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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.

Pigeon pea [Cajanus cajan (L.) Millspaugh] is a very important grain legume crop for food, cash and firewood in Malawi. However, its production is affected by Fusarium wilt (Fusarium udum Butler), which causes up to 100% yield loss. The deployment of resistant varieties would be an economical way to manage the disease, and for this, more information is needed on farmers' preferences for local landraces, how farmers and consumers can be involved in developing new varieties resistant to wilt disease, and the genetics of inheritance of resistance. This information would be used to devise a breeding strategy. A participatory rural appraisal was used in the southern region of Malawi to identify pigeonpea production and marketing constraints. Results showed that Fusarium wilt was the most prevalent and destructive disease of pigeonpea in the area. Other constraints included pests, flower abortion, low yields, and low soil fertility. Local landraces accounted for 84% of the pigeonpea production in Malawi. Local landraces were preferred due to their fast cooking time, taste, and the high prices they earn the farmer. Participatory variety selection was used to identify landraces with desirable traits that could be used in the breeding programme. Farmers and buyers selected ten local landraces which were used in the genetic improvement programme. Pigeonpea local landraces and international Crops Research Institute for the Semi-Arid Tropics (ICRISAT) genotypes were evaluated for wilt resistance, yield, and secondary traits at three sites over three seasons. Most of the landraces were susceptible to wilt and late maturing. However, AP10, a local landrace, was high yielding and resistant to wilt and outperformed ICRISAT varieties. This local landrace showed promise for use as a source material for Fusarium wilt resistance in other locally adapted farmer-preferred varieties lacking resistance. The local landraces needed genetic improvement in wilt resistance, yield, early maturity, number of branches and seeds pod. Laboratory and screenhouse studies were performed to develop a new Fusarium wilt screening technique. Grains of finger millet, sorghum, and wheat were tested as media for multiplying F. udum isolates. Pathogenicity tests were done on Bunda College and Bvumbwe Research Station isolates. The Bunda isolate was then used in an infested-seed inoculation technique against eight differential cultivars. The results showed that finger millet, sorghum and wheat were equally effective for rapid multiplication of F. udum isolates. Wheat grain showed the best results for pathogen multiplication and inoculation, due to the large seed size for easy handling. The inoculation process involved placing infested wheat grain on bruised pigeonpea roots and transplanting into soil in pots. The infested seed inoculation technique, which is the first of its kind for pigeonpea, was effective in screening pigeonpea for wilt resistance. The selected landraces were crossed with wilt resistant testers in a 12 lines x 4 testers mating scheme, and 48 F1 crosses were generated. These F1 crosses were evaluated for wilt resistance, yield, and secondary traits. The variations among F1 crosses for wilt and secondary traits were due to additive gene action in both parents and the dominance effects arising from the interactions of parents. Parental lines, with good combining ability effects for wilt resistance (AP2, AP3, and AP4), days to 50% flowering, seed pod, plant height, stem diameter, and number of primary and secondary branches were identified, while ICEAP00554 (tester) was a good general combiner for wilt resistance and days to 50% flowering. These lines would be useful in breeding for Fusarium wilt resistance in farmer-preferred pigeonpea genotypes in Malawi or similar environments. Specific F1 crosses were identified with significant SCAs for wilt resistance, days to 50% flowering, and secondary branches. The significance of GCA and SCA effects, which indicated importance of both additive and non-additive gene effects, respectively, suggested that both selection and hybridisation would be useful to improve the resistance in farmer-preferred varieties. Segregation analyses were conducted on F2 populations to determine the resistance to susceptibility phenotypic ratios. The Chi-square analyses showed that resistance to wilt was dominant over susceptibility in most F2 populations. The segregation ratios of 3:1, 13:3, 15:1, and 9:7 (R:S) indicated that either one dominant gene, or two inhibitory genes, or two independent dominant genes, or two complementary genes, respectively, were conferring wilt resistance in these crosses. Involvement of only a few genes governing wilt resistance suggested few complications, if any, in breeding for this trait in these locally adapted pigeonpeas. The Pedigree breeding method would be recommended for incorporating these traits. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2007.

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.

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.

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.