuur. Swamme wat by die siektekompleks betrokke is, kan in simptoomlose plantweefsel voorkom en daarom is die moontlikhede van die gebruik van molekulêre tegnieke vir swamidentifikasie in oënskou geneem. In Suid-Afrika is Pa. chlamydospora die dominante swam wat met Petriterugsterwing geassosieerword, gevolglik is verskillende fungisiedes vir die chemiese beheer van Pa. chlamydospora geëvalueer. Ses isolate van Pa. chlamydospora, versamel vanaf verskillende areas in die Wes-Kaap provinsie, is in dié studie gebruik. Benomyl, chlorothalonil, fenarimol, fosetyl-Al, iprodione, kresoxim-methyl, mancozeb, metalaxyl, prochloraz manganese chloride, quintozene, tebuconazole en thiram se effek op miselium inhibisie van Pa. chlamydospora is in vitro geëvalueer. Benomyl, fenarimol, kresoxim-methyl, prochloraz manganese chloride en tebuconazole was die mees effektiewe middels. Die effektiewe konsentrasie waarby 50% van die miselium groei geïnhibeer is (EKso),was tussen 0.01 en 0.05 ug/ml vir die mees effektiewe groep middels. Benomyl, fenarimol, kresoxim-methyl, prochloraz manganese chloride en tebuconazole het in vitro goeie potensiaal getoon, en verder toetse moet in vivo uitgevoer word. 'n Molekulêre studie is van Phaeoacremonium en Phaeomoniella isolate; verkry uit Suid-Afrikaanse wingerde, of vanaf die "Centraalbureau voor Schimmelcultures" (CBS) van Nederland; gedoen. Deur van die PKR tegniek gebruik te maak, is die basispaaropeenvolgingsdata van 33 isolate, van die ITSl, 5.8S, ITS2 rDNA area en die gedeeltelike B-tubullen geen verkry. Gekombineerde molekulêre data het die teorie ondersteun dat Phaeomoniella (Herpotrichiellaceae) gedistansieerd is van Phaeoacremonium (Magnaporthaceae). Pm. aleophilum en Pm. angustius was morfologies moeilik onderskeibaar, maar kon op grond van molekulêre data en kulturele eienskappe onderskei word. Pm. aleophilum se optimum groeitemperatuur was by 30°C en die swam besit die vermoë om by 35°C te groei. Pm. angus/ius se optimum groeitemperatuur was by 25°C, maar het nie by 35°C gegroei nie. 'n Studie van molekulêre en kulturele eienskappe het getoon dat Pm. angus/ius en Pm. viticola sinoniem is. 'n Nuwe spesie, Pm. mortoniae, wat uit wingerde van Kalifornie geïsoleer is, is beskrywe. Verder is Pm. aleophilum die eerste keer in Suid-Afrikaanse wingerde aangetref en Pm. tnflatipes isolate, wat vanuit wingerde geïsoleer is, is almal met molekulêre data gewys om Pm. aleophilum te wees. Hierdie bevindinge trek die rol van Pm. inflatipes in Petri-terugsterwing van wingerde in twyfel. Phaeomoniella chlamydospora IS m voortplantingsmateriaal en kwekerystokkies opgespoor. Omdat dit jare kan duur voordat siektesimptome ontwikkel, is dit belangrik om vroegtydig te weet of jong stokkies met Pa. chlamydospora geïnfekteer is. Pa. chlamydospora groei baie stadig en positiewe identifikasie van simptoomlose infeksies duur tot vier weke. Die toepassing van molekulêre tegnieke vir die vinnige identifikasie van Pa. chlamydospora in wingerde is dus ondersoek. Spesie-spesifieke oligonukleotiedes (PCU en PCL2) is vir Pa. chlamydospora ontwerp. Hierdie oligonukleotiedes is uiters sensitief en genomiese DNA van Pa. chlamydospora is van so laag as 16 pg geamplifiseer. Phaeoacremonium spp., verwante genera en algemene swamme vanuit wingerdmateriaal is met die oligonukleotiedes getoets, maar positiewe amplifikasie was slegs met Pa. chlamydospora moontlik. Die teenwoordigheid van Pa. chlamydospora is binne 24 uur in asimptomatiese wingerd weefselkultuurplantjies bevestig. Hierdie oligonukleotiedes identifiseer Pa. chlamydospora vinnig en akkuraat en toetsing op 'n groter skaal moet vervolgens met kwekerymateriaal onderneem word.

Thesis (MSc)--Stellenbosch University, 1992.<br>One copy microfiche.<br>ENGLISH ABSTRACT: The effects of synthetic pyrethroids on the spider mite, Tetranychus urticae Koch, and its natural enemies was investigated. The pyrethroid cyfluthrin, sprayed for the control of fruit weevil Phlyctinus callosus, did not cause mite outbreaks and was not harmful to the predators when sprayed before they appeared on the apple trees. This spray also gave good control of thrips, bollworm (Heliothis armigera) and fruit weevil (P. callosus). A late season spray of deltamethrin against codling moth, caused an increase in the phytophagous mite population. This was assosiated with a temporary reduction in the population level of the predatory beetle, Oligota fageli. The plants found on the orchard floor were examined. The plant diversity was low and plants that acted as hosts of phytophagous mites during the winter were scarce. However, Solanum nigum carried high mite populations and infected the adjacent apple trees. The plants were too few to have an effect on the whole orchard. Eight synthetic pyrethroids were tested for repellency in a leafdisk dip and spray bioassay using T. urticae. Deltamethrin and fenvalerate caused no walk-off. Cyfluthrin and fluvalinate caused moderate walk-off and bifenthrin, cyhalothrin and lambda-cyhalothrin caused high walk-off.<br>AFRIKAANSE OPSOMMING: Die invloed van sintetiese piretroIedes op die rooispinmyt, Tetranychus urticae Koch en hul natuurlike vyande is ondersoek. Die piretroIed, cyfluthrin, gespuit vir die beheer van snuitkewer, P4ctinus callosus, het the mytuitbrake veroorsaak the en was ook the skadelik vir predatore nadat dit vroeg in die seisoen, voordat myte in die appelbome verskyn het, gespuit is the. Die bespuiting het ook goeie beheer van blaaspootjies, bolwurm (Heliothis amdgera) en P. callosus gegee. Deltametrien wat later in die seisoen toegedien is vir die beheer van kodlingmot, Cydia pomonella, het 'n mytuitbraak veroorsaak. Dit word geassossieer met die tydelike onderdrukking van die populasievlak van die predatoriese kewer, Oligota fageli. Die plante in die dekgewas is ondersoek. Die plant-verskeidenheid was laag en plante wat kon dien as oorwinterings gashere vir fitofage myte kon dien was skaars. Solanum nigrum het egter groot bevolkings myte gedra wat die appelbome naaste aan hulle eerste besmet het. Daar was egter te min van hierdie plante om 'n invloed op die hele boord te he. Agt piretroIedes is vir afwering van T. urticae in 'n blaarskyf doop en spuit bioessal getoets. Deltamethrin en fenvalerate het geen afwering getoon the. Cyfluthrin en fluvalinate het 'n redelike afwering, terwyl bifenthrin, cyhalothrin en lamda-cyhalothrin het 'n hod mate van afwering getoon

Thesis (MSc)--Stellenbosch University, 2015.<br>ENGLISH ABSTRACT: The codling moth, Cydia pomonella (Lepidoptera: Tortricidae), is the key pest of apples and pears worldwide. The withdrawal of certain fundamental chemicals from codling moth management spray programmes, due to concerns about human, environmental and ecosystem health, has resulted in the search for softer, more environmentally friendly, and safer control measures. Entomopathogenic nematodes (EPNs), naturally occur in the soil, and actively search for hosts. The interest in using EPNs from the families Heterorhabditidae and Steinernematidae as a control measure was sparked in 1953, when an EPN was discovered in an insect. The aim is to incorporate EPNs in an integrated pest management (IPM) programme, to ensure minimal residue and eventually residue-free pome fruit production in South Africa. In order to ensure EPN success, both the environmental and technical factors influencing their efficacy, were investigated in this study. The biocontrol potential of three imported EPN isolates, being Steinernema feltiae and two isolates of Heterorhabditis bacteriophora (Hb1, Hb2), as well as a local isolate, Steinernema yirgalemense, were evaluated for the control of the codling moth under local conditions. All concentrations of S. yirgalemense, applied by immersion in a suspension of nematodes, gave > 98% control. The two formulated isolates of H. bacteriophora, Hb1-f and Hb2-f, gave < 30% control. When using the same nematode isolates, produced in vivo, S. yirgalemense still resulted in a higher codling moth control of > 90%, compared to 54% and 31% control of the H. bacteriophora Hb1 and Hb2 isolates, respectively. In follow up field trials, S. feltiae resulted in ≥ 80% control, and was thus more effective than both S. yirgalemense and the H. bacteriophora (Hb1) isolates, with 66% and 24%, and 24% and 9% control, for two separate trials, respectively. To validate the data obtained from the field trials, subsequent laboratory bioassays were conducted evaluating temperature regimes, following the same cycle as under natural conditions, with a constant humidity of 100%. Steinernema feltiae proved to be most effective, causing > 90% mortality, followed by S. yirgalemense, with 78% mortality. The two H. bacteriophora isolates (Hb1, Hb2) under the above-mentioned laboratory conditions, resulted in 73% and 59% control, respectively. Humidity thus seems to be the most important factor affecting EPN efficacy during above-ground applications. Steinernema feltiae proved to be a better candidate than S. yirgalemense for the control of the codling moth. The efficacy of different EPN isolates in controlling diapausing codling moth larvae at different temperatures was also evaluated, under local conditions, using spray application. Steinernema feltiae and two isolates of H. bacteriophora Hb1 and Hb2, including two local isolates, S. yirgalemense and Steinernema jeffreyense, were evaluated. The use of S. jeffreyense resulted in the most effective control, with 67% mortality, followed by H. bacteriophora (Hb1) with 42%, and then by S. yirgalemense with 41%. Laboratory bioassays simulating field conditions revealed that S. feltiae was most virulent to codling moth larvae, with 67% mortality by infection, followed by S. yirgalemense with 58%, the H. bacteriophora strain Hb1 with 48%, and the Hb2 strain with 24%. A comparison of the infection and penetration rate of two isolates of H. bacteriophora (Hb1, Hb2), S. feltiae and S. yirgalemense, which was carried out in multiwell plates at 14°C and 25°C, respectively, confirms the dramatic effect of temperature on EPN efficacy. At 14°C, all treatments with EPN species resulted in slower codling moth mortality than they did at 25°C, as after 48 h, < 15% mortality was recorded for all species, whereas at the warmer temperature, > 98% mortality was recorded for all species. After the exposure of washed, cool-treated larvae to 25°C for 24 h, the application of both S. feltiae and S. yirgalemense resulted in 100% mortality, whereas the application of the two H. bacteriophora isolates, Hb1 and Hb2, resulted in 68% and 54% control, respectively, over the same time period. At 14°C, S. feltiae had the highest average penetration rate of 20 IJs/insect, followed by S. yirgalemense with 14 IJs/insect, whereas S. yirgalemense had the highest penetration rate at 25°C, with 39 IJs/insect, followed by S. feltiae, with 9 IJs/insect. The two H. bacteriophora isolates had higher average penetration rates at the higher temperature. This study has highlighted the biocontrol potential of S. jeffreyense, as well as showing that S. feltiae is a cold-active nematode, whereas the other three EPN isolates prefer warmer temperatures. Stacked wooden fruit bins are regarded as preferred overwintering sites for codling moth diapausing larvae. Control strategies against the codling moth in South Africa have been hampered by the reinfestation of orchards by nearby stacked infested fruit bins or by the movement of bins between orchards. Worldwide, wooden fruit bins are systematically being replaced with plastic bins, which, in South Africa, will only be phased out over a few years. The objective of this study was to evaluate the potential of H. bacteriophora, S. feltiae, and S. yirgalemense, to disinfest miniature wooden fruit bins under controlled conditions in the laboratory. After dipping minibins in a suspension of 25 IJs/ml of all three EPN species, under optimum conditions of temperature and humidity, the highest percentage of control was obtained using S. feltiae (75%) followed by S. yirgalemense (57%), and then by H. bacteriophora (Hb1) (27%). The addition of adjuvants significantly increased (p < 0.001) S. feltiae infectivity to > 95%, whereas it did not result in a significant increase in H. bacteriophora or S. yirgalemense infectivity. The results indicated that H. bacteriophora would not be a suitable candidate to use for the control of the codling moth larvae in wooden fruit bins. The current preferred candidate for control would be S. feltiae, whose efficacy could be increased by means of the addition of an adjuvant. During winter, when the whole codling moth population are larvae and in diapause, no control measures are applied in orchards. This study has shown that EPNs can be sprayed in orchards to lower the codling moth cohort emerging after winter, as well as be included in an IPM programme. EPNs can act as a second line of defence, through supplementary control, and ensure effective control of the codling moth larvae which survived chemical spray applications, to safeguard against resistant codling moth populations in the next season.<br>AFRIKAANSE OPSOMMING: Kodlingmot, Cydia pomonella (Lepidoptera: Tortricidae), is ‘n belangrike plaag van appels en pere wêreldwyd. Die onttrekking van sekere fundamentele chemikalieë vanuit die kodlingmot beheerprogram weens die kommer oor menslike, omgewings en ekosisteemgesondheid, het gelei tot die soektog na sagter, meer omgewingsvriendelike en veiliger beheermaatreëls. Entomopatogeniese nematodes (EPNs) kom natuurlik in die grond voor en soek aktief na gashere. Die belangstelling in die gebruik van EPNs van die families Heterorhabditidae en Steinernematidae as 'n beheermaatreël is te danke aan die ontdekking van 'n EPN in ‘n insek in 1953. Die doel is om EPNs in 'n geïntegreerde plaagbeheerprogram (GPB) te inkorporeer om sodoende minimale residue te verseker en uiteindelik residu vrye produksie van kernvrugte in Suid-Afrika. Ten einde die sukses van EPNs te verseker, is beide die omgewings- en tegniese faktore wat hul doeltreffendheid beïnvloed in die studie ondersoek. Die biologiese beheer potensiaal van drie ingevoerde EPN isolate, Steinernema feltiae en twee Heterorhabditis bacteriophora (Hb1, Hb2) isolate, sowel as 'n plaaslike isolaat, Steinernema yirgalemense, is vir die beheer van kodlingmot onder plaaslike toestande geëvalueer. Alle konsentrasies van S. yirgalemense, wat deur indompeling in ‘n suspensie van nematodes toegedien is, het > 98% beheer tot gevolg gehad. Die twee geformuleerde isolate van H. bacteriophora, Hb1-f en Hb2-f, het < 30% beheer gegee. Met die gebruik van dieselfde nematode isolate, wat in vivo geproduseer is, het S. yirgalemense nog steeds 'n hoër kodlingmot beheer van > 90% opgelewer, in vergelyking met die 54% en 31% beheer van die H. bacteriophora Hb1 en Hb2 isolate, onderskeidelik. Steinernema feltiae het in opvolg veldproewe ≥ 80% beheer tot gevolg gehad en was dus meer effektief as beide S. yirgalemense en die H. bacteriophora (Hb1) isolate, met 66% en 24% en 24% en 9% beheer onderskeidelik in twee afsonderlike veldproewe. Om die resultate van die veldproewe te bevestig, is daaropvolgende laboratorium biotoetse uitgevoer en temperatuur regimes is geëvalueer deur die selfde siklus as onder natuurlike toestande te volg, met 'n konstante humiditeit van 100%. Die studie het bewys dat S. feltiae die mees doeltreffende isolate was met > 90% mortaliteit, S. yirgalemense het gevolg met 78% mortaliteit. Die twee H. bacteriophora isolate (Hb1, Hb2) het onderskeidelik onder bogenoemde laboratorium toestande 73% en 59% beheer tot gevolg gehad. Humiditeit blyk dus die belangrikste faktor te wees wat EPN se doeltreffendheid tydens bogrondse toediening affekteer. Die studie het bewys dat S. feltiae 'n beter kandidaat as S. yirgalemense vir die beheer van kodlingmot is. Die doeltreffendheid van verskillende EPN isolate vir die beheer van diapause kodlingmot larwes sowel as EPN se aktiwiteit by verskillende temperature is ook onder plaaslike toestande, deur bogrondse bespuitings, geëvalueer. Steinernema feltiae en twee isolate van H. bacteriophora (Hb1, Hb2), S. yirgalemense en ‘n ander plaaslike isolaat, Steinernema jeffreyense, is geëvalueer. Die gebruik van S. jeffreyense, het tot die mees effektiewe beheer gelei, met 67% mortaliteit, gevolg deur H. bacteriophora (Hb1) met 42%, en dan S. yirgalemense met 41%. Laboratorium biotoetse wat veldtoestande simuleer, het bewys dat S. feltiae die mees doeltreffend teen kodlingmot larwes is, met 67% mortaliteit tydens infeksie, gevolg deur S. yirgalemense met 58%, die H. bacteriophora Hb1 isolaat met 48%, en die Hb2 isolaat met 24%. 'n Vergelyking van die infeksie- en penetrasie tempo van twee isolate van H. bacteriophora (Hb1, Hb2), S. feltiae en S. yirgalemense wat in 12-put plate teen 14°C en 25°C uitgevoer is, het die dramatiese effek van temperatuur op EPN doeltreffendheid bevestig. By 14°C het alle EPN spesies behandelings stadiger kodlingmot mortaliteit as by 25°C na 48h tot gevolg gehad. ‘n Mortaliteit van < 15% is vir alle spesies aangeteken terwyl by die warmer temperature is > 98% mortaliteit vir alle spesies aangeteken. Na die blootstelling van afgespoelde, koel behandelde larwes aan 25°C vir 24 uur, het die toediening van beide S. feltiae en S. yirgalemense, 100% mortaliteit van larwes tot gevolg gehad terwyl die toediening van die twee H. bacteriophora isolate, Hb1 en Hb2, onderskeidelik 68% en 54% beheer tot gevolg gehad, oor dieselfde tydperk. By 14°C, het S. feltiae die hoogste gemiddelde penetrasie tempo van 20 ILs/ larwe, gevolg deur S. yirgalemense met 14 ILs/ larwe tot gevolg gehad, terwyl S. yirgalemense die hoogste penetrasie tempo getoon het by 25°C met 39 ILs/ insek, gevolg deur S. feltiae met 9 ILs/ insek. Die twee H. bacteriophora isolate (Hb1 en Hb2) het ook hoër gemiddelde penetrasie tempo by die hoër temperatuur getoon. Hierdie studie het die biobeheer potensiaal van S. jeffreyense beklemtoon, asook weereens bevestig dat S. feltiae ‘n koue-aktiewe nematode is, terwyl die ander drie EPN isolate warmer temperature verkies. Hout vrugtekratstapels, word beskou as ‘n ideale oorwintering skuiling vir kodlingmot diapause larwes. In Suid-Afrika word beheerstrategieë teen kodlingmot in die wiele gery deur die herbesmetting van boorde deur nabygeleë besmette hout vrugtekratte of deur die beweging van kratte tussen boorde. Hout vrugtekratte word wêreldwyd stelselmatig vervang met plastiek kratte. Dit sal egter eers oor ‘n aantal jare in Suid-Afrika uitgefaseer word. Die doel van hierdie studie was om die potensiaal van H. bacteriophora, S. feltiae, en S. yirgalemense te evalueer deur miniatuur hout vrugtekratte onder gekontroleerde toestande in die laboratorium te disinfekteer. Na die onderdompeling van die mini vrugtekratte in 'n nematode suspensie van 25 ILs/ml van al drie EPN spesies, onder optimale toestande van temperatuur en humiditeit, is die hoogste persentasie van beheer met die gebruik van S. feltiae (74,85% ± 3.64%) verkry. Die byvoeging van toevoegings middels het S. feltiae se vermoë om te infekteer betekenisvol (p <0,001) tot > 95% verhoog, maar dit het nie tot 'n betekenisvolle toename in die infektiwiteit van H. bacteriophora of S. yirgalemense gelei nie. Die resultate dui daarop dat H. bacteriophora nie 'n geskikte kandidaat is om te gebruik vir die beheer van kodlingmot larwes in besmette hout kratte nie. Die voorkeurkandidaat tans vir beheer is S. feltiae, waarvan die doeltreffendheid verhoog kan word deur middel van die byvoeging van 'n bymiddel. Gedurende die winter wanneer die hele kodlingmot populasie as larwes in diapause is, word geen beheer in boorde toegepas nie. Hierdie studie het getoon dat EPNs in boorde gespuit kan word om sodoende die opkomende kodlingmot populasie na die winter te verlaag en kan ook ingesluit word in 'n GPB program. Die EPNs kan as 'n tweede verdedigingslinie optree en doeltreffende beheer van kodlingmot larwes verseker wat chemiese bespuitings oorleef het, en sodoende beskerming teen weerstandige kodlingmot populasies in die volgende seisoen bied.

Thesis (MScAgric.)--Stellenbosch University, 2000.<br>ENGLISH ABSTRACT: Phomopsis cane and leaf spot disease of grapevine is an economically important disease in many of the vine-growing areas of the world. Four different Phomopsis spp. have previously been associated with this disease. The present study investigates the taxonomic significance of the different taxa found on grapevines in South Africa, as well as the endophytic growth and fungicide sensitivity of Phomopsis viticola isolates. The thesis is compiled of several different parts, which deal with specific, but related topics, and hence some duplication has been unavoidable. Understanding the epidemiology of a disease is important for the correct timing of disease control. To investigate the endophytic growth of P. viticola, asymptomatic shoots were collected at eight different growth stages. Nodes, internodes, leaf petioles, leaves, tendrils and bunch peduncles were investigated. Two Phomopsis spp., taxon 1 and 2 were identified in this study. The Phomopsis viticola-complex had a relative importance of 9% and accounted for 3% of the isolations. P. viticola (taxon 2) is mainly isolated from the nodes and internodes. Inoculations of healthy, young vine tissue confirmed taxon 2 to be a virulent pathogen, suggesting that it is a latent pathogen rather than an endophyte. In contrast, taxon 1 appeared to be a true endophyte, and did not seem to be an important pathogen on vines. The true identity of the causal organism of Phomopsis cane and leaf spot disease was investigated by collecting samples from 58 different vineyards in the grapevine growing areas of the Western Cape. P. viiicola occurred in grapevine material collected from Lutzville to Swellendam, but was not found in the Oudtshoorn and Orange River grapevine areas. Diaporthe perjuncta (taxon 1), P. vutcola (taxon 2), taxon 3 and a Phomopsis species commonly associated with shoot blight of peaches in the U.S.A., P. amygdali, were identified among the South African grapevine isolates. Examination of the Australian culture designated as taxon 4 found it to be a species of Libertella, thus excluding it from the P. viticola-complex. An Italian isolate was found to represent a species of Phomopsis not previously known from grapevines, and this was subsequently described as taxon 5. Species delimitation was based on morphological and cultural characteristics, stem inoculations and the formation of the teleomorph in vitro. The identity of each morphological taxon was confirmed by means of phylogenetic analyses of the nuclear ribosomal DNA internal transcribed spacers (ITS 1 and ITS2) and the 5' end partial sequence of the mitochondrial small subunit (mtSSU). P. amygdali, associated with peach shoot blight in the U.S.A., was isolated once only and appeared to be of lesser importance in this disease complex. Furthermore, taxa 1 (Diaporthe perjuncta) and 3 were also rarely encountered and proved to be non-pathogenic, indicating their non-functional role in Phomopsis cane and leaf spot disease. Taxon 2 (Phomopsis viticolas was common and widely distributed in diseased vineyards. This taxon was associated with the typical disease symptoms and proved to be pathogenic. Morphologically taxon 2 corresponded best with P. viticola, which was also neotypified in this study. Taxon 2 was mostly isolated from buds and nodes, indicating that these are important sites in which the fungus survives during winter. Molecular data indicated that taxon 3 and P. amygdali were not host specific to grapevine. The currently used foliar fungicides were compared to the new strobilurin fungicides. The effects of nine fungicides (azoxystrobin, flusilazole, folpet, fosetyl- Al+mancozeb, kresoxim-methyl, mancozeb, penconazole, spiroxamine and trifloxystrobin) were tested in vitro on inhibition of mycelial growth. The following EC50 (ug/ml) values were obtained: azoxystrobin (0.350), flusilazole (0.007), folpet (4.489), fosetyl-Al+mancozeb (3.925), kresoxim-methyl (1.665), mancozeb (2.891), penconazole (0.023), spiroxamine (0.321) and trifloxystrobin (0.051). Additionally, azoxystrobin, folpet, kresoxim-methyl, mancozeb, propineb and trifloxystrobin were tested for their ability to inhibit spore germination in vitro. The subsequent EC50 (ug/ml) values were obtained: azoxystrobin 0.123), folpet (0.510), kresoxim-methyl (0.0037), mancozeb (0.250), propineb (0.156) and trifloxystrobin (0.003). The results reported in part 4 showed that the strobilurin fungicides inhibited the mycelial growth and spore germination of P. viticola. However, further trials need to be conducted to verify these findings under field conditions. In the present study taxa 1, 3 and P. amygdali were infrequently isolated, suggesting that they played a less prominent role in the P. viticolacomplex.<br>AFRIKAANSE OPSOMMING: Streepvleksiekte van wingerd is 'n ekonomies belangrike siekte wat in die meeste wingerdproduserende gebiede van die wêreld voorkom. Vier Phomopsis spesies is in die verlede met dié siekte geassosieer. Hierdie studie ondersoek die taksonomiese belangrikheid van die verskillende taksa wat op wingerd in Suid Afrika gevind word, asook die endofietiese groei en fungisiedsensitiwiteit van die Phomopsis vitico/a isolate. Hierdie tesis bestaan uit verskeie dele met spesifieke, maar verwante onderwerpe wat tot onafwendbare duplisering lei. Dit is belangrik om die epidemiologie van 'n siekte te verstaan sodat korrekte en tydsberekende siektebeheer toegepas kan word. Die endofietiese groei van P. vitico/a is ondersoek deur simptoomlose lote by agt verskillende groei stadiums te versamel. Nodusse, internodusse, blaarstele, blare, rankies en trosstele is ondersoek. Twee Phomopsis spp., takson 1 en 2 is geïdentifiseer. Die Phomopsis vitico/a-kompleks het 3% van die isolasies uitgemaak en 'n relatiewe belangrikheid van 9% getoon. P. vitico/a (takson 2) is meestal uit die nodus en internodus geïsoleer. lnokulasies van gesonde, jong wingerdweefsel het bevestig dat takson 2 'n virulente patogeen is en dat die takson eerder 'n latente patogeen as 'n endofiet is. In teenstelling hiermee is takson 1 'n ware endofiet en 'n onbelangrike patogeen op wingerd. Die ware identiteit van die veroorsakende organisme van streepvlek is ondersoek deur plantmateriaal vanaf 58 verskillende wingerde in die wingerproduserende gebiede van die Wes-Kaap te versamel. P. vitico/a is in wingerdmateriaal vanaf Lutzville tot Swellendam aangetref, maar nie in die Oudtshoorn en Oranjerivier wingerd produserende gebiede nie. Diaporthe perjuncta (takson 1), P. vitico/a (takson 2), takson 3 en P. amygdali is in die Suid Afrikaanse wingerdisolate geïdentifiseer. P. amygdali word met lootverskroeiing van perske bome in die V.S.A. geassosieer. Die Australiese isolaat wat benoem is as takson 4, is met die huidige ondersoek gevind om 'n spesie van Libertella te wees. Takson 4 is daarvolgens uit die P. vitico/a-kompleks gelaat. 'n Italiaanse isolaat het 'n nuwe spesie van Phomopsis op wingerd verteenwoordig en is vervolgens as takson 5 beskryf. Spesie-onderskeiding is op morfologiese en kulturele eienskappe, staminokulasies en die vorming van die teleomorf in vitro gebaseer. Die identiteit vanelke morfologiese takson is met behulp van filogenetiese analises van die nukleêre ribosomale DNS intern transkriberende spasieerders (ITS 1 en ITS2) en die 5' punt gedeeltelike nukleotied volgorde van die mitochondriale klein subeenheid (mtSSU) bevestig. P. amygdali is slegs een keer geïsoleer en blyk van minder belang in die siektekompleks te wees. Takson 1 (Diaporthe perjuneta) en takson 3 het ook min voorgekom en is nie-patogenies, wat hul nie-funksionele rol in streepvleksiekte aandui. Takson 2 (P. viticola) is algemeen geïsoleer en kom wyd verspreid voor. Hierdie takson is geassosieer met die tipiese siektesimptome en is ook patogenies. Morfologies stem takson 2 met P. viiicola ooreen en is ook geneotipifiseer in hierdie studie. Takson 2 is meestal vanaf die ogies en nodusse geïsoleer, wat daarop dui dat hierdie belangrike setels is waar die swam tydens die winter oorleef. Die molekulêre data toon aan dat takson 3 en P. amygdali nie gasheerspesifiek tot wingerd is nie. Die swamdoders wat tans teen streepvlek gebruik word, is met die nuwe strobilurin swamdoders vergelyk. Die effek van nege swamdoders (azoksistrobin, flusilasool, folpet, fosetyl-Al + mancozeb, kresoxirn-metiel, mankozeb, penconasool, spiroksamien en trifloksistrobin) is in vitro op die inhibisie van miseliumgroei getoets. Die volgende EKso-waardes (g/ml) is verkry: azoxystrobin (0.350), flusilasool (0.007), folpet (4.489), fosetiel-Al + mankozeb (3.925), kresoxirn-metiel (l.665), mankozeb (2.891), penkonasool (0.023), spiroksamien (0.321) en trifloxystrobin (0.051). Azoxystrobin, folpet, kresoxim-rnetiel, mankozeb, propineb en trifloksistrobin is ook in vitro getoets vir hul inhibisie op spoorontkieming. Die volgende EKso-waardes is verkry: azoxystrobin (0.123), folpet (0.510), kresoxim-metiel (0.0037), mankozeb (0.250), propineb (0.156) en trifloxystrobin (0.003). Die resultate vervat in deel 4 toon dat die strobilurin swamdoders die miseliumgroei en spoorontkieming van P. viticola inhibeer. Toetsing in die veld word egter benodig om die effektiwiteit van die middels te bevestig. In hierdie studie is taksa I, 3 en P. amygdali selde geïsoleer, wat aangedui het dat hierdie taksa 'n minder belangrike rol in die P. viticola-kompleks speel.

In this study, we have demonstrated that inoculation of bean seeds with non-pathogenic binucleate Rhizoctonia (np-BNR) at sowing protected bean seedlings from infection of R. solani. Using quantitative real-time RT-PCR (QRT-PCR), transcript levels of defense genes encoding 1,3-beta-glucanase (GLUC), phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS) in one-week old bean seedlings was monitored during np-BNR and R. solani interaction. The results revealed that protection effect of np-BNR correspond to a systemic suppression of these three defense genes' expression from significant higher level elicited by R. solani to the level of non-infected plants. This indicates that bio-protection by np-BNR isolates is not correlated to activation of these three defense genes' expression. Similar suppression was achieved for pre-colonization of bean seedlings with arbuscular mycorrhizal (AM) Glomus introradices on GLUC gene expression, although the AM fungus did not significantly reduce rot symptoms. Possible mechanisms implicated in down-regulation during plant-pathogen and np-BNR or AM interaction are discussed.

Colombia covers an area of approximately 114 million hectares (ha) of which the potential forestly land has been estimated at 60.7 million ha, about 53% of the total area. Only 54 million ha are considered as natural forest, leaving approximately 29 million ha to be used for livestock and agriculture. In order to provide an alternative source of timber, Colombian groups have planted approximately 327 000 ha to different species of Pinus, Eucalyptus and native species. This clearly represents only a small proportion of the total area that might be used for forestry, which is set to grow in the future. In general, trees established in plantations have been grown as monocultures that allow for substantial productivity per management unit. But this is also a homogeneous genetic resource that is highly susceptible to damage caused by insect pests and diseases. It is therefore, surprising that relatively little work has been conducted on pests and pathogens negatively affecting plantation foresty in Colombia. In this regard, the studies presented in this thesis present the first comprehensive treatment of the topic for the country. It is consequently hoped that these will form a basis for the future management and sustainability of forestry in Colombia. In the first Chapter of this thesis, I provide a historical background and the current pest and disease situation for the forestry sector in Colombia. This includes an overview of the main pest and diseases affecting commercial nonnative trees, especially species of Pinus and Eucalyptus. A wide range of sites occurring in Colombia were considered and the document also includes the impact of different climatic conditions on the incidence and management of the various pests and diseases treated. Some of the most important defoliators in Colombia reside in the order Phasmatodea and one of these, Litosermyle ocanae, was treated in studies presented in chapter two. The overall aim was to contribute basic knowledge of L. ocanae including an understanding of the biology, egg population dynamics, and potential biological control assessments based on early detection of the insect. vi Chapter three includes the discovery of one of the members of the Geometridae that causes serious damage due to defoliation of Pinus and Eucalyptus plantations. This pest, known as Chrysomima semiluteraria, has been known in Colombia for many years and this study included a comprehensive evaluation of its biology and field monitoring contributing to an Integrated Pest Management strategy for it. The insect was studied under field and laboratory conditions and a special emphasis was placed on its biological control using the egg parasitoid Telenomus alsophilae. Pinus plantations in Colombia have been affected by numerous pests, including recently, the adelgid Pineus sp. To establish management strategies to assist commercial forestry operations, the life cycle of this insect and the susceptibility of different Pinus species were determined in Chapter four. In addition, the possible role of a Ceraeochrysa sp. as a biological control agent was investigated. In chapter five, Fusarium circinatum is recorded for the first time on Pinus spp. in Colombia. The discovery of this fungus, known to cause the disease Pitch Canker has had an important impact on forestry, especially with regard to management strategies aimed at reducing its impact. Studies in this chapter included those to evaluate the susceptibility of families and provenances of Pinus spp., which are currently used in forestry planting programs in Colombia. The first emergence of the foliage pathogen Dothistroma septosporum as an important constraint to pine forestry in Colombia is treated in chapter six. This fungus is a serious pathogen of many species of Pinus around the world. It was, however, not recognised as a serious threat to forestry in Colombia until it emerged as a serious source of damage to Pinus tecunumanii. In this chapter, the aim was firstly to confirm the identity of the pathogen based on DNA sequence data. Subsequently, the host range and distribution of the pathogen was established in different Colombian forestry areas. The impact of the disease and the susceptibility of different provenances of P. tecunumanii was also assessed. In chapter seven, the main objective was to described the susceptibility of E. grandis clones to a new species of Ceratocystis and to identify clones resistant to it. The fungus was described as Ceratocystis neglecta and management strategies are suggested for it. This thesis includes two Appendices treating new reports of insect pests and an important Eucalyptus pathogen. In Appendix 1, I report on the importance of three different pest insects. Appendix 2 includes a description of a new disease, as well as the potential risks that this disease poses for the Colombian forestry sector.<br>Thesis (PhD)--University of Pretoria, 2013.<br>gm2013<br>Forestry and Agricultural Biotechnology Institute (FABI)<br>Unrestricted

False codling moth (FCM), Thaumatotibia (=Cryptophlebia) leucotreta (Meyr) (Lepidoptera: Tortricidae), is one of the most important pests on citrus. The Cryptophlebia leucotreta granulovirus (CrleGV) has been developed into a successful biological control agent, registered under the name Cryptogran, and is currently the preferred product for the control of FCM on citrus in South Africa. A prerequisite to the continued success of Cryptogran as a means of controlling false codling moth is to understand the factors affecting field persistence of the virus, and to find ways to improve it. The aim of this study was to gain a clearer understanding of the product and the abiotic and biotic factors affecting its persistence in the field, and to investigate methods to improve this persistence. The effect of UV-irradiation on the virus was determined, and various products were tested as UV protectants in laboratory bioassays. Lignin was the most effective additive, and was tested in several field trials, where it also enhanced the efficacy of Cryptogran. Laboratory trials indicated that Cryptogran is rainfast. Cryptogran applications early in the season had a longer period of residual activity than sprays applied closer to harvest. Daytime applications were less effective that evening sprays. Sprays applied coinciding with peaks in pheromone moth trap catches were more effective than those applied between peaks. Biotic factors influencing persistence were investigated. Residual efficacy was longer when treatments were applied to blocks than as single tree treatments. Attempts were made to quantify the effect of the navel end of a navel orange on the field persistence of Cryptogran. Cryptogran was shown to be compatible with many agricultural chemicals used on citrus. Economic thresholds and various cost-benefit analyses are discussed. A list of practical recommendations to growers was drawn up, and possibilities for future research are presented.

A granulovirus isolated from Cryptophlebia leucotreta larvae was shown through restriction endonuclease analysis to be a novel strain (CrleGV-SA). No more than one isolate could be identified from a laboratory culture of C. leucotreta. However, a preliminary examination of restricted DNA profiles of isolates from different geographical regions indicated some minor differences. In surface dose bioassays on artificial diet, LC50 and LC90 values with neonate larvae were estimated to be 4.095 x 103 OBs/ml and 1.185 x 105 OBs/ml respectively. LT50 and LT90 values with neonate larvae were estimated to be 4 days 22 h and 7 days 8 h, respectively. Detached fruit (navel orange) bioassays with neonate larvae indicated that virus concentrations that are likely to be effective in the field range from 1.08 x 107 to 3.819 x 1010 OBs/ml. In surface dose bioassays with fifth instar larvae LC50 and LC90 values were estimated to be 2.678 x 107 OBs/ml and 9.118 x 109 OBs/ml respectively. LT50 and LT90 values were estimated to be 7 days 17 h and 9 days 8 h, respectively. A new artificial diet for mass rearing the host was developed. Microbial contamination of diet was significantly reduced by adding nipagin and sorbic acid to the diet and by surface sterilising C. leucotreta eggs with Sporekill. Almost 20 % more eggs were produced from moths reared on the new diet compared to moths reared on the old diet. A further 9 % improvement in egg production and a reduction in the labour required to produce eggs, was made with the development of a new oviposition cage attached to the moth eclosion box. Virus was mass produced in fifth instar C. leucotreta larvae by surface inoculating diet with the LC90. When 300 individuals were placed onto inoculated diet, 56 % of them were recovered six to 11 days later as infected larvae. Mean larval equivalents was 1.158 x 1011 OBs/larva. When larvae and diet were harvested together, highest yields of virus were achieved at eight days after inoculation. Microbial contamination in semi-purified preparations of CrleGV ranged from 176211 to 433594 (OB:CFU ratio). Half-life of CrleGV in the field was estimated to be less than 1 day on the northern aspect of trees and between 3 - 6 days on the southern aspect. Original activity remaining (OAR) of the virus dropped below 50 % after 5 days on the northern aspect of trees and was still at 69 % on the southern aspect of trees after 3 weeks. In field trials, CrleGV reduced C. leucotreta infestation of navel oranges by up to 60 % for a period of 39 days. CrleGV in combination with augmentation of the C. leucotreta egg parasitoid, Trichogrammatoidea cryptophlebiae, reduced infestation by 70 %. The integration of CrleGV into an integrated pest management (IPM) system for the management of C. leucotreta on citrus is proposed.

The false codling moth (FCM), Thaumatotibia (=Cryptophlebia) leucotreta (Meyrick) (Lepidoptera: Tortricidae) is a serious pest of citrus and other crops in Sub-Saharan Africa. The introduction of the Cryptophlebia leucotreta granulovirus (CrleGV-SA) Cryptogran and Cryptex (biopesticides) has proven to be very effective in the control of FCM. However, markedly lower susceptibility of some codling moth (CM), Cydia pomonella (L.) populations to Cydia pomonella granulovirus (CpGV-M), another granulovirus product used in the control of CM’s in Europe have been reported. Genetic differences between FCM populations in South Africa have also been established. It is therefore possible that differences in the susceptibility of these geographically distinct FCM populations to CrleGV-SA might also exist. To investigate this phenomenon, a benchmark for pathogenecity was established. In continuation of previous work with Cryptogran against the 1st and 5th instar FCM larvae, dose-response relationships were established for all five larval instars of FCM. In surface dose-response bioassays, the LC50 values for the 2nd, 3rd and 4th instars were calculated to be 4.516 x 104, 1.662 x 105 and 2.205 x 106 occlusion bodies (OBs)/ml, respectively. The LC90 values for the 2nd, 3rd and 4th instars were calculated to be 4.287 x 106, 9.992 x 106 and 1.661 x 108 OBs/ml, respectively. Susceptibility to CrleGV-SA was found to decline with larval stage and increase with time of exposure. The protocol was used in guiding bioassays with field collected FCM larvae. Laboratory assays conducted with Cryptogran (at 1.661 x 108 OBs/ml) against field collected FCM larvae from Addo, Kirkwood, Citrusdal and Clanwilliam as well as a standard laboratory colony, showed a significant difference in pathogenecity in only one case. This significant difference was observed between 5th instars from the Addo colony and 5th instars from the other populations. Four geographically distinct FCM colonies from Addo, Citrusdal, Marble Hall and Nelspruit were also established. Since Cryptogran and Cryptex are always targeted against 1st instar FCM larvae in the field, further comparative laboratory assays were conducted with the Addo colony and an old laboratory colony. Cryptogran was significantly more pathogenic than Cryptex against both the Addo and the old colony. However, a high level of heterogeneity was observed in responses within each population.

Includes bibliographical references (leaves 217-230). Details the biology and life history of the Quandong moth and investigates management strategies that would enable growers to manage the pest in an economically and environmentally sustainable program.

Thesis (MSc)--Stellenbosch University, 2003.<br>ENGLISH ABSTRACT: Paecilomyces Ii/acinus, ras 251 (geregistreer in terme van wet 36 van 1947 as Suid-Afrika se eerste natuurlike nematisiede en kommersieel beskikbaar as PI Plus) is as biologiese beheer agent getoets by aartappels en in geïntegreerde beheer programme by sitrus en wingerd teen respektiewelik Me/oidogyne species, Ty/enchu/us semipenetrans en verskeie ektoparasitiese nematodes. Die swam toon belofte vir die beheer van hierdie nematodes en het terselfdertyd nie 'n nadelige effek op nie-teiken, voordelige organismes in die grond nie. Veral in kombinasie met chemiese middels, as deel van geïntegreerde programme, kan dit lei tot verminderde gebruik van hoogs toksiese middels en dus meer omgewingsvriendelike landboupraktyke. Biological control of plant parasitic nematodes on potatoes, citrus and grapevine with the fungus, Paecilomyces liIacinus. Paecilomyces liIacinus, race 251 (registered in terms of act 36 of 1947 as South Africa's first natural nematicide, commercially available as PI Plus) was tested as a biological control agent on potatoes and in integrated control programs on citrus and grapevine against Me/oidogyne species, Ty/enchu/us semipenetrans and various ectoparasitic nematodes respectively. The fungus shows promise for the control of these nematodes, without having a harmful effect on non-target, beneficial organisms in the soil. Especially in combination with chemical products, as part of integrated programs, this can lead to less use of highly toxic compounds and thus to more environmentally friendly agricultural practices.<br>AFRIKAANSE OPSOMMING: Sedert die ontdekking van die swam, Paeci/omyces Ii/acinus (Thom.) Samson as 'n effektiewe eierparasiet van Meloidogyne incognita acrita en Globodera pal/ida (Jatala et al., 1979) het verdere veldproewe in Peru tot die effektiewe beheer van M. incognita en Tylenchulus semipenetrans gelei. Na verskeie suksesse in Peru is die swam onder verskillende klimaat- en grondkondisies in verskeie ander lande beproef. Die sukses behaal in die Filippyne het gelei tot die kommersiële produksie van die swam onder die handelsnaam Biocon. Anders as met chemiese middels vind die werking van biologiese agente stadig oor tyd plaas. Biologiese beheer sal nie chemiese beheer sonder meer kan vervang nie. Dit behoort egter deel te vorm van geïntegreerde nematode bestuur. Inkorporering van die natuurlike organismes, die oordeelkundige gebruik van chemiese nematisiedes, moontlik in kombinasie met die biobeheer agente, weerstand, en ander kulturele praktyke moet ernstig oorweeg word as ons hoop om die steeds groeiende wêreldbevolking te voed (Jatala, 1986). Paecilomyces liIacinus, ras 251, Suid-Afrika se eerste geregistreerde natuurlike nematisiede, kommersieel beskikbaar as PI Plus, is in die Olifantsrivier besproeiingsgebied geëvalueer vir die bestuur van ekonomies belangrike plantparasitiese nematodes by aartappels, sitrus en wingerd. Hierdie gewasse is belangrike bedryfstakke van die streek en is onderhewig aan skade deur nematodes wat die opbrengs nadelig beïnvloed. Chemiese beheer bied slegs 'n korttermyn oplossing vir nematode probleme en skadelike getalle word in 'n kort tyd weer opgebou. Boonop lei dié hoogs toksiese middels tot agteruitgang van die omgewing en sy waterbronne. Die toenemende besorgdheid hieroor en die groot potensiaal van biologiese beheer agente (Jatala, 1986) was die hoofrede vir die werk waaroor hier gerapporteer word.

Thesis (MscAgric (Horticulture))--University of Stellenbosch, 2005.<br>The occurrence of both pre- and postharvest leaf blackening in certain Protea species and cultivars is a problem that severely limits their marketability, vase life and transport options. This research focuses on : (I) The distribution of carbohydrates in inflorescence bearing stems of certain Protea cultivars from harvest, following pulsing with a 10 g.L-1 glucose solution until four weeks postharvest. Stems were held under a variety of postharvest conditions, and (II) The suppression of Protea postharvest leaf blackening with specific focus on the cultivar ‘Sylvia’ (P. eximia x P. susannae).

For a successful and economical integrated control program aimed at a particular disease, pertinent information, regarding the environmental conditions prevailing in the growing area, the crop itself and the pathogen, must be available. Recently, the control of powdery mildew disease on cashew has moved from the use of non-systemic fungicides with a wide range of action, to highly specific systemic ones. Such a shift requires a more effective integrated control system, whereby tolerant varieties in combination with fungicide unaffected biocontrol agents are timely used to ensure disease control and reduce the hazards associated with excessive fungicide applications. The purpose of this study was to understand the relationship between the disease epidemic and some climatic factors over time. Appropriate periods for management interventions were determined. The cellular host reaction to infection by Oidium anacardii Noack was studied with a view to rapidly identify disease tolerant host types. Potential antagonists were isolated, screened and compared with commercial biocontrol products using in vivo techniques and chemical control programs were finally evaluated. Electron microscopy elucidated that the powdery mildew tolerant cashew variety (H1) had a relatively higher consistency of cytoplasmic aggregates upon infection by O. anacardii when compared to the susceptible clone. Based on conidia and conidiophore morphology, conidial germination and conidiogenesis processes observed indicated that O. anacardii belongs to the subgenus Pseudoidium (Y.S. Paul&J.N. Kapoor) comb.Et. Stat. Nov. (Holomorph Erysiphe Sect. Erysiphe U. Braun). There was no direct relationship between the progress of the cashew powdery mildew epidemic and temperature, relative humidity or dew point over time. However, the epidemic did not start until conditions of average temperatures under the tree canopy were below 30°C, relative humidity was 80% and dew point was above 15. In vivo screening of 72 isolates, amongst them bacteria and fungi, from cashew leaves and florets showed that none were effective against O. anacardii, the causal agent of cashew powdery mildew. However, commercial antagonists, Candida saitoana, Bacillus subtilis and B. licheniformis, significantly reduced the growth and branching of primary hyphae. One antagonist, B. licheniformis, was as effective as the commercial fungicide triadimenol 25% EC (Bayfidan). Chemical fungicides were found to be effective against powdery mildew; however, the currently prevailing economic environment in Mozambique was found inappropriate for the use of expensive organic fungicides. Additional gain from the use of fungicides was found to be solely qualitative and thus did not represent a fair investment return ratio in terms of cashew nut prices and production costs. The use of integrated cashew management was finally recommended. Further studies should focus on development of integrated and cost effective disease management strategies.<br>Dissertation (MSc(Plant Pathology))--University of Pretoria, 2003.<br>Microbiology and Plant Pathology<br>unrestricted

Forecasts for the first fungicide application to manage cercospora blight of carrot (Daucus carota L. subsp. sativus (Hoffim) Arcang) caused by Cercospora carotae (Passerini) Solheim were evaluated during the summers of 1991 and 1992. In the forecast based on infection, the blight severity values (BSV) were calculated for each day using duration of leaf wetness, relative humidity and temperature. The first fungicide was applied when the cumulative blight severity value reached 14 (CBSV 14) and 18 (CBSV 18). The forecasts were compared with the 50% disease incidence threshold method where the first fungicide was applied when the middle leaf of 50% of plants was diseased, and the conventional method where the first fungicide application was made when the plant height reached 15 cm. For the threshold of CBSV 14 and CBSV 18, no loss was observed in yield and fewer fungicides were applied as compared to conventional and 50% disease incidence threshold methods. Three years of weather data from experimental plots and 155 commercial field data were used to study the behaviour of CBSV at 50% disease incidence threshold. Only in 3% of the fields, the disease incidence of 50% was observed before the CBSV reached 11. The increase from CBSV 11 to 20 was rapid. Depending on environmental conditions the first fungicide at a CBSV range of 11 to 20 is proposed rather than a critical threshold since no loss in yield was observed at 18 in the experimental plots. No fungicide is applied until the CBSV reaches 11 and the application must not be delayed after CBSV reaches 20.

Septoria blight of celery (Apium graveolens) caused by Septoria apiicola is a common disease. This work dealt with the development of a simple forecast model to time fungicide application to manage the blight. Two studies were conducted to examine the effect of relative humidity (65, 88, 92, 96%, and wet) temperature (15, 20, 25, and 28 C) and exposure period (6, 12, 24, 48, 72, and 96 hr) on spore production. Spores were produced at all levels of relative humidity and temperature tested. Overall spore production increased with increasing relative humidity. The greatest and the least were produced after a 48 hr wet period at 20 and 28 C, respectively. Two regression models were developed to predict spore production as functions of relative humidity and temperature. In order to validate the infection function model a field experiment was conducted in 1989 to quantify latent period. A mean latent period beginning of 12 days, a mean latent period 50% of 14 days, and a mean latent period ending of 19 days were established. Two regression models were developed to predict latent period beginning and latent period ending as functions of mean maximum and mean minimum temperatures. In order to develop a simple forecast model field experiments were conducted in 1989 and 1990. From these studies two simple forecast models are proposed to initiate fungicide applications: (1) Initial disease occurrence. (2) A cumulative disease severity value (CDSV) of 23. Using the forecasts proposed it is possible to reduce five to seven applications of fungicides during the celery growing season in Quebec.

This thesis was designed to address three seldom studied aspects of the relationship between herbivores and vegetational diversity. 1. Interactions between vegetational diversity and herbivore mortality due to predation were assessed by experimentally manipulating both the species diversity of plants and the densities of a common generalist predator, the spider Enoplagnatha ovata. 2. The importance of plant size to herbivore densities was examined by quantifying plant size (measured as plant height, width, leaf area and growth rates) and adjusting for it through covariate analysis. 3. Differences in population responses of several species of herbivores to both vegetational diversity and a predator, were compared by concurrently studying four lepidopterans. The main experiment used a two factor design, with two planting treatments and two predator treatments. The planting treatments consisted of plots planted with monocultures of brussels sprouts (Brassica oleraceae) and dicultures of brussels sprouts intercropped with peppermint (Mentha piperita). The natural enemy treatments involved augmentations of E ovata in some plots and untreated controls. Two of the herbivores studied, Plutella xylostella and Pieris rapae are monophagous lepidopterans, specializing on crucifers, while the others, Autographa californica and Mamestra configurata are polyphagous. For two species, P. xylostella and M. configurata responses to augmentations of the spider, E. ovata. were different in monocultures and dicultures. Reduced densities of these two species were found in monoculture plots with added spiders; in dicultures increased densities were found in plots with added spiders. This interaction effect points out that generalist predators can be effective in monocultures. I suggest that the importance of natural enemies in monocultures is often overlooked because only the initial colonization phase is being studied. By augmenting predator populations I was able to simulate densities equivalent to those in more established cropping systems. The increased herbivore densities in dicultures with added spiders might be explained by possible predation by E. ovata on other natural enemies of P. xylostella and M. configurata in dicultures but not in monocultures. Supporting evidence for this interpretation lies in the fact that percent parasitism of P. xylostella by the ichneumonid, Diadegma insulare was lower in plots with added spiders than in control plots. Furthermore, parasitism of P. xylostella by D. insulare increased with host density in diculture plots, but not in monoculture plots. Mamestra configurata was not subject to parasitism in this study, precluding assessment of a similar relationship. No A. californica larvae were found in plots with additional spiders. In contrast, P. rapae larvae were not affected by the experimental treatments. Plant size was a crucial determinant of both herbivore populations and percent parasitism of those herbivores. Most importantly, had plant size not been accounted for, the importance of vegetational diversity to both herbivore densities and percent parasitism would have been overestimated. For example, the incorrect conclusion, that vegetational diversity alone was important in determining the abundance of both of the generalist feeders would have been reached. The greater densities of A. californica in monocultures and M. configurata in dicultures were accounted for by plant size. Without plant size adjustments, percent parasitism of P. xylostella by D. insulare would have been misinterpeted as being greater in monocultures than dicultures. With plant size adjustments, the importance of E. ovata augmentations on lowering percent parasitismwas unmasked. All important interaction effects were discovered only after adjustments for plant size had been made. Despite the low densities of all herbivore species, significant responses to experimental treatments were found in three of the four species studied. Only P.l rapae was unaffected by any of the treatments. However, conclusions based on the feeding ng habits of the herbivores could not be made. The polyphagous feeders were affected by generalist predation as much as the crucifer specialist. Parasitism was found in only two of the species, P. xylostella and A. californica. Of these two species parasitism of the specialist, P. xylostella was affected by both vegetational diversity and generalist predation, whereas parasitism of A. californica was not. My study emphasizes multifaceted interactions between the size and diversity of a primary resource and several trophic levels of consumers. Multifactor models, involving several aspects of a cropping system, are required to uncover the important mechanisms behind variable herbivore responses to vegetational diversity.<br>Land and Food Systems, Faculty of<br>Graduate

Thesis (MSc)--Stellenbosch University, 2000.<br>Full text to be digitised and attached to bibliographic record.<br>ENGLISH ABSTRACT: The cultivation of citrus in the Vaalharts region is a fairly recent development. With the introduction of this crop, an insect pest complex has also developed in this region. The presence of these pests was studied in eleven orchards, planted with three citrus cultivars and of varying ages, distributed in the 300 square kilometer cultivation area. Each orchard was inspected for the presence of pests by making use of two weekly sampling techniques. Ten of the most important insect pests of citrus in the Vaalharts region are briefly described by refering to their general appearance, life cycles, feeding and pest status and economic threshold. For every pest various control options, including operational systems, crop cultivation, biological and chemical control, are discussed and, where applicable, illustrated by means of graphic presentations. The seasonal presence of the cirtrus pests in the Vaalharts region is tabulated and discussed individually. With these details as background, an insect pest management programme for citrus in the Vaalharts region is compiled.<br>AFRIKAANSE OPSOMMING: Die verbouing van sitrus in die Vaalhartsgebied is 'n redelik onlangse ontwikkeling. Gepaard met die nuwe gewas het daar ook 'n insekplaagkompleks in die gebied ontstaan. Die voorkoms van die plae is in elt .boorde, beplant met drie sitruskultivars en van verskillende ouderdomme, verspreid in die 300 vierkante kilometer verbouingsareaal, bestudeer. Elk van die boorde is weekliks ondersoek vir die aanwesigheid van plae deur van twee moniteringstegnieke gebruik te maak. Die tien belangrikste insekplae van sitrus in die Vaalhartsgebied word kortliks beskryf deur na hulle algemene voorkoms, lewenssiklus, voeding en plaagstatus en ekonomiese drempelwaardes asook die moniteringsmetodes wat gebruik is, te verwys. Vir elke plaag word beheeropsies, wat operasionele stelsels, gewasverbouing, bloloqlese en chemiese beheer insluit, bespreek wat, waar toepaslik, aan die hand van grafiese voorstellings gemustreer word. Die seisoenale aanwesigheid van die sitrusplae word in 'n tabel aangedui en individueel bespreek. Met die gegewens as agtergrond is 'n insekplaagbestuurprogram vir sitrus in die Vaalhartsgebied opgestel.

The within-field spatial distribution of F₁, F₂, and F₃ pink bollworm (PBW) (Pectinophora gossypiella Saunders) generations were modeled with Taylor's power law (TPL), Iwao's patchiness regression (IPR), and the geostatistical semivariogram. Kriging interpolation was used to grid data for the generation of isarithmic maps. Distributional patterns and movements within a field are displayed in a time series of three maps depicting density across the field. The sampling protocol was replicated in eight commercial cotton fields in south-central Arizona during 1985 and 1986. Permanent sample stations were situated throughout the fields on a regular grid pattern. Samples were collected during the peak larval population and handled so as to maintain the integrity of site-specific samples (spatially identified by X,Y coordinates). TPL and IPR could not be used satisfactorily to model the F₁ generation. TPL fit the observed F₂ and F₃ data better than IPR. Both methods predicted the F₂ to be more highly aggregated than the F₃. For a given precision, optimum sample size increased when TPL and IPR model parameters were incorporated into sample size formulae relative to a formula which assumed random distribution. Ninety-five percent of the modeled PBW distributions were autocorrelated in 2-dimensional space and shown to conform to regionalized variable theory by the successful application of geostatistics. The semivariogram models are in conceptual agreement with traditional models and represent a worthy alternative to traditional modeling methodology. The semivariogram models have a large nugget effect proportion (average = 67%) which, in combination with low PBW density in commercial fields, limits the applicability of geostatistics in this system. Isarithmic maps showed that F₁ larvae are either localized near a field edge or generally scattered throughout the field. No consistent inter-generational dispersal pattern was identified. The use of systematic grid sampling is most advantageous (relative to random sampling) when density and the spatial dependence of samples is high, or many samples can be taken. Systematic sampling and kriging estimation yielded more precise estimates than random sampling and classical statistics, but the advantage was buffered by low PBW densities and large nugget effect.

The purpose of this study was to assess the adoption response of cotton growers to Integrated Pest Management (IPM) programs, identify the extension strategies used to promote IPM, and provide descriptive comparison of IPM programs between the foremost eleven cotton-producing states in the United States. Integrated pest management is presented as an alternative agricultural production strategy referred to as sustainable agriculture. IPM programs for cotton were evaluated by use of a survey instrument and telephone interviews with the IPM coordinators for the eleven cotton-producing states. The most significant findings were that programs vary considerably between states; the most widely employed IPM techniques were selective use of chemicals, use of economic thresholds and sampling for pests; the Cooperative Extension Service worked closely with private consultants and grower associations; and adoption of a wide variety of IPM techniques has been extensive for cotton producers in the eleven foremost cotton-producing states.

The study examined the larval parasitoids of Cryptophlebia leucotreta (Meyrick) on citrus in South Africa and aimed to improve the existing rearing techniques of C. leucotreta with a view to mass rearing of biological control agents. The biological characteristics of Agathis bishopi Nixon (Hymenoptera: Braconidae) were studied, with an emphasis on parasitism rates in the field, host stage preference, developmental rate, life span and offspring sex ratios. Two larval parasitoids, A. bishopi and Apophua leucotretae (Wilkinson) (Hymenoptera: Ichneumonidae), and an egg parasitoid, Trichogrammatoidea cryptophlebiae Nagaraja (Hymenoptera: Trichogrammatidae), were recorded on C. leucotreta on citrus. A. bishopi was the more abundant of the larval parasitoids and exhibited density dependent parasitism. The highest parasitism rates were observed in December with up to 38% in Sundays River Valley and 34% in Gamtoos River Valley, at a time when the highest false codling moth infestations were observed. Agathis bishopi was recorded only in the Eastern Cape Province. The sex ratio of A. bishopi was biased towards females throughout the study (77% in Gamtoos River Valley and 72% in Sundays River Valley). Agathis bishopi is a solitary, koinobiont, larval-pupal endoparasitoid. The species showed a preference for 1st and 2"d instar hosts. Females regulate the sex of their progeny according to the size and larval stage of the host, ovipositing unfertilised eggs in younger, smaller larvae (1st instars) and fertilised eggs in older, larger larvae (2nd instars). The developmental rate of A. bishopi is in synchrony with that of the moth and adults emerge when adult moths that have escaped parasitism emerge. Agathis bishopi and T. cryptophlebiae compliment each other because they have different niches and strategies of attack. Integrating A. bishopi and T. cryptophlebiae into the management of citrus orchards has potential to suppress false codling moth. Larger rearing containers seemed ideal for large-scale rearing of false codling moth. A higher percentage of adults was obtained from larvae reared in larger containers than in smaller ones. The width of the sponges used as stoppers prevented escape of the larvae. Media prepared in larger containers are easier and simpler to prepare than in smaller ones, thus eliminating many precautions otherwise necessary to prevent contamination. Moth production was greatly reduced by the high concentration of Sporekill used for egg decontamination.

Thesis (MScAgric)--University of Stellenbosch, 2002.<br>ENGLISH ABSTRACT: The evaluation of fungicide efficacy in commercial vineyards can be influenced by the sporadic occurrence of Botrytis cinerea at various positions on vines, differences in bunch structure during bunch development and the phenomenon that symptom expression in shoots and bunches is governed by the resistance reaction of the various shoot and bunch parts. It has been postulated that, following air and water dispersal, infection by solitary conidia should playa prominent role in the epidemiology of B. cinerea on grapevine. The aim of this study was to determine (i) infection and (ii) fungicide efficacy at specific sites on shoots of vinelets and bunches (table grape cultivar Dauphine and the wine grape cultivar Merlot) inoculated with dry, airborne conidia of B. cinerea. Vinelets, prepared from cuttings, and bunches obtained from the vineyards at full bloom, pea size, bunch closure, véraison and harvest stages, were sprayed in a spray chamber at the recommended dosages with iprodione, pyrimethanil, cyprodinil/fludioxonil and fenhexamid or were left unsprayed. After 24 h the vinelets or bunches were dusted with dry conidia of Botrytis cinerea in a settling tower and incubated for 24 h at a high relative humidity (±93%). Following incubation, both the vinelets or bunches were divided into three groups. Vinelets and bunches of the one group were surface-sterilised, the others were left unsterile. Vinelets and bunches of one unsterile group were placed in dry chambers, kept for 14 days at 22°C with a 12 h photoperiod daily and monitored for symptom expression and the development of B. cinerea. Vinelets and bunches of the sterile group, and from one unsterile group were used for isolation. From each of these vinelets leaf blades, leaf petioles, shoots and inflorescences were removed. Sites used for isolation in bunch parts were rachises, laterals and pedicels, and sites on berries were the pedicel-end, cheek and style-end. The different parts and segments were placed in Petri dishes on Kerssies' B. cinerea selective medium, or on water agar medium supplemented with paraquat and incubated for 14 days at 22°C with a 12 h photoperiod daily. Infection and fungicide efficacy was determined by observing intact vinelets and bunches for symptom expression, and by estimating the amount of B. cinerea at the various sites on the vinelets and bunches with isolation studies. No symptoms of B. cinerea decay developed on sprayed and unsprayed vinelets that were kept in dry chambers during the 2 wk observation period. The isolation and incubation studies showed that the different fungicides were highly and nearly equally efficient in reducing superficial B. cinerea inoculum and latent infection. .In the case of leaf blades, which showed a high amount of B. cinerea on unsprayed vinelets under the two sterility regimes, decay was significantly reduced by each fungicide on both cultivars. This was not the case for the other parts, which yielded B. cinerea at low incidences under the two sterility regimes. The study with bunches showed that dry, airborne conidia, and the fungicide sprays, penetrated loose and tight clustered bunches from bloom to harvest and evenly landed on the various bunch parts. At full bloom, the amount of B. cinerea in unsprayed bunches was high on the laterals and pedicels, but low on the embryos. Unsprayed intact bunches at full bloom were highly susceptible to B. cinerea and developed symptoms of grey mould. The fungicides inhibited symptom expression at full bloom, but could not prevent infection. Unsprayed bunches inoculated at the other stages remained asymptomatic. The amount of B. cinerea was generally high in the rachises and laterals at pea size and bunch closure stages, and in the pedicel end of berries at harvest. Infection was constantly low in the berry cheek. The fungicides had a differential effect on infection at the various sites. In the case of rachises, the amount of B. cinerea was at each growth stage drastically reduced by each fungicide. In laterals, it was effectively reduced at pea size and bunch closure. However, at these two sites, significant differences were found between the fungicides in efficacy at stages when the amount of B. cinerea was high. This study showed that if these fungicides are applied properly to vine in commercial vineyards between budding and prebloom, during flowering, and at bunch closure, they should effectively prevent infection and symptom expression and thus the development of B. cinerea epiphytotics.<br>AFRIKAANSE OPSOMMING: INFEKSIE DEUR DROË, LUGGEDRAAGDE BOTRYTIS CINEREA KONIDIA EN DIE EFFEK VAN FUNGISlEDE OP VERSKILLENDE SETELS BINNE WINGERDTROSSE EN OP LOTE: Evaluering van fungisieddoeltreffendheid in kommersiële wingerde word beïnvloed deur die sporadiese voorkoms van Botrytis cinerea op verskeie posisies van wingerddele, verskille in trosstruktuur tydens trosontwikkeling, en die feit dat simptoomuitdrukking in lote en trosse deur die weerstandsaksie van die verskillende morfologiese dele van lote en trosse beheer word. In die natuur speel infeksie deur enkel konidia 'n prominente rol in die epidemiologie van B. cinerea van wingerd. Die doel van hierdie studie was om (i) infeksie en (ii) die effek van fungisiede op verskillende posisies op lote en trosse (tafeldruif kultivar Dauphine, wyndruif kultivar Merlot), wat met droë, luggedraagde konidia van B. cinerea geïnokuleer is, te bepaal. Lote, verkry vanaf steggies, en trosse versamel vanuit die wingerde tydens blom-, ertjiekorrel-, trostoemaak-, deurslaan- en oesstadium, is teen aanbevole dosisse met iprodione, pyrimethanil, cyprodinillfludioxonil of fenhexamid in 'n spuitkas bespuit, of is onbehandeld gelaat. Na 24 h is die lote en trosse met droë konidia van B. cinerea in 'n inokulasietoring geïnokuleer en daarna vir 24 h onder hoë humiditeit [±93% RH] geïnkubeer. Na inkubasie is die lote en trosse in drie groepe verdeel. Die een groep lote en trosse is oppervlakkig gesteriliseer om die patogeen op die oppervlakte te elimineer, en die ander twee groepe is onbehandeld gelaat. Die lote en trosse van een nie-steriele groep is vir 14 dae in droë voghokke by 22°C met 'n 12 uur daaglikse fotoperiode geplaas, en daagliks vir siekteuitdrukking en die ontwikkeling van B. cinerea gemonitor. Lote en trosse van die ander twee groepe is vir isolasiestudies gebruik. Vanaf elke loot is blaarskywe, blaarstele, internodes en ongeopende blomtrossies verwyder. Vanaftrosse is ragisse, laterale en korreisteie verwyder, en vanaf korrels is skilsegmente aangrensend aan die korrelsteel, die stempel-end, en die wang verwyder. Die dele en segmente is op B. cinerea selektiewe medium, en op paraquat medium in Petri bakkies geplaas en vir 14 dae by 22°C met 'n 12 uur daaglikse fotoperiode geïnkubeer. Infeksie en die fungisiedeffek is bepaal deur die intakte lote en trosse vir siekte- uitdrukking te monitor, en deur die hoeveelheid B. cinerea op verskeie posisies op lote en trosse te bepaal. Geen simptome het op enige posisie op bespuite en onbespuite lote, wat in droë hokke gehou is, ontwikkel nie. Die isolasie- en inkubasiestudies het getoon dat die verskillende fungisiede hoogs effektief op lote was, en inokulumvlakke van die patogeen doeltreffend verlaag het. In die geval van blaarskywe, wat hoë vlakke van B. cinerea op onbespuite steggies onder die twee steriliteitskondisies getoon het, is verrotting op beide kultivars betekenisvol deur die fungisiedes verlaag. Dit het egter nie vir die ander dele, waarop daar 'n lae voorkoms van B. cinerea onder die twee steriliteitskondisies was, gegeld me. Die studie met trosse het getoon dat droë, luggedraagde konidia en fungisiednewels beide oop en kompakte trosse vanaf blomstadium tot oes penetreer en eweredig op die verskillende dele land. Met blomstadium was die hoeveelheid B. cinerea in onbespuite trosse hoog op laterale en korrelstele, maar laag op die embrios. Onbespuite, intakte trosse was hoogs vatbaar vir B. cinerea by blomstadium en het simptome van vaalvrot ontwikkel. Die fungisiede het siekte-uitdrukking by blomstadium voorkom, maar kon nie infeksie voorkom me. Onbespuite trosse wat op ander stadia geïnokuleer is, het geen siekte-uitdrukking getoon me. Die hoeveelheid B. cinerea was hoër in die ragi, asook in laterale by ertjiekorrel- en trostoemaak stadium, en hoër in korreisteie by oesstadium. Infeksie was konstant laag in die korrelskil. Die fungisiede het 'n differensiële effek op infeksie by die verskillende posisies gehad. In die geval van ragi was die hoeveelheid B. cinerea drasties deur elke fungisied by alle groeistadia verlaag. In laterale was dit effektief by ertjiekorrel- en trostoemaakstadium verminder. By hierdie twee posisies waar die hoeveelheid B. cinerea hoog was, is daar egter betekenisvolle verskille in die doeltreffendheid van fungisiedes gevind. Hierdie studie toon dat as fungisiede behoorlik in kommersiële wingerde tussen botvorming en blomstadium, en tydens blom- en trostoemaakstadium toegedien word, infeksie en siekte-uitdrukking, en dus ook die epifitotiese ontwikkeling van B. cinerea, voorkom behoort te word.

Late blight, caused by Septoria apiicola Speg., is the most important disease affecting celery in Quebec. Biological control was investigated as an alternative to conventional chemical control of late blight. Two hundred and four bacterial isolates were collected from celery leaves, and muck and mineral soils of celery fields in south-western Quebec. Two experiments were conducted to screen the bacteria for antagonism toward Septoria apiicola: one on agar to test for inhibition of pycnidial formation, and the other on leaf disks to test for inhibition of germination of conidia. From these two experiments, 18 isolates were selected that prevented pycnidial formation in an inhibition zone $ ge$1.0 cm wide and reduced germination to below 30% of the control. These isolates were re-evaluated for inhibition of germination on leaf disks. A bacterial suspension (10$ sp7$ cells/ml) was incubated on leaf disks for 24 hours before a suspension of S. apiicola conidia (150,000 spores/ml) was applied, and the disks were incubated for 25 hours. Four isolates reduced germination to $ le$19% of the control. These isolates were tested on plants in a greenhouse. None of the isolates was able to reduce the number of late blight lesions compared to the control.

The false codling moth, Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae), is one of the major pests of citrus in South Africa, the others being mealybug, Mediterranean fruit fly, bollworm and some mites. Due to problems such as the expense of pesticides, insects evolving pesticide resistance (Hogsette 1999), chemical residue on the skin of export fruit and the negative impact of pesticides on the environment, it became necessary to find alternative methods for pest control (Viggiani 2000). Agathis bishopi (Nixon) (Hymenoptera: Braconidae), a larval parasitoid of false codling moth known only from the Sundays River Valley area (Sishuba 2003), offers a means of control for the pest. A total of 11 389 navel oranges were collected from various orchards in the Addo/Kirkwood area, and false codling moth larvae infested 36.09% of the fruit. A single parasitoid species, A. bishopi, was reared from these larvae. In 2006 the highest parasitism rate, 11.43%, was recorded in May and in 2007, the highest parasitism rate, 13.27%, was in April. Agathis bishopi parasitizes larvae in instars 2 and 3, possibly due to the accessibility of these younger instars to the female parasitoid and possibly due to the length of the life cycle of this koinobiont. Second instar hosts yielded the highest number of parasitoids, and there was no emergence of parasitoids from fifth instar larvae. Females of A. bishopi live for 18.5 days (n = 20; S.E. = 3.1) and males for 8.25 days (n = 20; S.E. = 1.23). Females produce an average of 23 offspring in a lifetime, while female false codling moths produce about 800 eggs each. A high number of parasitoids will be required per hectare to reduce the population of false codling moth. Captive rearing of A. bishopi proved difficult due to viral and fungal contamination. Agathis bishopi has potential for use in an integrated pest management programme once the hurdle of mass-rearing has been overcome.

Septoria blight of celery (Apium graveolens var. dulce), induced by Septoria apiicola Speg., is a destructive disease that requires fungicide applications for its management. A reliable method of inoculum production was established. Best sporulation was obtained on celery agar (CA) at a predicted optimum temperature of 22.4$ sp circ$C. A moderate inoculum density within a range of 17-35 conidia/cm$ sp2$ of leaf surface provided non-coalescing lesions necessary for rapid enumeration. Initial blight incidence thresholds of about 0.6 and 0.5% to time the first fungicide application were established based on the proportion of maximum plant weight equivalent to the cost of one fungicide application calculated plant weight and petiole number per plant, respectively, in the summers of 1990 and 1991. The IBI levels of 0 and 2% to time the first fungicide application provided non-significant results in terms of the final amount of blight and yield. (Abstract shortened by UMI.)

Confirm, Success and Proclaim were evaluated for lepidopterous insect control in lettuce with and without the addition of Mustang 1.5EW. Success and Proclaim used alone were highly efficacious toward cabbage looper and Heliothinae and did not appear to benefit greatly from the addition of Mustang. However, Confirm's activity towards Heliothinae was significantly improved by the addition of Mustang. Additionally, on large framed plants where coverage is difficult, Confirm benefitted from the addition of a pyrethroid for control of loopers.

Response of Direct-seeded Broccoli to Preemergence Oxyfluorfen and BAS 514 Preemergence treatments of oxyfluorfen at 0.14, 0.28, and 0.56 kg ai/ha caused a 40 to 99% reduction in stand of direct-seeded broccoli, while BAS 514 applied at the same rates caused no reduction in stand. All rates of oxyfluorfen completely controlled Venice mallow while only the 0.56 kg/ha rate of BAS 514 provided acceptable (>70%) control. Broccoli plants that survived preemergence applications of oxyfluorfen exhibited injury ranging from 32 to 97% 6 to 9 weeks after treatment. BAS 514 applied at 0.14, 0.28, 0.56 kg ai/ha provided excellent control of common ragweed with little or no visible crop injury. Common ragweed control with oxyfluorfen varied with soil moisture and organic matter. Applications of 0.28 and 0.56 kg/ha oxyfluorfen significantly reduced yield when compared to the cultivated check. Yields from plots treated with 0.14 kg/ha BAS 514 equalled or exceeded the cultivated check. Treatments of oxyfluorfen and BAS 514 to seeded broccoli caused approximately a one week delay in harvest. Nomenclature: oxyfluorfen, [2-chloro-1-(3-ethoxy-4- nitrophenoxy)-4-(trifluoromethyl)benzene]; BAS 514, quinclorac, (3,7-dicloro-8-quinoline-carboxylic acid); broccoli, [Brassica oleracea(L.) var botrytis]; common ragweed, (Ambrosia artemisifoliaL. #³ AMBEL); Venice mallow (Hibiscus trionum L. #3 HIBTR); Additional index words: Brassica oleracea (L.) botrytis, quinclorac. Weed Control and Crop Yield with Postemergence Applications of Oxyfluorfen, Pyridate, and BAS 514 to Direct-seeded Broccoli Postemergence applications of oxyfluorfen in field studies caused white chlorotic speckling on broccoli foliage. Plants treated with 0.07 kg/ha oxyfluorfen produced yields similar to the cultivated check. High rates of pyridate (4.03 kg/ha) caused yellow lesions on broccoli foliage. Total broccoli yields from plots treated with pyridate at 0.28 and 0.50 kg/ha were not significantly different from the cultivated check. BAS 514 caused moderate growth stunting to broccoli in field studies. At two out of four locations, injury increased with time. At these sites, total broccoli yield from plots treated with 0.28 and 0.56 kg/ ha BAS 514 was lower than the cultivated check. Total number of broccoli heads per hectare for plots treated with BAS 514 equaled or exceeded that of the cultivated check. Broccoli treated with BAS 514 plus the experimental adjuvant BAS 090 showed increased injury and decreased yield with increasing rates when compared to applications of BAS 514 alone. Broccoli developed foliar veinal chlorosis and whitish elongated heads following applications of BAS 514 plus BAS 090. High rates of BAS 514 (0.56 kg/ha) with or without BAS 090 reduced average head weight. Little to no stand reduction or delay in harvest was observed with any herbicide treatment. A two fold safety margin for herbicide rate was observed for oxyfluorfen at 0.07 kg/ha, pyridate at 0.50 kg/ha and BAS 514 at 0.14 kg/ha. In greenhouse studies, oxyfluorfen at 0.28 kg/ha provided >90% control of common ragweed and common lambsquarters, and 77% control of common pigweed. Pyridate at 2.02 kg/ha provided excellent control of horseweed, and greater than 70% control of prickly sida, common ragweed and common lambsquarters. All rates of pyridate controlled common pigweed. Applications of BAS 514 at 0.14 to 0.56 kg/ha provided 88 to 100% control of horseweed and 76 to 82% control of common ragweed. Greater than 90% control of large crabgrass was observed with BAS 514 plus BAS 090 at 0.28 kg/ha. Nomenclature: oxyfluorfen, 2-chloro-1-(3-ethoxy-4-nitrophenoxy)-4-(trifluoromethyl) benzene; pyridate 6-chloro-3-phenylpyridazine-4-yl-S-octyl carbonothionate; BAS 514, quinclorac, 3,7-dichloro-8-quinoline-carboxylic acid; broccoli Brassica oleracea L. var. botrytis; common ragweed Ambrosia artemisiifolia L. #3 AMBEL; redroot pigweed Amaranthus retroflesus L. #AMARE; common lambsquarters Chenopodium album L. #CHEAL; Prickly sida Sida spinosa L. #SIDSP; Horseweed Conyza canadensis L. #ERICA; Large crabgrass Digitaria sanguinalis L. #DIGSA; Additional index words. quinclorac, BAS 090, Brassica oleracea L. botrytis.<br>Master of Science

Thesis (MScAgric)--University of Stellenbosch, 2006.<br>ENGLISH ABSTRACT: Phaeomoniella chlamydospora, Eutypa lata, Phomopsis, Phaeoacremonium, and Botryosphaeria spp. are important trunk disease pathogens that cause premature decline and dieback of grapevine. Previous research has focused primarily on wine grapes and the incidence and symptomatology of these pathogens on table grapes were largely unknown. A survey was therefore conducted to determine the status and distribution of these pathogens and associated symptoms in climatically diverse table grape growing regions. Fifteen farms were identified in the winter rainfall (De Doorns, Paarl and Trawal) and summer rainfall (Upington and Groblersdal) areas. Samples were taken in July and August 2004 from Dan-ben-Hannah vineyards that were 8 years and older. Distal ends of arms were removed from 20 randomly selected plants in each vineyard. These sections were dissected and isolations were made from each of the various symptom types observed: brown or black vascular streaking, brown internal necrosis, wedge-shaped necrosis, watery necrosis, esca-like brown and yellow soft wood rot, as well as asymptomatic wood. Fungal isolates were identified using molecular and morphological techniques. Pa. chlamydospora was most frequently isolated (46.0%), followed by Phaeoacremonium aleophilum (10.0%), Phomopsis viticola (3.0%), Botryosphaeria obtusa (3.0%), B. rhodina (2.2%), B. parva (2.0%), Fusicoccum vitifusiforme (0.6%), B. australis, B. dothidea and an undescribed Diplodia sp. (0.2% each), while E. lata was not found. Most of these pathogens were isolated from a variety of symptom types, indicating that disease diagnosis can not be based on symptomatology alone. Pa. chlamydospora was isolated from all areas sampled, although most frequently from the winter rainfall region. Pm. aleophilum was found predominantly in Paarl, while P. viticola only occurred in this area. Although B. obtusa was not isolated from samples taken in De Doorns and Groblersdal, it was the most commonly isolated Botryosphaeria sp., being isolated from Upington, Paarl and Trawal. B. rhodina occurred only in Groblersdal and B. parva in Paarl, Trawal and Groblersdal, while B. australis was isolated from Paarl only. The rest of the isolates (33%) consisted of sterile cultures, Exochalara, Cephalosporium, Wangiella, Scytalidium, Penicillium spp. and two unidentified basidiomycetes, which were isolated from five samples with yellow esca-like symptoms from the Paarl area. These findings clearly illustrate that grapevine trunk diseases are caused by a complex of fungal pathogens, which has serious implications for disease diagnosis and management. Protection of wounds against infection by any of these trunk disease pathogens is the most efficient and cost-effective means to prevent grapevine trunk diseases. However, previous research on the effectiveness of chemical pruning wound protectants has mostly focused on the control of Eutypa dieback only. Fungicide sensitivity studies have been conducted for Pa. chlamydospora, P. viticola and Eutypa lata, but no such studies have been conducted for the pathogenic Botryosphaeria species from grapevine in South Africa. Ten fungicides were therefore tested in vitro for their efficacy on mycelial inhibition of the four most common and/or pathogenic Botryosphaeria species in South Africa, B. australis, B. obtusa, B. parva and B. rhodina. Iprodione, pyrimethanil, copper ammonium acetate, kresoxim-methyl and boscalid were ineffective in inhibiting the mycelial growth at the highest concentration tested (5 μg/ml; 20 μg/ml for copper ammonium acetate). Benomyl, tebuconazole, prochloraz manganese chloride and flusilazole were the most effective fungicides with EC50 values for the different species ranging from 0.36-0.55, 0.07-0.17, 0.07-1.15 and 0.04-0.36 μg/ml, respectively. These fungicides, except prochloraz manganese chloride, are registered on grapes in South Africa and were also reported to be effective against Pa. chlamydospora, P. viticola and E. lata. Results from bioassays on 1-year-old Chenin Blanc grapevine shoots indicated that benomyl, tebuconazole and prochloraz manganese chloride were most effective in limiting lesion length in pruning wounds that were inoculated with the Botryosphaeria spp after fungicide treatment. The bioassay findings were, however, inconclusive due to low and varied re-isolation data of the inoculated lesions. Benomyl, tebuconazole, prochloraz manganese chloride and flusilazole can nonetheless be identified as fungicides to be evaluated as pruning wound protectants in additional bioassays and vineyard trials against Botryosphaeria spp. as well as the other grapevine trunk disease pathogens.<br>AFRIKAANSE OPSOMMING: Phaeomoniella chlamydospora, Eutypa lata, Phomopsis, Phaeoacremonium, en Botryosphaeria spesies is die mees belangrikste stamsiekte patogene wat agteruitgang en vroeë terugsterwing van wingerd veroorsaak. Voorafgaande navorsing het hoofsaaklik gefokus op wyndruiwe en die voorkoms en simptomatologie van hierdie patogene op tafeldruiwe is dus grootliks onbekend. ‘n Opname is gevolglik gedoen in verskillende klimaaatsareas waar tafeldruiwe verbou word om die voorkoms en verspreiding, asook die simptome geassosieer met hierdie patogene, te bepaal. Vyftien plase is geïdentifiseer in die winter- (De Doorns, Paarl en Trawal) en somer-reënval (Upington en Groblersdal) streke. Wingerde (8 jaar en ouer) met die kultivar Dan-ben-Hannah is gekies vir opname en monsters is gedurende Julie en Augustus 2004 geneem. Die distale deel van ‘n arm is verwyder vanaf 20 lukraak gekose plante in elke wingerd. Hierdie dele is ontleed en isolasies is gemaak vanuit elke simptoomtipe wat beskryf is, naamlik bruin en swart vaskulêre verkleuring, bruin interne nekrose, wig-vormige nekrose, waterige nekrose, esca-geassosieerde bruin en geel sagte houtverrotting en asimptomatiese hout. Identifikasie van die swamagtige isolate is gedoen op grond van morfologiese eienskappe en molekulêre tegnieke. Pa. chlamydospora is die meeste geïsoleer (46.0%), gevolg deur Phaeoacremonium aleophilum (10.0%), Phomopsis viticola (3.0%), Botryosphaeria obtusa (3.0%), B. rhodina (2.2%), B. parva (2.0%), Fusicoccum vitifusiforme (0.6%), B. australis, B. dothidea en ‘n onbeskryfde Diplodia sp. (0.2% elk), terwyl E. lata nie geïsoleer is nie. Hierdie patogene is elk geïsoleer vanuit ‘n verskeidenheid simptoomtipes, wat daarop dui dat siektediagnose nie alleenlik op simptomatologie gebaseer kan word nie. Pa. chlamydospora is geïsoleer vanuit al die gebiede, alhoewel die patogeen opmerklik meer voorgekom het in die winter-reënval area. Pm. aleophilum het hoofsaaklik voorgekom in Paarl, terwyl P. viticola slegs in hierdie area voorgekom het. Alhoewel B. obtusa nie voorgekom het in die De Doorns en Groblersdal areas nie, was dit die mees algemeen geïsoleerde Botryosphaeria sp. en het in Upington, Paarl en Trawal voorgekom. B. rhodina het slegs in Groblersdal voorgekom, B. parva in Paarl, Groblersdal en Trawal en B. australis het slegs in Paarl voorgekom. Die res van die isolate (33%) het bestaan uit steriele kulture, Exochalara, Cephalosporium, Wangiella, Scytalidium, en Penicillium spesies asook twee onbekende basidiomycete isolate, geïsoleer vanuit vyf monsters met geel eska-geassosieerde simptome vanuit die Paarl area. Hierdie resultate illustreer dus die feit dat wingerdstamsiektes deur ‘n kompleks van swampatogene veroorsaak word, wat belangrike implikasies het vir die bestuur en diagnose van hierdie siektes. Wondbeskerming teen infeksie van enige van hierdie stamsiekte patogene is die mees doeltreffende en koste-effektiewe manier om wingerdstamsiektes te voorkom. Vorige navorsing aangaande die effektiwiteit van chemiese wondbeskermingsmiddels het egter slegs gefokus op die beheer van Eutypa terugsterwing. In vitro swamdoder sensitiwiteitstoetse is gedoen vir Pa. chlamydospora, P. viticola en Eutypa lata, maar geen studies is al gedoen ten opsigte van die patogeniese Botryosphaeria spesies op wingerd in Suid-Afrika nie. Tien swamdoders is dus getoets vir inhibisie van in vitro miseliumgroei van die vier mees algemene en/of patogeniese Botryosphaeria spesies wat in Suid-Afrika voorkom, naamlik B. australis, B. obtusa, B. parva en B. rhodina. Iprodione, pyrimethanil, koper ammonium asetaat, kresoxim-metiel en boscalid was oneffektief by die hoogste konsentrasies getoets (5 μg/ml; 20 μg/ml vir koper ammonium asetaat). Benomyl, tebuconasool, prochloraz mangaan chloried en flusilasool was die mees effektiewe swamdoders met EC50 waardes tussen 0.36-0.55, 0.07-0.17, 0.07-1.15 en 0.04-0.36 μg/ml, onderskeidelik vir die verskillende spesies. Hierdie fungisiedes, behalwe prochloraz mangaan chloried, is geregistreer op druiwe in Suid-Afrika en is ook effektief gevind teenoor Pa. chlamydospora, P. viticola en E. lata. Resultate van biotoetse op 1-jaar-oue Chenin Blanc wingerd lote het getoon dat benomyl, tebuconasool en prochloraz mangaan chloried die effektiefste was om die lengte van letsels in snoeiwonde, geinokuleer met Botryosphaeria spesies na die aanwending van swamdoder behandelings, te verminder. Die bevindinge was egter onbeslis as gevolg van die lae en variërende her-isolerings data. Benomyl, tebuconasool, prochloraz mangaan chloried en flusilasool kan egter geïdentifiseer word as swamdoders wat verder geevalueer kan word as snoeiwond beskermingsmiddels teen Botryosphaeria spesies asook ander wingerd stamsiekte patogene in verdere biotoetse en wingerdproewe.

O mofo cinzento é causado pelo fungo Botrytis cinerea (Pers.: Fr.), causador de grandes perdas econômicas em várias culturas. Este patógeno é de difícil controle, devido à ampla gama de hospedeiros, sua atividade saprófita e por formar estruturas de resistência (escleródios). Uma das formas de controle deste patógeno é a utilização de produtos químicos, no entanto, principalmente na pós-colheita de frutos, a tolerância por resíduos químicos nos alimentos, é cada vez menos desejável do ponto de vista ecológico e de saúde pública. Os tratamentos alternativos, como a utilização de extratos de plantas, vêm sendo pesquisados na fitopatologia, para reduzir o uso dos fungicidas sintéticos. A canola (Brassica napus) é uma planta que possui compostos biocidas, com potencial de controle de pragas e doenças. Este trabalho teve como objetivos avaliar o efeito de diferentes extratos da canola (alcoólico, macerado, aquoso sem tempo de reserva e infusão) no controle de Botrytis cinerea in vitro e em pós-colheita de morangos. Foram realizados dois experimentos in vitro, sendo um para avaliar o crescimento micelial e outro a germinação de conídios. O delineamento experimental para os dois experimentos foi inteiramente casualizado, em esquema fatorial 4 x 5, sendo o fator modos de extração de extratos e concentrações (0, 3, 6, 9 e 12%), em 4 repetições. No ensaio de crescimento micelial, a unidade experimental foi uma placa de Petri, e um tubo de ensaio no teste de germinação de conídios. Em pós-colheita foram testados os quatro tipos de extratos na concentração de 16%. O delineamento foi inteiramente casualizado, com 4 repetições por tratamento, sendo a parcela composta por 10 frutos/bandeja. Os parâmetros físico-quimicos avaliados foram podridões, perda de massa, firmeza de polpa e acidez titulável. As análises bioquímicas avaliadas foram proteínas totais, antocianinas e flavonóides, e a atividade da enzima fenilalanina amônia-liase (FAL) e peroxidases. Os resultados obtidos permitiram concluir que houve redução do crescimento micelial e da germinação de conídios, em função das concentrações, mas não ocorreram diferenças entre os extratos. A maior eficiência dos extratos ocorreu na concentração de 8,31%, na avaliação com 96horas. Para a germinação o comportamento foi linear decrescente, ou seja, o aumento das concentrações influenciou na menor germinação dos conídios. Os extratos: alcoólico, maceração e infusão reduziram as podridões causadas por B. cinerea em pós-colheita de morangos. Os extratos atuaram na alteração do teor de acidez dos frutos, e no comportamento das peroxidases, mas não apresentaram efeito sobre sólidos solúveis totais (SST), firmeza de polpa, perda de massa, antocianinas, flavonoides,e atividade da FAL. Os resultados obtidos neste trabalho, comprovam o potencial da canola no controle do mofo cinzento em pós-colheita de morangos, bem como do fungo Botrytis cinerea.<br>The gray mold is caused by the fungus Botrytis cinerea (Pers.: Fr.), causer of great economic losses in various cultures. This pathogen is difficult to control, due to the wide range of hosts, its saprophytic activity and by forming structures of resistance (esclerodios). One of the ways to control this pathogen is the use of chemical products, however, mainly in post-harvest fruit, tolerance by chemical residues in foods is increasingly less desirable from an ecological and public health point of view. The alternative treatments, such as the use of plant extracts, are being probed in phytopathology, to reduce the use of synthetic fungicides. The canola (Brassica napus) is a plant that has biocidal compounds, with potential to control pests and diseases. This study aimed to evaluate the effect of different extracts of canola (alcoholic, macerate, aqueous without reserve time and infusion) in the control of Botrytis cinerea in vitro and in post-harvest of strawberries. Two experiments were conducted in vitro, being one to evaluate the mycelial growth and another the germination of conidia. The experimental design for the two experiments was completely randomized, in a 4 x 5 factorial scheme, and the modes of extraction of extracts and concentrations (0, 3, 6, 9 and 12 %), in 4 repetitions. In the trial mycelial growth, the experimental unit was a Petri dish, and a test tube in germination test of conidia. In post-harvest were tested four types of extracts at a concentration of 16 %. The experimental design was completely randomized, with 4 repetitions per treatment, being the plot composed by10 fruits/tray. The physico-chemical parameters evaluated were rotting, weight loss, firmness and titratable acidity. The biochemical analyzes were evaluated total proteins, anthocyanins and flavonoids, and the activity of the enzyme phenylalanine ammonia-lyase (PAL) and peroxidases. The results obtained allowed us to conclude that there was a reduction of mycelial growth and germination of conidia, as a function of the concentrations, but there were no differences between the extracts. The greater efficiency of extracts occurred at a concentration of 8.31 %, in evaluation with 96 hours. For germination, the behavior was decreasing linear, in other words, the increase of concentrations influenced the lower germination of conidia. The extracts: alcoholic, maceration and infusion reduced the rot caused by B. cinerea in post-harvest of strawberries. The extracts operate in fruits acidity modification, and in the behavior of peroxidases, but they had no effect on total soluble solids (TSS), pulp firmness, weight loss, anthocyanins, flavonoids,and activity of the PAL. The results obtained in this work, have demonstrate the potential of canola in the control of gray mold on post-harvest of strawberries, as well as the fungus Botrytis cinerea.

Rhizoctonia solani and Pythium spp. are aggressive soil-borne fungal pathogens responsible for seed rot and seedling damping-off of many crops. With increased environmental and public concern over the use of chemicals, biological control of these diseases has been attracting more attention. However, success with this strategy depends on the development of effective antagonists, which requires repeated in vitro and in vivo tests. Bacillus spp. were isolated from a soil sample obtained from a field where sorghum and tef had been grown for at least two years. Potential Bacillus isolates were screened for their ability to inhibit in vitro growth of R. solani and Pythium sp. Among 80 isolates tested, endospore forming Bacillus spp. H44 and H51 gave highest antifungal activity against the two test-pathogens in three consecutive tests. Results demonstrated that both H44 and H51 have potential as biocontrol agents against diseases caused by these two pathogenic fungi. The interaction between three isolates of Trichoderma (T. harzianum Eco-T, Trichoderma spp. SY3 and SY4) and Pythium sp. were investigated using in vitro bioassays together with environmental scanning electron microscopy (ESEM). Visual observation on the dual culture tests revealed that hyphal growth of Pythium was inhibited by these antagonists soon after contact between the two organisms within 3-4 days of incubation. The ESEM investigations showed that all three isolates of Trichoderma grew toward the pathogen, attached firmly, coiled around and penetrated the hyphae of the pathogen, leading to the collapse and disintegration of the host's cell wall. Degradation of the host cell wall was postulated as being due to the production of lytic enzymes. Based on these observations, antibiosis (only by Eco-T) and mycoparasitism (by all three isolates) were the mechanisms of action by which in vitro growth of Pythium sp. was suppressed by these Trichoderma isolates. The reduction of seedling diseases caused by R. solani and a pythium sp. were evaluated by applying the antagonists as seed coating and drenching antagonistic Bacillus spp. (B81, H44 and H51) and Trichoderma (T. harzianum Eco-T and Trichoderma spp. SY3 and SY4). On both crops, R. solani and Pythium sp. affected stand and growth of seedlings severely. With the exceptions of H51, applications all of isoltes to seeds reduced damping-off caused by R. solani in both crops. Application of Eco-T, H44 and SY3 to sorghum controlled R. solani and Pythium sp. effectively by yielding similar results to that of Previcur®. On tef, biological treatments with Eco-T and SY4 reduced seedling damping-off caused by R. solani and Pythium sp., respectively, by providing seedling results similar to the standard fungicides, Benlate® and Previcur®. Most other treatments gave substantial control of the two pathogens on tef. Overall, Bacillus sp. H44 and T harzianum Eco-T were the best biocontrol agents from their respective groups in reducing damping-off by the two pathogens. In all instances, effects of application method on performance of biocontrol agents and adhesive on emergence and growth of seedlings were not significant. A field trial was conducted at Ukulinga Research Farm at the University of Natal, Pietermaritzburg, South Africa, to determine efficacy of biological and chemical treatments on growth promotion and reduction of damping-off incited by R. solani and Pythium sp., and to evaluate the effects of a seed coating material, carboxymethyl cellulose (CMC), on seedling emergence and disease incidence. Seeds of sorghum and tef were treated with suspensions of antagonistic Bacillus H44 or T harzianum Eco-T, or sprayed with fungicides, Benlate® or Previcur®. Application of Benlate® and Previcur® during planting significantly increased the final stand and growth of sorghum seedlings. Seed treatments with both H44 and Eco-T substantially controlled damping-off caused by Pythium, resulting in greater dry weights of seedlings than the standard fungicide. However, they had negative effects when they were tested for their growth stimulation and control of R. solani. The CMC had no significant effect on germination and disease levels. These results showed that these antagonists can be used as biocontrol agents against Pythium sp. However, repeated trials and better understanding of the interactions among the antagonists, the pathogens, the crop and their environment are needed to enhance control efficiency and growth promotion of these antagonists. Some of these biocontrol agents used in this study have the potential to diseases caused by R. solani and Pythium sp. However, a thorough understanding of the host, pathogen, the antagonist and the environment and the interactions among each other is needed for successful disease control using these antagonists.<br>Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2003.

Cephalosporium stripe of wheat (Triticum aestivum), caused by the soilborne fungus Cephalosporium gramineum, results in significant yield reductions in dryland winter wheat crops of the U.S. Pacific Northwest. The development of resistant cultivars offers the best hope for disease control. Breeding for resistance is hampered by the long trial times inherent in screening adult plants, and by cultivar x environment interactions in field tests. The principal objective of this research was to develop and test a procedure for screening wheat seedlings in controlled environments for resistance to Cephalosporium stripe. Wheat seedlings were raised hydroponically in growth chambers, and the fungus was increased in large fermentation tanks. The seedlings were inoculated at about 12 days post-germination. Disease severity was assessed approximately seven days later using a chlorophyll meter to measure the symptoms of chlorosis and striping. In three trials, five soft white cultivars from the Pacific Northwest and four hard red cultivars from the Southern Great Plains with known levels of field resistance were tested with a Pacific Northwest fungal isolate. With one exception, chlorophyll readings ordered the cultivars appropriately, with moderately resistant cultivars ranking above susceptible cultivars. Three other moderately resistant cultivars from the Pacific Northwest also appeared in one or two trials, and were ranked properly by chlorophyll level. Chlorophyll levels of uninoculated plants were assayed to determine if differences in chlorophyll content were innate in the cultivars. The chlorophyll levels of uninoculated and inoculated seedling treatments were only significantly correlated when the cultivar Madsen, which ranks high both in resistance and in chlorophyll content, was included. In adult plants, flag-leaf chlorophyll level corresponded to intensity of Cephalosporium stripe symptoms where disease was present, and was independent of known field resistance in undiseased cultivars. The seedling screening technique was used to investigate pathogenic variability in C. gramineum. In two experiments, a total of eight cultivars from the Pacific Northwest and the Southern Great Plains were tested with three fungal isolates from each region. No evidence of virulence/vertical resistance was found. There was also no significant adaptation of isolates to greater virulence on cultivars from the same region.<br>Graduation date: 1998

With an increasing realization that many agrochemicals are hazardous to animals and humans, came the desire to replace these chemical agents with biological approaches that are more friendly to the environment and human health. Microorganisms play an important role in plant disease control, as naturally occurring antagonists. Microorganisms may also have beneficial effects on plant development when applied to plant roots. Research efforts worldwide have recorded successes in biological control and growth stimulation on many crops, particularly when using members of the genera Bacillus and Trichoderma. Their use on citrus rootstock could be advantageous to nurserymen and growers in reducing the incidence of seedling mortality and increasing production. To achieve these objectives, laboratory and tunnel experiments were conducted to develop effective biocontrol agents for citrus seedlings and cuttings. Nineteen 0 ut 0 f 23 Trichoderma isolates tested in vitro against Phytophthora p arasitica sp showed antagonistic activity by hyperparasitism and four out of eight Bacillus isolates resulted in antagonism by forming inhibition zones. The positive in vitro activity of Trichoderma and Bacillus isolates on Phytophthora provided motivation step for further trials in the greenhouse to evaluate their biological control activity on citrus seedlings and cuttings. A greenhouse trial was carried out to evaluate the biological control potential of 23 Trichoderma isolates (drenched at 5 x 105 spores / rnI) and two Bacillus isolates (drenched at 1 X 106 or 1 X 108 colony forming units (CFU) / rnI) to suppress Phytophthora parasitica sp. of rough lemon (Citrus jambhirini Lush.) seedlings. Five isolates ofTrichoderma (AA12, AA5, Trichoderma harzianum (AA16), SY3F and Eco-T~ were highly effective in suppressing Phytophthora root rot, with AA12 providing the best control. The Bacillus isolates also suppressed the pathogen but were not as effective as the Trichoderma isolates. This trial was used to test for growth stimulation activity by some of the biocontrol agents. To verify these results, a further trial was carried out to evaluate growth stimulation capabilities in the absence of any pathogen. Trichoderma Isolates AA13 and AA17 caused no 111 change in seedling growth, while other Trichoderma and Bacillus isolates had an inhibitory effect on the seedling growth. This trial indicated that the biocontrol activity was affected by inoculum densities, and as a result in vitro sporulation capacity was evaluated. TrichodermaIsolate AA16 was the largest spore producer, followed by Eco-T®. Spore production was lowest from Trichoderma isolates AA4 and AA12. Growth stimulation responses of Trichoderma Isolates AA4, AA16, Eco-TID and SYN6 were further studied at four different doses (1 X 103, 1 X 104, 5 X 105 or 1 X 106 spores / ml) on rough lemon and trifoliate orange seedlings. Trifoliate oranges responded positively to 1 X 104 and 5 X 105 spores / ml of Eco-TID, but rough lemon responded negatively to all dosages of the Trichoderma isolates applied. This indicates that the inoculum density responses may be host specific. Higher population density of 1 X 106 spores / ml of all tested Trichoderma isolates had a stunting effect on seedling growth of both species. Based on t he positive results 0 f individual applications of some Trichoderma and Bacillus isolates, of the biological control agents on rough lemon seedlings against Phytophthora parasitica in an earlier greenhouse trial, their combined effect in the control of the pathogen was performed. Before carrying out a greenhouse trial, activities of the isolates to be combined were evaluated in vitro. This trial showed that Trichoderma Isolates AA16 and Eco-T®were compatible. Trichoderma isolates AA16 and Eco-T®were also found to be compatible with Bacillus Isolates B77, B81 and PHP. As a result, further in vivo trials were conducted. The tunnel trials were carried out as two separate experiments: In the first experiment, a combination of two Trichoderma Isolates A A 16 and Eco-T®was conducted assayed at 5 X 105 or 1 X 106 spores / ml, on rough lemon seedling, and cuttings and trifoliate orange and sour orange seedlings. A combination of Trichoderma isolate AA16 and Eco-T®at 5 X 105 spore / ml increased significantly the new flush biomass of rough lemon cuttings compared to AA16 alone, but was not different from Eco-TID alone. The combination of AA16 and Eco-T® achieved no change of biomass of rough lemon and trifoliate orange seedlings. The combination of AA16 and Eco-TID did not increase the root biomass of sour orange compared to AA16 or Eco-r® alone. The combination of AA16 and Eco-r® at higher doses (1 x 106 spores / ml) showed significantly better suppression of Phytophthora root rot of rough lemon cuttings but did not show disease suppression in all seedling species verities tested. In a second experiment, individual and combined effects of Trichoderma isolates (drenched at 5 X 105 spores / ml) with Bacillus isolate (drenched at 1 X 106 colony forming units (CFU) / ml) for suppression of Phytophthora root rot on rough lemon and trifoliate orange seedlings was performed. The combination of Trichoderma Isolate AA16 and Bacillus Isolate B81 increased root biomass on rough lemon seedlings compared to the combination of Trichoderma AAI6 or Bacillus PHP but was not significantly different to Trichoderma AA16 alone. Bacillus PHP combined with Trichoderma AA16 or singly had no effect on rough lemon seedlings. Combining Trichoderma Eco--r® and with Bacillus B8I or PHP did not increase biomass of rough lemon seedlings compared to Trichoderma Isolate Eco--r® alone. There was no statistically significant differences in the effects of the combinations of the Trichoderma and Bacillus isolates compared to their individual applications on the biomass of trifoliate oranges. This study established the antagonistic potential of several South African isolates of Trichoderma and Bacillus as a viable alternative to agrochemicals for controlling Phytophthora parasitica. The growth stimulation capabilities of Trichoderma isolates in terms of seedling development was also demonstrated.<br>Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

Aphids are a widespread family of plant pests, whose abilities to suppress shoot and root growth are well documented for many terrestrial plants. Only a few studies have been conducted on conifer aphids of the genus Cinara. Cinara pseudotsugae are known to attack Douglas-fir seedlings, an important crop in the Pacific Northwest. Douglas-fir are most susceptible to aphid damage as seedlings, especially in nurseries where conditions favor aphid outbreaks. A parasitoid wasp (Pauesia sp.) attacks C. pseudotsugae, and may be useful as a biological control agent. Studies of its natural history and host interactions are needed to assess its potential as a bio-control agent. This study examined the effects of an experimental range of aphid densities on the growth of total shoot and root volume and biomass and shoot morphology of Douglas-fir seedlings. Eighteen-week tests explored short term effects of different aphid feeding intensities in both the greenhouse and field. Long term effects were tested by exposing greenhouse seedlings to 16 months of aphid feeding. The ability of the plants to recover was tested by allowing one set of seedlings to grow aphid free for one year, after being exposed to aphid feeding for 18 weeks. The success rate of parasitoids over an 18 week period was compared to aphid density in both greenhouse and field tests. Increasing aphid destiny was significantly related to decreasing root and shoot dry weights in greenhouse tests. Growth suppression increased slightly during the second year of testing, regardless of whether or not aphid feeding continued. The results for root and shoot volumes were highly variable. However, root tissue density was significantly reduced after the second year of testing. Few shoot characteristics showed consistently significant aphid effects among the trials. Stem diameter and height decreased and needle density of new buds increased significantly with aphid feeding in most tests. Root and shoot growth of field plants did not show any significant aphid effects. Percentage of parasitoid success was independent of aphid density except at the lowest aphid densities. There was a block effect on parasitoid success in the field test, that may have been a result of varying environmental conditions. These results indicate that even short term aphid feeding can have long lasting effects on plant growth and structure. The effect on shoot and root growth was small, but there were no signs of recovery. The long term effects of the reduced root tissue density on Douglas-fir is unknown.<br>Graduation date: 1998

The increasing concerns about chemical pesticides that are environmentally hazardous and the continuous development of resistance by palhogens to chemical pesticides have led to this study. Many studies have shown that some Gram-negative bacteria, such as Pseudomonas flouresens, control plant diseases and promote plant growth. In this study Gram positive bacteria, Bacillus sp., were chosen because of their ability to produce endospores. Endospores can be used in stable, dry formulations. The advantage of using endospores is their ability to survive harsh conditions such as droughts and high temperatures, which give a long shelf life to the biological control agent. Bacillus isolates were recovered from the rhizosphere of 12 different crops, and were subsequently screened in vitro for their antimicrobial activity. Of 130 isolates, 87 exhibited antimicrobial activity against the test organisms: Rhizoctonia solani, Pythium sp., Phytophthora cinnamoni, Fusarium sp., and single representatives of Gram negative and Gram positive bacteria, namely, Erwinia carotovora and Staphylococcus aureus respectively. The Bacillus isolates B77, B81 and B69 inhibited all the test organisms investigated, which suggests that they produced broad spectrum antimicrobial compounds or more than one antimicrobial compound. Of the isolates that showed antimicrobial activity, 78 of them did not inhibit Trichoderma harzianum K D, which is a registered biological control agent; indicating their potential for combined application. Selected Bacillus isolates were tested for the biological control of R. solani under greenhouse conditions in wheat, cabbage, tomato, maize, and cucumber seedlings. Bacillus isolates were applied as seed treatments, and the inoculated seeds were planted in R. solani infested speedling trays. Shoot dry weight measurement of seedlings indicated that 12 out of 19 Bacillus isolates showed significantly different shoot dry weight in wheat whereas all the isolates tested in tomato and cucumber gave significantly different shoot dry weight. No significantly different shoot dry weight was obtained for maize or cabbage. Seed emergence findings indicated that none of the Bacillus isolates gave significantly different emergence percentage on wheat, cabbage, tomato, and maize but all of them showed significantly different emergence percentage on cucumber. The results indicate that both the pathogen and the biological control agents exhibited varying levels of specificity on each crop tested. The biological control potential of the best Bacillus isolates was tested on bean and maize crops in the field. Green bean and maize seeds were coated with the selected Bacillus isolates and then sown under field conditions. For each isolate, four replicate treatment plots were established, with and without a R. solani inoculum. Percentage emergence, plant survival levels to harvesting and yield of maize cobs and green beans pods were measured. For all parameters measured the positive and negative controls were not significantly different thereby rendering the results for the entire field study inconclusive. However, Bacillus isolates B77, BII, R5 and R7 improved green bean pod yield and Bacillus Isolate B8I increased maize yield, indicating their potentials as plant growth promoting rhizobacteria (PGPR).<br>Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2003.

Various Trichoderma and Bacillus spp. have been documented as being antagonistic to a wide range of soilborne plant pathogens, as well as being plant growth stimulants. Successes in biological control and plant growth promotion research has led to the development of various Trichoderma and Bacillus products, which are available commercially. This study was conducted to evaluate the effect of six Trichoderma spp. and three Bacillus spp. and their respective combinations, for the biological control of Rhizoctonia solani damping-off of cucumber and plant growth promotion of dry bean (Phaseolus vulgaris L.). In vivo biological control and growth promotion studies were carried out under greenhouse and shadehouse conditions with the use of seed treatment as the method of application. In vitro and in vivo screening was undertaken to select the best Trichoderma isolates from 20 Trichoderma isolated from composted soil. For in vitro screening, dual culture bioassays were undertaken and assessed for antagonisms/antibiosis using the Bell test ratings and a proposed Invasive Ability rating based on a scale of 1-4 for possible mycoparasitic/hyperparasitic activity. The isolates were further screened in vivo under greenhouse conditions for antagonistic activity against R. solani damping-off of cucumber (Cucumis sativus L.) cv. Ashley seedlings. The data generated from the in vivo greenhouse screening with cucumber plants were analysed and grouped according to performance of isolates using Ward‟s Cluster Analysis based on a four cluster solution to select the best isolates in vivo. Isolates exhibiting marked mycoparasitism of R. solani (during ultrastructural studies) viz, T. atroviride SY3A and T. harzianum SYN, were found to be the best biological control agents in vivo with 62.50 and 60.06% control of R. solani damping-off of cucumber respectively. The in vitro mode of action of the commercial Trichoderma product, Eco-T®, and Bacillus B69 and B81 suggested the production of antimicrobial substances active against R. solani. In vitro interaction studies on V8 tomato juice medium showed that the Trichoderma and Bacillus isolates did not antagonise each other, indicating the possibility of using the two organisms together for biological control and plant growth promotion studies. Greenhouse studies indicated that combined inoculation of T. atroviride SYN6 and Bacillus B69 gave the greatest plant growth promotion (43.0% over the uninoculated control) of bean seedlings in terms of seedling dry biomass. This was confirmed during in vivo rhizotron studies. However, results obtained from two successive bean yield trials in the greenhouse did not correlate with the seedling trials. Moreover, no increase in protein or fat content of bean seed for selected treatments was observed. In the biological control trials with cucumber seedlings, none of the Trichoderma and Bacillus combinations was better than single inoculations of Eco-T®, T. atroviride SY3A and T. harzianum SYN. Under nutrient limiting conditions, dry bean plants treated with single and dual inoculations of Trichoderma and Bacillus isolates exhibited a greater photosynthetic efficiency that the unfertilized control plants. Bacillus B77, under nutrient limiting conditions, caused 126.0% increase in dry biomass of bean seedlings after a 35-day period. Nitrogen concentrations significantly increased in leaves of plants treated with Trichoderma-Bacillus isolates. However, no significant differences in potassium and calcium concentrations were found. Integrated control (i.e. combining chemical and biological treatments) of R. solani damping-off of cucumber seedlings proved successful. In vitro bioassays with three Rizolex® concentrations, viz., 0.01g.l-1, 0.1g.l-1 and 0.25g.l-1 indicated that the selected Trichoderma isolates were partly sensitive to these concentrations whereas the Bacillus isolates were not at all affected. In a greenhouse trial, up to 86% control was achieved by integrating 0.1g.l-1 Rizolex® with T. harzianum SYN, which was comparable to the full strength Rizolex® (1g.l-1) application. Irrespective of either a single or dual inoculations of Trichoderma and/or Bacillus isolates used, improved percentage seedling survival as achieved with the integrated system, indicating a synergistic effect. The results presented in this thesis further reinforce the concept of biological control by Trichoderma and Bacillus spp. as an alternative disease control strategy. Furthermore, this thesis forms a basis for Trichoderma-Bacillus interaction studies and proposes that the two organisms could be used together to enhance biological control and plant growth promotion.<br>Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

碩士<br>國立臺灣大學<br>園藝學研究所<br>100<br>The Populus spp. in Taiwan encountered a great difficutlies of a vareity of diseases and pests in the cultivation. Recently, we have analyzed the survey based on the diseases and pests of Poplar sp.. We further identy those survey, and provide the the updated information of our diseases and pests’ investigation from our collected data. In addition, we select those rust and anthracnose that caused more serious hazards to proceed the screening test of control chemicals and essential oils. This study hopes to investigate the issue of diseases and pests of Popalr sp. in Taiwan as the basis of the prevention in the future. We hope that this study can come out with some solutions to solve the current problem of Populus spp. in Taiwan in its cultivation and enhance the production of Populus spp.’s planting quality while facing the industry’s issue. The investigated areas in this study included the National Taiwan University Highland Experimental Farm, Sihhu Township of Yunlin County, Dongshih Township of Yunlin County and Yuanchang Township of Yunlin County. The investigation involved the phenomenon and categorization of Populus spp. in Taiwan endangered of diseases and pests. We collect types of pests from the occurrence of diseases location and the hazards part of the Populus spp. Furthermore, those collections are being separated by its diseases-affecting leaves and roots to proceed the pathogenic isolation, purification and culture. Beside identify the pathogenic form, we also add the step to identify its molecular and inoculation to complete the Koch’s postulate. After we investigate the pests, we identify the pests that full of hazards are: Giant beetles, Zeuzera coffeae Niether, Jacobiasca formosana Paoli, Erthesina fullo Thunberg, Orgyia postica Walker, Clania preyeri Leech, Coptotermes gestroi Wasmann, Aleurodicus disperses hazards and Phyllocnistis citrella Staintain. The giant alternatus species that’s still under identification. On the other hand, we found Rust, Alternaria tenuis Nees, Ophiostoma quercus (Georgev.) Nannf, Anthracnose, Algal spot of poplar, Leaf spot, Sterm blight and Botrytis blight under the diseases identification based on its morphological characteristics, the separation of fungal tissue and molecular identification. We select 29 kinds of bactericides that’s easily pruchased on the market and 23 kinds of essential oils of plant’s leaves and woods in Taiwan to focus on the rust pathogen, Melampsora larici-populina (Kleb) and Colletotrichum gloeosporioides (Penz.) for the fungicides and essentail oils screening procedure. We test about the spre germination rate of rust pethogen, the growth inhibition rate and spore germination rate of anthrancnose pathogen mycelial growth all separetly. The pharmaceucal ones that has best control response reaction to the rust pathogen spore germinication inhibition are: Kresoxim-methyl, Boscalid + Kresoxim-methyl, Oxine-copper, Potassium hydrogen carbonate, Pyraclostrobin +Boscalid and essential oil which includes: Leaf Essential oil from Eucalyptus robusta, Leaf Essential oil from Melaleuca alternifolia, Tree Essential oil from Chamaecyparis obtuse, Tree Essential oil from Chamaecyparis formosensis, Leaf Essential oil from Cinnamomum osmophloeum, Leaf Essential oil from Cinnamomum zeylanicum and Leaf essential oil from Cinnamomum micranthum; and the ones that are best response to the anthrancnose spore germination inhibition are: Kresoxi m-methyl, Hymexazol, Fluazinam, Prochloraz, Mancozeb and essential oils included: Leaf Essential oil from Eucalyptus robusta, Leaf Essential oil from Melaleuca alternifolia, Tree Essential oil from Chamaecyparis obtuse, Tree Essential oil from Chamaecyparis formosensis, Leaf Essential oil from Cinnamomum osmophloeum, Leaf Essential oil from Cinnamomum zeylanicum, Leaf Essential oil from Juniperus chinensis for further screening of seedling control chemicals and essential oils test. There are 5 kinds of pharmaceutical and 8 kinds of essential oils all perform the best prevent reaction to the rust preventive effect. They are: Kresoxim-methyl, Tree Essential oil from Chamaecyparis formosensis and Leaf Essential oil from Cinnamomum osmophloeum that perform good prevent effect on rust inhibition. Fluazinam, Mancozeb, and Leaf Essential oil from Eucalyptus robusta can help with prevent the occurrence of anthrax. Fluazinam, Prochloraz, Mancozeb, Leaf Essential oil from Melaleuca alternifolia, Tree Essential oil from Chamaecyparis obtuse, Tree Essential oil from Chamaecyparis formosensis and Leaf Essential oil from Juniperus chinensis can help with prevent the anthrax inhibitory effect.