Tesi sul tema "Fusarium diseases of plants taxonomy"

Fusarium root rot is one of the most important diseases of pulse crops, with numerous Fusarium spp. comprising the disease complex. Fusarium solani and F. avenaceum have been reported to be major pathogens in the pea root rot complex, and all commonly grown varieties are susceptible. Greenhouse methods to evaluate peas for resistance to Fusarium root rot resulted in inconsistent disease severity across varieties. In 2015, F. avenaceum infested field plots were more heavily damaged based on emergence and yield than F. solani infested plots, and opposite trends were observed in 2016. Differences in root rot severity between years could be due to F. solani infestation causing more damage under warmer temperatures, while plots infested with F. avenaceum caused more damage under cooler temperatures. These results highlight the difficulties observed when screening for soil-borne pathogens, and the increased difficulties when a pathogen complex and changing environmental conditions are involved.

Thesis (Ph. D.)--Ohio State University, 2005.
Title from first page of PDF file. Document formatted into pages; contains xiii, 118 p.; also includes graphics Includes bibliographical references (p. 114-118). Available online via OhioLINK's ETD Center

A new disease of sugar beet, named Fusarium yellowing decline, was recently found in in the Red River Valley of MN and ND. This disease is caused by a novel pathogen named Fusarium secorum. Pathogens such as F. secorum secrete proteins during infection called ?effectors? that help establish disease. Since pathogenicity and disease development may depend on effector proteins produced by F. secorum during infection, effector protein identification furthers our understanding of the biology of this important pathogen. A list of 11 candidate effectors was generated previously. In this study, to characterize putative effectors, we developed a transformation system using polyethylene glycol?mediated transformation. Several mutant lines were created with an effector deleted from the genome using a split-marker knock-out strategy. To explore their role in pathogenicity, mutant strains have been inoculated to sugarbeet and compared to WT F. secorum.

Tubers of Russet Burbank were surface disinfested and 3 mm diameter by 3 mm deep wounds were made with cork borer. The holes were inoculated with 20muL of 104 macroconidia/ml suspension of (Fusarium sambucinum) or 20muL of 104 sporangia/ml suspension (Pythium ultimum) and incubated under mist. For infection studies, the inoculated tubers were exposed to 0--48 h of mist at 4--20ºC, dried and stored at 16ºC and 95% RH in growth chamber with forced air for 15 d (F. sambucinum), whereas stored at 12ºC and 95% RH for 30 d (P. ultimum). For lesion expansion studies tubers exposed to 24 h wet at 16ºC were stored in growth chambers at 4, 8, 12, and 16ºC for 15--90 d. At the end of storage tubers were cut and the volume of diseased area was measured. Models explained 94.2% of the variation in infection and 99.7% in lesion expansion for F. sambucinum . Whereas, models explained 96.7% of the variation in infection and 99.6% lesion expansion for P. ultimum. (Abstract shortened by UMI.)

An evaluation of nineteen barley lines using three artificial inoculation methods concluded that spray inoculation was the most reproducible method and provided the greatest discrimination of resistance. Six of the nineteen barley lines were used for proteomic studies to identify defense responses following F. graminearum infection. All lines responded by inducing an oxidative burst and pathogenesis-related proteins. Differences in response magnitude and the proteins activated could be attributed to varying levels of FHB resistance amongst the barley lines. RNA microarray profiling and iTRAQ technology were used to study the interaction between two barley lines under five different treatments testing the effect of the fungus, trichothecene, and their interaction. Resistance was differentiated by the early induction of defense-related genes and the activation of the JA and ethylene defense pathways in Chevron, compared to the induction of a less efficient defense pathway in Stander; observed intra- and inter-cultivar differential responses are discussed.
xvii, 196 leaves : ill. ; 29 cm.

Three independent studies were conducted for spot blotch (Bipolaris sorokiniana), Fusarium head blight (FHB) (Fusarium graminearum), and root rot diseases (Fusarium species and B. sorokiniana). Histopathology of compatible and incompatible interactions between different pathotypes of B. sorokiniana and different genotypes of barley was examined with red fluorescent protein-tagged fungal isolates. The fungus penetrated the host cell wall and developed multicellular globular infection hyphae (IH) in the lumen of epidermal cells, but infected epidermal cells appeared to be alive till 16 hours post-inoculation (HPI). In the susceptible plants, the tip of IH was found to grow ahead of the dead tissue and invade the surrounding live mesophyll cells, whereas growth of IH in the resistant plants was restricted to the dead tissue after 20 HPI. The amount of H2O2 accumulation and the fungal biomass were also significantly higher in the susceptible hosts than in the resistant hosts. To map resistance to FHB, two populations consisting 130 doubled haploid lines from the cross Grandin ? PI277012 and 237 recombinant inbred lines from the cross Bobwhite ? ND2710 were phenotyped and genotyped. QTL for Type I resistance were identified on chromosomes 1A, 2B, 4B, 5B and 6B in the GP population. These QTL explained 10.7-19 % of the total phenotypic variation. With the BN population, QTL for Type I resistance were identified on chromosomes 2A, 5A and 6B, explaining 6.2-13.7% of the total phenotypic variation. To assess the prevalence, incidence and severity of wheat crown rot (CR) and common root rot (CRR) in ND, wheat root samples were collected from fields across the state in 2012, 2013, and 2014. Fungal isolations indicated that B. sorokiniana was most frequently recovered in all sampled years. Seedling tests on ten spring wheat lines showed that Glenn was the least susceptible while Steele-ND was the most susceptible to one F. culmorum isolate and one B. sorokiniana isolate tested. Evaluation of 20 spring wheat genotypes for reaction to CRR at the adult plant stage showed that Freyr and RB07 were more resistant while Len and Briggs were more susceptible to CRR compared to other wheat genotypes evaluated.
North Dakota Wheat Commission,
Minnesota Wheat Research and Promotion Council
ND State Board of Agricultural Research and Education
Triticeae-CAP project (2011-68002-30029) of the US Department of Agriculture National Institute of Food and Agriculture
U.S. Wheat and Barley Scab Initiative (USWBSI)

The wheat stem sawfly is the most destructive insect pest in both winter wheat and spring wheat production in the northern Great Plains. The sawfly is univoltine, and spends all immature stages within protective wheat stems, which explains the difficulty in controlling populations. However, the almost continuous inhabitation of stems also makes larvae more vulnerable to invasion by microorganisms colonizing both living stems and postharvest stubble. Fusarium spp. were frequently isolated from fungalcolonized larval cadavers, and were found to be the major lethal factors for overwintering larvae in both laboratory emergence experiments and field surveys over three years. The pathogenicity of single isolates of three Fusarium spp., including F. pseudograminearum, F. culmorum, and F. acuminatum, was evaluated against overwintering larvae in sawflycut stems and against actively-feeding larvae in growing winter wheat plants over two years. The tested Fusarium isolates caused twenty to sixty percent mortality in overwintering larvae, and caused forty to eighty percent mortality in actively-feeding larvae. The Fusarium isolates also caused decomposition of sawfly-cut stems and disease in wheat plants, which reflected the versatility of Fusarium isolates acting as saprophytes, entomopathogens, and phytopathogens. Deoxynivalenol was detected in wheat stem tissues colonized by the Fusarium isolates from two years of field experiments, and deoxynivalenol caused toxicity and inhibited the growth of second and third-instar actively-feeding larvae in laboratory bioassays. Wheat grower observations of greater sawfly infestation in dryland wheat fields led to assessment of larval mortality from Fusarium infection in both dryland and irrigated wheat fields. This was studied using cage experiments or field surveys at different locations for three years. Parasitoid attack and fungal infection, mainly by Fusarium spp., were found to be the major lethal factors for developing and overwintering larvae, respectively. There was no difference in sawfly survival in dryland or irrigated wheat fields. As ubiquitous soil microorganisms and plant pathogens, Fusarium spp. impact wheat stem sawfly populations in the field each year.

Responses from pedigree selection for resistance to gibberella ear rot were assessed in four maize (Zea mays L.) populations, two selected after inoculation of Fusarium graminearum (Schwabe) macroconidia into the silk channel and two selected after inoculation into developing kernels. Responses were significant in both populations selected for silk resistance and in one of the populations selected for kernel resistance. Selection was more effective in later generations and genetic gains were associated with among-family selection but not with within-family selection. Results obtained here indicate that responses to selection could be more efficiently obtained by applying high selection intensities in advanced generations, by managing earlier generations as bulks and by reducing the number of plants per family. In another experiment, a wide sample of Argentine maize germplasm was evaluated for silk and kernel resistance to gibberella ear rot and to fusarium ear rot (caused by F. verticillioides (Saccardo) Nirenberg [=F. moniliforme (Sheldon)]. Several entries exhibited disease resistance in comparison with local check hybrids, particularly for fusarium ear rot, the most prevalent ear rot in Argentina. Results obtained in this study suggested the presence of general mechanisms controlling silk and kernel resistance to both diseases. In a supplementary study, viral diseases were surveyed in maize fields from the provinces of Ontario and Quebec in 1999 and 2000. Barley yellow dwarf was found in 1999. Sugarcane mosaic, maize dwarf mosaic and wheat streak mosaic were found in 2000. These diseases were not important for grain-maize planted in May, the most prevalent kind of maize crop in these provinces. Some of these diseases, such as sugarcane maize mosaic and maize dwarf mosaic were found important only in maize fields planted during or after the month of June, and this is of commercial relevance only for sweet corn.

Thesis (DSc (Biochemistry))--Stellenbosch University, 2009.
The fumonisin mycotoxins are known to be the causative principle for several animal diseases and are associated with the development of liver and oesophagus cancer and neural tube defects in humans. The thesis focuses mainly on the characterisation of the compounds from maize cultures of the fungus Fusarium verticillioides, isolated from maize, the toxicological effects in animals, mechanism involved in hepato- and nephrocarcinogenicity and discussing the major differences and contradictions in the literature together with their impact on setting relevant risk assessment parameters to safeguard human health. Controversies include the importance of non-genotoxicity vs genotoxicity in the development of cancer, the role of threshold effects in carcinogenesis and the establishment of realistic risk assessment parameters that will also be applicable in developing countries. Recent approaches suggest that thresholds should also apply for genotoxic carcinogens as interaction with the DNA is only one event in the multi-step process of cancer development and therefore could not be taken as the basis for applying a no-effect threshold for genotoxins. It would appear that a carcinogen such as fumonisin, whether it is labeled genotoxic or non-genotoxic per se, exhibits some degree of risk at any level due to additive or synergistic interactions with other xenobiotics and/or dietary constituents. The underlying mechanisms of fumonisin-induced carcinogenicity includes the disruption of sphingolipid, phospholipids and fatty acid metabolism, which plays a major role in the modulation of apoptotic and cell proliferative pathways related to cancer development. Interactive responses between arachidonic acid and ceramide affect downstream cell signal transduction pathways and depending on the cell type the disruption of these pathways could either stimulate or inhibit cell proliferation which eventually will determine the induction of apoptosis and hence affect cell survival. The modulating roles of dietary constituents such as vitamins, protein and the South African herbal teas are also highlighted as they affected the outcome of toxicological assays, thus determining thresholds of the adverse effects in specific target organs that will impact risk assessment parameters. Regulation of the fumonisins in food and the associated risk are debated from many perspectives. In developing countries there is a lack of quality control implying that maize highly contaminated with mycotoxins may directly enter the food chain of adults and children as control of mycotoxins is difficult or in some cases totally absent. The interaction of politics, economy and technology will eventually determine the impact on health as the regulation of fumonisin in food differs between countries. Knowledge about the biological effects of the fumonisins is currently playing an important role in the development of simple and inexpensive methods to reduce the levels of the fumonisin in maize by targeting specific populations at risk.

Thesis (PhD(Agric))--University of Stellenbosch, 2000.
ENGLISH ABSTRACT: The purpose of this study was to characterize the race and vegetative compatibility of Fusarium oxysporum f. sp. melonis (FOM) isolates collected in the major melon producing areas, to report on their geographical distribution, and their possible relatedness to isolates from other countries. Seventy two FOM isolates obtained from 30 fields in 17 melon producing regions were race-typed using the differential cultivars Topmark (susceptible to all races), Doublon (Fomi), CM 17187 (Fom2) and Perlita (Fom3) and grouped by means of vegetative compatibility. All isolates belonged to vegetative compatibility group 0134, indicating a high degree of genetic homogeneity among the South African FOM population. Fifty four isolates were identified as race 0, eight as race 1, and 10 as race 2. Race 0 occurred in 15 of the regions whereas race 1 was sporadically recovered. Race 2, on the other hand, was obtained only from four fields located in one geographical region. Perlita plants (carrying the gene Fom3) inoculated with local isolates ofrace 0 and race 2 and reference isolates of race 0 became stunted, their leaves turned yellow, and became thickened and brittle. These results suggested that Fom3 in Perlita confers a tolerant reaction compared to the resistant reaction of gene FornI in Doublon. The disease reaction of cultivar Perlita to FOM was therefore reinvestigated. Twenty isolates, including the four FOM races (0, 1, 2, and 1,2) obtained from different countries, were used. The differential cultivars were included to verify virulence of the isolates. Perlita plants inoculated with three isolates of race 2 remained asymptomatic. The remaining race 2 and 0 isolates, induced severe stunting of Perlita plants, but mean percentage stunting values did not differ significantly (P = 0.05) and ranged between 25.1 and 50.0. Leaves of stunted plants were chlorotic, thickened and brittle. Disease reaction of Perlita was verified at a lower inoculum concentration with two race 2 (pipette method) and two race 0 isolates (root dip method). Results proved that Fom3 does not confer similar resistance towards race 0 and some race 2 isolates as FornI in Doublon. Cultivars possessing Fom3, should therefore be considered tolerant to FOM races 0 and 2. The ability of a nit mutant isolate, generated from FOM race 0 which belongs to VCG 0134, to change its virulence during infection of melon plants, was investigated under quarantine. Seedlings of melon cultivars Imperial 45 and Early Sweet (no resistance genes), Amber (Fom2) and Fiata (FomI, Fom2) were consecutively grown in two cement troughs in a gauzehouse. Each planting was terminated when plants had advanced Fusarium wilt or after the fruit were harvested. In the first planting, Imperial 45 seedlings were transplanted and artificially inoculated with the nil mutant isolate. In the consecutive plantings, seeds were sown in the infested soil to enable natural infection. For each crop, representative plants showing Fusarium wilt were selected for isolation. All F. oxysporum isolates recovered were single-spored and their nit mutant and VCG status verified. Virulence of the labelled isolates was determined using differential cultivars. In trough A, all plants of the susceptible cultivars Imperial 45 and Early Sweet crops showed Fusarium wilt. The labelled isolates recovered from the selected plants were all designated race O. In the first crop (planting No.5) of the resistant cultivar Amber, 6.7% of the plants developed Fusarium wilt. In the second Amber crop the disease incidence increased to 56.6%, and to 81.8% in the final crop. Contrary to the susceptible cultivars, only race 2 isolates were obtained from the symptomatic Amber plants. Similar data were found with the susceptible cultivar Imperial 45 and the resistant cultivar Amber in trough B. Planting of Fiata caused a dramatic reduction in Fusarium wilt incidence in trough B. However, 1.2% of plants were affected by Fusarium wilt in the first Fiata crop (planting No.6), whereas 4% of the plants were symptomatic in the final planting. From these symptomatic Fiata plants only race 1,2 isolates were obtained. These findings, and the fact that the symptomatic plants represented a substantial proportion of the first Amber (approximately 7-15%) and Fiata (approximately 2%) crops, provedthat changes in the race structure of this fungal pathogen occurred rapidly when confronted with a resistant cultivar. The potential of RAPD analysis to differentiate between the isolates displaying virulence changes was evaluated. Four F. oxysporum f. sp. niveum isolates were included as an outgroup. A histopathological study was conducted to verify whether these isolates retain their ability to behave as true vascular pathogens. The three primers used clearly distinguished the 12 FOM isolates from the four F. oxysporum f. sp. niveum isolates. However, the primers showed a highly conserved and characteristic banding pattern for the FOM isolates which represented three physiological races (race 0, race 2, race 1,2), indicating that RAPD analysis cannot detect race-specific groupings in FOM. Disease reactions on the three differential cultivars confirmed the virulence of FOM isolates. The histopathological data furthermore proved that the two FOM races (race 2, race 1,2), which derived from the race 0 parent isolate, retained their ability to behave as true vascular pathogens.
AFRIKAANSE OPSOMMING: DIE KARAKTERISERING EN PATOGENESITEIT VAN SUID-AFRIKAANSE ISOLATE VAN FUSARIUMOXYSPORUMF. SP.MELONIS Die doel van die studie was om Fusarium oxysporum f. sp. melonis (FOM) isolate wat in die hoof spanspekproduserende gebiede versamel is, volgens ras en vegetatiewe verenigbaarheid te karakteriseer, en hul geografiese verspreiding en verwantskap met isolate van ander lande aan te dui. Twee en sewentig FOM isolate afkomstig vanaf 30 landerye wat 17 spanspekproduserende areas verteenwoordig, is gebruik. Die differensiële kultivars Topmark (vatbaar vir alle rasse), Doublon (Forni), CM 17187 (Fom2) en Perlita (Fond) is gebruik om die rasbepalings te doen asook om die vegetatiewe verenigbare groepe (VVG) te bepaal. Al die isolate is as VVG 0134 geklassifiseer, wat 'n hoë mate van genetiese homogenesiteit binne die Suid-Afrikaanse populasie aandui. Vier en vyftig isolate is as ras 0, agt as ras 1 en 10 as ras 2 geklassifiseer. Ras 0 is vanaf 15 gebiede afkomstig, terwyl ras 1 sporadies voorgekom het. Ras 2 is vanuit vier landerye binne dieselfde geografiese gebied verkry. Plante van die kultivar Perlita wat met plaaslike isolate van ras 0 en 2, asook verwysings-isolate van ras 0 geïnokuleer is, het verdwerg voorgekom. Die blare van die plante het vergeel, verdik en bros voorgekom. Hierdie siekte reaksie het aangedui dat Fond in Perlita toleransie bewerkstellig in teenstelling met die weerstandbiedende reaksie van geen Fomi in Doublon. Die siekte reaksie van Perlita teenoor FOM is dus verder ondesoek. Hiervoor is 20 isolate wat al vier FOM rasse insluit (0, 1, 2, en 1,2), en van verskillende wêrelddele afkomstig is, gebruik. Die virulensie van die isolate is met die differensiële kultivars bevestig. Drie van die ras 2 isolate het geen siektesimptome op Perlita veroorsaak nie. Die ander ras 2 isolate, en al die ras 0 isolate, het egter die Perlita plante aansienlik verdwerg en die blare vergeel en verdik. Laasgenoemde groep isolate het 'n gemiddelde verdwergingsindeks van tussen 25.1% en 50.0% veroorsaak. Die siekte reaksie by Perlita is verder bevestig deur plante teen 'n laer inokulumdigtheid van twee ras 2 (pipet metode), en twee ras 0 (wortel-doop metode) isolate, te inokuleer. Uit die resultate was dit duidelik dat die weerstand wat Fom3 teenoor ras 0 en sommige ras 2 isolate verskaf, van FornI verskil. Kultivars wat oor die weerstandsgeen Fom3 beskik moet dus as tolerant beskou word. 'n Ondersoek is geloods na die vermoë van 'n nil mutant isolaat, genereer vanaf die wilde ras 0 isolaat van FOM (VVG 0134), om onder kwarantyn sy virulensie gedurende infeksie van spanspekplante te verander. Saailinge van die spanspekkultivars Early Sweet (geen weerstandsgene), Amber (Fom2) en Fiata (FornI, Fom2) is opeenvolgens in twee sement trêe in 'n gaashuis verbou. Die afsonderlike aanplantings is beëindig sodra gevorderde Fusarium-verwelksimptome verkry is, of nadat vrugte ge-oes is. Vir die eerste aanplanting is oorgeplante Imperial 45 saailinge kunsmatig met die nil mutant isolaat geïnokuleer. Tydens die opeenvolgende aanplantings is saad direk in die besmette grond gesaai ten einde natuurlike infeksie te verkry. Met elke aanplanting is isolasies gedoen vanaf verteenwoordigende plante wat Fusarium-verwelksimptome getoon het. Alle F. oxysporum isolate wat verkry is, is ge-enkelspoor en hul nit mutant status en VVG is bevestig. Virulensie van die gemerkte isolate is bepaal deur inokulasie van die differensiële kultivars. Alle plante van die vatbare Imperial 45 en Early Sweet kultivars wat in trog A geplant is, het Fusarium-verwelksimptome getoon. Die gemerkte isolate wat vanaf die verteenwoordigende plante verkry is, is almal as ras 0 geklassifiseer. Tydens die eerste aanplanting van die weerstandbiedende kultivar, Amber (aanplanting No.5), het 6.7% van die plante Fusarium-verwe1ksimptome ontwikkel. Tydens die tweede en derde aanplanting van Amber het die frekwensie van siektevoorkoms verhoog na 56.6% en 81.8 %, onderskeidelik. In teenstelling met die vatbare kultivars, is slegs ras 2 vanuit die Amber plante met siektesimptome verkry. Soortgelyke resultate is met Imperial 45 en Amber in trog B verkry. Aanplanting van kultivar Fiata het egter 'n dramatiese verlaging in die voorkoms van Fusarium-verwelk bewerkstellig. Tydens die eerste Fiata aanplanting (aanplanting No.6) het 1.2% plante Fusarium-verwelksimptome ontwikkel, en 4% tydens die laaste aanplanting. Vanaf hierdie plante is slegs ras 1,2 isolate verkry. Hierdie bevindings, en die feit dat 'n aansienlike hoeveelheid van die Amber (ongeveer 7-15%) en Fiata plante (ongeveer 2%) siektesimptome getoon het, bewys dat FOM vinnig van virulensie verander wanneer die patogeen 'n weerstanbiedende kultivar infekteer. Die vermoë van RAPD analise om tussen isolate wat in virulensie verander het, te onderskei, is ondersoek. Vier isolate van F. oxysporum f. sp. niveum is as 'n buite-groep ingesluit. Om te bevestig dat die isolate wat van ras verander het wel egte vaskulêre patogene is, is 'n histopatologiese ondersoek gedoen. Die drie inleiers wat gebruik is, het die 12 FOM isolate duidelik van die vier F. oxysporum f. sp. niveum isolate onderskei. Die 12 FOM isolate wat drie fiosologiese rasse (ras 0, ras 2, ras 1,2) verteenwoordig het, is egter saam gegroepeer, wat aandui dat hierdie metode nie tussen rasse van FOM kan onderskei nie. Inokulasiestudies met die differensiële kultivars het die virulensie van die isolate bevestig. Die histopatologiese ondersoek het verder bewys dat beide FOM rasse (ras 2, ras 1,2) wat vanaf die wilde tipe ras ° isolaat ontstaan het, hul vermoë behou het om as egte vaskulêre patogene op te tree.

Common barley diseases observed in North Dakota include net blotch, spot blotch, leaf and stripe rust, bacterial leaf streak, and Fusarium head blight. The first objective of this research was to determine the effect of variety and fungicide timing on disease development of barley under conventionally tilled systems. Five field trials were performed in 2016-2017 to test the effect of common varieties and fungicide applications on foliar disease of barley. Overall, varietal selection had a greater effect on the level of foliar disease observed than fungicide application. The second objective focused on the efficacy and timing of adepidyn and prothioconazole + tebuconazole on Fusarium head blight. An inoculated greenhouse experiment was performed the fall of 2017 to determine the effectiveness of fungicide timing at half-spike, full-spike, and five days after full-spike. The protectant capabilities of the fungicides were greater than their curative properties.

Fungicides have been developed to protect plants against diseases and pests, which cause serious problems such as the loss of germination and vigour. The aim of this study was to test the germination and vigour of maize (Zea mays L.) seeds treated with several fungicides Apron® Star 42 WS (difenoconazole, thiamethoxam, and metalaxyl-m), Apron® XL (mefenoxam), Celest® XL (fludioxonil, mefenoxam) and thiram in the laboratory. In the greenhouse, the efficacy of fungicide treatment was evaluated in soil inoculated with Fusarium graminearum. The control consisted of untreated seeds. Germination and vigour were evaluated according to the International Seed Testing Association (1ST A) rules. The results from the standard gennination tests showed that all the fungicide treated seeds did not differ to the untreated control. The conductivity of solute leakage was read following slow and fast imbibition. Maize seeds treated with Apron® Star 42 WS, Celest® XL, Apron® XL and thiram improved or maintain vigour, which was indicated by a reduced or equivalent solute leakage following fast imbibition when compared with the untreated control. The good performance of fungicide treated seed expressed during conductivity test after fast imbibition correlated with the tetrazolium. All the fungicide treated seeds maintained the same viability as the untreated control following fast imbibition. After 6 h after fast imbibition, Apron® Star 42 WS, Celest® XL and Apron® XL treated seeds maintained similar germination percentages when compared to the untreated control with the exception of thiram treated seeds that exhibited a decline in seed viability. There was reduction in vigour in all the fungicide treated seeds fo llowing 24 and 40 h fast imbibition as illustrated by the reduction in germination percentage below the acceptable level (70%) when compared with the untreated control. The greenhouse study showed that all the fungicide treated seeds maintained the same emergence percentage in both inoculated and uninoculated soil with the exception of thiram treated seeds, where emergence improved in inoculated soil when compared to the untreated control. Apron® Star 42 WS and Celest® XL reduced the disease caused by F. graminearum in the inoculated soil. This study also revealed that the application of Apron® Star 42 WS, Celest® XL and thiram to seeds improved both the shoot and root dry mass of plants in the inoculated soil.
Dissertation (MInstAgrar)--University of Pretoria, 2011.
Microbiology and Plant Pathology
unrestricted

Les maladies du blé causées par Fusarium, comme la brulure de l’épi (FHB) et la pourriture de la tige (FCR), entrainent une réduction de production, de la qualité du blé et des problèmes de sécurité alimentaire liés à la présence de mycotoxines affectant la santé de l’Homme et des animaux: la plus représentée étant le déoxynivalénol (DON). Le DON est un inhibiteur de la synthèse protéique qui agit durant l’infection comme un facteur de virulence. La glycosylation du DON en D3G (DON-3-O-glicoside) catalysée par des UDP-glycosyltransférases (UGTs) est le principal mécanisme de protection des plantes contre sa toxicité. Dans ce travail, nous avons démontré que la détoxification du DON par l’UGT confère une résistance à large spectre contre les champignons produisant DON F.graminearum et F.culmorum. Nous avons produit des plants de blé dur exprimant de manière constitutive le gène HvUGT13248 (Ubi-UGT) et des plants de blé panifiables exprimant ce gène au niveau du tissu floral (Lem-UGT). Les plants Ubi-UGT ont montré une réduction significative des symptômes de FHB durant les stades précoces et médians de l’infection, et de FCR à tous les stades de l’infection. De plus, les plants Lem-UGT ont montré une corrélation entre les niveaux d’expression de l’UGT et de protection observée. Finalement, nous avons démontré que la pyramidation des gènes associés à des mécanismes de résistance différents peut renforcer la résistance de l’hôte à l’infection. Des plants de blé ont été générés exprimant à la fois l’enzyme HvUGT13248, et des inhibiteurs de glycosidases: AcPMEI ou PvPGIP2, impliqués dans la dégradation de la paroi cellulosique, et qui ont montré une résistance accrue à la FHB
Fusarium diseases, including Fusarium head blight (FHB) and Fusarium crown rot (FCR) represent major agricultural problems worldwide, causing reduction of grain yield and quality and food safety. In particular, grain contamination by Fusarium mycotoxins, mainly deoxynivalenol (DON), is responsible for health problems in humans and animals. DON is a protein synthesis inhibitor, acting as a virulence factor during pathogenesis. The principal mechanism involved in enhancing plant tolerance to DON is glycosylation, forming DON-3-β-D-glucoside (D3G), performed by specific UDP-glucosyltransferases (UGTs). In this work, we demonstrated that DON-detoxification by UGT confers a broad-spectrum resistance against the DON-producing fungi F. graminearum and F. culmorum, characterized by different time of infection and target organs. We produced transgenic durum wheat plants (Ubi-UGT) constitutively expressing the barley HvUGT13248 and bread wheat plants (Lem-UGT) expressing HvUGT13248 in flower tissues. Ubi-UGT plants revealed significant reduction of FHB symptom, during early-mid stages of infection, and of FCR symptom, throughout the infection timing. The floral-specific expression highlighted a dose-dependent efficacy of the UGT detoxification mechanism. In addition, we demonstrated that pyramiding of genes controlling different resistance mechanisms can further reinforce the host response by stacking transgenes controlling the DON-to-D3G conversion and the inhibition of cell wall degrading enzymes by glycosidase inhibitors in the same wheat genotype. We obtained plants expressing HvUGT13248 and AcPMEI or HvUGT13248 and PvPGIP2, which exhibited increased FHB resistance

Thesis (PhD)--Stellenbosch University, 2013.
ENGLISH ABSTRACT: Maize (Zea mays L.) is used for human and animal consumption in diverse forms, from specialised foods in developed countries, to staple food in developing countries. Unfortunately, maize is prone to infection by different Fusarium species that can produce harmful mycotoxins. Fusarium verticillioides is capable of asymptomatic infection, where infected kernels show no sign of fungal growth, but are contaminated with mycotoxins. If fungal contamination is not detected early on, mycotoxins can enter the food chain. Rapid and accurate methods are required to detect, identify and distinguish between pathogens to enable swift decisions regarding the fate of a batch or consignment of cereal. Near infrared (NIR) hyperspectral imaging and multivariate image analysis (MIA) were evaluated to investigate the fungal development in maize kernels over time. When plotting principal component (PC) 4 against PC5, with percentages sum of squares (%SS) 0.49% and 0.34%, three distinct clusters were apparent in the score plot and this was associated with degree of infection. Prominent peaks at 1900 nm and 2136 nm confirmed that the source of variation was due to changes in starch and protein. Variable importance plots (VIP) confirmed the peaks observed in the PCA loading line plots. Early detection of fungal contamination and activity (20 h after inoculation) was possible before visual symptoms of infection appeared. Using NIR hyperspectral imaging and MIA it was possible to differentiate between species of Fusarium associated with maize. It was additionally applied to examine the fungal growth kinetics on culture media. Partial least squares discriminant analysis (PLS-DA) prediction results showed that it was possible to discriminate between species, with F. verticillioides the least correctly predicted (between 16-47% pixels correctly predicted). For F. subglutinans 78-100% and for F. proliferatum 60-80% pixels were correctly predicted. Three prominent bands at 1166, 1380 and 1918 nm were considered to be responsible for the differences between the growth zones. Variations in the bands at 1166 and 1380 nm were correlated with the depletion of carbohydrates as the fungus grew while the band at 1918 nm was a possible indication of spore and new mycelial formation. By plotting the pixels from the individual growth zones as a function of time, it was possible to visualise the emergence and interaction of the growth zones as separate growth profiles. The microstructure of fungal infected maize kernels was studied over time using high resolution X-ray micro-computed tomography (μCT). The presence of voids and airspaces could be seen in two dimensional (2D) X-ray transmission images and in the three dimensional (3D) tomograms. Clear differences were detected between kernels imaged after 20 and 596 h of inoculation. This difference in voids as the fungus progressed showed the effect of fungal damage on the microstructure of the maize kernels. Imaging techniques are important for rapid, accurate and objective evaluation of products for quality and safety. NIR hyperspectral imaging offers rapid chemical evaluation of samples in 2D images while μCT offers 3D microstructural information. By combining these image techniques more value was added and this led to a comprehensive evaluation of Fusarium infection in maize.
AFRIKAANSE OPSOMMING: Mielies (Zea mays L.) word in verskeie vorms deur mens en dier verbruik, van gespesialiseerde voedsel in ontwikkelde lande, tot stapelvoedsel in ontwikkelende lande. Ongelukkig is mielies onderhewig aan besmetting deur verskeie Fusarium spesies wat skadelike mikotoksiene kan produseer. Fusarium verticilloioides is in staat tot asimptomatiese infeksie waar die besmette pit geen teken van fungusgroei toon nie, maar (reeds) met mikotoksiene besmet is. Indien fungusbesmetting nie vroegtydig opgespoor word nie, kan mikotoksiene die voedselketting betree. Vinnige en akkurate metodes word benodig om patogene op te spoor, te identifiseer en ook om onderskeid tussen patogene te tref om sodoende (effektiewe) besluite aangaande die gebruik van ‘n lot of besending graan te neem. Naby-infrarooi (NIR) hiperspektrale beelding en meerveranderlike beeld ontleding (MIA) is geëvalueer om fungusontwikkeling in mieliepitte oor tyd te ondersoek. Wanneer hoofkomponent (PC) 4 teenoor PC5 gestip word, met persentasies som van kwadrate (%SS) 0.49% en 0/34%, is drie afsonderlike groepein die telling grafiek waargeneem. Dit is geassosieer met die graad van besmetting. Prominente pieke by 1900 nm en 2136 nm het bevestig dat veranderinge in stysel en proteïene die bron van die variasie was. Veranderlike belangrikheidsgrafieke (VIP) het die pieke wat in die PCA beladingslyngrafieke waargeneem is, bevestig. Vroegtydige opsporing (bespeuring) van fungusbesmetting en aktiwiteit (20 h na inokulasie) was moontlik voor visuele besmettingsimptome verskyn het. Onderskeid tussen Fusarium spesies wat met mielies geassosieer word, was moontlik deur gebruik te maak van NIR hiperspektrale beelding en MIA. Dit is bykomend toegepas om fungusgroeikinetika op kwekingsmedia te bestudeer. Parsiële kleinste kwadrate diskriminantanalise (PLS-DA) voorspellingsresultate het getoon dat dit moontlik was om tussen spesies te onderskei, met F. verticillioides die minste korrek voorspel (tussen 19-47% beeldelemente korrek voorspel). Vir F. subglutinans is 78-100% en vir F. proliferatum is 60-80% beeldelemente korrek voorspel. Drie prominente bande by 1166, 1380 en 1918 nm is oorweeg as oorsaak vir die verskille tussen die groeisones. Variasies in die bande by 1166 en 1380 nm is gekorreleer met die vermindering van koolhidrate soos die fungus groei, terwyl die band by 1918 nm ‘n moontlike aanduiding van spoor en nuwe miseliale vorming is. Deur die beeldelemente van die individuele groeisones as ‘n funksie van tyd te stip, was dit moontlik om die verskyning en interaksie van die groeisones as aparte groeiprofiele te visualiseer. Hoë-resolusie X-straal mikro-berekende tomografie (μCT) is gebruik om die mikrostruktuur van fungusbesmette mieliepitte oor tyd te ondersoek. Die voorkoms van leemtes en lugruimtes kon in die twee-dimensionele (2D) X-straal transmissie beelde en in die drie-dimensionele (3D) tomogramme gesien word. Duidelike verskille is waargeneem tussen pitte wat na 20 en 596 h na inokulasie verbeeld is. Hierdie verskil in leemtes soos die fungus vorder, het die effek van fungusskade op die mikrostruktuur van mieliepitte getoon. Beeldingstegnieke is belangrik vir vinnige, akkurate en objektiewe evaluasie van produkte vir kwaliteit en veiligheid. NIR hiperspektrale beelding bied vinnige chemiese evaluering van monsters in 2D beelde, terwyl μCT 3D mikrostrukturele inligting gee. Meer waarde is toegevoeg deur hierdie beeldingstegnieke te kombineer en dit het gelei tot ‘n omvangryke evaluering van Fusarium besmetting in mielies.

Fusarium subglutinans sensu lato is a complex of fungi, which are the causal agents of important diseases on a wide variety of plants. Two important diseases caused by F. subglutinans sensu lato are pitch canker and mango malformation. F. subglutinans sensu lato isolates causing pitch canker on pine trees have been described as a separate species, F. circinatum. whereas F. subglutinans sensu lato isolates associated with mango malformation have not been formally described. The objective of study was to clarify the taxonomy and population genetics of the pitch canker and mango malformation fungi residing in the Gibberellafujikuroi complex. The introductory chapter of this thesis provides a review of the taxonomic classifications used for Fusarium spp. in the G. fujikuroi complex. In addition, the current knowledge pertaining to the population structure of the pitch canker and mango malformation fungi is discussed. In the second chapter the occurrence of F. circinatum was investigated in Mexico. Fusarium isolates were collected from pine trees in Mexico and identified as F. circinatum. Morphology, sexual compatibility studies, pathogenicity tests and histone H3-RFLPs were used to identify and characterize this fungus. The pitch canker fungus, F. circinatum and its teleomorph, G. circinata has been recently described. However, the name G. circinata is invalid, because insufficient information was provided to characterize the type specimen in the description. Additional information and a selection of F. circinatum isolates were, therefore, obtained and studies were undertaken in order to validate the description of G. circinata. The teleomorph G. circinata was validated and morphological criteria were shown to clearly distinguish F. circinatum from other F. subglutinans sensu lato isolates. Chapter four presents a study aimed at better understanding relationships between populations of F. circinatum from different geographical areas. For this study co¬dominant molecular markers were developed. These were then used to determine the genetic diversity, genetic distance and migration between different F. circinatum populations. Analysis revealed a high diversity in the Florida population and a low diversity in the South African population. Genetic analysis also indicated that the South African F. circinatum population originated in Mexico. In chapter five, the influence of sexual reproduction on the F. circinatum populations sampled over ten years in South Africa were studied. Sexual compatibility, vegetative compatibility and allelic diversity that were determined using polymorphic markers, developed in chapter four, were used. These results suggested that sexual reproduction is occurring more frequently in the more recently collected populations than in the initial population. Mango malformation is an important disease in mango growing areas. The study presented in chapter six indicated that this disease is associated with two distinct Fusarium spp. in the section Liseola. The two new Fusarium spp. are thus described as F. mangiferae and F. sterilihyphosum using morphological criteria In chapter seven, the distribution and vegetative compatibility of both F. mangiferae and F. sterilihyphosum was determined for the South Africa populations. Results revealed that each of these species differ in their distribution in South Africa. Vegetative compatibility tests also suggest that both species represent single genets in South Africa. Fusarium subglutinans sensu lato isolates associated with pme and mango are economically important fungi. The focus of the studies presented in this thesis has been on the taxonomy and population genetics of these fungi, with special reference to their occurrence in South Africa. Each of the chapters will contributes towards a better understanding of the taxonomy, population genetics and biology of these fungi.
Thesis (PhD)--University of Pretoria, 2006.
Microbiology and Plant Pathology
Unrestricted

Submitted in partial fulfillment for the Degree of Master of Technology: Biotechnology, M. L. Sultan Technikon, 2002.
Fusarium species produce toxic mycotoxins that are known to exert adverse health effects in humans and animals. No attempts have been made to establish mycotoxin-producing capabilities of isolates of Fusarium species from bananas exhibiting symptoms of crown rot. Crown rot is one of the most serious post harvest problems in banana and the disease is caused by different fungal species, principally Fusarium species. Banana, which is of great economic significance in growing countries (i.e. Costa Rica, Cameroon, Ecuador) is seriously affected by crown rot and is a major cause of fruit loss
M

Fusarium circinatum is a fungal pathogen that has had a serious impact on pine production throughout the world. It attacks most Pinus species including Pinus elliottii, Pinus patula and Pinus radiata. Infections in South Africa (SA) are largely on seedlings, and result in fatal seedling wilt. Accurate and quick detection systems suitable for field use are needed to monitor the spread of the disease and optimize fungicide applications. Detection of F. circinatum is currently based on visual observations of typical symptoms. However, symptoms are not unique to the pathogen and can be caused by other biotic and abiotic stress factors. Nucleic acid-based identification techniques using PCR are available for different fungal species. These are sensitive and accurate, but they are expensive and require skilled biotechnologists to conduct the assays. In this study an enzyme-linked immunosorbent assay (ELISA) was developed to identify F. circinatum in infected seedlings. This optimized ELISA is able to discriminate between F. circinatum and two other fungi that frequently affect pine. This method has advantages over other assays because of its ease of operation and sample preparation, sensitivity and the ability to run multiple tests simultaneously. Mycelium-soluble antigens from Diplodia pinea (=Sphaeropsis sapinea), F. circinatum and F. oxysporum were prepared in nutrient broth. Analysis of these antigens on SDS-PAGE indicated the presence of common antigens between the different fungal pathogens. Some antigens were expressed more by some isolates than by others. Separate groups of chickens were immunised with mycelium-soluble antigens from D. pinea, F. circinatum and F. oxysporum and exo-antigen from F. circinatum prepared in nutrient broth. A 34 kDa protein purified from SDS-PAGE specific for D. pinea was also used for immunisation. Five sets of antibodies were obtained including anti-D. pinea, anti-F. circinatum, anti-F. oxysporum, anti-F. circinatumexo and anti-D. pinea 34 kDa antibodies, respectively. Reactivity of these antibodies was evaluated against antigens prepared in nutrient broth using western blotting and ELISA. Western blot analysis indicated that immuno-dominant antigens for F. circinatum were larger than 34 kDa and their reactivity was not the same between different isolates. Each of the antibodies prepared using mycelium-soluble antigens showed increased reactivity when detecting its own specific pathogen, but cross-reactivity was observed. Anti-D.pineaantibodies showed minimal cross-reactivity with antigens from F. circinatum and F. oxysporum. Anti-F. circinatum antibodies cross-reacted with antigens from F. oxysporum but showed little cross-reactivity with D. pinea antigens. Anti-F. oxysporum antibodies showed more cross-reactivity towards antigens from F. circinatum than those from D. pinea. No reactivity was observed when anti-F. circinatum-exo antigen and anti-D. pinea 34 kDa antibodies were used in immuno-blotting analysis. Evaluation of antibody reactivity using indirect ELISA showed patterns similar to those observed on western blotting, where anti-D. pinea, anti-F. circinatum and anti-F. oxysporum antibodies showed the same cross-reactivity relationships. Anti-F. circinatum and anti-F. oxysporumantibodies showed a significant difference when reacting with antigens isolated from other pathogens including D. pinea, F. circinatum, F. oxysporum, F. solani, F. graminearum and F. culmorum (P = 0.001). No significant difference was observed when the antigens were detected with anti-D. pinea antibodies. Reactivity of anti-F. circinatum-exo and anti-D. pinea34 kDa antibodies was mostly similar to that of non-immune antibodies and showed no significant difference between detection of different antigens. Pine seedlings were artificially infected with the three fungal pathogens using a spore concentration of 1 – 1 x 106conidiaml-1.Infection was monitored using scanning electron microscopy. Results showed increased levels of mycelium growth on the stem and roots of the F. circinatum and F. oxysporum infected seedlings and on the leaves and stem in the case of D. pinea infected seedlings. These plant parts were used in ELISA tests for the detection of antigens. Isolation of antigens from the plant materials involved crushing plant parts in buffer and centrifugation of the suspension. The supernatant obtained was directly used in the assay. ELISA tests prepared in this study were sensitive enough to detect infection caused by 1 conidium ml-1at two weeks post inoculation. A positive reaction for detection of F. circinatum and F. oxysporum was indicated by an ELISA reading above an optical density at 405 nm. The plant material used in ELISA tests were further analysed using PCR. Results indicated that there was no cross-infection between seedlings and served as a confirmation of the disease-causing pathogen. This indicated that cross-reactivity observed was due to other factors such as common epitopes on the major antigens. Use of an ELISA dip-stick or ELISA using these antibodies should provide an easy, fast field test to identify infections of pine, discriminating between F. circinatum, F. oxysporum and D. pinea.
M.Sc.Agric. University of KwaZulu-Natal, Pietermaritzburg 2013.

In the genus Fusarium are many important plant pathogens. The diversity of hosts the genus attacks, the number of pathogenic taxa and the range of habitats in which they cause disease are the greatest in plant pathology. One important species complex within the genus Fusarium is Fusarium oxysporum Schlecht. This species complex consists more than 80 pathogenic forma specialis and is particularly difficult to control. The fungi can survive in soil for decades as specialized spores, known as chlamydospores. Interestingly, however, this species complex also contains beneficial non-pathogenic forms that can be exploited to manage Fusarium wilt diseases. In this study, the ability of non-pathogenic F. oxysporum strains, Trichoderma harzianum Rifai Eco-T®, soluble silicon, and their combination was evaluated on two important crops, banana (Musa sp. L.) and beans (Phaseolus vulgaris L.), for their potential to suppress pathogenic strains of F. oxysporum. The ability of these crops to take up and accumulate silicon in their organs, and its effect on low temperature stress was also investigated. Several endophytic fungi, mainly Fusarium spp. and bacteria, were isolated from healthy mature banana plants. After preliminary and secondary in vivo screening tests against F. oxysporum f.sp. phaseoli on beans (Phaseolus vulgaris L.) cv. Outeniqua, two non-pathogenic F. oxysporum strains were selected for further testing. These two non-pathogenic F. oxysporum strains were found to colonize banana (Musa sp.) cv. Cavendish Williams and bean plants, and to suppress Fusarium wilt of these crops. In order to improve the efficacy of these biocontrol fungi, soluble silicon was introduced. The relationship between plant mineral nutrition and plant diseases have been reported by several authors. Plants take up silicon equivalent to some macronutrients, although it is not widely recognized as an essential element. In this study, we established that roots, the target plant organ for soilborne plant pathogens, accumulated the greatest quantity of silicon of any plant organs when fertilized with high concentrations of silicon. On the other hand, the corm and stem accumulated the least silicon. Such observations contradict the concept of passive uptake of silicon via the transpiration stream in these plants as the only uptake mechanism. The prophylactic properties of silicon have been documented for many crops against a variety of diseases. In vitro bioassay tests of silicon against these wilt pathogens showed that silicon can be toxic to Fusarium wilt fungi at high concentrations (>7840 mg .-1), resulting in complete inhibition of hyphal growth, spore germination and sporulation. However, low concentrations of silicon (<490 mg .-1) encouraged hyphal growth. Silicon fertilization of banana and beans significantly reduced disease severity of these crops by reducing the impact of the Fusarium wilt pathogens on these crops. However, it could not prevent infection of plants from the wilt pathogens on its own. Synergistic responses were obtained from combined applications of silicon and non-pathogenic F. oxysporum strains against Fusarium wilt of banana. Combinations of silicon with the non-pathogenic F. oxysporum strains significantly suppressed disease severity of Fusarium wilt of banana, caused by F. oxysporum f.sp. cubense (E.F. Smith) Snyder & Hansen, better than applications of either control measure on their own. Banana production in the subtropical regions frequently suffer from chilling injury, and from extreme variations between night and day temperatures. Such stress predisposes banana plants to Fusarium wilt disease. Silicon, on the other hand, is emerging as important mineral in the physiology of many plants, ameliorating a variety of biotic and abiotic stress factors. We established that silicon fertilization of banana plants significantly reduced chilling injury of banana plants. Membrane permeability, lipid peroxidation (MDA level) and proline levels were higher in silicon-untreated plants than the treated ones, all of which demonstrated the stress alleviating effect of silicon. Low temperatures damage the cell membrane of susceptible plants and cause desiccation or dehydration of these cells. Levels of sucrose and raffinose, recognized as cryoprotectants, were significantly higher in silicon-amended banana plants than unamended plants.
Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.

The fungus Fusarium sacchari (Butler) Gams causes stem rot in sugarcane especially in association with the stem borer Eldana saccharina Walker. Sugarcane plants tolerant to F. sacchari PNG40 were obtained by chemical mutagenesis and in vitro selection during somatic embryogenesis and plantlet regeneration on media containing F. sacchari culture filtrates (CF), followed by selection in the greenhouse. Somaclonal variants tolerant to F. sacchari PNG40 CF were established by treatment of calli with ethyl methanesulphonate (EMS) and various selection treatments. Investigations were conducted to test the effect of varying CF concentrations and the culture developmental stages (embryo maturation, embryo germination and plantlets) that were most effective in screening calli and plants. Incorporation of CF (0-100 ppm) in the media, at either embryo maturation or germination stages, resulted in significant callus necrosis, and consequent decreased plantlet yield. The highest callus necrosis of 95.55 ± 0.9 % and the lowest plant yield of 1.4 ± 0.45 plants/0.2 g were obtained after inclusion of 100 ppm CF in the germination medium compared with 61.5 ± 3.8 % and 43.8 ± 5.6 plants/0.2 g in the maturation medium, respectively. Exposure of whole plants with trimmed roots to 0-1500 ppm CF resulted in inhibition of root re-growth, with the 1500 ppm CF treatment having the greatest negative effect. Subsequent treatments involved immersing in vitro plantlets in varying concentrations of F. sacchari conidial suspensions. This resulted in 33.3 % and 100 % mortality with 103 and 105 conidia/ml treatments, respectively. Control and EMS-treated calli and potentially tolerant regenerated plants were selected using the established CF and inoculation treatments. Plants from EMS treatments displayed more varying root length. More plants with increased root growth, in the presence of CF, were produced from these treatments than from non-EMS treatments, indicating the ability of EMS to induce somaclonal variation. These putative tolerant plants were inoculated with PNG40 and those selected using CF in vitro were symptomless whilst the positive controls (plants unexposed to CF) were symptomatic. Re-isolation of Fusarium from the inoculated plants and identifying isolates as PNG40 using ISSR analysis confirmed tolerance of the asymptomatic plants and the fungus as the causal agent of the observed symptoms. This confirmed that tolerance to CF correlates to tolerance to F. sacchari PNG40. Future work includes testing stability of tolerance in the field and after sexual reproduction, and use of this protocol to produce plants that permit endophytic PNG40 colonisation towards biological control of E. saccharina.
Thesis (M.Sc.)-University of KwaZulu-Natal, Westville, 2012.

Thesis (Ph.D. (Chemistry)) -- University of Limpopo, 2021
Fusarium species are among pathogenic organisms responsible for massive yield and quality losses in crop production. They cause crop diseases in the field and during storage, and some species are capable of producing mycotoxins which contaminate products and threaten consumer s’ health. Conventional synthetic fungicides are available for the control of Fusarium pathogens, however, their applications have been restricted or discouraged due to their harmful effect on the environment, livestocks and human health. There are also reports about fungal-resistance to available fungicides. Moreover, the synthetic chemicals are not affordable to smallholder farmers and to some extent, they are not recommended for applications in organic farming. As an alternative to these fungicides, selected medicinal plant species were investigated as sources of natural chemicals or compounds with potential to be developed into plant-based fungicides to control Fusarium pathogens. This study aimed to identify antifungal extracts among the selected medicinal plant species which could be used to develop plant-based fungicides to control Fusarium diseases. It also focused on isolation and characterization of antifungal compounds from selected medicinal plant species. Thirteen medicinal plant species (Combretum erythrophyllum (Burch.) Sond , Melia azedarach L, Solanum mauritianum Scop, Nicotiana glauca Graham, Schotia brachypetala Sond, Lantana camara L, Combretum molle R. Br. ex G. Don, Quercus acutissima Carruth, Olea europaea L, Vangueria infausta Burch, Withania somnifera (L.) Dunal, Harpephyllum caffrum Bernh and Senna didymobotrya (Fresen.) H.S. Irwin & Barneby) were selected from literature based on their reported strong antimicrobial activity against human and/or animal pathogens. The leaves of these plant species were collected, shade-dried and extracted with water, petroleum ether, ethyl acetate and acetone. Extractant yield was recorded and each extract was evaluated for antifungal activity using a micro-dilution assay against nine Fusarium pathogens (Fusarium verticillioides, Fusarium proliferatum, Fusarium subglutinans, Fusarium graminearum, Fusarium solani, xxvii Fusarium oxysporum, Fusarium semitectum, Fusarium chlamydosporum and Fusarium equiseti). Similar solvent extracts from different plant species that demonstrated MIC value of less than 0.1 mg/ml against the same pathogen were combined and evaluated for antifungal activity. The interation effect of combined extracts was determined by calculating their fractional inhibitory concentration index (FICI) in order to determine their possible synergistic, additive, indifference or antagonistic antifungal activity against tested pathogens. Plant extracts demonstrating synergistic and or additive interaction were further evaluated in combination and individually for in vivo antifungal activity against maize seed Fusarium pathogens. At least, one of the extracts obtained from these medicinal plant species showed strong antifungal activity with minimum inhibitory concentration (MIC) of less than 0.1 mg/ml against at least one of the tested pathogens. Of the four solvent extracts evaluated, acetone and ethyl acetate extracts showed stronger antifungal activity compared to petroleum ether and water extracts. Of the nine pathogens tested, F. proliferatum was the most susceptible and was strongly inhibited (MIC < 0.1 mg/ml) by 41 plant extracts whilst F. equisite was found to be resistant with MIC < 0.1 mg/ml by only three plant extracts. In total, each pathogen was tested against 52 plant extracts. There were 17, 16 and 15 extracts from C. erythrophyllum, S. mauritianum and Q. acutissima, respectively, with MIC values less than 0.1 mg/ml. These species were the most active when tested individually. Schotia brachypetala was found to be the least active medicinal plant with only seven extracts demonstrating very strong activity (MIC < 0.1 mg/ml) against the tested pathogens. Minimum inhibitory dilution (MID) or total activity was also calculated and it was found that water and acetone extracts had the highest MID, followed by ethyl acetate extracts while petroleum ether extracts recorded the lowest. Of all plant extracts tested against the nine pathogens, 59 plant extracts demonstrated MID values of more than 1000 ml/g. Out of the 348 extract combinations evaluated, 116 and 87 extract combinations demonstrated synergistic and additive antifungal activity, respectively. The strongest activity xxviii recorded for the combined extracts resulted from synergistic interaction with MIC value of 0.001 mg/ml against F. proliferatum and F. verticilloides. Combined acetone extract of C. erythrophyllum and Q. acutissima was very active (95.75% inhibition) against F. verticilloides inoculated on maize seeds while individual preparation from M. azedarach acetone extract demonstrated 97.10% inhibition against F. proliferatum. The extracts showing good antifungal activity (≥ 50% inhibition) were further tested for phytotoxicity on maize seed germination and the lowest recorded seed germination was 86.25%, resulting from Q. acutissima ethyl acetate extract. Combined acetone extract of C. erythrophyllum and Q. acutissima did not significantly affect maize seedling growth when compared to negative control (water treatment). All plant extracts that showed strong activity (MIC < 0.1 mg/ml) when tested using micro-dilution assay were spotted on thin layer chromatography (TLC) bioautographic assay to establish and determine the number of active compounds or bands. The white spots observed on the chromatograms indicated the presence of antifungal compounds. Combretum erythrophyllum, W. somnifera and L. camara exhibited the presence of antifungal compounds against 7, 5 and 4 pathogens, respectively. Hence, these plant species were selected for isolation of antifungal compounds where open column chromatography and preparative TLC were used for compound purification. At least, three isolated fractions from the three plant species were found to be active (MIC values ranging from 0.0098 to 0.625 mg/ml) against more than five pathogens. The fractions were also found to contain different levels of phytochemicals such as glycosides, flavonoids, steroids, and terpernoids. The structures of isolated compounds or fractions were determined using nuclear magnetic resonance (NMR) and mass spectroscopic (MS) techniques. A mixture of apeginin (4′,5,7-trihydroxyflavone) and salvigenin (5-hydroxy-6,7,4'-trimethoxyflavone) isolated from the leaves of C. erythrophyllum showed strong antifungal activity (MIC values ranging from 0.01 mg/ml to 0.63 mg.ml) against 5 tested Fusarium pathogens. Also isolated from C. erythrophyllum was a derivative of maslinic acid and it has xxix shown antifungal activity with MIC values ranging from 0.08 mg/ml to 0.63 mg/ml against 6 tested pathogens. On the other hand, lantadene A (22- angeloyloxy-9-hydroxy-3-oxo-olean-12-en-28-oic acid), boswellic acid (11-keto-β-boswellic acid) and boswellic acid glycoside isolated from the leaves of Lantana camara showed good activity (MIC values ≤ 0.63 mg/ml) against one or more Fusarium pathogens. Withaferin A (4β,27-dihydroxy-1-oxo-5β,6β-epoxywitha-2-24-dienolide) glycoside isolated from the leaves of Withania somnifera showed antifungal activity with MIC value of 0.16 mg/ml against F. verticilloides. This study demonstrated potential applications of medicinal plant extracts as cheap, accessible and sustainable source of eco-friendly pesticides for fighting crop diseases in organic and smallholder farming. The extracts can be used as treatment agents to control maize seed spoilage during post-harvest storage. Additionally, characterised antifungals may serve as scaffold compounds during commercial synthesis of plant-based fungicides.
Agricultural Research Council (ARC) and National Research Foundation (NRF)

A necrosis at the tip of cones was observed on hop (Humulus lupulus), cultivar "Nugget", grown in Oregon in the early 1990's. Fusarium sambucinum and F. avenaceum were recovered from symptomatic cones in 1998 and preliminary inoculation experiments suggested both Fusarium species could cause hop cone necrosis. Studies were carried out to (1) examine pathogenicity and demonstrate Koch's postulates for hop cone tip blight using isolates of F. avenaceum and F. sambucinum obtained from hop cones; (2) examine isolates of F. avenaceum and F. sambucinum derived from other diseased plant hosts, and other Fusarium species derived from hop cones, for ability to cause cone necrosis; and (3) survey commercial fields to determine Fusarium populations on 'Nugget' cone parts. Isolates ofF. avenaceum and F. sambucinum recovered from diseased hop cones were used for pathogenicity experiments. In addition, cone inoculations were done with single isolates of F. avenaceum and F. sambucinum from diseased sweet corn roots, one isolate of F. sambucinum recovered from a diseased potato tuber, individual isolates of F. equiseti and F. oxysporum from hop cones. Cones of two hop cultivars, 'Nugget' and 'Willamette', were collected from three different farms on three sampling dates and inoculated with spore suspensions of hop-derived F. avenaceum and F. sambucinum at concentrations of 10��, 1O���, and 10��� conidia/ml to examine dosage effects. Necrosis was evaluated 2, 4, 6, and 8 days after inoculation. Percent cone necrosis decreased as inoculum concentration of either F. avenaceum or F. sambucinum decreased, and was lowest on water-treated cones, for all three sampling dates. The respective Fusarium species were recovered from symptomatic cones. Cone necrosis developed following cone inoculation with F. avenaceum or F. sambucinum from potato or corn. Hop cones inoculated with F. equiseti or F. oxysporum also developed necrosis, but at relatively lower levels compared to the other Fusarium species used for inoculations. For the surveys in commercial hop fields, burr and cone material were collected on five different dates. Fusarium sambucinum was recovered most frequently, but F. avenaceum was also found. Both Fusarium species were recovered from asymptomatic burr and cone materials throughout the growing season. In general, Fusarium species, F. equiseti, F. oxysporum, F. culmorum, F. solani, plus F. avenaceum and F. sambucinum were found more frequently early in the season on stigmatic tissue, and Fusarium recovery decreased as the season progressed. Fusarium prolferatum and F. monilforme were recovered rarely.
Graduation date: 2004

The main objective of this thesis was the elucidation of the host-pathogen interaction between Pinus patula and Fusarium circinatum. This was accomplished by studying differential gene expression at the molecular level. Therefore, the first chapter reports the use of PCR-based methods in gene discovery and transcriptome analysis. The use of these techniques in the identification of novel transcripts in host-pathogen interactions addressed. These examples illustrate the differences and strong features of each technique. Chitinases are linked to defence responses in plants. In chapter tw0, the induction of chitinases in P. patula was assessed at both the protein and genetic level. Western blot analysis and enzyme activity assays indicate that chitinase enzyme is not detected a part of the defence response by P. patula after infection by F. circinatum. This was further confirmed by the lack of significant induction of two Pinus chitinase genes, LP6 and PSCHI4, as determined by RT-PCR analysis. Partial DNA sequence homologues for the LP6 and PSCH14 genes were determined and compared with a variety of plant chitinases. The low levels of detectable chitinase induction in P. patula might explain the high levels of susceptibility to the pitch canker fungus observed in seedlings of this tree. Pinus patula, the most widely planted species in South Africa, is highly susceptible to infection by F. circinatum. In chapter three, suppression subtractive hybridisation was used to elucidate the changes taking place at the molecular level early on in this interaction. Most of the identified transcripts shared homology to both biotic and abiotic stress in plants. The induction of one fragment, displaying homology to phytocyanin proteins, as followed through RT-PCR. Induction levels for this fragment differed significantly between less and more susceptible plants. Although most of the sequences isolated in this study can be Iiked to stress, most have not been linked with specific plant-pathogen interactions. This raises questions in regard to the function of these genes in host-pathogen interactions. Further research identify the function of these sequences in the defence response will be needed. These sequences can also be tested against a family of Pinus trees to ascertain if they will be useful in marker assisted selection. A molecular analysis of culture degeneration and pathogenicity of F. circinatum was attempted in chapter four. In this chapter, the differential induction of transcripts in F. circinatum was determined against several other Fusarium spp. Several of he identified fragments shared homology with stress related proteins. One transcript shared homology to a polyketide synthase, FUM5, that could be linked to fumonisin production in other Fusarium spp. ELISA detected no fumonisin production, although the FUM5 transcripts were detected. The identification of all the transcripts could provide a basis for more intensive gene discovery studies in F. circinatum and other Fusarium spp. The induction of these sequences in different isolates needs to be studied to prove their function in F. circinatum. This study also complements several other studies that studied the morphological characteristics of culture degeneration. Resistance gene analogues have been reported from a diverge set of plant species. In chapter five, degenerate PCR amplification was used to isolate TI-NBS-LRR like resistance gene analogues from a range of Pinus species. These sequences w re further characterised through comparative analysis with previously reported Pinus resistance gene analogues. Through motif analysis, several of the known conserved motifs found in NBS domains were identified and conservation with other plant NBS motifs is indicated. The P-Ioop and GLPL motifs displayed a high level of conservation on amino acid level with other plant NBS motifs. However, slight differences in several of the conserved regions were observed when the Pinus analogues were compared with Arabidopsis thaliana. The identification of differences between angiosperm and gymnosperm NBS sequences indicates that design of new degenerate probes and primers for the isolation of more ancient NBS sequences is needed. Further, phylogenetic and structural analyses of these sequences will also aid in understanding the relationship between angiosperm and gymnosperm NBS sequences. The knowledge gained from such a study will highlight the similarities between angiosperm and gymnosperm defence responses. This study represents the first detailed report on RGA in Pinus.
Thesis (PhD)--University of Pretoria, 2006.
Genetics
Unrestricted

Sweet potato is a popular food security crop in South Africa and has a considerable commercial value. Fusarium wilt (FW), caused by the fungal pathogen Fusarium oxysporum formae speciales (f. sp.) batatas, has been reported worldwide and is widespread in sweet potato production areas in South Africa. Preliminary molecular identification of South African isolates from diseased sweet potato plants indicated that there are other formae speciales besides F. oxysporum f. sp. batatas associated with FW. The objectives of the study were to conduct a field survey and to characterise the isolates of the Fusarium oxysporum species complex (FOSC) using phylogenetic analyses, morphological characterisation and DNA barcoding. Phylogenetic analyses revealed two other formae speciales, namely F. oxysporum f. sp. tuberosi and F. oxysporum f. sp. vanillae that were associated with FW. This study has contributed in understanding and knowledge of FOSC associated with FW of sweet potato in South Africa.
Life and Consumer Sciences
M. Sc. (Life Sciences)

Fusarium root rot (FRR) disease, caused by the fungus Fusarium solani f. sp. phaseoli (FSP), is an important soil-borne disease reducing common bean (Phaseolus vulgaris L.) yields, and hence food security, in Uganda and elsewhere in developing countries where the crop is grown without fungicides. The key aim of this study was to elucidate the significance of bean root rot (BRR), appraise methods for screening germplasm for resistance to FRR, determine the genotypic variability of resistance, and the inheritance of resistance to FRR in common bean. This information was deemed useful in devising an appropriate strategy for breeding FRR resistance in beans. A participatory rural appraisal (PRA) was conducted in south-western and eastern Uganda to ascertain farmers’ awareness of BRR and their influence on preferred bean varieties. Bean root rot is considered to be the most devastating and most recognised disease, especially in south-western Uganda. Control measures for BRR were very minimal, and in some cases, non-existent. Use of resistant varieties to control the disease was not evident, because the most popular varieties were susceptible to the disease. The resistant bean varieties currently available have undesirable characteristics such as small seed size, black seed and late maturity. Large-seeded bean varieties, even though cited as being more susceptible to BRR than the small-seeded varieties, are still very popular. The study highlighted the need for breeding FRR resistance in the large-seeded bean varieties that are highly preferred by farmers. Four isolates of FSP (FSP-1, FSP-2, FSP-3 and FSP-4) were tested for pathogenicity under screenhouse and laboratory conditions. In addition, three methods of storing and maintaining the viability of FSP isolates were appraised. The isolate FSP-3, was found to be the most pathogenic, resulting in 100% disease incidence on all bean varieties tested, with high severity scores. The potato dextrose agar (PDA) slants stored at 5oC were found to be the best method of storage for pathogenic isolates. The FSP-3 isolate was subsequently utilised for screening bean lines for resistance to FRR. The influence of soil composition, irrigation frequency, and inoculation technique on the severity of FRR was studied on six bean lines. Interactions of irrigation frequency, soil composition, and bean lines were not significant. The 50% swamp soil:50% forest soil composition and forest soil alone categorized the varieties most distinctly according to their reaction to FRR. Also, the best distinct classification for the varieties was obtained under treatments that were watered daily and once in a week. Based on economic considerations, the standard forest soil and daily irrigation were subsequently adopted for screening bean germplasm for resistance to FRR. It was also found that sorghum seed as a medium for pathogen inoculation was better than the agar slurry medium. One hundred and forty seven common bean varieties were evaluated for resistance to FRR (isolate FSP-3) under screenhouse conditions. In order to confirm this resistance, 46 common bean lines selected from the screenhouse trial were further evaluated using natural inoculum in a BRR-infested field. Forty-four varieties comprising ten large-seeded, four medium-seeded and 30 small-seeded varieties showed moderate resistance to FRR; but none were resistant or immune to the disease. Based on adaptability, eight moderately resistant varieties were selected for use as parents in the study of inheritance of resistance to FRR. A 12 x 12 diallel mating design was utilised to develop 66 F1 and F2 populations, plus their reciprocal crosses, with the aim of studying the mode of inheritance of resistance to FRR. The F1 and F2 progeny evaluations showed that FRR resistance was mainly governed by additive genes in most populations. However, there were a few crosses which displayed highly significant specific combining ability (SCA) effects, implying that dominant effects were important in some populations. Maternal effects were also highly significant at both the F1 and F2 generations, suggesting that resistance was modified by cytoplasmic genes. The non-maternal effects were also significant in some populations, suggesting that the cytoplasmic genes were interacting with nuclear genes. The number of genes governing resistance to FRR varied from two to nine among the eight sources of resistance. The allelism test of resistant x resistant populations, and the observation of continuous distributions of severity scores, suggested the presence of many loci governing FRR resistance in beans. Broad sense heritability of disease resistance varied from 0.22-0.69, while heritability in the narrow sense was estimated as 0.35-0.49 in the populations. These results suggested that selection and backcrossing to both parents would be the best breeding procedures for improving resistance in the popular large-seeded bean varieties in Uganda. However, there could be complications in breeding for resistance to FRR in beans, because resistance was modified by cytoplasmic gene effects and their interaction with nuclear genes in some of the populations.
Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.

Potential vegetative and reproductive increases in yield, as well as the biological efficacy against Fusarium verticillioides and F. proliferatum causing ear and stem rot in maize crops of commercially-formulated micro-organism formulation T-Gro (Trichoderma harzianum isolate DB103 WP) combined with Spartacus (Beauveria bassiana isolate DB 105 WP), T-Gro combined with Armenius (Bacillus subtilis isolate DB 109 WP), T-Gro combined with Maximus (Bacillus subtilis isolate DB 108 WP), T-Gro combined with Shelter (Bacillus subtilis isolate DB 101), T-Gro combined with Bismarck (Microbacterium maritypicum isolate DB 107 WP), as well as individual treatments of T-Gro, Armenius, Bismarck, Maximus and Shelter, were investigated when applied to maize seed and soil under field conditions. All the micro-organism treatments were compared with Thiram 750WP (750g/kg thiram WP) and an untreated control. The micro-organism treatments showed an increase in vegetative as well as reproductive yields when compared to the reference product Thiram 750 WP and the untreated control. There were no observations of adverse effects on the germination of maize seed in all the treatments that were applied. The three isolates B. subtilis, T. harzianum, and M. maritypicum, showed a significant reduction in vascular tissue discolouration of the main and ear stems, indicating a potential to be used in the reduction and control of diseases caused by Fusarium spp. Results also showed poor to very good increases of stem and foliage biomass as well as cob yield per plant produced by the micro-organism treatments when compared to the untreated control. The highest cob yield per plant that differed significantly from the untreated control was produced by T-Gro and Shelter. No phytotoxicity of any kind was observed with the application of the micro-organism formulations and they could therefore be deemed suitable to be used for the treatment of maize seed. The micro-organism formulations containing fungal and bacterial biological control agents have the potential to be used in commercial maize production to increase vegetative and reproductive yields and reduce the severity of ear and stem rot in maize.
Agriculture Animal Health & Human Ecology
M.Sc. (Agriculture)

Thesis (M. Sc.)--University of Alberta, 2009.
Title from PDF file main screen (viewed on Oct. 20, 2009). "A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Master of Science in Plant Science, Department of Agricultural, Food and Nutritional Science, University of Alberta." Includes bibliographical references.

Thesis (M.Sc. (Agriculture)) --University of Limpopo, 2018
Tomato is second most cultivated crop globally and in South Africa it is planted by both commercial and smallholder farmers. However, the crop is susceptible to a number of diseases including those caused by fungal pathogens. Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici and seedling damping-off caused by Rhizoctonia solani, are known to cause serious yield loss in tomato production. Their management is mainly based on the application of synthetic fungicides and cultural practices. However, both methods have limitations which result in their inefficiency. Synthetic fungicides also have negative impact on the environment and human health. The ability of fungal pathogens to develop resistance to fungicides has also resulted in their reduced application. These challenges have led to a need to identify novel methods using plant extracts and biological control agents which can be used to manage these diseases. The objectives of this study were therefore to, firstly determine the efficacy of both plant extracts on mycelial growth of F. oxysporum f. sp. lycopersici and R. solani under laboratory conditions and secondly, to evaluate the effectiveness of both plant extracts as well as antagonistic fungi Trichoderma harzianum against Fusarium wilt and damping-off of tomato under greenhouse conditions. Food poisoning assay was used to investigate the efficacy of M. burkeana and M. oleifera extracts in vitro. Six (0, 2, 4, 6, 8, 10 g/ml) treatments were arranged in a completely randomised design and replicated four times. After 7 days of incubation at 25 °C, radial growth colony was measured. For the greenhouse xp im nt, Fusa ium wilt was t st d on cv. ‘HTX14’ as th most susc ptibl cultiva whilst seedling damping-off was t st d on cv. ‘Mon y-make ’. Aqu ous xt acts were prepared by decocting different concentrations of M. burkeana (4, 6, 8 g/ml) xiv and M. oleifera (2, 4 and 6 g/ml) in 100 ml of distilled water at 100 °C for 15 minutes then left to cool before filtering and applying as a treatment. Trichoderma harzianum as a treatment was applied 7 days after inoculating the soil-borne pathogens. In-vitro M. burkeana treatments concentrations had the highest mycelia growth suppression against both F. oxysporum f. sp. lycopersici at 10 g/ml (76 %) whilst suppression on R. solani was at 8 g/ml (71 %) relative to control. Moringa oleifera xt acts’ highest pathogen suppression for both F. oxysporum f. sp. lycopersici and R. solani were respectively 35 % and 60 % relative to control at concentration 6 g/ml. Under greenhouse conditions shoot disease severity had highest suppression at 0.6 g/ml of M. burkeana and 0.4 g/ml of M. oleifera treatment concentrations resulting to 32 and 49 % relative to control. Whereas, treatment 0.8 g/ml of M. burkeana and 0.4 g/ml of M. oleifera suppressed stem and root discoloration by 39 and 54 % respectively. Trichoderma harzianum significantly (P ≤ 0.05) reduced shoot severity and root and stem discolouration contributing the highest suppression of 49 % relative to control. In damping-off treatments, both plant extracts and T. harzianum also significantly duc d (P ≤ 0.05) pre- and post-emergence damping-off incidence with M.burkeana recording the highest suppression at 78 % followed by M. oleifera at 64 %. Trichoderma harzianum reduced incidence of damping-off by 60 % relative to untreated control on both M. burkeana and M. oleifera experiments. The results of this study showed that M. burkeana, M. oleifera extracts and T. harzianum can be highly suppressive to both tested plant diseases. However, further studies should be conducted to determine their mode of action, application method and their effect on other soil microorganisms. Keywords: Damping-off, Fusarium wilt, Plant extracts, T. harzianum, Tomato plant

Maize ear rots are among the most important impediments to increased maize production in Africa. Besides yield loss, they produce mycotoxins in their host whose contamination has been linked to several human and animal mycoses. The main objectives of the studies reported on in this thesis were (i) to investigate farmer perceptions of maize ear rot disease and prospects for breeding for host plant resistance in Zambia; and (ii) to establish the levels of incidence and extent of maize ear rot infection as well as the level of mycotoxins in the maize crops of smallholder farms in central and southern Zambia; (iii) to appraise the field inoculation techniques and assess them for their suitability for the Zambian environmental conditions, (iv) to determine the combining ability of Zambian maize populations for resistance to ear rot and investigate the genetic basis of this resistance; and (v) to investigate both direct and indirect responses to full-sib selection for ear rot resistance in Zambian maize populations. A participatory rural appraisal (PRA) was conducted in four communities, involving a total of 90 farmers. Participatory methods were used, such as focused group discussions, group interviews, participant scoring and ranking. Farmers ranked and scored the various constraints affecting their maize production in general and the maize ear rots in particular. Ear rots were ranked as the third most important biotic stress and it was evident that although farmers were aware of the disease, they were not aware of mycotoxins. This was reflected in the way they disposed of rotten maize: either by feeding livestock or eating it in periods of hunger. The survey of ear rots and mycotoxins was carried out in the Southern and Central Provinces of Zambia. A total of 114 farms were covered in the survey: maize samples were collected and both ear rot fungi and mycotoxins were isolated. Fusarium and Stenocarpella were the most frequently isolated fungi from smallholder farms. The levels of fumonisins on these farms ranged from 0.05 to 192 ppm, while those of aflatoxins were between 1.5 and 10.6 ppb. In 50% of the farmsteads surveyed, the mycotoxins, i.e. fumonisins and aflatoxins, exceeded the recommended FAO/WHO 1limits of 2 ppm and 2 ppb, respectively. Five field inoculation techniques namely, colonised toothpick, leaf whorl placement, ear top placement, spore suspension spray, and silk channel injection, were evaluated over three seasons in a series of experiments. It was found that the leaf whorl placement of inoculums, followed by colonized toothpick method, gave a constant ranking of genotypes across locations and years compared to the other three methods. In addition, the use of a mixture of ear rots as inoculum was as effective as its principal single species constituents. In the population diallel analysis, five broad-based maize populations were crossed in a diallel and evaluated under artificial ear rot inoculation using an inoculum mixture of three ear rot fungi, Aspergillus flavus, Fusarium verticilloides and Stenocarpella maydis at four locations in Zambia. The purpose was to estimate general (GCA) and specific combining ability (SCA) and investigate genotype x environment interaction. GCA effects were found not to be significant for disease severity but were significant for grain yield across environments. Populations with a strong GCA effect for disease severity across sites included PRA783244c3, Pop25, MMV600, and ZUCASRc2. Across sites, the F1 combinations, MMV600 x Pop25, ZUCASRc2 X Pop25, and Pop25 x PRA783244c2 had strong SCA effects for root lodging, ear drooping, husk cover and ear insect damage. In a related diallel analysis of 10 full-sib families derived from these populations, it was observed that resistant x susceptible families and their reciprocal crosses performed better than their resistant parents, suggesting an over dominant expression of resistance. Both maternal and non maternal effects were observed to be influencing resistance to ear rots. There was a preponderance influence of non-additive gene action. A response to full-sib recurrent selection was conducted in four locations in Central Zambia. Out of the 343 families created in 2005/6 season, 10% were selected from each population and recombined to create five new populations. These, with the original populations, were evaluated in four sites during the 2007/8 season. There was a net reduction in ear rot incidence and rot severity in the new synthetic population. Pop10 had the largest reduction in disease severity. The predicted gain per cycle was -4.1% and realized gain was -2.5% for disease incidence, and 0.19% and 19.4% for grain yield. Genetic variability was maintained though with low heritability estimates. Negative but at times strong association between grain yield and ear rot disease severity was detected suggesting that in general selecting for ear rot resistance would enhance grain yield in the five populations. Overall the importance of the ear rots and mycotoxins in compromising yield and health of the communities in Zambia, respectively, were confirmed and support the call to improve maize varieties for resistance to ear rots. The results indicate that the five populations could be enhanced for ear rot resistance through population improvement procedures such reciprocal recurrent selection that exploit both additive and non-additive variation. Selection might be compromised by the large genotype x environment interaction effects, and large reciprocal effects and their interaction with the environments. To enhance repeatability genotypes should be artificially inoculated, by placing the inoculum in the leaf whorl followed by colonized toothpick inoculation, and screened in many environments to identify genotypes with stable resistance to ear rots.
Thesis (Ph.D) - University of KwaZulu-Natal, Pietermaritzburg, 2009.

Summary: The future mass propagation of elite families of Pinus patula by cuttings is a realistic method of deployment if the short-term performance of cuttings and seedlings are confirmed at harvesting. This will impact significantly on the future outlook of forestry in South Africa as softwood yields are improved substantially through the introduction of material of high genetic value in commercial plantings. This, however, will require significant changes in future silviculture and other management practices as foresters and plantation staff learn to regenerate, maintain, and schedule the harvesting of cutting stands according to a different set of demands as a result of the change in plant type. Contrary to operational experience, cutting survival was similar to seedling survival in all field studies. This indicates that factors other than those that were studied and reported on, such as planting techniques, may be contributing to mortality. Also, due to the different root structure of cuttings they may be more fragile. The similar survival observed in these trials, therefore, may have been due to the close supervision given to the planting operations by the research staff. Although survival was similar, both plant types survived unacceptably poorly in the majority of studies with an average stocking of approximately 50% at one year. It is therefore anticipated that commercial stands will require several blanking operations in order to achieve an acceptable stocking in excess of 85% by the following planting season. The reduction in expected profitability as a result of blanking costs, delayed establishment, and the loss of improved genetic plant material, indicates that this is an area that still requires further research irrespective of what plant type is being planted. The pathogen, Fusarium circinatum, was commonly isolated from the planting stock before and after planting in two studies. Due to its virulent nature, it was assumed that mortality on the trees on which F. circinatum was isolated was principally due to this pathogen. At planting all plants were observed to be healthy and free of disease indicating that this pathogen maybe carried from the nursery to the field in a cryptic form, either inside or outside the plant tissue , which results in the death of the newly planted tree. In two field studies, where F. circinatum was commonly isolated, the application of Benomyl fungicide and to some extent the biological control agent Trichoderma harzianum at planting appeared to improve survival although this improvement was not significant. Laboratory studies, designed to determine alternatives to Benomyl fungicide, indicated that three fungicides (Octave, Folicur and Tilt), three sterilants (Sporekill®, Prasin®and Citex®) , as well as a biological control agent (T.harzianum), were all highly successful in controlling F. circinatum colony growth in vitro. It is recommended that these products undergo nursery testing , where the plant material is inoculated with F. circinatum spores, in order to test their efficacy and possible phytotoxicity in vivo before commercial application. Post-planting survival was also affected by site climate . Greater temperature extremes, as well as lower humidity and less rainfall resulted in poor survival. Plant dimension at planting was found to interact with site quality where it was a significant factor on a poor quality site. Optimal cutting dimensions at planting was a root collar diameter of 2.8 - 3.2 mm, and a stem height greater than 7 cm at planting for cuttings produced in cavities 90 ml in volume. Optimal seedling dimensions at planting were a root collar diameter of 1.8 - 2 mm, and a stem height of 10 - 15 cm for seedlings produced in cavities 80 ml in volume. In a separate study, plant morphological criteria influenced medium-term growth, where greater root mass and thicker cutting root collar diameters at planting improved field growth performance for seven years after planting. A greater root mass at planting was achieved by raising cuttings in containers that could support greater medium volume. From the study it was concluded that cuttings should be raised for an approximate period of 9 months in container cavities no smaller than 80 ml in volume and possess an oven-dry root mass of 0.3 - 0.5 g at planting. In addition to similar survival, the cuttings in this study grew either similarly to, or in some cases out-performed, the seedlings that were used as a control. Several other published studies indicate that hedge maturation poses the greatest threat to the success of softwood cutting deployment. This is especially true in clonal forestry and methods to maintain juvenility, such as cold storage of shoots and cryopreservation, require further research before clonal plantations of P. patula can be realised. In the studies carried out on family hedges in this report, the effect of donor hedge maturation was found to influence nursery management practice and the characteristics of rooted cuttings. The nursery data indicates that rooting efficiency, root system quality, and stem size and form, all decline with increasing hedge age particularly from two years after the date of sowing. A decline in root system quality was particularly apparent and was observed prior to a decline in rooting efficiency. If field trials indicate poorer performance from older hedges , it may be necessary to determine whether the causes are purely ontogenetic, morphological, or both before drawing final conclusions about hedge longevity. Until such results are known, it is recommended that P. patula cuttings should be propagated from seedling donors maintained as hedges , approximately 15 cm high, for a period not more than three years from the date of sowing.
Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.