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.

Reduced banana yield owing to black Sigatoka Mycosphaerella fijiensis Morelet is a threat to the livelihoods of Ugandan subsistence farmers who depend entirely on the banana crop for food security. The objectives of this investigation were to: (i) assess farmers’ knowledge of black Sigatoka disease in central Uganda; (ii) document the qualities farmers would desire in the banana genotypes to be developed for black Sigatoka resistance; (iii) appraise the methods for assessing black Sigatoka resistance in diploid banana populations; (iv) determine the phenotypic variation for black Sigatoka resistance and agronomic traits in diploid and tetraploid bananas; (v) determine the influence of tetraploid and diploid parents on the black Sigatoka resistance and agronomic traits in the triploid progenies; and (vi) evaluate 2x by 2x banana progenies for yield and black Sigatoka resistance. A survey that focused on low and medium banana production zones in Uganda established that there was limited awareness of black Sigatoka disease as a constraint on banana production in the areas surveyed. It was also established that farmers liked local bananas because of their superior taste, early maturity, and marketability. There were farmers who had been exposed to new black Sigatoka resistant materials but never liked these new banana materials because of poor taste and lack of market. Farmers desired new banana materials with good taste on cooking, heavy bunches, resistance to pests and diseases, drought tolerance, and early maturing capacity in that order. The results indicated that the banana farmers in Uganda attached more importance to food quality attributes than to production attributes especially when considering new banana materials. This suggested that farmers mainly grow bananas for consumption. Three black Sigatoka assessment methods, youngest leaf spotted, disease development time and area under disease progress curve (AUDPC) were appraised using a diploid population. All the three methods were able to classify the diploid accessions into resistant and susceptible clones. The cultivar rankings of AUDPC correlated strongly with the rankings of disease development time. The cultivar rankings of AUDPC correlated positively with the rankings of youngest leaf spotted method. The youngest leaf spotted at flowering and AUDPC predicted significantly total number of leaves at flowering (R2 = 0.53). Overall AUDPC had the highest coefficient of determination (R2=0.84) in assessment of banana diploids for black Sigatoka resistance indicating that it accounted for the highest variation in disease response observed among the diploid clones. From this investigation it was recommended that AUDPC should be used to assess resistance on black Sigatoka in Musa species. A phenotypic analysis on the diploid and synthetic tetraploids, and a molecular analysis using RAPD markers on the tetraploid population were conducted. Results indicated that the diploid population had significant (P<0.001) variation for plant height, plant girth, days from flowering to harvest, bunch weight, number of suckers, youngest leaf spotted, total leaves at flowering, area under disease progress curve, and number of functional leaves at harvest. Principal component analysis showed that plant height and girth explained most of the variation observed in the diploid population. In the tetraploid population, significant differences were observed for plant height, plant girth, and number of suckers (P<0.05). In the tetraploids principal component analysis, indicated that youngest leaf spotted and total leaves at flowering had higher loadings on principal component one. Genetic distances computed from RAPD markers indicated limited genetic variability in the tetraploid population. Another investigation was also carried out to determine the influence of tetraploid and diploid parents on black Sigatoka resistance and agronomic traits in the triploid progenies generated from tetraploid-diploid crosses. The results indicated that diploids transferred black Sigatoka resistance to triploid progenies as measured by disease development over time, the number of functional leaves at flowering and at harvest. On the other hand, the female synthetic tetraploids influenced plant height and bunch weight in the triploid progenies generated from tetraploid-diploid crosses as observed from triploid progeny correlations and parent-offspring regressions. Therefore, it is important to select tetraploids with heavy bunch weights to generate high yielding triploids in tetraploid-diploid crosses. Lastly, this thesis investigated the relationship between bunch weight and black Sigatoka resistance traits in 2x by 2x progenies generated using a random polycross design. Phenotypic correlations revealed strong positive relationships between bunch weight with total leaves at flowering, youngest leaf spotted, plant girth, and days from planting to flowering among the 2x by 2x progenies. Linear regression analysis indicated that girth, total fingers and finger length significantly predicted bunch weight (R2=0.67). However, days from planting to flowering, and total leaves at flowering had strong indirect effects on bunch weight via plant girth. The results imply that selection for parents with good combining ability for girth, finger length and total fingers can improve bunch weight in a diploid population. / Thesis (Ph.D.) - University of KwaZulu-Natal, Pietermaritzburg, 2009.

Maize (Zea mays L.) production is constrained by a number of stresses, amongst the most important are gray leaf spot (GLS) caused by a fungus Cercospora zeaemaydis Tehon and E.Y. Daniels and Phaeosphaeria leaf spot (PLS) caused by Phaeosphaeria maydis (Henn.). The diverse germplasm comprising farmer collections and exotic material used in the medium and highland altitudes maize breeding programmes in western Kenya has not been improved for resistance to the two diseases. Heterotic patterns of germplasm from this region have also not been studied. Therefore, the objectives of this study were to (i) assess the prevalence, importance, and farmers’ perceptions of GLS and PLS, (ii) characterize maize germplasm collections into their heterotic groups and (iii) improve four maize populations for GLS and PLS resistance through recurrent selection. The participatory rural appraisal (PRA) was conducted at three sites in western Kenya during the 2005/2006 cropping season. Data was generated using a checklist in group discussions with 109 male and 123 female farmers as well as key informants. Constraints were identified and prioritised. The five most limiting, in order of importance, were low soil fertility, poor varieties and seed, drought, Striga, pests and diseases (GLS and PLS). Gray leaf spot and PLS were reported in all sites but farmers did not know the causes of these diseases. Farmers preferred local varieties Tiriki, Anzika and Kipindi due to their greater resistance to diseases than commercial hybrids. Farmer criteria for variety selection were low fertilizer, Striga and disease resistance, drought tolerance, closed tips, and high yield potential. Due to the high cost of hybrid seed farmers selected and planted their own seed from advanced generations from previous seasons. Across all the sites, yield gap between on-farm and expected yield potential was estimated as ranging from 4.73t ha-1 to 5.3t ha-1 mainly due to the identified constraints. Therefore maize breeding should focus on addressing important maize production constraints and farmers’ preferences identified in this study in developing varieties that will increase maize yields on-farm. During 2005/2006, seventy 77 testcrosses were developed through crossing 47 germplasm collections with four population testers, Kitale synthetic II (KSII), Ecuador 573 (EC 573), Pool A and Pool B. Crosses and testers were evaluated at Kakamega during 2006/2007 in a 9 x 9 triple lattice design. Significant (p < 0.05) differences in grain yield, ear height, days to 50% anthesis, GLS and PLS resistance were observed. Both general and specific combining ability effects (GCA and SCA, respectively) were significant (p < 0.01), with SCA accounting for more than 50% of the variation for GLS, PLS and yield and less than 50% for ear height, days to 50% anthesis and silk. This indicated that both additive and non-additive gene effects were important but non-additive gene effects were more important in conditioning these traits. High SCA effects indicated high heterosis between collections and populations. Both yield heterosis and SCA were used to study heterotic patterns, but percentage yield heterosis data was used to classify these materials into heterotic groups. Based on significance (p < 0.05) of percentage yield heterosis as a primary factor for classification, seven collections were classified to Pool A, 17 to Pool B, 12 to KSII and 6 to EC 573 heterotic groups. The study indicated that germplasm collections belong to distinct heterotic groups therefore they can be infused into these populations (Pool A, Pool B, KSII and EC 573). Four populations, KSII, EC 573, Pool A and Pool B were subjected to one cycle of reciprocal recurrent selection (RRS) and two cycles of simple recurrent selection (SRS) during the 2004-2006 cropping seasons at Kakamega. Response to selection was assessed by evaluating C0, C1 and C2 and four commercial checks in a randomised complete block design in three replications at Kakamega and Kitale during 2007. All cycles except C0 of Pool A were more resistant to GLS than the three checks, H623, KSTP94 and PHB3253. Response to selection for GLS was significant (p < 0.01) in the desired direction. Gains ranged from -32.2% to 6.4% cycle-1 for RRS and 0.0% to -61.3% cycle-1 for SRS. Heritability estimates of between 59% and 76.3% for GLS and 39% and 80% for PLS were observed indicating that both GLS and PLS can be improved through selection. Significant negative correlations between GLS and yield were observed in Pool A C0 (r = -0.947, p < 0.01) and between yield and PLS in Pool A C0 (r = -0.926, p < 0.01). These indicated gain in yield as GLS and PLS were selected against. Generally, SRS out performed RRS method both in genetic gain and time, as indicated by gain of -61% for SRS and -32.2% for RRS, respectively. Two cycles of selection were achieved in two years with SRS as compared to only one with RRS. These results clearly demonstrated that it is possible to improve for GLS resistance using simple and reciprocal recurrent selection methods. The main constraints to maize production in Western Kenya were low soil fertility, Striga, drought, lack of seed and diseases. Farmers preferred varieties that can do well under the constraints mentioned. Local collections belonged to distinct heterotic groups with good resistance to GLS and PLS and were highly heterotic to four maize population testers with both SCA and GCA effects being important in conditioning GLS and PLS resistance. Recurrent selection methods were found to improve maize resistance to GLS and PLS. Breeding should therefore, focus in development of hybrids and improvement of populations using these local collections by employing SRS and RRS selection methods with identified constraints and farmer preferences in mind. / Thesis (Ph.D.) - University of KwaZulu-Natal, Pietermaritzburg, 2007.

Powdery mildew (PM) is an important foliar disease of many crops, occurring under both greenhouse and field conditions. The application of biological control and soluble silicon (Si) against PM has received increasing acceptance as a result of increased environmental and public concern over the use of fungicides for disease management, and because many key fungicides are no longer effective because of resistance problems. However, success with these control options depends on the development of effective antagonists and understanding how best to use Si in agriculture. Potential antagonists of PM were isolated from naturally infected leaves of different plants. A total of 2000 isolates were tested in a preliminary screening on detached leaves of zucchini. The best 30 isolates showing consistent results were further tested under greenhouse conditions for their efficacy against PM of zucchini. In a greenhouse trial, 23 isolates provided disease control to levels of 30 to 77%. Application of 29 isolates resulted in significant reductions in values of area under disease progress curve (AUDPC). The best five isolates were identified as Clonostachys rosea (Link) Schroers, Samuels, Seifert & Gams (syn. Gliocladium roseum) (Isolate EH), Trichothecium roseum (Pers.) Link (syn. Cephalothecium roseum) (Isolate H20) and Serratia marcescens (Bizio) (Isolates B15, Y15 and Y41). Three adjuvants (Break-ThruR (BK), PartnerR (PR) and Tween-80R (T-80)) were compared for their ability to improve efficacy of spray application of silicon (Si) and biocontrol agents (BCAs) against PM. Both BK and PR improved the efficacy of Si significantly (P < 0.05). Microscopic studies showed that BK affected PM fungi directly and enhanced the deposition of BCAs on the pathogen. Break-ThruR was only toxic to the pathogen mycelia when used at > 0.25 m. .-1, but phytotoxic to zucchini plants when used at > 0.45m. .-1. However, it did not affect the c.f.u. of bacterial BCAs. Use of BK at 0.2-0.4 m. .-1 can be recommended to assist spray application of Si (at 750 mg .-1) or BCAs for improved control of PM. The effect of concentration, frequency of application and runoff of Si sprays applied to the foliage was evaluated for control of PM of zucchini. Silicon (250-1000 mg .-1) + BK (0.25 m. .-1), was sprayed onto zucchini plants at frequencies of 1-3 wk-1. Spraying Si reduced the severity of PM significantly (P < 0.05). Regardless of the concentration of Si, the best results were obtained when the frequency of the treatment was increased, and when spray drift or spray runoff were allowed to reach the rhizosphere of the plants. When Si was applied onto leaves, direct contact between the spray and the pathogen resulted in mycelial death. Part of the spray (i.e., drift and runoff) was absorbed by plant roots, and subsequently played an important role in the health of the plants. If affordable, soluble Si should be included in nutrient solutions of hydroponics or supplied with overhead irrigation schemes when PM susceptible crops are grown. Under greenhouse conditions, application of BCAs, with or without Si, reduced the severity and development of PM significantly (P < 0.001). Application of Si significantly reduced the severity and AUDPC values of PM (P < 0.05 for both parameters). Silicon alone reduced the final disease level and AUDPC values of PM by 23-32%, and improved the efficacy of most BCAs. In the course of the investigation, antagonistic fungi consistently provided superior performances to bacterial isolates, providing disease control levels of up to 90%. Higher overall disease levels reduced the efficacy of Si against PM, but did not affect the efficacy of BCAs. Under field conditions, Si alone reduced disease by 32-70%, Isolate B15 reduced disease by 30-53% and Isolate B15 + Si reduced disease by 33-65%. Other BCAs applied alone or together with Si reduced the disease level by 9-68%. Most BCAs reduced AUDPC values of PM significantly. For most antagonists, better efficacy was obtained when Si was drenched into the rhizosphere of the plant. However, efficacy of some of the BCAs and Si were affected by environmental conditions in the field. Repeated trials and better understanding of how to use Si and the BCAs, in terms of their concentration and application frequency, and their interactions with the plant and the environment, are needed before they can be used for the commercial control of PM. Elemental analysis was conducted to determine the impact of differing application levels of silicon (Si) in a form of potassium silicate (KSi) in solution in terms of Si accumulation and selected elements in different tissues of zucchini and zinnia and growth of these plants, and to study the effect of PM on the levels of selected elements in these two plant species. Plants were grown in re-circulating nutrient solutions supplied with Si at different concentrations and elemental composition in different parts were analysed using EDX and ICP-OES. Increased levels of Si in the solution increased the levels of Si in leaves and roots of both plants without affecting its distribution to other plant parts. In zucchini, the roots accumulated the highest levels of Si, substantially more than in the shoots. In contrast with zinnia, accumulation of Si was highest in the leaves. Accumulation of potassium (K) in shoots of both plants increased with increased levels of KSi in the nutrient solution. However, K levels in flower of zinnia, fruits of zucchini and roots of both plants remained unaffected. Increased level of Si reduced accumulation of calcium (Ca) in both plants. Adding Si into the nutrient solution at 50 mg .-1 resulted in increased growth of zucchini and increased uptake of P, Ca, and Mg by both plant species. However, application of higher levels of Si did not result in any further biomass increase in zucchini. Levels of Si in the nutrient solution had no effects on elemental composition and characteristics of the fruits of zucchini. In both plant species, the presence of PM on the leaves of plants resulted in these leaves accumulating higher levels of Si and Ca, but less P, than leaves of uninfected plants exposed to the same levels of soluble Si. The highest concentrations of Si were observed in leaf areas infected with PM, and around the bases of trichomes. For optimum disease control and maximum accumulation of different elements in these two plants, hydroponic applications of Si at 50-150 mg .-1 is recommended. Five selected biocontrol agents and potassium silicate, used as source of soluble Si, were tested under hydroponic conditions at various concentrations against PM of zinnia (Glovinomyces cichoracearum (DC) Gelyuta, V.P.). Application of BCAs resulted in reductions in final disease level and AUDPC values of PM by 38-68% and 30-65%, respectively. Both severity and AUDPC values of PM were reduced by 87-95% when plants were supplied with Si (50-200 mg .-1). It is proposed that the provision of a continuous supply of Si and the ability of this plant species to accumulate high levels of Si in its leaves were the major reasons for the good response of zinnia to Si treatments against PM. Silicon played a protective role before infection and suppressed development of PM after infection. The combination of the best selected BCAs and Si can be used as an effective control option against PM of zinnia when grown in hydroponic system. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.

The majority of farmers in Tanzania have not yet adopted modern maize varieties and still cultivate landraces and open pollinated varieties (OPVs) with low production potential and susceptible to diseases like maize streak virus (MSV), grey leaf spot (GLS) and northern corn leaf blight (NLB). The NLB disease is among the major causes of low yield and has been reported in all 21 maize growing regions in Tanzania. Breeding for host plant resistance with high yielding potential and involving the community in the breeding process is expected to address the problem of low yield, NLB disease susceptibility and low rate of F1 hybrid adoption. Therefore, the study was conducted to obtain additional sources of resistance to NLB disease, high yielding cultivars with community acceptable traits adapted to Tanzanian conditions. The main objective was to contribute to increased maize productivity in the western zone of Tanzania. The specific objectives of this study were therefore to : 1) investigate maize production limiting factors for smallholder farmers in western Tanzania, 2) identify farmers and stockist perceptions, opinions and maize variety selection criteria in western Tanzania, 3) establish NLB disease status in farmers’ fields of western Tanzania, 4) determine the genetic relationships among landraces and assess maize landraces as sources of breeding materials, 5) determine the combining ability and heterosis for NLB disease resistance of eleven maize inbred lines adapted to Tanzanian conditions, and 6) determine the gene action and inheritance of resistance to NLB disease in five maize inbred lines adapted to Tanzanian conditions. The study was conducted from 2008-2011 in three diverse environments which represent all the maize growing regions in the country The participatory rural appraisal (PRA) was conducted in three districts to investigate farmers’ and stockists preferred traits for maize selection in western Tanzania, determine maize production constraints facing farmers and assess NLB disease prevalence in the same area. A focus group of 30 farmers was selected in each of the three villages. Transect walks, wealth ranking and historical profiles were used in an informal survey. One hundred and fifty questionnaires were used in a formal survey. The recorded yield was only 1 t haˉ¹. Thirteen major maize production constraints, 13 insect pests and vermin and, 11 diseases were recorded. The NLB disease was reported to be increasing in severity in all farmers’ fields. Farmers’ preferred traits included resistance to abiotic and biotic stresses, early maturity, preferred milling qualities, high storage qualities and high yielding potential. Stockists mentioned 12 preferred maize variety traits which included high yielding, disease and insect pest resistance, heavy grain, large cob size and large grain sizes. Similarity between farmers and stockist variety preference ranking were found to exist. The occurrence and distribution of northern leaf blight (NLB) disease study was conducted to assess the incidence and severity of NLB disease in farmers’ fields in seven districts. The study was conducted for two seasons. In each season, 175 fields with 5600 plants were sampled. There were sixteen varieties grown with wide NLB disease reaction variation. Gembe, a landrace, was among the three observed resistant varieties. The NLB disease has changed its distribution pattern affecting all districts of the western zone. The disease incidence in season two (2009/2010) significantly increased from season one (2008/2009) t= -3.25 (348), P= 0.001. About 30% of both means of blight incidence and severity were recorded in the area. Characterization and screening of maize landraces for northern leaf blight disease resistance was conducted to determine the genetic relationships among landraces, assess maize landraces as sources of NLB disease resistance and assess important agronomic traits for future maize improvement. Ninety breeding materials consisting of 71 landraces and 19 commercial varieties were evaluated. The average yield of landraces under research management was 2.3 t haˉ¹. Landrace TZA 3075 was identified as NLB disease resistant. Yield potential, dent grain texture, white endosperm and husk cover were important agronomic traits observed among landraces. There were high variations in terms of morphology and NLB disease resistance among the landraces. Five principal components contributed to 71.98 % of total variation. Clusters analysis revealed five distinct groups of landraces. Leaves/plant, infested leaves/plant, lesion number, lesion length, lesion width and NLB disease incidence traits highly contributed to variation and grouping of landraces. Combining ability analysis for northern leaf blight disease resistance was conducted to estimate the combining ability for NLB disease resistance of 11 maize inbred lines adapted to Tanzanian conditions, determine maternal effects which are involved in NLB disease resistance in maize germplasm, and determine the heterosis in the F1 hybrids. A full 11 x 11 diallel cross was performed. All top ten experimental hybrids in each of the three sites had negative midparent heterosis for NLB disease severity. The overall mid-parent heterosis means for yield across sites was 152%. The mean sum of squares for GCA was highly significant (P< 0.001) on disease severity indicating additive gene action effects. Mean sum of squares for SCA were highly significant for disease severity and yield implying non-additive gene action effects. The mean squares for reciprocal effects were highly significant on yield and non-maternal sum of squares had significant effect (P<0.05) on yield. The GCA contribution was high for disease severity (91%) and lesion number (85%). Almost, all GCA effects for NLB disease resistance were negative implying contribution to disease resistance. Due to preponderance of the additive gene action, recurrent selection could be used to improve the resistance of inbred lines while the non-additive gene action could be exploited in breeding for disease resistant hybrids. Generation mean analysis of northern leaf blight disease resistance was conducted to determine the mode of gene action involved in the inheritance of resistance to NLB disease in five inbred lines adapted to Tanzania at contrasting environments, estimate heterosis and heritability in five tropical inbred lines. Generation mean analysis was conducted using a six parameter model comprising P1, P2, F1, F2, BCP1 and BCP2 generation progenies. The mean sum of squares for environment, replication with the nested environment, generations, generations x environment interactions were highly significant (P<0.001). The full model of additive, dominance, additive x additive and additive x dominance epistatic effects was highly significant (P<0.001). Nonetheless, the additive gene effects were predominant ranging between 57% and 89% which was matched by large heritability (54%-85%). The average degree of dominance ranged between -0.52 and 0.88 supporting observations of partial dominance. The NLB disease severity showed a continuous distribution in all three sets for F2, BCP1 and BCP2 populations which is an indication of quantitative nature of inheritance and additive gene effects. The mid parent heterosis ranged from -19 to 1%. Therefore, resistance to NLB disease could be improved through selection by exploiting the additive gene effects. The epistatic gene effects would cause less complications because they were negligible (<25%). The client oriented breeding for maize northern leaf blight disease resistance was carried out to perform farmers and stockists assessment on the 110 F1 experimental maize hybrids and compare them with breeders selection criteria. Breeders selection criteria ranked 10 top high yielding experimental hybrids. Farmers developed 14 while stockists developed 13 selection criteria. The most preferred hybrids by farmers were VL 05616 x CML 159, CML 159 x KS03- 0B15-47 and EB04-0A01-304 x CML 442 while stockists preferred VL 05616 x CML 395, EB04-0A01-304 x CML 442 and VL 05616 x CML 159. Two F1 experimental hybrids EB04- 0A01-304 x CML 442 and CML 159 x CML 442 appeared in all top five ranked hybrids by breeders, farmers and stockists. Generally, findings showed that, farmers, stockists and breeders coincide in some selection criteria but also differ in other cases. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

The Sugarcane Industry contributes approximately 400 000 jobs and ZAR 8 billion annually to South Africa’s economy. Due to climate change and the subsequent threat posed by disease, these figures have been on the decline. Brown rust, a contributor to this decline is caused by the basidiomycete Puccinia melanocephala Syd. and P. Syd., which previously resulted in 50% yield losses in susceptible varieties. This highlighted the need for improved screening and breeding techniques which will result in the replacement of susceptible varieties. The objectives of this study were to: a) Adopt and optimise a glasshouse whorl inoculation screening technique applicable for mass screening of large populations. b) Develop a rapid and cost effective rust resistance screening technique using detached leaves. c)Utilise two flanking marker sets (R12H16 and 9O20-F4-PCR primers) for the rust resistance Bru1 gene in a diagnostic polymerase chain reaction (PCR) to identify rust resistant genotypes lacking Bru1 and possessing either quantitative resistance or an alternative major qualitative resistance gene. d) Correlate rust phenotypic data to AFLP marker data for the Linkage Disequilibrium (LD2) mapping population. e) Utilise suppression subtractive hybridization (SSH) profiling on rust challenged genotypes to discover differentially expressed genes between susceptible and resistant (susceptible Bru1 negatives taken away from resistant Bru1 negatives); and resistant genotypes (resistant Bru1 positives taken away from resistant Bru1 negatives). 4 Results from the glasshouse whorl inoculation trials showed the technique could be reliably used to screen large populations, as two independently conducted pot trials showed highly correlated rust ratings. A visually assessed detached leaf assay (DLA) was developed using selected genotypes. Chlorophyll fluorescence and SPAD readings were used in the DLA to determine the leaf photochemical efficiency (PIABS) with relation to chlorophyll content, resulting in reduced assessment time of at least two days. PCR diagnostics revealed 31% of LD2 did not possess either flanking marker, 8% had one or the other marker, and 61% had both markers. The overall rust phenotypic ratings (rating scale of 0-10) and Bru1 status of the genotypes was used to group the population, with the Bru1 negative genotypes containing all three rating categories (resistant 0-3.5; intermediate 3.51-6.5; susceptible 6.51-10); while the Bru1 positive genotypes were all resistant. The phenotypic data was correlated to AFLP data using the Pearson product-moment correlation coefficient and stepwise multiple linear regression employed to build marker based models to use for predicting non-Bru1 mediated resistance. SSH analysis was then subsequently conducted on genotypes selected on the basis of Bru1 status and AFLP correlation data. Two subtraction cDNA libraries were constructed and the cDNA inserted into electro-competent Escherichia coli cells. PCR on transformed cells revealed cDNA inserts ranging from 200- 1300bp. BLAST analysis of the cDNA sequences indicated the presence of high proportions of disease and drought stress related sequences in the libraries. Analysis of the sequences in both libraries showed that the resistant Bru1 negative genotypes contained oxidative stress related sequences which were however absent in the Bru1 positive resistant genotypes. The library comparing the Bru1 negative resistant genotypes against the Bru1 negative intermediate and susceptible genotypes showed a higher proportion of differentially expressed sequences coding for putative disease resistance proteins, highlighting their presence in the resistant genotypes. Both subtraction libraries also contained high proportions of a leucine rich repeat protein coding cDNA which contained a conserved domain homologous to that of a disease resistance protein conferring resistance to Pseudomonas syringae in Arabidopsis thaliana. The outcomes of this study will subsequently enable an improved understanding of sugarcane-rust resistance mechanisms and improved breeding and screening techniques for sugarcane by identifying SSH and AFLP markers linked to rust resistance QTLs or alternative R genes. / Thesis (M.Sc.Agric)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

The first serious outbreak of brown rust of sugarcane caused by Puccinia melanocephala Syd. & P. Syd. was reported in India in 1907. It was first reported in South Africa (SA) in 1941 on the variety Co301 and is now present in almost all the sugarcane growing areas of the world. In SA, it is now described as an important disease of sugarcane, causing yield losses of up to 26% in susceptible varieties. Within the SA sugar industry, rust is controlled through the use of resistant varieties as it is the most economical method of control. However, most of the newer varieties that are being released have an intermediate resistance rating for rust. An integrated management approach for the control of rust is therefore being investigated. Aspects investigated in this study included environmental conditions required for development of the disease i.e. epidemiology, the use of silicon (Si) as a cultural control method against brown rust and identification of gene sequences expressed in response to brown rust infection. For the epidemiology study, inoculated plants were incubated in a dew chamber at different temperatures and leaf wetness periods. The choice of leaf wetness duration and temperature was based on urediniospore germination studies. The optimum temperature for urediniospore germination and disease development at > 98% relative humidity was found to be between 20 and 25°C with nine hours of leaf wetness. Silicon has been shown to reduce the incidence of diseases and pests in a number of crops. The ability of sugarcane to accumulate Si and the location of Si deposition was established using two uptake and deposition trials. Different concentrations of Si were applied to the plant and accumulation in the roots, stalks, old leaves and young leaves was determined using inductively coupled plasma optical emission spectrometry, with accumulation found to be roots > old leaves > stalks > young leaves. Silicon deposition in the leaves was determined using energy dispersive X-ray mapping on freeze dried specimens and significant differences were found between the upper epidermis, lower epidermis and mesophyll with the most Si being deposited in the lower epidermis. For disease severity, plants were naturally infected with rust and rated weekly. A significant decrease in disease severity and area under disease progress curve was noted when the Si concentration increased, indicating that Si has potential in reducing rust incidence. Currently, the most reliable and economical method of managing brown rust is with the use of resistant varieties. Identification of resistance within breeding lines is therefore important. For this part of the study, suppression subtractive hybridization was used as a tool to identify differentially expressed genes between a susceptible and resistant variety and a susceptible and intermediate variety, in response to brown rust infection. Two efficient subtracted cDNA libraries were generated and differentially expressed sequences were identified within each library. The results of this study show potential for the development of molecular markers which could be used for the early identification of brown rust resistance during the breeding process. This study forms a firm basis on which an integrated management strategy, for the management of brown rust in the SA sugar industry, could be designed. The cDNA sequences identified could be further investigated and used to develop molecular markers to select for rust resistant varieties, the epidemiology results together with further field data could be used to develop a disease prediction model and Si has potential in the field to reduce brown rust severity. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

No description available.

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.

Soybeans, Glycine max, are an economically and strategically important crop in South Africa (SA). In order to meet local demands, large imports of soybeans are required, e.g., in the 2005/2006 soybean production period, 842 107 tonnes of oilcake were imported. Due to an increase in soybean production throughout the world, diseases that affect this crop have also increased in incidence and severity. Sclerotinia sclerotiorum, the causal organism of sclerotinia stem rot (SSR), is an important yield limiting disease of soybeans, as well as numerous other crops. The pathogen was first reported in SA in 1979. However, it was only in 2002 that this fungus was considered a major pathogen of soybeans in SA. The research reported in this thesis was conducted to investigate the epidemiology of S. sclerotiorum and examine numerous potential control methods for this pathogen, i.e., resistant cultivars, biocontrol, chemical control and seed treatments. A S. sclerotiorum isolate was obtained from sunflowers in Delmas, Mpumulanga, SA, in the form of sclerotia. This isolate was cultured and sent for identification and deposition in the Plant Protection Research Institute collection. This isolate, in the form of mycelia, was used for the duration of the study. For epidemiology studies, the effect of temperature, leaf wetness duration (LWD) and relative humidity (RH) were examined for their effect on rate of pathogen development. Twenty four combinations of temperature (19°C, 22°C, 25°C and 28°C), LWD (24, 48 and 72 hr) and RH (85 and 95%) were investigated. No interaction between temperature, LWD and RH was found. Temperature alone was the only factor that affected disease development. At 22°C, the rate of pathogen development (0.45 per unit per day) was significantly higher than all other temperatures, indicating that this temperature is optimum for disease development. Thirteen different soybean cultivars, i.e., LS6626RR, LS6710RR, LS666RR, LS555RR, LS6514RR, LS678RR, Prima 2000, Pan 626, AG5601RR, AG5409RR, 95B33, 95B53 and 96B01B, commercially grown in SA were investigated for their reaction to S. sclerotiorum. Prima 2000, 96B01B, 95B33 and AG5409RR were considered to be the least susceptible as they showed a significantly low rate of pathogen development (0.28, 0.28, 0.24, 0.23 per unit per day, respectively) and produced a significantly low number of sclerotia (3.03, 3.42, 3.21, 2.38, respectively). LS6626R and LS666RR may be considered most susceptible because of their significantly high rate of pathogen development (0.45 and 0.42 per unit per day, respectively) and high sclerotia production (8.16 and 7.50, respectively). Regression analysis showed a positive correlation coefficient (R2=0.71) between rate of growth of the pathogen and number of sclerotia produced, indicating that a higher rate is associated with a higher number of sclerotia. In vitro dual culture bioassays were performed to identify the biocontrol mechanisms of the biocontrol agents, EcoT® (a seed treatment) and Eco77® (a foliar treatment), against hyphae and sclerotia of S. sclerotiorum. Ultrastructural studies revealed that mycoparasitism is the probable mode of action as initial signs of hyphae of EcoT® and Eco77® coiling around hyphae of S. sclerotiorum were observed. Surface colonization of sclerotia by hyphae of EcoT® and Eco77® was also observed. In vitro antagonism of EcoT® against S. sclerotiorum on soybean seed was performed to determine pre-emergence and post-emergence disease. There was no significant difference in percentage germination between seeds treated with EcoT® and plated with the pathogen, untreated seeds and no S. sclerotiorum, and the control (i.e. no EcoT® and no pathogen). However, percentage non infected seedlings from seeds not treated with EcoT® was significantly lower, suggesting that EcoT® may be successfully used as a seed treatment for the control of SSR. In vivo trials were performed to investigate the effect of silicon (Si) alone, and in combination with Eco77®, on the effect of the rate of disease development. Plants treated with Eco77® had a significantly lower rate of disease development (0.19 per unit per day for plants treated with Eco77® and S. sclerotiorum and 0.20 per unit per day for plants treated with Eco77®, S. sclerotiorum and Si), compared to plants not treated with Eco77® (0.29 per unit per day for plants treated with S. sclerotiorum and 0.30 per unit per day for plants treated with S. sclerotiorum and Si), regardless of the application of Si. Similarly, plants treated with Eco77® had a significantly lower number of sclerotia (0.46 for plants treated with Eco77® and S. sclerotiorum and 0.91 for plants treated with Eco77®, S. sclerotiorum and Si), compared to plants not treated with Eco77® (3.31 for plants treated with S. sclerotiorum and 3.64 for plants treated with S. sclerotiorum and Si). The significantly lower rate of disease development coupled with a significant reduction in sclerotia showed that Eco77®, and not Si, was responsible for reducing the severity of SSR. A strong positive correlation between rate of disease development and the number of sclerotia produced (R2=0.79) was observed. For the investigation of various fungicides for the control of S. sclerotiorum, in vitro trials to determine the potential of three different fungicides at different rates, i.e., BAS 516 04F (133 g a.i. ha-1), BAS 516 04F (266 g a.i. ha-1), BAS 512 06F (380 g a.i. ha-1) and Sumisclex (760 g a.i. ha-1) were initially conducted. The control (non-amended PDA) had a significantly higher area under mycelial growth curve (243.0) than all fungicides tested. BAS 516 04F (at both concentrations) and BAS 512 06F completely inhibited the mycelial growth of S. sclerotiorum. Sumisclex inhibited the fungus by 89.07%. For in vivo trials, preventative treatments, i.e., BAS 516 04F (133 g a.i. ha-1), BAS 516 04F (266 g a.i. ha-1), BAS 512 06F (380 g a.i. ha-1), curative treatment, i.e. Sumisclex (760 g a.i. ha-1) and a combination preventative/curative treatment, i.e., BAS 512 06F (380 g a.i. ha-1)/Sumisclex (570 g a.i. ha-1) were investigated. No significant difference in disease severity index (DSI) was found between fungicide treatments and the inoculated control. BAS 512 06F and BAS 512 06F/Sumisclex had significantly lower grain yields (6.09 g and 5.96 g, respectively) compared to all other treatments. There was a positive correlation coefficient (R2=0.76), between DSI and grain yield, indicating that a high DSI is correlated with low grain yield. Trials to evaluate the effect of commercially available and currently unregistered seed treatments for the control of S. sclerotiorum on soybean seeds in vivo and in vitro were performed. Seed germination tests were performed to determine if seed treatments had any negative effects on seed germination in vitro. All seed treatments tested, i.e., BAS 516 03F (8, 16 and 32 ml a.i. 100 kg-1 seed), BAS 512 00F (7.5, 15 and 32 ml a.i. 100 kg-1 seed), Celest XL (100, 125, 200 and 250 ml a.i. 100 kg-1 seed), Sumisclex (5 and 10 ml a.i. 100 kg-1 seed), Benomyl (150 g a.i. 100 kg-1 seed), Captan (240 ml a.i. 100 kg-1 seed), Thiulin (180 g a.i. 100 kg-1 seed) and Anchor Red (300 ml a.i. 100 kg-1 seed), showed no negative effect on seed germination. For in vivo trials, BAS 516 03F (16 and 32 ml a.i. 100 kg-1 seed), BAS 512 00F (7.5, 15 and 32 ml a.i. 100 kg-1 seed), Celest XL (100, 125, 200 and 250 ml a.i. 100 kg-1 seed), Sumisclex (5 and 10 ml a.i. 100 kg-1 seed), Benomyl and Anchor Red had significantly similar percent germination and percent seedling survival as the untreated/uninoculated control. These seed treatments should be recommended for the control of S. sclerotiorum, as they protected seed during germination and subsequent seedling development. BAS 516 03F (8 ml a.i. 100 kg-1 seed) should not be recommended for the control of SSR, as it gave the lowest percent germination and percent seedling survival. The results presented in this thesis have helped to identify optimal environmental conditions for the development of S. sclerotiorum, which is important for the development of forecasting models for disease control. The least and most susceptible cultivars of those tested have been identified. Biocontrol using Eco77® as a foliar application showed great potential. The effect of Si needs to be further investigated, including the testing of more frequent applications and higher concentrations. The fungicides tested in this research did not show any potential for the control of SSR. However, the spray programme tested is for the control of soybean rust (Phakopsora pachyrhizi), and was investigated for its potential for the control of SSR. The spray programme, fungicide application and rating scale needs to be modified, to determine the true potential of these fungicides for the control of SSR. Numerous seed treatments have shown potential for the control of seed infection by SSR. Due to difficulties in producing ascospores, which are the primary source of inoculum for this pathogen in the field, all studies in this research were conducted with mycelia and not ascospores. The production, collection and storage of ascospores needs to be thoroughly investigated, and research conducted with ascospores. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2007.