Tesis sobre el tema "Citrus Diseases and pests"

6 pp.<br>Low Desert Citrus Varieties; Irrigating Citrus Trees<br>Diagnosing Home Citrus Problems includes information on all the commonly encountered dooryard citrus problems encountered in Arizona. Problems/disorders are grouped into three catagories: problems with fruit, problems with leaves, and problems with stems, branches and entire tree. Symptoms, causes and control measures are given for each disorder.

Thesis (MScAgric (Horticulture))--University of Stellenbosch, 2010.<br>ENGLISH ABSTRACT: Creasing, also known as albedo breakdown, is a preharvest disorder that affects the albedo of citrus fruit causing creases on the surface of the fruit. It is a recurrent problem in Navel and Valencia oranges and can cause individual orchard losses which often exceed 50%. Although the contributing factors are known, the physiological basis of creasing development is unresolved and the current control measures do not prevent creasing satisfactory. Hence, better control measures and further understanding of the physiology of creasing development is required. The objective of this two-year study was to determine if the position of fruit in a tree, light and carbohydrate manipulation techniques, and albedo mineral nutrients influence creasing development. Furthermore, the most effective application timing of gibberellic acid (GA3) with the least negative effect on fruit rind colour development and the effectiveness of cytokinins, other products and different root biostimulants to reduce creasing incidence were evaluated. The position of fruit in the tree and light influenced the development of creasing and the distribution of mineral nutrients in the albedo. Creasing incidence was higher on the south side than on the north side of the tree and fruit from the inside sub-sectors had a greater creasing incidence compared to fruit from the outside sub-sectors. The shady part of outside fruit was more creased compared to the sunny part of the fruit and covering fruit with brown paper bags increased creasing severity. The light manipulation techniques used on the leaves and fruit increased the nitrogen (N), phosphorus (P), potassium (K) and manganese (Mn) concentrations in the albedo and differences in the albedo mineral nutrients amongst the sub-sectors evaluated were observed, but creasing severity or creasing incidence was not significantly correlated with the albedo mineral concentrations at harvest. Albedo mineral concentrations earlier in the season may play a role in creasing development, as creasing severity was significantly correlated with copper (Cu), K, and Mn concentrations in the albedo during stage II of fruit development. Creasing incidence and albedo mineral concentrations were not affected by any of the carbohydrate manipulation techniques used in this study. The incidence and severity of creasing was significantly reduced, with a minor negative effect on fruit rind colour development, by the application of GA3, from mid November to mid January. Localised fruit application of CPPU [N-(2-chloro-4-pyridyl)-N-phenylurea], MaxCel (6- Benzyladenine) and CPPU in combination with calcium after physiological fruit drop reduced the incidence and severity of creasing, although creasing incidence was not significantly different from the control. The application of Messenger®, AVG (aminoethoxyvinylglycine) and different root biostimulants did not reduce creasing incidence. The results showed that cytokinins could reduce creasing incidence and justify further studies on application and uptake efficiency. The use of different root biostimulants are not recommended, but it is suggested that treatment effects may be more pronounced over a longer period.<br>AFRIKAANSE OPSOMMING: Kraakskil is ‘n vooroes abnormalitiet wat die albedo van sitrusvrugte affekteer, deur krake op die oppervlak van vrugte te veroorsaak. Dit is ‘n algemene probleem in Navel en Valencia lemoene en kan boordverliese van tot 50% of soms hoër veroorsaak. Alhoewel die bydraende faktore bekend is, is die fisiologiese basis van kraakskil ontwikkeling onopgelos en die beskikbare beheermaatreëls is nie bevredigend nie. Dus, beter beheermaatreëls en ‘n beter begrip van die fisiologie van kraakskil ontwikkeling is nodig. Die doel van die twee-jaar studie was om te bepaal of die posisie van vrugte in ‘n boom, lig en koolhidraat manipulasie tegnieke en minerale elemente in die albedo, kraakskil ontwikkeling beïnvloed. Die mees effektiewe toedieningstyd van gibberelliensuur (GA3) sonder ‘n negatiewe effek op vrugkleur is bepaal en die effektiwiteit van sitokiniene, ander produkte en verskillende wortel biostimulante om kraakskil voorkoms te verminder, is geëvalueer. Die posisie van vrugte in ‘n boom en lig het kraakskil ontwikkeling en die verspreiding van minerale element in die albedo beïnvloed. Kraakskil voorkoms was hoër aan die suidekant van die boom as aan die noordekant en vrugte in die binnekant van die boom het ‘n groter kraakskil voorkoms as vrugte in die buitekant van die boom gehad. Die skadukant van buitevrugte het meer kraakskil gehad as die sonkant en die toemaak van vrugte met ‘n bruin papiersak het die graad van kraaksil verhoog. Die lig manipulasie tegnieke wat op die blare en vrugte gebruik is, het die stikstof (N), fosfaat (P), kalium (K) en mangaan (Mn) konsentasies in die albedo verhoog en verskille in die albedo minerale elemente tussen sub-sektore is waargeneem, maar betekenisvolle korrelasies is nie tussen die graad en voorkoms van kraakskil en die albedo minerale element konsentrasies by oestyd waargeneem nie. Albedo minerale element konsentrasies vroeër in die seisoen mag ‘n rol speel by kraakskil ontwikkeling, omdat die graad van kraakskil betekenisvol gekorreleer was met albedo koper (Cu), K, en Mn konsentrasies tydens fase II van vrugontwikkeling. Kraakskil voorkoms en albedo minerale element konsentrasies is nie deur enige van die koolhidraat manipulasie tegnieke geaffekteer nie. Die voorkoms en graad van kraakskil is betekenisvol verlaag, met ‘n geringe negatiewe effek op vrugkleur, deur die toediening van GA3 vanaf mid November tot mid Januarie. Gelokaliseerde vrugtoedienings van CPPU [N-(2-chloro-4-piridiel)-N-phenielureum], MaxCel (6- Bensieladenien) en CPPU saam met kalsium na fisiologiese vrugval het die voorkoms en graad van kraakskil verlaag, alhoewel kraakskil voorkoms nie betekenisvol van die kontrole verskil het nie. Die toediening van Messenger®, AVG (amino etoksievinielglisien) en veskillende wortel biostimulante het nie kraakskil voorkoms verlaag nie. Die resultate het getoon dat sitokiniene kraakskil voorkoms kan verlaag en verdere studies op die toediening en opname effektiwiteit word aanbeveel. Die gebruik van verskillende wortel biostimulante word nie aanbeveel nie, maar die effek behoort meer sigbaar te wees na ‘n langer periode van behandeling.

False codling moth is a known pest of economic importance to many cultivated crops in South Africa and Africa south of the Sahara, and is particularly severe on citrus. If the fruit is infested just before harvest the chances of detecting signs of infestation are very low. As a result, the risk of packaging infested fruit and exporting them as healthy fruit is high. It is therefore a priority to develop a post-harvest technique for detection of False codling moth in citrus fruit at different levels of infestation in order to reduce phytosanitary risk. Compounds released and detected were indicative of infestation and were not insect produced but naturally produced fruit volatiles emitted at higher levels as a result of the insect within the fruit. Five major volatile compounds of interest were released by the infested oranges. These major volatile compounds include D-limonene, 3,7-dimethyl-1,3,6-octatriene, (E)-4,8-dimethyl-1,3,7-nonatriene, caryophyllene and naphthalene. Limonene was one of the most abundant volatile compounds released by the infested citrus fruit. Naphthalene, which is possibly produced due to larval feeding and development within the fruit maintained higher concentrations than controls throughout the infestation within the fruit. Naphthalene would be a good indicator of False codling moth infestation, however, not primarily for early infestation detection. A significantly higher concentration of D-limonene, 3,7-dimethyl-1,3,6-octatriene, (E)-4,8-dimethyl-1,3,7-nonatriene and naphthalene was detected using the SEP over the SPME technique. The application of an SPME procedure and the utilization of this method for detection of volatiles present in the headspace of intact infested fruit are evaluated and the possible volatile compounds diagnostic of Thaumatotibia leucotreta infestation of orange fruit and differences in volatile compound response in different orange varieties is discussed.

Includes bibliographical references (leaves 152-164) Microscopy investigations into the oil glands, localisation of the rind oils and the development of oleocellosis have been carried out in Washington navel orange (Citrus sinensis [L.] Osbeck). Aims to develop an improved understanding of the physiological basis of the rind disorder.

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.

Mealybugs (Planococcus citri) and thrips (Scirtothrips aurantii) are common and extremely damaging citrus crop pests which have proven difficult to control via conventional methods, such as chemical pesticides and insect growth regulators. The objective of this study was to determine the efficacy of entomopathogenic fungi against these pests in laboratory bioassays. Isolates of Metarhizium anisopliae and Beauveria bassiana from citrus orchards in the Eastern Cape, South Africa were maintained on Sabouraud Dextrose 4% Agar supplemented with Dodine, chloramphenicol and rifampicin at 25°C. Infectivity of the fungal isolates was initially assessed using 5th instar false codling moth, Thaumatotibia leucotreta, larvae. Mealybug bioassays were performed in 24 well plates using 1 x 107 ml-1 conidial suspensions and kept at 26°C for 5 days with a photoperiod of 12 L:12 D. A Beauveria commercial product and an un-inoculated control were also screened for comparison. Isolates GAR 17 B3 (B. bassiana) and FCM AR 23 B3 (M. anisopliae) both resulted in 67.5% mealybug crawler mortality and GB AR 23 13 3 (B. bassiana) resulted in 64% crawler mortality. These 3 isolates were further tested in dose-dependent assays. Probit analyses were conducted on the dose-dependent assays data using PROBAN to determine LC₅₀ values. For both the mealybug adult and crawlers FCM AR 23 B3 required the lowest concentration to achieve LC₅₀ at 4.96 x 10⁶ conidia ml-1 and 5.29 x 10⁵ conidia ml-1, respectively. Bioassays on adult thrips were conducted in munger cells with leaf buds inoculated with the conidial suspensions. Isolate GAR 17 B3 had the highest mortality rate at 70% on thrips while FCM AR 23 B3 resulted in 60% mortality. Identification of the isolates, FCM AR 23 B3, GAR 17 B3 and GB AR 23 13 3, were confirmed to be correct using both microscopic and molecularly techniques. ITS sequences were compared to other sequences from GenBank and confirmed phylogenetically using MEGA6. Mealybug infection was investigated using scanning electron microscopy, mycosis was confirmed but the infection process could not be followed due to the extensive waxy cuticle. These results indicate that there is potential for the isolates FCM AR 23 B3 and GAR 17 B3 to be developed as biological control agents for the control of citrus mealybug and thrips. Further research would be required to determine their ability to perform under field conditions.

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 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.

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 South African Citrus Thrips, Scirtothrips aurantii Faure (Thysanoptera: Thripidae) has been a serious pest of the citrus industry of Southern Africa for over 70 years. It is indigenous to Africa and has no recorded parasitoids and, in most citrus-growing regions, predators are not economically effective. Firstly, in this study, the general ecology of thrips was reviewed along with the recorded history of S. aurantii and its control. Host plant relationships of S. aurantii were then examined and wild hosts were not found to be important in promoting citrus thrips outbreaks in the orchard after flowering. In addition, Scirtothrips dorsalis Hood, which attacks citrus in Asia, was collected for the first time in Africa, but from Castor Oil plant. It is therefore a potential pest of citrus here. Notes on its appearance compared to that of S. aurantii were prepared. Weekly sampling of S. aurantii adults was carried out from June 1984 to May 1990. Population fluctuations were then correlated with phenology of the citrus trees and the direct and indirect effects of weather. Temperature and rainfall were not found to be significantly directly correlated with thrips numbers recorded. However, the indirect effects of rainfall were important in promoting a large winter thrips population in certain years. Relatively high rainfall during March and April compared to that of the previous January and February stimulated atypical flushing of the citrus trees during autumn and the setting of out-of-season fruit. S. aurantii then exploited this unusual food source and high numbers were subsequently recorded in the following winter; as well as in spring. Thus the mild winter climate alone could not suppress thrips numbers at Letaba. Dispersal/Emergence traps, which are used in the U.S.A. for the monitoring of Scirtothrips citri, were evaluated over 24 months, and were effective in recording population peaks of S. aurantii. The emergence rate of adults in relation to second instar larvae trapped was 43.7%. 35.7% of adults caught after emerging from the soil were males and 64.3 were females. In contrast, yellow traps had recorded 59.1% males and 40.9% females over the same period. The yellow traps were subsequently found to be biased towards male catches when young fruit and/or soft flush was present on the citrus trees. Relative inhibition of female flight activity during times of food abundance is known in other thrips species. During times of food scarcity, the bias in the yellow trap results disappeared. At Letaba Estates, availability of food rather than the direct effects of weather was seen to be the most important factor in governing S. aurantii numbers. Because of the importance of soft flush in the promotion of S. aurantii population increase, it is recommended that more attention should be given to the control of thrips on, as well as conducting surveys for, this food source. As the climate at Letaba is so favourable for citrus thrips, even during winter, it is further recommended that trapping for S. aurantii is carried out throughout the year. Furthermore, males and females should be recorded separately in trapping results as the sex ratio gives an indication of the phenological state of the tree.

The initial phase in the development of a biological control strategy is screening of biological control agents. Secondary to this phase is the establishment of accurate, effective application techniques. However, successful control requires a thorough understanding of all factors affecting the relationship between host plant, pathogen and other microbes. The purpose of this study was to screen and identify potential bacterial antagonists against Alternaria, a fungal citrus pathogen, attachment of the antagonists to bees, and bee dissemination of the antagonist to citrus flowers. A total of 568 bacterial epiphytes were screened on agar plates for antagonism against Alternaria. Only eight of these isolates, which were identified as Bacillus subtilis, B licheniformis, B. melcerons, B. polymyxa, B. thermoglycodasius, B. sphaericus, B. amiloliquefaciens, and B. coagulans, showed inhibitory effects on the growth of Alternaria. The most effective isolates were B. subtilis and B. licheniformis. Further screening was done with B. subtilis and B. subtilis commercial powder (Avogreen). These bacteria were sprayed on citrus flowers for colonisation studies. Mean populations of B. subtilis and the commercial powder recovered from the flowers were 104 and 103 cfu/stamen respectively. The organisms colonised the styler end and ovary of the flowers when observed under scanning electron microscope (SEM). Avogreen was placed in an inoculum dispenser, which was attached to the entrance of the hive. Honeybees emerging from the beehive acquired 104 cfu/bee. The powder attached to the thorax and thoracic appendages, as revealed by SEM. One active beehive was placed in an enclosure with fifteen flowering citrus nursery trees in pots for dissemination trials. Mean populations of commercial B. subtilis recovered from the flowers visited by bees were 104 cfu/stamen. Electron microscope studies revealed that the antagonist was colonising the styler end and ovary of the flowers. Field dissemination studies were unsuccessful due to low yields.<br>Dissertation (Magister Institutiones Agrariae)--University of Pretoria, 2006.<br>Plant Production and Soil Science<br>unrestricted

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.

Control of the citrus pest, false codling moth (FCM), Thaumatotibia leucotreta Meyrick (Lepidoptera: Tortricidae) is crucial for the South African citrus industry. The economic losses and phytosanitary status of this pest, coupled with increased consumer awareness and demands, has created a need for effective, IPM-compatible control measures for use against the soil-dwelling life stages of FCM. Promising developments in the field of microbial control through the use of entomopathogenic fungi (EPF) and entomopathogenic nematodes (EPNs) have highlighted the need for research regarding pupation biology, behaviour and survival of FCM, as a good understanding of biology of the target organism is an important component of any biological control programme. The aim of this study was to improve the current understanding of FCM pupation habits through the manipulation of soil texture class, ground cover, shading, soil compaction, air temperature, and soil moisture in the laboratory. These findings would then be used to aid the biological control programmes using EPF and EPNs against FCM in the soil. Three soil texture classes (sandy loam, silt loam and silty clay loam) were obtained from orchards for use in the study. FCM larvae were allowed to drop into the soil of their own accord and the pupation behaviour that followed was then captured on film with pupae formed in the soil being kept in order to measure adult eclosion. In general, very few abiotic factors had a clear influence on FCM pupation. Larval wandering time and distance was short, but also variable between individuals. Distance did increase when soils were moist. Pupation depth was shallow, with pupal cocoons generally being formed on the soil surface. Depth of pupation was less than one centimetre for all abiotic conditions, with little burrowing into soil. Eclosion success was higher for sandier soils when these were dry and uncompacted, but the addition of both moisture and soil compaction increased FCM eclosion success. FCM was sensitive to desiccation when the soils were dry and temperature limits of 15 °C and 32 °C had a strongly negative impact on eclosion success. Preferences for particular abiotic conditions were limited to only certain moisture conditions when interacting with soil texture class and a preference for pupating in soil when it is available. Limited preference was found for particular soil textures despite this having a strong influence on eclosion success, but individuals did appear to pupate in close proximity to one another. Viable direct habitat manipulation for FCM control could not be identified. These results and all of the abiotic variables measured have important implications for EPF and EPN application, survival and persistence in the soil in order to improve the ability of these biological control agents to control FCM. These are discussed in each chapter.

Thesis (MScAgric)--Stellenbosch University, 2012.<br>ENGLISH ABSTRACT: Planococcus citri (Risso) (Hemiptera: Pseudococcidae), the citrus mealybug, is a highly destructive pest of citrus in South Africa. The ability of mealybugs to develop resistance to chemicals, as well as their cryptic nature and protective wax coverings, all individually and combined, impair the ability of insecticides to control them. Furthermore, chemicals deplete natural enemy populations that usually establish control over mealybug populations well before harvest in March to May. The inability of chemicals to control P. citri successfully and the growing public awareness of the detrimental environmental effects and health risks that are associated with pesticides are pressuring citrus growers to find alternative methods to chemical control. Entomopathogenic nematodes of the Rhabditida order, belonging either to the families Heterorhabditidae or Steinernematidae, have proven to be valuable biocontrol agents of a variety of insect pest species. These nematodes are, however, not yet commercially available in South Africa. Various laboratory bioassays were conducted to determine the potential of entomopathogenic nematodes to control P. citri. Adult female P. citri were screened for susceptibility to six indigenous nematode species. Planococcus citri was found to be most susceptible to Steinernema yirgalemense and Heterorhabditis zealandica, causing 97% and 91% mortality, respectively. Both H. zealandica and S. yirgalemense completed their life cycles inside adult female P. citri. Under optimal conditions, H. zealandica and S. yirgalemense, at an application rate of 170 infective juveniles (IJs) / insect (LD90), were able to control P. citri efficiently with a discriminating dosage of 11(LD50). The water activity (aw) bioassay indicated that S. yirgalemense was two times more tolerant to lower levels of free water than H. zealandica, with aw50 = 0.96 and aw90 = 0.99, compared to H. zealandica with aw50 = 0.98 and aw90 = 1.01. Furthermore, S. yirgalemense proved able to locate and infect P. citri at a faster rate than H. zealandica. Nematode activity was not significantly affected when exposed to 15°C, 20°C and 25°C. Results of the exposure trial also showed that the first 2−4 h post-application was the most decisive time for establishing successful infection of mealybugs. Further experiments were conducted to determine the ability of two polymer products, Zeba® and Xanthan gum, and a surfactant, Nu-Film-P®, to improve nematode application suspensions. Despite not being as effective as Xanthan gum, Zeba® still retarded sedimentation significantly. The addition of 0.02% Xanthan gum did not improve the ability of nematodes to control P. citri at 60% and 80% relative humidity containing H. zealandica and 0.03% Zeba® increased mortality by 14% at 60% RH and by 22% at 80% RH. The same polymer formulation was tested with S. yirgalemense, with mortality being found to have increased by 21% at 60% RH and by 27% at 80% RH. The combined addition of Nu-Film-P® and Zeba® was able to retard sedimentation significantly, increasing the average number of nematodes deposited on 2-cm2 leaf discs. To illustrate those factors that should be taken into consideration before applying nematodes in an integrated pest management (IPM) programme for citrus, the compatibility of H. zealandica and S. yirgalemense with biological control agents and agrochemicals to which they are most likely to be exposed was determined. Bioassays showed the coccinellid beetle, Cryptolaemus montrouzieri, to be susceptible to both nematode species. Beetle larvae proved to be highly susceptible, with H. zealandica obtaining 80%, and S. yirgalemense obtaining 92% control. Adult beetles were found to be twice as susceptible to S. yirgalemense, with 64% mortality recorded, as to H. zealandica, with 30% mortality recorded. Tolerance of H. zealandica and S. yirgalemense infective juveniles to the affect of aqueous solutions of an insecticide (Cyperphos 500 E.C.®), two biopesticides (CryptogranTM and HelicovirTM), and two adjuvants (Nu-Film-P® and Zeba®) on the infectivity and survival of H. zealandica and S. yirgalemense was evaluated. Heterorhabditis zealandica proved to be highly compatible with all products tested. Significant increase in mortality of S. yirgalemense was recorded after 12 h exposure to CryptogranTM, HelicovirTM and Cyperphos 500 E.C.®, and after 6 h exposure to Nu-Film-P®. However, there was no decrease in the ability of S. yirgalemense to infect the insect hosts. Towards selecting the appropriate nematode species for field studies, the ability of S. yirgalemense and H. zealandica to control P. citri was first evaluated under less harsh simulated glasshouse conditions. The ability of both the above-mentioned nematode species to control P. citri, with and without the addition of 0.03% Zeba® and 0.06% Nu-Film-P®, was evaluated in a growth chamber at 75 ± 8% relative humidity and 22°C. All treatments resulted in significantly higher mortality of adult female P. citri than did the control. The addition of 0.03% Zeba® and 0.06% Nu-Film-P® to an H. zealandica and an S. yirgalemense suspension increased resultant mortality from 26% to 30%, and from 34% to 45%, respectively. This increase in mortality was, however, not significant. The ability of this formulation to prolong the ability of S. yirgalemense to infect P. citri and to prolong nematode survival was also evaluated under the same conditions and showed the formulation to improve both infectivity and survival for up to 2–3 h post-application. In the semi-field trial, S. yirgalemense was unable to obtain significant control of P. citri without the addition of 0.03% Zeba® to nematode application suspensions. However, with the addition of 0.03% Zeba® S. yirgalemnse was able to obtain up to 53% control. The study established that the polymer product Zeba® improves the ability of S. yirgalemense to infect P. citri by retarding desiccation and by buffering nematodes from suboptimal environmental conditions.<br>AFRIKAANSE OPSOMMING: Planococcus citri (Risso) (Hemiptera: Pseudococcidae), die sitrus witluis, is ʼn baie skadelike pes van sitrus in Suid Afrika. Die vermoë van witluise om weerstand teen chemiese middels te ontwikkel, hul kriptiese lewenswyse en die beskermende waslaag wat hul liggame omhul inhibeer gesamentlik en individueel die vermoë van insektisiede om witluis bevolkings te beheer. Chemiese middels verminder ook die natuurlike vyande wat gewoonlik witluis bevolkings beheer voor die sitrus oestydperk in Maart tot Mei. Die onvermoë van chemiese middels om P. citri suksesvol te beheer en verhoogde bewustheid van die publiek rond om die vernietigende omgewings impak en gesondheidsrisiko’s verbonde aan chemiese insek beheer, noodsaak sitrus produsente om alternatiewe beheermetodes te ontwikkel. Entomopatogeniese nematodes, van die orde Rhabditida wat aan die families Heterorhabditidae of Steinernematidae behoort, is bekend as effektiewe biologiese beheeragente van ʼn verskeidenheid insek pes spesies. Hierdie nematodes is egter tans nie kommersieel beskikbaar in Suid- Afrika nie. Om ten einde die vermoë van plaaslike nematode spesies te bepaal om P. citri te beheer, is verskeie biotoetse in die laboratorium uitgevoer. ʼn Vinnige siftings proses is uitgevoer om vas te stel watter plaaslike nematode spesies die hoogste persentasie mortaliteit van P. citri wyfies veroorsaak. Daar is bevind dat P. citri die mees vatbaarste is vir Steinernema yirgalemense en Heterorhabditis zealandica wat 97% en 91% mortaliteit respektiewelik veroorsaak het. Die ontwikkeling van beide H. zealandica en S. yirgalemense na die infektering van volwasse P. citri wyfies is gevolg en daar is bevind dat beide nematode spesies hul lewensiklusse kon voltooi in insek kadawers. Onder optimale toestande was H. zealandica en S. yirgalemense in staat om P. citri effektief te beheer, as hul teen ʼn konsentrasie van 170 infektiewe larwes (JIs) per insek (LD90) toegedien word, met ʼn diskriminerende toedienings dosis van 11 (LD50). ʼn Water aktiwiteit biotoets het gewys dat S. yirgalemense twee keer so verdraagsaam is teenoor laer vlakke van vrye water as wat H. zealandica is met aw50 = 0.96 en aw90 = 0.99, in vergelyking met H. zealandica met aw50 = 0.98 en aw90 = 1.01. Resultate van ʼn blootstellings toets het ook gewys dat S. yirgalemense die vermoë het om P. citri vinniger op te spoor en te infekteer as H. zealandica. Die blootstellings toets was uitgevoer teen 15°C, 20°C en 25°C en daar is bevind dat die aktiwiteit van H. zealandica nie beduidend verhoog het met ʼn verhoging in temperatuur nie. Resultate vir die blootstellings toets het ook gewys dat die eerste 2 tot 4 uur na toediening van nematodes die mees kritieke tyd is om suksesvolle infektering van witluise te bevestig. Verdere eksperimente is uitgevoer om te bepaal of die byvoeging van twee polimeer produkte nl. Zeba® en Xanthan gum en ʼn benatter Nu-Film-P®, nematode toedienings suspensies kan verbeter. Deur 0.02% Xanthan gum by nematode toedienings suspensies te voeg is die vermoë van nematodes om P. citri by 60% en 80% relatiewe humiditeit (RH) te beheer nie beduidend verhoog nie, terwyl die byvoeging van 0.03% Zeba® wel ʼn beduidende verhoging in mortaliteit veroorsaak het. ʼn Suspensie van H. zealandica en 0.03% Zeba® het beheer met 14% by 60% RH en met 22% by 80% RH verhoog. Die invloed van dieselfde polimeer formulasie was ook getoets op S. yirgalemense en mortaliteit het verhoog met 21% by 60% RH en met 27% by 80% RH. Ten spyte daarvan dat “Xanthan gum” nie so effektief was om afsakking van nematodes te verhoed nie, het die byvoeginging van 0.03% Zeba® by toedienings suspensies steeds ʼn beduidende invloed gehad. Die gekombineerde byvoeging Nu-Film-P® en Zeba® was in staat om die gemiddelde aantal nematodes gedeponeer op 2-cm2 blaar skyfies te verhoog. Om te wys watter faktore in ag geneem moet word voor EPNs as deel van ʼn geïntegreerde pes beheer program toegedien word, is die verenigbaarheid van H. zealandica en S. yirgalemense vir biologiese beheer agente en landbouchemikalieë, waaraan hulle heel waarskynlik blootgestel gaan word, bepaal. Biotoetse het gewys dat die liewenheersbesie, Cryptolaemus montrouzieri, vatbaar is vir beide nematode spesies. Die larwale fase was hoogs vatbaar met mortaliteit van 80% en 92% verkry deur die toedeining van H. zealandica en S. yirgalemense respektiewelik. Daar is ook bepaal dat volwasse besies twee maal so vatbaar is vir S. yirgalemense wat 64% mortaliteit veroorsaak het, as vir H. zealandica met 30% mortaliteit aangeteken. Verdraagsaamheid van IJs van H. zealandica en S. yirgalemense met oplossings van ʼn insektisied (Cyperphos 500 E.C.®), twee bio-plaagdoders (CryptogranTM en HelicovirTM), en twee byvoegmiddels (Nu-Film-P® en Zeba®), vir infektiwiteit en oorlewing is bepaal. Resultate het gewys dat H. zealandica hoogs verenigbaar is met alle produkte wat getoets is. ʼn Beduidende verhoging in mortaliteit van S. yirgalemense is aangeteken na 12 ure se blootstelling aan CryptogranTM, HelicovirTM en Cyperphos 500 E.C.®, en na 6 ure se blootstelling aan Nu-Film-P®. Resultate het egter ook gewys dat die vermoë van S. yirgalemense om om gashere te infekteer nie deur beinvloed word nie. Om ten einde te bepaal watter een van H. zealandica of S. yirgalemense die beter isolaat sou wees om te gebruik in veldproewe, is hul vermoë om P. citri te beheer eers evalueer onder minder ongunstige gesimuleerde glashuis toestande. Die vermoë van beide nematode spesies om P. citri te beheer is bepaal in ʼn groeikamer by 75 ± 8% (RH) met en sonder die byvoeging van 0.03% Zeba® en 0.06% Nu-Film-P®. Mortaliteit verkry deur alle behandelings was beduidend hoër as die kontrole. Deur 0.03% Zeba® en 0.06% Nu-Film-P® by toedienings suspensies van H. zealandica en S. yirgalemense te voeg is die gevolglike mortaliteit van P. citri verhoog van 26% na 30% en van 34% na 45%, respektiewelik. Hierdie verhoging in mortaliteit was egter nie beduidend nie. Die vermoë van die bo-genoemde formulasie om oorlewing van S. yirgalemense en infeksie tydperk van P. citri te verleng was bepaal onder dieselfde toestande en daar is gevind dat die formulasie beide die infektiwiteit en oorlewing van nematodes met 2 tot 3 uur na toediening verleng het. Tydens die semi-veldproef was S. yirgalemense nie in staat om beduidend hoër mortaliteit van P. citri te verkry, in vergelyking met die kontrole, sonder die byvoeging van 0.03% Zeba® by toedienings suspensies nie. Met die byvoeging van Zeba® is daar tot 53% beheer verkry. Die studie het onweerlegbaar gewys dat die polimeer produk Zeba® die vermoë van S. yirgalemense om P. citri te infekteer verbeter deur die uitdroging van nematodes te vertraag en deur hul te beskerm teen ongunstige omgewings toestande.

The false codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae), is an important pest of citrus in South Africa and challenging to manage due to its inconspicuous nature. An effective method currently being employed for the area-wide suppression of the FCM is the Sterile Insect Technique (SIT) and the effective dispersal of sterile moths is very important for success with SIT. This study was conducted in the Addo area of the Sundays River Valley (Eastern Cape) where the programme is commercially used. In this study, sterile male moths were released in different orchards on a citrus farm, and in nearby veld at different times of the year, and their dispersal was monitored through the use of pheromone traps. Various climatic factors were monitored. This provided insight into the local dispersal of sterile male FCM adults in response to abiotic cues (particularly climatic factors). The movement of the FCM in four citrus cultivars, namely lemons, navel and Valencia oranges and mandarins and in the nearby veld (open field), was determined at six different stages of the year. Results clearly indicated that sterile FCM movement is concentrated within citrus orchards, as very few moths were trapped beyond 30 m from the release point, particularly in navel and Valencia orchards. Of the climatic factors measured, minimum and maximum temperatures had the most significant influence on FCM dispersal, and based on the results, various recommendations are made for the releases of sterile FCM in an area-wide SIT management programmes on citrus. A better understanding of the dispersal capabilities of the FCM in an agricultural system, under different conditions and at different times of the year, is invaluable not only in improving release strategies in an SIT programme but in planning future control strategies against the FCM.

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.

Thesis (MScAgric)--University of Stellenbosch, 2005.<br>ENGLISH ABSTRACT: Fruit and Leaf Spot Disease (FLSD) of citrus, caused by Phaeoramularia angolensis, is found only in 18 countries in Africa, the Comores Islands in the Indian Ocean and Yemen in the Arabian peninsula. The major citrus export countries in Africa are Morocco, South Africa, Swaziland, and Zimbabwe. Zimbabwe is the only country affected by FLSD. FLSD is a disease of major phytosanitary and economic importance and its devastating effect on citrus is highlighted by the fact that the damage is cosmetic, which renders the fruit unmarketable. Total crop losses are not uncommon in Kenya. The aims of the present study, therefore, was was to determine the occurrence of P. angolensis in Zimbabwe and neighbouring Mozambique, to compare these isolates with the Cercospora Fresen. isolates from Swaziland and South Africa, to determine the epidemiology of the pathogen and to implement an effective control strategy to prevent the spread of FLSD. Leaf samples with citrus canker-like lesions collected in the early 1990’s in Zimbabwe were found to be infected by the fungus, Phaeoramularia angolensis. Surveys were undertaken to determine the spread and intensity of FLSD in Zimbabwe and Mozambique. In Zimbabwe, P. angolensis was limited to an area above the 19° south latitude, predominantly the moist areas and not the low-lying drier parts of the country. In Mozambique, no P. angolensis symptoms were found. Observations during the survey indicated that no proper management systems were implemented by Zimbabwean growers. A cercosporoid fungus causing a new Fruit and Leaf Spot Disease on Citrus in South Africa was identified. From morphological and rDNA sequence data (ITS 1, 5.8S and ITS 2), it was concluded that the new disease was caused by Cercospora penzigii, belonging to the Cercospora apii species complex. The genera Pseudophaeoramularia and Phaeoramularia are regarded as synonyms of Pseudocercospora, and subsequently a new combination was proposed in Pseudocercospora as P. angolensis. Cercospora gigantea was shown to not represent a species of Cercospora, while Mycosphaerella citri was found to be morphologically variable, suggesting that it could represent more than one taxon. A control strategy for the control of FLSD was evaluated in the study. The data showed that P. angolensis in Zimbabwe can be managed successfully by the removal of all old and neglected orchards, and on timely fungicide applications. Trifloxystrobin + mancozeb + mineral spray oil (20 g + 200 g + 500 ml/100 l water) applied in November, January and March was the most effective treatment. Three applications of benomyl + mancozeb + mineral spray oil (25 g + 200 g + 500 ml/100 l water) applied during the same period, was the second most effective treatment, and two applications (November and January) of trifloxystrobin + mineral spray oil (20g + 500 ml/100 l water) and difenoconazole (40 g) per 100 l/water applied twice in November and January, the third most effective treatment. The spore trap and weather data showed that P. angolensis needs high moisture and temperatures in excess of 25°C for disease development. It is concluded that P. angolensis in Zimbabwe can be managed successfully by implementing a holistic approach, which should be supported by the authorities, organised agriculture and all technical personnel involved in citrus production.<br>AFRIKAANSE OPSOMMING: Blaar- en vrugvleksiekte (BVVS) op sitrus, veroorsaak deur Phaeoramularia angolensis, kom in 18 lande in Afrika voor asook die Comores Eilande in die Indiese Oseaan en Yemen op die Arabiese skiereiland. Marokko, Suid Afrika, Swaziland en Zimbabwe is die belangrikste uitvoerders van sitrus in Afrika. Van dié lande het slegs Zimbabwe blaar en vrugvleksiekte op sitrus. Hierdie siekte is van fitosanitêre en ekonomiese waarde en die nadelige effek van die siekte, wat slegs kosmetiese van aard is, is venietigend aangesien vrugte onbemarkbaar is. Totale opbrengsverliese is nie ongewoon in lande soos Kenya nie. Die doelwitte van die studie was dus om die voorkoms van P. angolensis in Zimbabwe te bepaal, om die Cercospora Fresen. isolate vanaf Swaziland en Suid-Afrika met mekaar te vergelyk, om die epidemiologie van die siekte vas te stel en om ‘n effektiewe beheermaatreël teen die siekte te ondersoek. Blaarmonsters met kankeragtige letsels wat in die vroeë 1990’s in Zimbabwe gevind is, het getoon dat die blare geinfekteer is met die swam, Phaeoramularia angolensis. Ondersoeke is geloots om die verspreiding en intensiteit van BVVS in Zimbabwe en Mosambiek te bepaal. In Zimbabwe was gevind dat P. angolensis beperk was tot gebiede bo die 19° Suid breedtegraad, wat die hoër vogtiger gebiede insluit eerder as die droeër, laagliggende gebiede. Geen P. angolensis simptome kon in Mosambiek gevind word nie. Tydens die opnames was dit duidelik dat geen geskikte beheerstrategieë toegepas word deur Zimbabwe se produsente nie. ‘n Nuwe cercosporoid swam, wat blaar en vrugvleksiekte op sitrus is in Suid Afrika veroorsaak is geidentifiseer. Morfologiese en rDNA volgorde (ITS 1, 5.8S en ITS 2) data het getoon dat die siekte veroorsaak word deur Cercospora penzigii wat tot die Cercospora apii spesie kompleks behoort. Die genus Pseudophaeoramularia kan as sinoniem van Pseudocercospora beskou word en ‘n nuwe kombinasie word voorgestel in Pseudocercospora as P. angolensis. Cercospora gigantea het getoon dat dit nie ‘n spesie van Cercospora kon verteenwoordig nie terwyl Mycosphaerella citri varieërend voorkom en meer as een takson kan verteenwoordig. ‘n Beheerstrategie vir die beheer van BVVS is ondersoek. Die data wys dat P. angolensis in Zimbabwe doeltreffend beheer kan word deur die uitroeiing van ou en verwaarloosde bome, en deur goed beplande fungisied bespuiting. Trifloxystrobin + mancozeb + minerale spuitolie (20 g + 200 g + 500 ml/100 l water), wat in November, Januarie en Maart toegedien is, was die mees effektiefste behandeling. Drie bespuitings van benomyl + mancozeb + minerale spuitolie (25 g + 200 g + 500 ml/100 l water) wat oor dieselfde tydperk toegedien is, was die naas beste behandeling. Trifloxystrobin (20 g) + minerale spuitolie (500 ml) per 100 l/water en difenoconazole (40 g) per 100 l/water, beide as twee bespuitings toegedien in November en Januarie, het die derde beste resultaat opgelewer. Die spoorlokval en klimatologiese data het getoon dat P. angolensis vogtige toestande en temperature hoër as 25°C benodig vir siekteontwikkeling. Die afleiding uit die studie is dat P. angolensis suksesvol beheer kan word indien ‘n holistiese benadering gevolg word en alle rolspelers naamlik die owerheid, georganiseerde landbou en tegniese personeel die proses ondersteun.

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.

Made available in DSpace on 2014-06-11T19:25:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-06-22Bitstream added on 2014-06-13T18:53:21Z : No. of bitstreams: 1 sumares_jap_me_arafcf.pdf: 902012 bytes, checksum: 885db9e62dd6f2315568900f3dbc8407 (MD5)<br>Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)<br>Universidade Estadual Paulista (UNESP)<br>O cancro cítrico asiático, causado pela bactéria Xanthomonas citri subsp. citri (Xac), é uma das doenças mais importantes dos Citrus e de ocorrência mundial. Esta doença constitui um grande risco para a citricultura brasileira uma vez que os sintomas podem diminuir o valor de mercado das frutas, e, por vezes, até impedir a sua comercialização. Neste trabalho determinamos a influência da temperatura no crescimento e fisiologia de Xac, com especial atenção à temperatura da morte bacteriana e sua influência sobre a competência infecciosa da bactéria. Descobrimos que a taxa de crescimento de Xac é de forma geral influenciada por altas temperaturas. Notamos uma diminuição do crescimento quando bactérias foram submetidas a temperaturas superiores àquela considerada ótima para cultivo controlado (30 ºC). Bactérias expostas a temperaturas de até 37 ºC foram perfeitamente competentes para infectar o hospedeiro citros, além de recobrarem perfil de crescimento padrão quando retornadas a temperatura de 30 ºC. Tratamentos com temperaturas superiores a 40 ºC promovem alterações fisiológicas irreversíveis e acima de 42 ºC não foi possível resgatar células capazes de induzir sintomas de cancro cítrico, sendo, pois, designadas temperatura de morte. Temperaturas diferentes de 30 ºC promoveram aumento de tamanho celular, sugestivo de perturbação de processos essenciais de replicação/segregação cromossômica e/ou divisão celular. Para investigar a influência do tratamento térmico em tais processos, submetemos linhagens de Xac com marcação para centrômero e septo bacterianos a tratamentos com diferentes temperaturas, seguindo-se avaliação por microscopia de fluorescência. Tratamento...<br>Asiatic citrus canker, caused by Xanthomonas citri subsp. citri (Xac), is one of the most important Citrus diseases and occurs worldwide. This disease poses a major risk to the Brazilian citrus industry since the symptoms may decrease the market value of the fruit, and restrict its commercialization. In this work we have determined the influence of temperature on growth and physiology of Xac, with special attention to the temperature of bacterial death and its influence on the bacterial infectious competence. We have found that the growth rate of Xac was generally affected by high temperatures. We have noticed a decrease of bacterial growth when exposed to temperatures higher than that considered optimal for controlled cultivation (30 ºC). Bacteria exposed to temperatures up to 37 ºC were fully competent to infect the citrus host, and regain ability to grow when temperature was returned to 30 ºC. Treatments at temperatures above 40 ºC promote irreversible physiological chances. Above 42 ºC it has not been possible to recover viable cells that could produce symptoms of citrus canker, and therefore, it was designated death temperature. Temperatures different from 30 ºC caused an increase in cell size, suggesting disturbance on essential biological processes, as chromosomal replication/segregation and/or cell division. To investigate the influence of heat treatment in such processes, we have submitted mutant Xac strains, marked for bacterial centromere and septum, to treatments at different temperatures, followed by fluorescence microscopy analysis. Treatment at 37 ° C for 6 hours did not affect... (Complete abstract click electronic access below)

False codling moth (FCM), Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is considered the most important indigenous pest of citrus in southern Africa. Major concerns such as progressive insecticidal resistance, the negative impact of insecticides on the environment, as well as the influence of consumers opposed to chemical residues on fruit, created opportunities for biological control methods such as Sterile Insect Technology (SIT). This technology is now established in the Western and Eastern Cape provinces of South Africa as an effective, sustainable alternative to conventional FCM control methods. Due to the prevalence of the pest in all citrus producing areas of South Africa, potential for SIT to expand is enormous. Success of an SIT programme is highly dependent on efficient application of the technology to achieve its objectives in a timeous manner. The aim of this study was to advance the application of SIT for control of FCM on citrus in South Africa, by investigating the effect of certain critical stages in the process. The effect of long-distance transportation on fitness of irradiated FCM was determined, showing reduced performance with cold-immobilized transport. A significant decrease in flight ability and longevity of irradiated FCM was found, although critically, realized fecundity was not affected. The effect of two different insecticides in the pyrethroid and organophosphate chemical groups were investigated for their residual effect on mortality of released irradiated FCM, to determine if these pest control programmes could be integrated. Both chlorpyrifos and tau-fluvalinate were effective in killing irradiated FCM for a number of days after application, after which degradation of the active ingredient rendered it harmless. This effect was found to be similar for irradiated and non-irradiated males, consequently ratios of sterile : wild male FCM should be retained regardless of whether sprays are applied or not. The modes for release of sterile FCM in an SIT programme were investigated. Efficacy of ground and aerial release platforms were tested by evaluating the recovery of released irradiated male FCM in these orchards. More irradiated FCM were recovered in orchards released from the ground compared to air. However, an economic analysis of both methods shows application of irradiated insects over a large geographical area is more cost-effective by air. Depending on the terrain and size of the target area, a combination of both methods is ideal for application of SIT for control of FCM in citrus. Development of application technology for advance of the programme is discussed and recommendations for future research and development are offered.

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.

Made available in DSpace on 2014-06-11T19:26:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-22Bitstream added on 2014-06-13T18:29:32Z : No. of bitstreams: 1 santos_m_me_jabo.pdf: 333325 bytes, checksum: 9f90954b7831d0ab7d554fea50c99a27 (MD5)<br>O huanglongbing (HLB ou greening) é uma doença altamente destrutiva que ataca a maioria das espécies de citros. Trabalhos e observações recentes sugerem que os porta-enxertos podem ter influência na incidência e severidade da doença. O presente projeto teve como objetivo monitorar a reação de 16 variedades de porta-enxertos sob laranjeira „Valencia‟ frente ao Candidatus Liberibacter asiaticus por meio da técnica de q-PCR, e suas respostas biométricas, tais como: diâmetro 2 cm a baixo e a cima da linha da enxertia, altura do colo da planta ao enxerto e do enxerto a poda, comprimento do lançamento, peso fresco da parte aérea e do sistema radicular, volume do sistema radicular, e análise foliar de macro e micronutrientes. Os porta-enxertos empregados foram os principais em uso comercial: trifloliateiro „Flying Dragon‟, limoeiro „Cravo‟, „Poncirus trifoliata’, tangerineira „Sunki‟ e citrumelo „Swingle‟, testados experimentalmente com bons resultados: citrangeiros „Troyer‟ e „Carrizo‟ tetraplóides, HRS 643, HRS 702, HRS 849 e HRS 812, em fase de experimentação: „Clementina x trifoliata‟ (1615), „Cleópatra x Swingle‟ (1614), „Cleópatra x Rubidoux‟ (1600) e „Changsha x English Large‟, além de um híbrido „Rhode Red + Volkameriano‟, também em fase experimental. O experimento foi conduzido no Centro de Citricultura Sylvio Moreira, em Cordeirópolis. Cada associação copa e porta-enxerto apresentaram sete plantas, cinco delas foram inoculadas e duas não inoculadas. Para analisar a evolução da bactéria nas plantas foram realizadas três amostragens (8, 12, 16 meses após a inoculação), de pecíolos e nervuras centrais...<br>The Huanglongbing (HLB or greening) is a highly destructive disease that affects most citrus cultivars, species and hybrids. Recent works and researches suggest that rootstock variety influenced disease severity and incidence. This work aimed to characterize variation in the reaction of 16 cultivars of rootstocks grafted with a sweet orange „Valência‟ scion in response to Candidatus Liberibacter asiaticus by q-PCR and biometric characteristics such as: stem diameter 2 cm above and below the grafting, collar height of the plant to the graft, collar height of the graft to the pruning, length of the first bud after pruning, fresh weight of aerial part of the plant, fresh weight of root system, root volume and leaf analysis of macro and micronutrients. The rootstocks used in the work were the major commercial cultivars: trifoliate orange „Flying Dragon‟, Rangpur lime, „Poncirus trifoliata‟, mandarin „Sunki‟ and citrumelo „Swingle‟, tested experimentally with good results, citrange „Troyer‟ and „Carrizo‟ tetraploid, HRS 643, HRS 702, HRS 849 and HRS 812, in experimental stage, „Clementina x trifoliata‟ (1615), „Cleópatra x Swingle‟ (1614), „Cleópatra x Rubidoux‟ (1600) and „Changsha x English Large‟, plus a hybrid „Rhode Red + Volkameriano‟. The experiment was conducted at Centro de Citricultura Sylvio Moreira in Cordeirópolis. Each cultivar had seven plants, five were inoculated and two were used as control plants. To evaluate the growth of bacteria in plants, three sampling were done at (8, 12, 16 months after inoculation) by visually infected... (Complete abstract click electronic access below)

Orientador: Eduardo Sanches Stuchi<br>Coorientador: Helvécio Della Coletta Filho<br>Banca: Antonio Baldo Geraldo Martins<br>Banca: Simone Rodrigues da Silva<br>Resumo: O huanglongbing (HLB ou greening) é uma doença altamente destrutiva que ataca a maioria das espécies de citros. Trabalhos e observações recentes sugerem que os porta-enxertos podem ter influência na incidência e severidade da doença. O presente projeto teve como objetivo monitorar a reação de 16 variedades de porta-enxertos sob laranjeira „Valencia‟ frente ao Candidatus Liberibacter asiaticus por meio da técnica de q-PCR, e suas respostas biométricas, tais como: diâmetro 2 cm a baixo e a cima da linha da enxertia, altura do colo da planta ao enxerto e do enxerto a poda, comprimento do lançamento, peso fresco da parte aérea e do sistema radicular, volume do sistema radicular, e análise foliar de macro e micronutrientes. Os porta-enxertos empregados foram os principais em uso comercial: trifloliateiro „Flying Dragon‟, limoeiro „Cravo‟, „Poncirus trifoliata', tangerineira „Sunki‟ e citrumelo „Swingle‟, testados experimentalmente com bons resultados: citrangeiros „Troyer‟ e „Carrizo‟ tetraplóides, HRS 643, HRS 702, HRS 849 e HRS 812, em fase de experimentação: „Clementina x trifoliata‟ (1615), „Cleópatra x Swingle‟ (1614), „Cleópatra x Rubidoux‟ (1600) e „Changsha x English Large‟, além de um híbrido „Rhode Red + Volkameriano‟, também em fase experimental. O experimento foi conduzido no Centro de Citricultura Sylvio Moreira, em Cordeirópolis. Cada associação copa e porta-enxerto apresentaram sete plantas, cinco delas foram inoculadas e duas não inoculadas. Para analisar a evolução da bactéria nas plantas foram realizadas três amostragens (8, 12, 16 meses após a inoculação), de pecíolos e nervuras centrais... (Resumo completo, clicar acesso eletrônico abaixo)<br>Abstract: The Huanglongbing (HLB or greening) is a highly destructive disease that affects most citrus cultivars, species and hybrids. Recent works and researches suggest that rootstock variety influenced disease severity and incidence. This work aimed to characterize variation in the reaction of 16 cultivars of rootstocks grafted with a sweet orange „Valência‟ scion in response to Candidatus Liberibacter asiaticus by q-PCR and biometric characteristics such as: stem diameter 2 cm above and below the grafting, collar height of the plant to the graft, collar height of the graft to the pruning, length of the first bud after pruning, fresh weight of aerial part of the plant, fresh weight of root system, root volume and leaf analysis of macro and micronutrients. The rootstocks used in the work were the major commercial cultivars: trifoliate orange „Flying Dragon‟, Rangpur lime, „Poncirus trifoliata‟, mandarin „Sunki‟ and citrumelo „Swingle‟, tested experimentally with good results, citrange „Troyer‟ and „Carrizo‟ tetraploid, HRS 643, HRS 702, HRS 849 and HRS 812, in experimental stage, „Clementina x trifoliata‟ (1615), „Cleópatra x Swingle‟ (1614), „Cleópatra x Rubidoux‟ (1600) and „Changsha x English Large‟, plus a hybrid „Rhode Red + Volkameriano‟. The experiment was conducted at Centro de Citricultura Sylvio Moreira in Cordeirópolis. Each cultivar had seven plants, five were inoculated and two were used as control plants. To evaluate the growth of bacteria in plants, three sampling were done at (8, 12, 16 months after inoculation) by visually infected... (Complete abstract click electronic access below)<br>Mestre

Made available in DSpace on 2014-06-11T19:26:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-12-15Bitstream added on 2014-06-13T19:33:26Z : No. of bitstreams: 1 dallacqua_fc_me_jabo.pdf: 321125 bytes, checksum: 095ba41139dc507b26139c5c4897dd6c (MD5)<br>Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)<br>As bactérias pertencentes ao gênero Xanthomonas constituem um dos grupos de fitopatógenos mais importantes na natureza, com capacidade de infectar aproximadamente 120 tipos diferentes de plantas monocotiledôneas e 270 dicotiledôneas. A bactéria Xanthomonas citri subsp. citri (Xcc) é o agente causal do cancro cítrico, uma das principais doenças dos citros. Restrita à Flórida, a mancha bacteriana dos citros, causada por Xanthomonas alfalfae subsp. citrumelonis (Xacm), afeta principalmente o citrumelo ‘Swingle’. A bactéria Xanthomonas fuscans subsp. aurantifolii (Xfa) é o agente causal das cancroses B e C, encontradas apenas na América do Sul. As técnicas mais utilizadas na diferenciação e caracterização dos patógenos bacterianos, hibridização DNA-DNA (DDH) e sequenciamento da região do DNA que codifica o RNA ribossomal 16S (rRNA 16S), são onerosas, trabalhosas, lentas e de resultados restritos, não permitindo a comparação dos resultados obtidos entre diferentes estudos laboratoriais. Por isso, a técnica de análise de sequências multilocus (MLSA), considerada rápida, barata e confiável, tem sido recomendada para a caracterização e delimitação de espécies de patógenos bacterianos, tais como as do gênero Xanthomonas, permitindo o estabelecimento das relações filogenéticas por meio de genes housekeeping. Portanto, o objetivo do trabalho foi caracterizar os isolados de Xanthomonas (Xcc-A, Xfa-B, Xfa-C e Xacm) por meio da técnica MLSA. Foram utilizados 27 isolados de Xanthomonas patogênicas a citros, sendo quatro de X. citri subsp. citri (Xcc), três de X. fuscans subsp. aurantifolli B (Xfa-B), dezenove de X. fuscans subsp. aurantifolli C (Xfa-C) e um de X. alfalfae subsp. citrumelonis (Xacm). Os genes atpD, dnaK e fusA foram utilizados como genes housekeeping e para a reação da PCR...<br>Bacteria belonging to the genus Xanthomonas are one of the main groups of phytopathogens in nature, capable to infecting about 120 different monocots and 270 dicots plants. Xanthomonas citri subsp. citri (Xcc) is the causal agent of bacterial citrus canker, one of the major diseases of citrus. Restricted to Florida, citrus bacterial spot, caused by Xanthomonas alfalfae subsp. citrumelonis (Xacm), affects mainly citrumelo 'Swingle'. Xanthomonas fuscans subsp. aurantifolii (Xfa) is the causal agent of canker B and C, found only in South America. The most widely used techniques for the differentiation and characterization of bacterial pathogens, DNA-DNA hybridization (DDH) and analysis of DNA sequences coding for small subunit ribosomal RNAs (16S rRNA), are costly, cumbersome, slow and give limited results, do not allowing the interlaboratorial comparisons of the results. Therefore, the technique of multilocus sequence analysis (MLSA), which is considered fast, cheap and reliable, has been recommended for characterization and delineation of species of bacterial pathogens, such as the genus Xanthomonas, allowing the establishment of phylogenetic relationships between them through housekeeping genes. Therefore, the objective of this study was to characterize strains of Xanthomonas (Xcc-A, Xfa-B, Xfa-C and Xacm) using the MLSA technique. We used 27 strains of Xanthomonas pathogenic to citrus, being four Xcc, three Xfa-B, 19 Xfa- C and one Xacm. The PCR reaction was performed using the specific primers to amplify the atpD, dnaK and fusA genes. PCR products were purified, sequenced and the sequences were analyzed using the software CodenCode Aligner version 3.7.1. Phylogenetic analysis was performed using Maximum Likelihood based on the model of Tamura-Nei and using the software MEGA 5. Analyses ... (Complete abstract click electronic access below)

Thesis (MSc (Botany and Zoology))--University of Stellenbosch, 2010.<br>ENGLISH ABSTRACT:Chapter 1 - Mealybugs are tiny, soft-bodied insects which constitute the second largest scale insect family Pseudococcidae (Downie & Gullan 2004). The family comprises approximately 2000 species in 300 genera (Ben-Dov 1994), of which 20 species are pests of cultivated plants in South Africa (Annecke & Moran 1982). In South Africa, approximately 109 species of mealybugs have been recorded from 50 genera (Millar 2002). Chapter 2 - The effect of constant temperatures on the development, survival and fecundity of the oleander mealybug, Paracoccus burnerae on citrus was determined. Developmental time, rate of development, fecundity and survival were investigated at five constant temperatures and a 16L: 8D light: darkness regime. The rate of development increased linearly with an increase in temperature for the egg, 1st nymphal and pupal stages as well as the entire biological cycle (egg – adult), but was nonlinear for the 2nd and 3rd nymphal stages. Survival decreased with an increase in temperature. P. burnerae required 666.7 degree-days above a lower threshold of 8.7°C to complete one generation. The highest mean number of 68 eggs per female was reached at 22°C. A sex ratio of 0.52:0.48 (male:female) was obtained from the life table. The net reproductive rate (Ro) was >1 at all five temperatures, an indication that it is capable of increasing its population numbers despite the high mortality experienced in the 1st and 2nd nymphal stages. Chapter 3 - The oleander mealybug, Paracoccus burnerae (Brain) is a pest of citrus in South Africa. This study was carried out to determine the effect of temperature on development rate of P. burnerae and to investigate whether development rate is the reason why P. burnerae is out competing the citrus mealybug, Planococcus citri (Risso), in the Eastern and Western Cape Provinces of South Africa. The influence of temperature on life history traits of P. burnerae was determined at 20, 22, 25 and 27°C and compared with corresponding data for P. citri. The rate of development increased linearly with an increase in rearing temperature in the embryonic, first nymphal and pupal stages but reached a climax at 26.13 and 28.6°C in the second nymphal stage of both species, respectively. P. citri exhibited lower developmental thresholds except in first instar, shorter degree-days and higher developmental rates than P. burnerae. Results of the current study indicated that the dominance of oleander mealybug over the citrus mealybug is neither linked to developmental rates nor sum of effective temperatures. Chapter 4 - The importance of Paracoccus burnerae has risen over the years to an extent where it is now regarded as a quarantine pest for citrus fruit from South Africa. The field biology of P. burnerae on citrus in the Western Cape Province of South Africa was studied through periodic sampling of leaves from twigs enclosed in sleeve cages. The species composition and abundance of natural enemies was investigated. Both adult and immature stages attained maximum population peaks in March and P. burnerae had four generations. The highest level of mortality was experienced in the immature stages. Climate and an unidentified fungus were the key mortality factors. The level of abundance of the two observed predators, the harlequin beetle, Harmonia axyridis and the green lacewing, Chrysoperla sp. was relatively low. Although parasitism occurred in some cages, the level was low ranging between 1.62 to 9.43%. If biocontrol is the preferred method of controlling P. burnerae, suitable candidate parasitoids for inoculative biocontrol are Acerophagus sp., Leptomastix sp. and Microterys nietneri. The oleander mealybug does not share the same parasitoids with Planococcus citri, Pseudococcus calceolariae and Pseudococcus longispinus except the parasitoid Coccophagus sp. The most popular species of parasitoids used in the biolological control of mealybugs, Anagyrus sp. and Coccixenoides sp. were insignificant in the case of P. burnerae. Chapetr 5 - Biological control programs of mealybug species have relied on sprouting potatoes, pumpkins and butternut for rearing of both mealybugs and their natural enemies. In this study, the suitability of sprouting potatoes, butternuts and citrus as mass rearing substrates for the oleander mealybug, Paracoccus burnerae was investigated. Developmental times, rate and fecundity on each substrate were determined and compared at three different temperatures. The developmental time on sprouting potatoes was shorter than on citrus. P. burnerae was unable to complete its life cycle on butternut. The rate of development increased linearly with an increase in temperature on both sprouting potatoes and citrus. P. burnerae required 666.7 degree-days on citrus and 434.8 degree-days on sprouting potatoes above lower developmental thresholds of 7.6°C and 10.4°C respectively to complete one generation. The mean number of eggs per female was higher on sprouting potatoes (121.3) than on citrus (68), but declined with an increase in temperature from 22 to 27°C. Despite the shorter shelf life, sprouting potatoes are the preferred host for mass rearing of the oleander mealybug. Chapter 6 - general conclusions Chapter 7 - Researchers often present impressive results of their studies on the biology of the Coccoidea without mentioning the problems they came across and had to solve. In this paper the practical problems encountered during a study of the biology of the oleander mealybug, Paracoccus burnerae (Brain), an endemic pest of citrus in South Africa, are discussed.<br>AFRIKAANSE OPSOMMING: Geen opsomming beskikbaar.

Orientador: Henrique Ferreira<br>Coorientador: José Belasque Júnior<br>Banca: Franklin Behlau<br>Banca: Luís Octávio Regasini<br>Resumo: O cancro cítrico asiático, causado pela bactéria Xanthomonas citri subsp. citri (Xac), é uma das doenças mais importantes dos Citrus e de ocorrência mundial. Esta doença constitui um grande risco para a citricultura brasileira uma vez que os sintomas podem diminuir o valor de mercado das frutas, e, por vezes, até impedir a sua comercialização. Neste trabalho determinamos a influência da temperatura no crescimento e fisiologia de Xac, com especial atenção à temperatura da morte bacteriana e sua influência sobre a competência infecciosa da bactéria. Descobrimos que a taxa de crescimento de Xac é de forma geral influenciada por altas temperaturas. Notamos uma diminuição do crescimento quando bactérias foram submetidas a temperaturas superiores àquela considerada ótima para cultivo controlado (30 ºC). Bactérias expostas a temperaturas de até 37 ºC foram perfeitamente competentes para infectar o hospedeiro citros, além de recobrarem perfil de crescimento padrão quando retornadas a temperatura de 30 ºC. Tratamentos com temperaturas superiores a 40 ºC promovem alterações fisiológicas irreversíveis e acima de 42 ºC não foi possível resgatar células capazes de induzir sintomas de cancro cítrico, sendo, pois, designadas temperatura de morte. Temperaturas diferentes de 30 ºC promoveram aumento de tamanho celular, sugestivo de perturbação de processos essenciais de replicação/segregação cromossômica e/ou divisão celular. Para investigar a influência do tratamento térmico em tais processos, submetemos linhagens de Xac com marcação para centrômero e septo bacterianos a tratamentos com diferentes temperaturas, seguindo-se avaliação por microscopia de fluorescência. Tratamento... (Resumo completo, clicar acesso eletrônico abaixo)<br>Abstract: Asiatic citrus canker, caused by Xanthomonas citri subsp. citri (Xac), is one of the most important Citrus diseases and occurs worldwide. This disease poses a major risk to the Brazilian citrus industry since the symptoms may decrease the market value of the fruit, and restrict its commercialization. In this work we have determined the influence of temperature on growth and physiology of Xac, with special attention to the temperature of bacterial death and its influence on the bacterial infectious competence. We have found that the growth rate of Xac was generally affected by high temperatures. We have noticed a decrease of bacterial growth when exposed to temperatures higher than that considered optimal for controlled cultivation (30 ºC). Bacteria exposed to temperatures up to 37 ºC were fully competent to infect the citrus host, and regain ability to grow when temperature was returned to 30 ºC. Treatments at temperatures above 40 ºC promote irreversible physiological chances. Above 42 ºC it has not been possible to recover viable cells that could produce symptoms of citrus canker, and therefore, it was designated death temperature. Temperatures different from 30 ºC caused an increase in cell size, suggesting disturbance on essential biological processes, as chromosomal replication/segregation and/or cell division. To investigate the influence of heat treatment in such processes, we have submitted mutant Xac strains, marked for bacterial centromere and septum, to treatments at different temperatures, followed by fluorescence microscopy analysis. Treatment at 37 ° C for 6 hours did not affect... (Complete abstract click electronic access below)<br>Mestre

Two ant species, Pheidole megacephala (Fabricius) and Anoplolepis custodiens (Smith) reach pest status in citrus orchards through precipitating outbreaks of homopterous pests. However, predacious ants, including these two ant species, play an important role in pest suppression in agroecosystems and are therefore beneficial to these systems. If A. custodiens and P. megacephala are important natural control agents in citrus, using ant bands to break the mutualism between the ants and the Homoptera as a method of ant control is preferable to poisoning. Ant communities were sampled by pitfall trapping in three experimental subunits of 2-, 4-, 15- and 30-year-old citrus orchards, in the Grahamstown area of the Eastern Cape. In one subunit in each orchard, populations of P. megacephala and A. custodiens were suppressed by poison applications. In a second subunit, trees were banded with trunk barriers so that ants were prevented from foraging in the trees and a third subunit served as the untreated control. Bait pupae of bollworm, false codling moth and fruit fly were planted in bait trays in all of the subunits to investigate predation on these citrus pests in the relative absence of predacious ants and where they were excluded from the trees. Pheidole megacephala dominated exclusively in all of the plots. Community composition did not change dramatically with increasing age of the trees, but species diversity and species abundance did. Rank-abundance curves showed that community diversity was highest in the 2-year-old plots and lowest in the 30-year-old plots. The Simpson and Shannon-Wiener diversity indices and their evenness measures indicated that diversity and equitability were highest in the poisoned subunits and lowest in the banded subunits. Principle component analysis revealed that the poisoned subunits were similar and distinct in species composition, that there was significant monthly variation in species composition and that community stability increases with an increase in orchard age. The presence of P. megacephala was significantly negatively correlated (rs = -0.293; p < 0.001) with pest pupal survival. Pupal survival was significantly higher for bollworm (p < 0.001), FCM (p < 0.001) and fruit fly (p < 0.001) in the poisoned subunits, than in the banded and control subunits. There was a general trend for survivorship to increase with an increase in the age of the trees. A significant difference (p < 0.001) was found between the months in which the trials were carried out. Pupal survival was significantly lower (p < 0.001) for FCM than for bollworm and fruit fly. In citrus orchards, ant communities are organised by ecological processes and interactions and are influenced by methods of ant control. Ant bands are preferable to poisoning as a method of ant control, so that beneficial species are left on the ground to prey on pests that pupate in the soil. Maintaining high ant species diversity in citrus orchards would be beneficial as predation on the pupae was more effective where ant species diversity was higher.

Thaumatotibia leucotreta (Meyrick) (Lepidoptera: Tortricidae) is one of the major citrus pests of economic importance for South Africa’s citrus industry. It is endemic to Africa, and therefore a phytosanitary pest with zero tolerance by most export markets. The cryptic nature of T. leucotreta makes visual inspection an inefficient method for detecting neonate larvae in fruit in the packhouse. Therefore, a more accurate method for sorting infested fruit at the packhouse, particularly for newly infested fruit could ensure market access. A recent study showed that fruit infested by T. leucotreta emit a chemical profile different from that of a healthy fruit. Several studies provide evidence that parasitoids locate their hosts feeding on fruit by exploiting the novel chemical profiles produced due to host herbivory. The aim of this study was to evaluate the potential of using the naturally occurring behaviour of a larval parasitoid Agathis bishopi (Nixon) (Hymenoptera: Braconidae) for detection of T. leucotreta infested fruit, by determining which compound in infested fruit is attractive to parasitoids. Ytube olfactometer and flight-tunnel bioassays with healthy and T. leucotreta infested fruit showed a significantly stronger response of A. bishopi female parasitoids to infested fruit. Among the volatile compounds associated with T. leucotreta infested fruit, D-limonene elicited the strongest attraction to A. bishopi female parasitoids. Attraction of mated A. bishopi female parasitoids to T. leucotreta infested fruit and D-limonene significantly increased after oviposition experience. Behavioural responses of A. bishopi female parasitoids that were associated with T. leucotreta infested fruit were investigated to determine which behaviours are distinct and interpretable. Probing and oviposition behaviours were the most noticeable and were only elicited on infested fruit when parasitoids contacted T. leucotreta frass, indicating that chemical compounds in frass are short-range cues used for final host location. Since production of D-limonene by fruit is elevated due to herbivory by different pests including mechanical injury on fruit, response of A. bishopi female parasitoids to compounds in frass offers a more specific and potentially useful mechanism for development of a detection system for T. leucotreta infested fruit. Chemical analysis of T. leucotreta frass and conditioning A. bishopi parasitoids to respond behaviourally to compounds in frass is proposed.

The biology of key citrus pests Thaumatotibia leucotreta Meyrick (Lepidoptera: Tortricidae), Ceratitis capitata Wiedemann (Diptera: Tephritidae) and Ceratitis rosa Karsch (Diptera: Tephritidae) includes their dropping from host plants to pupate in the soil below citrus trees. Since most EP fungi are soil-borne microorganisms, the development and formulation of alternative control strategies using these fungi as subterranean control agents, targeted at larvae and pupae in the soil, can potentially benefit existing IPM management of citrus in South Africa. Thus, a survey of occurrence of entomopathogenic fungi was undertaken on soils from citrus orchards and natural vegetation (refugia) on conventionally and organically managed farms in the Eastern Cape Province in South Africa. A method for baiting soil samples with citrus pest T. leucotreta and C. capitata larvae, as well as with the standard bait insect, Galleria mellonella Linnaeus (Lepidoptera: Pyralidae), was implemented. Sixty-two potentially useful entomopathogenic fungal isolates belonging to four genera were collected from 288 soil samples, an occurrence frequency of 21.53%. The most frequently isolated entomopathogenic fungal species was Beauveria bassiana (Balsamo) Vuillemin (15.63%), followed by Metarhizium anisopliae var. anisopliae (Metschnikoff) Sorokin (3.82%). Galleria mellonella was the most effective insect used to isolate fungal species (χ2=40.13, df=2, P≤ 0.005), with a total of 45 isolates obtained, followed by C. capitata with 11 isolates, and T. leucotreta with six isolates recovered. There was a significantly (χ2=11.65, df=1, P≤ 0.005) higher occurrence of entomopathogenic fungi in soil samples taken from refugia compared to cultivated orchards of both organically and conventionally managed farms. No significant differences were observed in the recovery of fungal isolates when soil samples from both farming systems were compared. The physiological effects and host range of 21 indigenous fungal isolates obtained in the Eastern Cape were investigated in the laboratory to establish whether these isolates could be effectively used as biological control agents against the subterranean life stages of C. rosa, C. capitata and T. leucotreta. When these pests were treated with a fungal concentration of 1 x 10⁷ conidia ml⁻¹, the percentage of T. leucotreta adults which emerged in fungal treated sand ranged from 5 to 60% (F=33.295; df=21; P=0.0001) depending on fungal isolate and the percentage of pupae with visible signs of mycosis ranged from 21 to 93% (F= 96.436; df=21; P=0.0001). Based on fungal isolates, the percentage adult survival in C. rosa and C. capitata ranged from 30 to 90% and 55 to 86% respectively. The percentage of C. rosa and C. capitata puparia with visible signs of mycosis ranged from 1 to 14% and 1 to 11% respectively. Deferred mortality due to mycosis in C. rosa and C. capitata adult flies ranged from 1 to 58% and 1 to 33% respectively, depending on fungal isolate. Entomopathogenic fungal isolates had a significantly greater effect on the adults of C. rosa and C. capitata than they did on the puparia of these two fruit fly species. Further, C. rosa and C. capitata did not differ significantly in their response to entomopathogenic fungi when adult survival or adult and pupal mycosis were considered. The relative potency of the four most virulent Beauveria isolates as well as the commercially available Beauveria bassiana product, Bb Plus® (Biological Control Products, South Africa), were compared against one another as log-probit regressions of mortality against C. rosa, C. capitata and T. leucotreta which all exhibited a dose-dependent response. Against fruit flies the estimated LC50 values of all five Beauveria isolates ranged from 5.5 x 10¹¹ to 2.8 x 10¹² conidia/ml⁻¹. There were no significant differences between the relative potencies of these five fungal isolates. When T. leucotreta was considered, isolates: G Moss R10 and G 14 2 B5 and Bb Plus® were significantly more pathogenic than G B Ar 23 B3 and FCM 10 13 L1. The estimated LC₅₀ values of the three most pathogenic isolates ranged from 6.8 x 10⁵ to 2.1 x 10⁶ conidia/ml⁻¹, while those of the least pathogenic ranged from 1.6 x 10⁷ to 3.7 x 10⁷ conidia/ml⁻¹. Thaumatotibia leucotreta final instar larvae were exposed to two conidial concentrations, at four different exposure times (12, 48, 72 and 96 hrs) and showed an exposure time-dependant relationship (F=5.43; df=3; P=0.001). At 1 x 10⁷conidia/ml⁻¹ two Beauveria isolates: G Moss R10 and G 14 2 B5 were able to elicit a response in 50% of test insects at 72 hrs (3 days) exposure. Although a limited amount of mycosis was observed in the puparia of both fruit fly species, deferred adult mortality due to mycosis was high. The increased incidence of adult mortality suggests that post emergence mycosis in adult fruit flies may play a more significant role in field suppression than the control of fruit flies at the pupal stage. The increased incidence of pupal mortality, as well as the relatively low concentrations of conidia required to elicit meaningful responses in T. leucotreta pupae may suggest that pre-emergent control of false codling moth will play a more significant role in field suppression than the control of adult life stages using indigenous isolates of entomopathogenic fungi. Various entomopathogenic fungal application techniques targeted at key insect pests within integrated pest management (IPM) systems of citrus are discussed.

Orientador: Jesus Aparecido Ferro<br>Coorientador: José Belasque Júnior<br>Banca: Lúcia Maria Carareto Alves<br>Banca: Franklin Behlau<br>Resumo: As bactérias pertencentes ao gênero Xanthomonas constituem um dos grupos de fitopatógenos mais importantes na natureza, com capacidade de infectar aproximadamente 120 tipos diferentes de plantas monocotiledôneas e 270 dicotiledôneas. A bactéria Xanthomonas citri subsp. citri (Xcc) é o agente causal do cancro cítrico, uma das principais doenças dos citros. Restrita à Flórida, a mancha bacteriana dos citros, causada por Xanthomonas alfalfae subsp. citrumelonis (Xacm), afeta principalmente o citrumelo 'Swingle'. A bactéria Xanthomonas fuscans subsp. aurantifolii (Xfa) é o agente causal das cancroses B e C, encontradas apenas na América do Sul. As técnicas mais utilizadas na diferenciação e caracterização dos patógenos bacterianos, hibridização DNA-DNA (DDH) e sequenciamento da região do DNA que codifica o RNA ribossomal 16S (rRNA 16S), são onerosas, trabalhosas, lentas e de resultados restritos, não permitindo a comparação dos resultados obtidos entre diferentes estudos laboratoriais. Por isso, a técnica de análise de sequências multilocus (MLSA), considerada rápida, barata e confiável, tem sido recomendada para a caracterização e delimitação de espécies de patógenos bacterianos, tais como as do gênero Xanthomonas, permitindo o estabelecimento das relações filogenéticas por meio de genes housekeeping. Portanto, o objetivo do trabalho foi caracterizar os isolados de Xanthomonas (Xcc-A, Xfa-B, Xfa-C e Xacm) por meio da técnica MLSA. Foram utilizados 27 isolados de Xanthomonas patogênicas a citros, sendo quatro de X. citri subsp. citri (Xcc), três de X. fuscans subsp. aurantifolli B (Xfa-B), dezenove de X. fuscans subsp. aurantifolli C (Xfa-C) e um de X. alfalfae subsp. citrumelonis (Xacm). Os genes atpD, dnaK e fusA foram utilizados como genes housekeeping e para a reação da PCR... (Resumo completo, clicar acesso eletrônico abaixo)<br>Abstract: Bacteria belonging to the genus Xanthomonas are one of the main groups of phytopathogens in nature, capable to infecting about 120 different monocots and 270 dicots plants. Xanthomonas citri subsp. citri (Xcc) is the causal agent of bacterial citrus canker, one of the major diseases of citrus. Restricted to Florida, citrus bacterial spot, caused by Xanthomonas alfalfae subsp. citrumelonis (Xacm), affects mainly citrumelo 'Swingle'. Xanthomonas fuscans subsp. aurantifolii (Xfa) is the causal agent of canker B and C, found only in South America. The most widely used techniques for the differentiation and characterization of bacterial pathogens, DNA-DNA hybridization (DDH) and analysis of DNA sequences coding for small subunit ribosomal RNAs (16S rRNA), are costly, cumbersome, slow and give limited results, do not allowing the interlaboratorial comparisons of the results. Therefore, the technique of multilocus sequence analysis (MLSA), which is considered fast, cheap and reliable, has been recommended for characterization and delineation of species of bacterial pathogens, such as the genus Xanthomonas, allowing the establishment of phylogenetic relationships between them through housekeeping genes. Therefore, the objective of this study was to characterize strains of Xanthomonas (Xcc-A, Xfa-B, Xfa-C and Xacm) using the MLSA technique. We used 27 strains of Xanthomonas pathogenic to citrus, being four Xcc, three Xfa-B, 19 Xfa- C and one Xacm. The PCR reaction was performed using the specific primers to amplify the atpD, dnaK and fusA genes. PCR products were purified, sequenced and the sequences were analyzed using the software CodenCode Aligner version 3.7.1. Phylogenetic analysis was performed using Maximum Likelihood based on the model of Tamura-Nei and using the software MEGA 5. Analyses ... (Complete abstract click electronic access below)<br>Mestre

O Brasil é o maior produtor de citros, sendo um dos seus principais produtos de exportação da cadeia agroindustrial. Um dos fatores que afetam a qualidade e a produção do produto final obtido é a Mancha Marrom de Alternaria, causado pelo fungo Alternaria alternata. A doença provoca desfolha, seca de ramos e queda de frutos em tangerinas e seus híbridos. Para controle dessa doença são utilizados fungicidas, porém, a mancha marrom é uma doença de difícil controle podendo ser necessárias de 7 a 10 aplicações do fungicida, aparecimento de resistência dos fungos, intoxicação nas pessoas e contaminação do meio ambiente devido ao emprego de fungicidas. Métodos alternativos como o controle biológico com aplicações de micro-organismos antagonistas e a nutrição das plantas são opções de menor impacto ao meio ambiente. Sendo assim, o objetivo deste trabalho foi avaliar o potencial de Trichoderma spp., Bacillus subtilis e fertilizante organomineral, no controle da Mancha Marrom de Alternaria em plantas cítricas de tangerina Ponkan. O experimento foi realizado com delineamento casualizado, constituído de oito tratamentos com quatro repetições. Os experimentos foram avaliados quanto a incidência da doença. Os dados mostraram que o melhor controle foi o tratamento químico, porém os tratamentos com Trichoderma spp., Trichoderma spp. + fertilizante e Trichoderma spp. + Bacillus subtilis apresentaram ótimas taxas de controle, podendo inclusive substituir o tratamento químico no controle desta doença.<br>Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2017-06-13T14:28:19Z No. of bitstreams: 1 Dissertacao Bianca Luzardo Porto.pdf: 1070166 bytes, checksum: 4e890666fce3c2b0d8ce25ef2c71241f (MD5)<br>Made available in DSpace on 2017-06-13T14:28:19Z (GMT). No. of bitstreams: 1 Dissertacao Bianca Luzardo Porto.pdf: 1070166 bytes, checksum: 4e890666fce3c2b0d8ce25ef2c71241f (MD5) Previous issue date: 2017-06-13<br>Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES.<br>Brazil is the largest citrus producer, being one of its main exports of agro-industrial chain. One of the factors that affect the quality and production of the final product is the Brown Spot Alternaria, caused by the fungus Alternaria alternata. The disease causes leaf drop, dry branches and fruit drop in tangerines and their hybrids. To control this disease fungicides are used, however, the brown spot is a difficult to control disease, may be required from 7 to 10 applications of fungicide, appearance of resistance of fungi, intoxication in people and environment contamination owing the application of fungicides. Alternative methods such as biological control with applications antagonistic microorganisms and plant nutrition are options of smaller impact to the environment. Therefore, the objective of this work was to evaluate the potential of Trichoderma spp., Bacillus subtilis and fertilizer, in control of Alternaria Brown Spot in citrus plants Ponkan tangerine. The experiment was carried out in a randomized design, consisting of eight treatments with four replicates. The experiments were evaluated for the incidence of the disease. The data showed that the best control was the chemical treatment, but the treatments with Trichoderma spp., Trichoderma spp. + Fertilizer and Trichoderma spp. + Bacillus subtilis presented excellent control rates, and may even replace chemical treatment in the control of this disease.

Includes bibliographical references (leaves 102-114) The highly successful biological control of the citrophilous mealybug Pseudococcus calceolarie (Maskell) (CM) by the parasitic wasp Coccophagus gurneyi Compere in several countries led to the release of this parasitoid in the Riverland of South Australia as part of an integrated pest management program. However CM has not been successfully controlled in this region. The results of this study may help to explain the lack of effective biological control of CM in Riverland citrus.

False codling moth (FCM), Thaumatotibia leucotreta is an extremely important pest of citrus in South Africa and with the shift away from the use of chemicals, alternate control options are needed. One avenue of control which has only recently been investigated against the soil-borne life stages of FCM is the use of entomopathogenic fungi (EPF). In 2009, 12 entomopathogenic fungal isolates collected from South African citrus orchards showed good control potential during laboratory conducted bioassays. The aim of this study was to further analyse the potential of these isolates through concentration-dose and exposure-time response bioassays. After initial re-screening, concentration-dose response and exposure-time response sandconidial bioassays, three isolates were identified as exhibiting the greatest control potential against FCM in soil, Metarhizium anisopliae var. anisopliae (G 11 3 L6 and FCM Ar 23 B3) and Beauveria bassiana (G Ar 17 B3). Percentage mycosis was found to be directly related to fungal concentration as well as the amount of time FCM 5th instar larvae were exposed to the fungal conidia. LC50 values for the three isolates were not greater than 1.92 x 10⁶ conidia.ml⁻ₑ and at the LC₅₀, FCM 5th instar larvae would need to be exposed to the fungus for a maximum of 13 days to ensure a high mortality level. These isolates along with two commercially available EPF products were subjected to field persistence trials whereby net bags filled with a mixture of autoclaved sand and formulated fungal product were buried in an Eastern Cape citrus orchard. The viability of each isolate was measured on a monthly basis for a period of six months. All isolates were capable of persisting in the soil for six months with the collected isolates persisting far better than the commercially used isolates. Two of the isolates, G 11 3 L6 and G Ar 17 B3, were subjected to small scale laboratory application trials. Two formulations were investigated at two concentrations. For each isolate, each formulation and each concentration, FCM 5th instar larvae were applied and allowed to burrow into the soil to pupate before fungal application or after fungal application. Contact between fungi and FCM host is essential as, in contrast to pre-larval treatments, percentage mortality in post-larval treatments was low for both formulations and both isolates. For isolate G Ar 17 B3, a conidial suspension applied as a spray at a concentration of 1 x 10⁷ conidia.ml⁻ₑ obtained the highest percentage mortality (80 %). For isolate G 11 3 L6 however, both formulations performed equally well at a high, 1 x10⁷ conidia.ml⁻ₑ concentration (conidial suspension: 60 %; granular: 65 %) The results obtained thus far are promising for the control of FCM in citrus, but if these EPFs are to successfully integrate into current FCM control practices more research, some of which is discussed, is essential

Os modelos de crescimento econômico mostram que a única forma de promover o crescimento sustentado da economia é por meio do progresso tecnológico. Porém, a manutenção de um fluxo crescente de inovações tecnológicas requer uma forte estrutura de pesquisa e desenvolvimento (P&D), o que implica em uma série de gastos por parte de ambos os setores, público e privado. Conseqüentemente, as instituições de pesquisa são crescentemente cobradas quanto aos resultados dos recursos nelas investidos, o que torna interessante a realização de estudos que procurem avaliar os retornos de tais investimentos e determinar quão proveitosos estes vêm-se revelando para a sociedade. Encontram-se na literatura diversos estudos objetivando medir os retornos econômicos provenientes dos investimentos em P&D na agricultura em geral, porém, existem poucos trabalhos para setores específicos. Desta forma, o presente estudo teve como principal objetivo medir o retorno econômico dos investimentos em pesquisa na citricultura paulista. A justificativa para a escolha do setor analisado é que o Brasil, além de representar sozinho cerca de 80% do total de suco de laranja concentrado e congelado comercializado no mercado internacional, responde por 28% da produção mundial de laranja. O Estado de São Paulo, por sua vez, responde por cerca de 80% da produção nacional de laranja e 98% das exportações brasileiras de suco concentrado. A relevância econômica dessa atividade no Estado tem estimulado inúmeras pesquisas, especialmente em relação aos aspectos fitossanitários, uma vez que a citricultura paulista é fortemente atacada por uma série de pragas e doenças, desde o seu surgimento. A metodologia utilizada para a estimativa do retorno aos investimentos na pesquisa citrícola foi o cálculo da produtividade total dos fatores (PTF), através do Índice de Tornqüist, para posterior comparação com os gastos em pesquisa. Além disso, utilizou-se o método do valor presente líquido (VPL) para comparação entre diferentes cenários com relação à contaminação dos pomares, de modo a estimar o dano econômico evitado pela descoberta e pelo controle das doenças. Dentre os principais resultados encontrados verificou-se que para cada R$1,00 investido na pesquisa citrícola obtém-se um aumento de R$ 13,67 no valor da produção de laranja no Estado de São Paulo. Com relação às perdas de produtividade evitadas graças ao controle das principais doenças, verificou-se que se o cancro cítrico não fosse rigorosamente controlado através da erradicação, o prejuízo teria sido ao redor de R$ 2 bilhões nos últimos 35 anos. Finalmente, a partir da simulação de diferentes níveis de contaminação pelo greening os resultados sugeriram que o prejuízo causado pela possível disseminação desta doença nos pomares paulistas seria bastante significativo nos próximos 20 anos, especialmente se estiverem associados a um cenário de manejo integrado do cancro cítrico, ao invés da erradicação. Em outras palavras, os resultados do estudo corroboram a importância dos investimentos em P&D na citricultura paulista.<br>The economic growth models show that the only way to promote a sustained growth of the economy is through technological progress. However, keeping a growing technological innovation flow requires a strong research and development - R&D structure, demanding expenses from both public and private sectors. Consequently, the research institutions have been increasingly charged about the outcomes from applied funds, which turn necessary to conceptualize and evaluate such R&D investment returns, as well as to highlight what benefits they can reveal to society. There are several studies aiming to assess the whole agriculture R&D investments returns, whereas only few studies has been doing towards to specific sectors. Thus, the general objective of this study is to assess the economic return from R&D investments in São Paulo State's citrus sector over the last 35 years. Brazil represents 80% of the total frozen concentrated orange juice - FCOJ commercialized in the international market, and 28% of the world orange production. São Paulo State is responsible by 80% of national orange production and 98% of Brazilian FCOJ exportation. The economic importance of this industry has been stimulating several research activities, especially related to plant disease aspects, given that São Paulo's citrus sector is strongly threatened by pests and diseases. It was used the Tornqüist Index Method to estimate the citrus industry total productivity of factors - TPF, in order to compare the outcomes with citrus R&D investments. In addition, the net present value - NPV method was used to compare different scenarios, in relation to citrus canker and greening outbreak in São Paulo's groves, to estimate the economic losses avoided due to diseases control. For every R$1 invested in the citrus industry R&D, it was found that R$13,67 is risen on the São Paulo State orange production value. In relation to avoided economic losses due to diseases control, if the citrus canker had not been controlled through tree eradication, the economic losses would have been approximately R$2 billion throughout the past 35 years. Finally, from different levels of citrus greening outbreak simulation, the results suggest that the damage caused by the possible citrus greening outbreak in São Paulo's orchards would be quite significant for the next 20 years, especially if the levels are associated with canker integrated management, regardless of eradication program. Summarizing, the results achieved from this work reinforces the remarkable importance of São Paulo's citrus sector R&D investments.

Made available in DSpace on 2014-06-11T19:28:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-26Bitstream added on 2014-06-13T19:37:24Z : No. of bitstreams: 1 gabia_aa_me_botfca.pdf: 416606 bytes, checksum: 894930ec055a3224888bc51077855bce (MD5)<br>Tylenchulus semipenetrans é o principal nematoide em citros, causando perdas que variam de 10 a 30%. Está amplamente disseminado nos pomares citrícolas do Brasil. Para determinar o nível populacional desse nematoide, que varia durante o ano, diferentes variáveis são utilizadas. Sendo assim o objetivo deste trabalho foi avaliar a flutuação populacional do nematoide T. semipenetrans em pomar de laranja ‘Pera Rio’ enxertada em limoeiro ‘Cravo’, correlacionar a população do nematoide no final de cada estação com a produção e correlacionar o número de fêmeas na raiz com o número de juvenis encontrados nas amostras de solo e raiz. As amostragens foram realizadas no final de cada estação climática, durante a safra 2011/2012, numa profundidade de 0-30 cm, na projeção da copa de 10 plantas sem sintomas, 10 com sintomas e as adjacentes das sintomáticas, totalizando 60 plantas. Os nematoides presentes no solo foram extraídos pela metodologia de flutuação em centrifuga e das raízes pela técnica do liquidificador e centrifugação. Os frutos de cada planta foram colhidos separadamente e pesados. Para contagem das fêmeas, 1g de raiz de cada amostra foi submetida à coloração para facilitar a contagem. O número de nematoides no solo e na raiz foram plotados em gráficos e correlacionados com dados de temperatura e precipitação pluviométrica pela análise de Pearson; os dados de peso de fruto por planta e número de nematoides em solo...<br>Tylenchulus semipenetrans is the main nematodes in citrus trees, causing losses ranging from 10 to 30%. It is widespread in the citrus orchards of Brazil. To determine the level of nematode population which varies over the year, different variable are used. Therefore the objective of this study was to evaluate the fluctuation of nematode T. semipenetrans on acreage with orange ‘Pera Rio’ grafted on ‘Rangpur’, correlate the nematode population at the end of each season with the production and correlate the number of females in the root with the number of juveniles found in soil samples and root . Samples were taken at the end of each season climate, during the 2011/2012 season, at a depth of 0-30 cm, the crown projection of 10 plants without symptoms, 10 with symptoms and symptomatic adjacent plant, totaling 60 plants. The nematodes in the soil were extracted by centrifugal flotation method and the technique of the roots of the blender and centrifugation. The fruits of each plant were harvested and weighed separately. To count the number of females in the roots, 1g of each root sample was colored and subsequently dried. The number of nematodes in soil and roots were plotted on graphs and correlated with data on temperature and rainfall by Pearson analysis, the data for fruit weight per plant and the number of nematodes in soil and roots, on each season, were transformed and analyzed by Spearman correlation, considering both the significance level of 5%. The data on soil nematode, root nematode, sum of soil and root nematode and females per gram of root were analyzed by Pearson correlation analysis and subsequent to the correlation matrix. In the region of Botucatu, central western state of São Paulo, the largest population level of the citrus nematode occurred in late winter and lowest in late spring, and the nematodes... (Complete abstract click electronic access below)

Orientador: Silvia Renata Siciliano Wilcken<br>Banca: Sarita Leonel<br>Banca: Claudio Marcelo Gonçalves de Oliveira<br>Resumo: Tylenchulus semipenetrans é o principal nematoide em citros, causando perdas que variam de 10 a 30%. Está amplamente disseminado nos pomares citrícolas do Brasil. Para determinar o nível populacional desse nematoide, que varia durante o ano, diferentes variáveis são utilizadas. Sendo assim o objetivo deste trabalho foi avaliar a flutuação populacional do nematoide T. semipenetrans em pomar de laranja 'Pera Rio' enxertada em limoeiro 'Cravo', correlacionar a população do nematoide no final de cada estação com a produção e correlacionar o número de fêmeas na raiz com o número de juvenis encontrados nas amostras de solo e raiz. As amostragens foram realizadas no final de cada estação climática, durante a safra 2011/2012, numa profundidade de 0-30 cm, na projeção da copa de 10 plantas sem sintomas, 10 com sintomas e as adjacentes das sintomáticas, totalizando 60 plantas. Os nematoides presentes no solo foram extraídos pela metodologia de flutuação em centrifuga e das raízes pela técnica do liquidificador e centrifugação. Os frutos de cada planta foram colhidos separadamente e pesados. Para contagem das fêmeas, 1g de raiz de cada amostra foi submetida à coloração para facilitar a contagem. O número de nematoides no solo e na raiz foram plotados em gráficos e correlacionados com dados de temperatura e precipitação pluviométrica pela análise de Pearson; os dados de peso de fruto por planta e número de nematoides em solo... (Resumo completo, clicar acesso eletrônico abaixo)<br>Abstract: Tylenchulus semipenetrans is the main nematodes in citrus trees, causing losses ranging from 10 to 30%. It is widespread in the citrus orchards of Brazil. To determine the level of nematode population which varies over the year, different variable are used. Therefore the objective of this study was to evaluate the fluctuation of nematode T. semipenetrans on acreage with orange 'Pera Rio' grafted on 'Rangpur', correlate the nematode population at the end of each season with the production and correlate the number of females in the root with the number of juveniles found in soil samples and root . Samples were taken at the end of each season climate, during the 2011/2012 season, at a depth of 0-30 cm, the crown projection of 10 plants without symptoms, 10 with symptoms and symptomatic adjacent plant, totaling 60 plants. The nematodes in the soil were extracted by centrifugal flotation method and the technique of the roots of the blender and centrifugation. The fruits of each plant were harvested and weighed separately. To count the number of females in the roots, 1g of each root sample was colored and subsequently dried. The number of nematodes in soil and roots were plotted on graphs and correlated with data on temperature and rainfall by Pearson analysis, the data for fruit weight per plant and the number of nematodes in soil and roots, on each season, were transformed and analyzed by Spearman correlation, considering both the significance level of 5%. The data on soil nematode, root nematode, sum of soil and root nematode and females per gram of root were analyzed by Pearson correlation analysis and subsequent to the correlation matrix. In the region of Botucatu, central western state of São Paulo, the largest population level of the citrus nematode occurred in late winter and lowest in late spring, and the nematodes... (Complete abstract click electronic access below)<br>Mestre

15 pp.<br>Originally published: 2000<br>This publication discusses some diseases that are sufficiently important to the citrus in Arizona. Topics include: -Parasitic Diseases -fungi / virus diseases / virus or virus-like diseases -Mycoplasma Diseases -Nematode Diseases -Nonparasitic Diseases

Stony corals (Order Scleractinia) are susceptible to a variety of diseases, which can occur from abiotic or biotic factors, or a combination of both. Public aquaria provide opportunities to study coral disease. Because coral mucus is the first line of defense against disease, the Biolog EcoplateTM is a useful tool to detect differences in microbial assemblages in the surface mucopolysaccharide (mucus) layer when comparing healthy and diseased corals. Histological examination is essential to document structural changes in coral tissue in response to diseases. This study identifies and characterizes diseases in captive corals through visual recognition, characterization of carbon utilization by microbial assemblages in coral mucus samples, and histological examination. In March 2010, surveys were sent to public aquaria throughout the United States that house corals. If the survey was returned indicating that the aquarium had diseased coral specimens present, sample kits were sent to the aquarium to acquire photographs, mucus samples for microbial carbon utilization analysis, and tissue samples for histological examination. Eighteen aquaria participated in the survey and 25 sets of samples were provided. The gross lesions from diseased samples fit into six categories: discoloration associated with darkening of the tissue or with color loss (bleaching), growth anomalies, and tissue loss associated with pests, with brown jelly, or with no obvious cause. Seven categories of possible contributing factors were reported: addition of inadequately quarantined corals to the tank, damage during transport, change of location, manual stress, and variations in light, salinity, or temperature. Introduction of inadequately quarantined specimens was the most common possible contributing factor to pest introduction. Significant differences in carbon source utilization were found between tank-water samples and mucus from both healthy and diseased areas of sampled corals. Although mucus samples from healthy and diseased samples did not differ in carbon source utilization overall, D-mannitol was used by 52% of microbial assemblages from mucus from diseased areas compared with only 17% of microbial assemblages from healthy mucus samples. Histologically, the most commonly observed features across all samples were healthy zooxanthellae, endolithic organisms, and nematocysts, all of which are normal features that can be influenced by stress factors. Brown granular material and ciliates were found associated with some anomalies, primarily the three categories of tissue loss. The presence of dense aggregates of zooxanthellate-engorged ciliates in corals afflicted with brown jelly was highly similar to histological observations of brown band syndrome, previously described from natural coral reefs.

Experiments were initiated to evaluate chemical disease management tools for Alternaria fruit rot on navel orange and Coniophora brown wood rot on lemon trees, examine the possible effect of branch diameter on development of Coniophora wood rot on lemon trees and continue evaluations of relative resistance of rootstocks to root rot and stem canker development when inoculated with P. citrophthora and P. parasitica. Rovral or Kocide did not significantly reduce the amount of Alternaria fruit rot on navel orange trees occurring in late summer and early autumn when applied during the preceding winter or spring months. Of several chemical treatments tested, only Nectec paste inhibited the development of Coniophora brown wood rot on inoculated lemon branches. The size of wood decay columns on branches 10 mm (0.5 inch) in diameter were significantly smaller than those developing on branches 50-70 mm (2.0-2.75 inches) in diameter. In extensive trials evaluating root rot caused by Phytophthora citrophthora and P. parasitica, some relatively tolerant rootstocks were found among the group of new potential rootstocks as well as currently used rootstocks such as rough lemon, C. macrophylla and Troyer citrange. C. volkameriana was relatively tolerant to the development of root rot by P. citrophthora but demonstrated variable tolerance to P. parasitica. Comprehensive evaluation of stem canker development on citrus rootstocks inoculated with P. citrophthora or P. parasitica revealed that rough lemon is usually highly susceptible to both pathogens, while C. volkameriana was at times less susceptible (more tolerant) than rough lemon to both pathogens. Some of the new potential rootstocks were highly tolerant or resistant to infection of stem tissue by P. citrophthora or P. parasitica.

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.