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.

Sorghum breeders have not made much yield improvement in new sorghum varieties in Kenya since Serena in was released in the late 1960s. KARI Mtama-1 which was released in 1993 has no yield advantage over Serena. A yield plateau for sorghum in Kenya is apparent. A new breeding approach was adopted to break that yield barrier. Development of hybrid sorghum was proposed and is expected to break the yield barrier and also deliver cultivars that meet farmers' main requirements. The objectives of the study were to (1) identify farmers' requirements in sorghum cultivars, constraints to sorghum production and why improved cultivars from research are not being adopted, (2) characterize male and female parents and establish if genetic distance could identify superior parent populations for hybrid production (3) estimate genetic variance components and determine the possibility of using GCA and SCA estimates in choosing parents for use in hybrid production, (4) test hybrids and open pollinated variety (OPV) parental lines for stress tolerance and identify tolerant hybrids for further testing and, (5) compare single cross hybrids and OPV varieties in yield performance. Participatory rural appraisal in Kitengei and Nzambani areas of Kambu showed that sorghum was especially important in semi-arid parts of Kenya. Food, trade, feed, nursing food and thatching were the most important uses of sorghum. High grain and stover yield, large grain size, early maturity, drought tolerance, pest and disease resistance, coloured grain and intermediate plant height were the major requirements of farmers. Fifty-three pollinators and forty-one male sterile parents were introduced from four sources and screened together with 27 pollen parents from Kenya. Parents and hybrids were tested in 4 environments: high and low plant density, in high and low moisture regimes laid out in a triple square lattice design in Kenya, with parents having two additional tests in South Africa. Males, females, sexes and parental sources differed significantly in head weight. There were sex x country and sex x environment interactions for head weight. Genetically distant parents' populations had higher chances of superior heterosis. Parents showed significant additive genetic variance in head weight. The regression of non-additive to additive genetic variance was roughly one and significant. Three female and five male parents were suitable for production of hybrids adapted to multiple environments. Hybrids and OPV lines significantly varied in head weight. Hybrids were superior to OPV lines in most agronomic traits. Economic superiority of the hybrids was sufficient to cover cost of hybrid production and distribution in Kenya. Hybrids and OPV lines varied significantly for plant density stress. Hybrids were less sensitive to stress and more productive than OPV lines under population density stress. KARI varieties were sensitive to plant density stress. In general low sensitivity to stress was beneficial and hybrids had superior yield to inbred varieties. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.

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.

Sorghum is the fifth most important cereal crop in the world and ranks third in Africa, and it is potentially the number one cereal for the semi-arid environments in sub-Saharan Africa. Sorghum varieties have been developed specifically for grain, fodder or stem sugar but not for dual-purpose combining grain and stem sugar. Such varieties could be beneficial to the resource-poor farmers by providing grain for food and sugar rich stalks that can be sold for bioethanol production. However, there are no suitable dual-purpose cultivars on the market. There is also limited information about the combining ability, gene action and genetic effects and relationships between stem sugar and grain yield which is required in devising appropriate strategies for developing dual-purpose sorghum varieties. Furthermore, there is also lack of information about the perceptions of resource-poor, small-scale farmers and other important stakeholders on the potential of dual-purpose sorghum production and the value chain. Therefore, the objectives of this study were to: (i) investigate the awareness of the farmers, industry and other stakeholders on the dual-purpose sorghum varietal development and its feasibility, (ii) screen germplasm for use as source materials useful for grain yield and stem sugar traits, (iii) investigate the inheritance and heterosis levels attainable in grain yield components and stem sugar traits in dual-purpose sorghums, (iv) determine the relationships between stem sugar traits and grain yield components in dual-purpose sorghums, and (v) investigate the fertility restoration capacities of selected male-fertile lines used as male parents through the evaluating seed set in experimental dual-purpose hybrids. Two surveys were conducted to establish stakeholders’ level of awareness and perceptions on the potential and feasibility of developing and utilising dual-purpose sorghums in Southern Africa. One survey was carried out in the semi-arid tropical lowlands in Zimbabwe under the conditions of small-scale and resource-poor farmers while the other, which targeted sugar industries, plant breeders, engineers, political leaders, economists and extension workers, was conducted in South Africa and Zimbabwe. Data were analysed using SPSS computer package. Results showed that both farmers and the non-farmer stakeholders were in agreement on the view that dual-purpose sorghum would be a viable enterprise that could alleviate poverty, enhance food security, create rural employment and boost rural development in southern African countries. Farmers were willing to adopt the cultivars if they were made available. The stakeholders also suggested mechanisms to overcome the infrastructural, economic and technical challenges associated with the technology. Screening of regional and international germplasm collection held at the University of KwaZulu- Natal in South Africa revealed high genetic variability for grain yield, stem brix and stem biomass yield that can be exploited in dual-purpose sorghum cultivar development. Ten lines were selected for inclusion as parents in the dual-purpose sorghum breeding programme. The selections were crossed to eight cytoplasmic male-sterile lines originating from the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) in accordance with a North Carolina Design II mating scheme. The 18 parents, together with the 80 experimental hybrids generated and two check varieties were evaluated for grain yield and stem sugar traits in six tropical low- and mid-altitude environments in Mozambique, South Africa and Zimbabwe. Stem sugar concentration and stem biomass were measured at the hard dough stage of each entry due to maturity differences between the genotypes. Grain yield was measured and adjusted to 12.5% moisture content. Data were analysed in GenStat computer package following a fixed effects model. Both additive and non-additive gene effects were important in controlling stem brix, stem biomass, grain yield and the associated traits in dual-purpose sorghum. This showed that breeding progress can be achieved through hybridisation and selection. Cultivars showing high stability, and high standard and better-parent heterosis for the three traits were identified implying that breeding for general adaptation was an option and that productivity could be enhanced by breeding hybrid cultivars. The relationships between traits were estimated using correlation and path-coefficients analysis. Grain yield was found to be negatively and significantly associated with stem brix but was positively and significantly associated with stem biomass. This implied that breeding for high stem brix might compromise grain yield but selection for high stem biomass improved grain yield. Stem biomass and stem brix were not significantly correlated. The general negative relationship between grain and stem brix was attributed to the predominance of entries with contrasting performances for the two traits. However, the relationship between grain yield and stem brix of the top 20 performing entries showed a non-significant relationship between stem brix and grain yield suggesting that the traits were independent of each other. This finding was confirmed by the presence of crosses that combined high performance for both stem brix and grain yield as well as stem biomass among the hybrids. The relationships between stem brix and stem biomass for the top 20 performers remained non-significant while that between stem biomass and grain yield became stronger, positive and significant. Direct selection for stem brix and grain yield was shown to be more important than indirect selection, while selection for stem biomass improves grain yield but had no effect on stem brix. Therefore, it is possible to breed dualpurpose sorghum cultivars and the identification of genotypes combining the desirable traits is prudent in addition to general relationships information. The study on fertility restoration capacities as evaluated through hybrid seed set showed that fertility restoration was under the control of genes with both additive and non-additive action. Since restoration is conferred by a single dominant gene (Rf1), this could have arisen from the action of the modifier genes that have been previously reported to influence it. This showed that fertility restoration can be improved through breeding. Hybrid combinations showing complete seed set and high performance for grain, stem brix and stem biomass were identified and are potential dual-purpose sorghum cultivars. Overall, the study showed that development of dualpurpose sorghum cultivars would be feasible and genotypes identified as potential cultivars in this study will be forwarded for further testing across many sites and seasons in the target environments. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Viral diseases are a major limiting factor to cowpea production in many countries of Africa. In Uganda, studies indicated that the cowpea aphid-borne mosaic virus (CABMV) is common and a potential threat to cowpea production in the region. There have been no efforts to develop cowpea cultivars with resistance to CABMV in Uganda. This work focused on the development of cultivars resistant to CABMV. Production of cowpea in Uganda is constrained by several factors, including a lack of awareness of diseases among the majority of farmers. A participatory rural appraisal (PRA) was conducted to elicit farmers’ indigenous knowledge of cowpea production and also to gain insight into their understanding of viral diseases affecting cowpea in Uganda. PRA tools such as group discussions, transect walks, problem listing and ranking were used to gather information. Insect pests, diseases, low yielding cultivars and the high cost of pesticides were perceived to be the most important production constraints. Farmers were not aware of the problem of virus diseases, but provided descriptive names of symptoms. Only three cowpea cultivars (Ebelat, Ecirikukwai and Blackcowpea) were produced in the area. Seed size and colour were seen as important traits in new varieties. Information about the occurrence, distribution and identity of cowpea viruses is limited in Uganda. The objective of this study was to identify the important cowpea virus diseases occurring naturally in the major cowpea growing regions of Uganda. Surveys were conducted to determine the incidence and severity of virus symptoms in four districts (Soroti, Kumi, Pallisa and Tororo) in 2004 and 2005. The incidence ranged from 40.5 to 94.4% and severity ranged from 15.0 to 30.6% (for Kumi and Pallisa districts, respectively) during the 2004 surveys. In 2005, the incidence ranged from 55.9 to 85.4% and severity ranged from 4.7 to 14.5% (for Tororo and Soroti districts, respectively). The CABMV, cowpea mild mottle virus (CPMMV), cowpea severe mosaic virus (CPSMV) and cucumber mosaic virus (CMV) were serologically detected by double antibody sandwich enzyme- linked immunosorbent assay (DAS-ELISA). Fifty four improved cowpea genotypes were screened for resistance to CABMV during the first season of 2004 at Serere Agricultural and Animal Production Research Institute in Uganda. Further screening was conducted in the second season of 2004 using 27 genotypes. The genotypes were planted in single rows between the rows of the susceptible cultivar, Ebelat. This was to provide high pressure of aphid vector (Aphis craccivora Koch) and CABMV inoculum. In addition, the test genotypes were artificially inoculated with a CABMV extract on fully expanded primary leaves of fourteen day-old seedlings. The CABMV incidence and severity was assessed. Disease severity was assessed on a 0-60% visual estimation scale where 0 = with no symptoms and 60 = with severe symptoms. Serological analysis was conducted using DAS-ELISA. Five genotypes showed good levels of resistance to CABMV, namely MU-93, IT82D-889, IT82D-516-2, IT85F-2841 and SECOW-2W. These resistant lines were crossed with three susceptible local landraces, namely Ebelat, Ecirikukwai and Blackcowpea in a North Carolina II mating design. The F1, F2 and BC1F1 populations and the parents were evaluated in the field to assess the response to CABMV and to study the inheritance of resistance to CABMV. The general combining ability (GCA) and specific combining ability (SCA) effects were significant, indicating that both additive and non-additive genetic factors are important in determining the control of CABMV in cowpea. The proportions (%) of the sum of squares for crosses attributable to GCA and SCA for CABMV severity were 51.4% for GCA due to females, 8.4% for GCA due to males and 40.2% for the SCA. The narrow-sense heritability estimates, obtained by regressing F1 on mid-parents was 0.87 and 0.84, F2 on F1 progenies 0.49 and 0.48, and F2 progenies on mid-parents 0.63 and 0.79, for AUDPC and final disease severity, respectively. Single gene conditioned resistance in seven populations, but resistance was quantitatively inherited and involved many genes in eight populations. Observation of transgressive segregation and moderate to high heritability suggests a quantitative mode of gene action and the importance of additive effects. The predominance of GCA variance, high heritability estimates and observation of transgressive segregation suggested that resistance could be improved by selection. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2007.

Finger millet (Eleusine coracana (L.) Gaertn. ssp. coracana) is an important food, food security and cash crop in eastern and southern Africa where small-scale farmers grow it in low input farming systems. The crop has food security, nutritional, cultural, medicinal, and economic value with high industrial potential. Little research and hardly any breeding have been done on the crop leading to low yields and low production. A project was therefore implemented in western Kenya during 2004-2007 seasons to investigate the possible breeding contributions to enhance productivity and production of the crop. The research comprised a social survey, germplasm evaluation, appraisal of ethrel as a chemical hybridising agent (CHA), genetic analysis of yield, and resistance to blast and Striga, and breeding progress in developing new finger millet varieties. A participatory rural appraisal (PRA) was conducted in three districts during 2006 to position finger millet (FM) in the farming systems, production constraints, and variety diversity and farmer preferences. The PRA established the high rating the peasant farmers gave to finger millet among crop enterprises, using it for food, cash, brewing, ceremonies and medicinal purposes. Farmers cultivated many varieties ranging from five to nine in a district, but each district had its own popular variety. Farmers used the following criteria to select new cultivars: high yield potential; early maturity; resistance to blast disease, Striga, birds, drought, and lodging; large head size, dark grain colour, and good taste. This probably indicated the willingness of farmers to adopt new varieties. Farmers identified constraints to production as blast disease, Striga, wild FM, birds, rats, termites, lack of market, labour shortage, and low yield. The farmers’ variety selection criteria and production constraints underscored the need to improve finger millet varieties. Evaluation of 310 accessions for trait variability and association conducted during 2005 long rain (LR) season at two sites revealed wide variation among the accessions for yield and secondary traits. The best accessions grain yield was above the yield potential of 5,000- 6,000kg ha-1 reported in other environments. Accessions KNE 072 (7,833kg ha-1), GBK 028463 (7,085kg ha-1), GBK 029661 (6,666kg ha-1) and FMBT ACC#42 (6,566kg ha-1) were outstanding. The data showed the opportunity to select for yield directly because of its wide variability but indirect selection could also be used to exploit seedling vigour as shown by its high correlation to yield and direct and indirect positive effects on yield through plant height and single plant yield in path analysis. The wide genetic variability among the genotypes for several traits indicated high potential to breed new and better finger millet varieties. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.

Maize (Zea mays L.) yields in the smallholder (SH) farming sector in Southern Africa have remained low. despite the availability of many improved varieties. Among the major constraints contributing to tow yields and threatening food security in the region are diseases which include grey leaf spot (GLS). common rust, northern corn leaf blight (NLB) and Phaeosphaeria leaf spot (PLS). These diseases are highly unpredictable in their occurrence every season, making them difficult to control. In addition, the majority of SH farmers cannot afford to control the diseases due to limited access to chemicals. Therefore, maize cultivars with high levels of disease resistance and tolerance to abiotic stresses would provide a long-term solution to addressing the problem of low yields, especially in the smallholder-farming sector. The objectives of this study were therefore to: i) establish farmers' perceptions on diseases, key limiting production constraints and preferred traits of maize cultivars. ii) screen germplasm adapted to tropical environments for resistance to PLS, iii) determine gene action for resistance to PLS and GLS, iv) estimate combining ability effects for resistance to PLS, GLS, NLB and common rust diseases, and v) determine grain yield stability of F, hybrids derived from crosses among selected tropical advanced maize inbred lines. These studies were conducted from 2006/7 to 2008/9 seasons at various sites in South Africa, Zimbabwe, Zambia and Uganda. Structured surveys and participatory rural appraisal (PRA) conducted in Obonjaneni. Busingatha and Okhombe villages of Amazizi district in the Northern Drakensberg established maize as the principal crop grown in the area. All the farmers who participated grew the local variety (landrace) they called Natal-8- row or Is/Zulu. The adoption of hybrids and improved open pollinated varieties (OPVs) was tow. Farmers preferred the local variety ahead of hybrids and improved OPVs mainly for its taste, tolerance to abiotic stresses and yield stability. Characteristics of maize varieties preferred by the farmers included: inexpensive seed, high yield, early maturity and tow input costs. Pests/d is eases and drought were not ranked highly, as farmers planted early to escape diseases and drought. Abiotic stresses were amongst the top four constraints faced by the farmers. The local varieties exhibited high yield potential and genetic variability fordisease resistance. Evaluation of maize germplasm adapted to tropical and subtropical environments of Africa for PLS resistance indicated significant (PS0.05) variation among the inbreds. populations and hybrids. In general, 63% of the inbreds/populations were resistant to PLS. Regionally important inbred lines; SC and N3 and CIMMYT's most successful lines such as CML395. CML444. CML202. CML312. and CML488 were resistant to PLS. Fifty- four percent of the single-cross experimental hybrids were also resistant to PLS. Correlation coefficients for area under disease progress curve (AUDPC) values for disease severity with PLS final disease severity scores were significant (P<0.001) and positive, implying that ranking of the genotypes for AUDPC and final PLS disease severity score was by and large similar. Forty five F, hybrids generated by crossing ten advanced maize inbred lines in a half diallel mating scheme were evaluated in two to six environments to determine combining ability, gene action and heterosis estimates for grain yield and resistance to PLS. GLS. NLB and common rust diseases. Highly significant (PS0.001) general combining ability (GCA) and specific combining ability (SCA) effects were observed for PLS. GLS. NLB. common rust, grain yield and other agronomic traits. The GCA effects were more important than SCA effects, indicating the predominance of additive over non-additive gene action for all the traits studied in these inbred lines. The inbred lines with good GCA for PLS resistance were: A12204, N3. A16. MP18 and CML488. and for GLS resistance were A1220-4. CZL00009. CZL00001. CML205 and CML443. Lines A16 and CML443 had good GCA for NLB and common rust resistance, lines A1220-4, N3, CML205, A16, and CML443 contributed towards high yield. Lines A1220-4 and A16 were late maturing, whereas CZL00009 displayed eariy maturity. High mid-parent and better-parent heterosis for high grain yield and resistance to all the diseases were observed. Generation mean analysis was used to deteimine the inheritance of PLS and GLS resistance in populations involving six tropical advanced maize inbred lines. Reciprocal crosses and backcross progenies were generated among inbreds A1220-4, A15, B17 (resistant. R), CML445 (moderately resistant. MR). CML441 and CZL00001 (susceptible. S) for PLS inheritance, and among inbreds A1220-4. A15, CML441 (resistant. R). and N3 and B17 (susceptible. S), for GLS inheritance. Results indicated highly significant additive effects (P<0.001) for PLS and GLS resistance, with dominance effects accounting forSH%of the variation in all the crosses for PLS and only A15 x B17 cross for GLS. Epistasis and cytoplasmic gene effects in favour of PLS resistance in F, crosses when the more susceptible parent was used as female were significant. For GLS resistance, epistasis was observed only in CML441 x N3 and A1220-4 x B17 crosses, while no cytoplasmic gene effects were detected. Resistance for PLS was medium to highly heritable and conditioned by less than four genes which exhibited incomplete dominance. In general resistance to GLS was controlled by two to three genes exhibiting zero to partial dominance and was moderate to highly heritable. Stability analysis of the hybrids was done over 11 environments using the additive main effects and multiplicative interaction (AMMI) and the genotype and genotype by environment (GGE) biplot analyses. Both AMMI and GGE biplot analyses selected hybrids H21 (CZL00009 x A16). H14 (A1220-4 x A16). S63 (SeedCo hybrid check). N72 (MP72/N3) and H26 (CZL00001 x A16) as stable and high yielding. Hybrids H1 (CML445 x A1220-4), H44 (CZL00009 x CML443) and H18 (CZL00009 x CZL00001) were identified by both methods as unstable but high yielding. AMMI and GGE biplot analyses identified ZAM08, C108, RAO9 and C09 as the most representative environments which were high yielding and relatively stable. In general, the study has revealed that based on the farmers ranking of the constraints in their area, breeding opportunities do exist for incorporating tolerance to both biotic and abiotic stresses in their varieties. It also identified maize lines resistant to the main foliar diseases, with good combining ability and heterosis for resistance and high grain yield. Hybrids with wide adaptation and high yields across environments were also observed. The experimental hybrids that exhibited high levels of resistance can be recommended for further testing and release. On the whole, highly significant additive effects and moderate to high heritability estimates observed for all the diseases and grain yield implied progress would be made through selection, although significant epistasis and dominance could slow progress. Dominance effects towards resistance and high yield could be exploited in developing single cross maize hybrids among these inbreds when only one parent is resistant. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Black wattle (Acacia mearnsii), which belongs to the genus Acacia, is one of the many species of trees or hardwoods grown commercially in South Africa. Black wattle is a species indigenous to Australia and was introduced into South Africa by the van der Plank brothers in 1864. These trees are grown in South Africa because of its tannin-rich bark, the extract of which is used by the leather tanning industry. Black wattle is also grown for its timber, timber products and pulp. The introduction and cultivation history of black wattle suggests that the South African plantations contain limited genetic variation with relatedness amongst groups estimated to be high, thus implying a narrow genetic base in the South African black wattle population. In this investigation, Random Amplified Polymorphic DNA (RAPD) was used to estimate the genetic variation between seven different black wattle groups. A total number of 34 individuals obtained from different areas in South Africa were examined; Piet Retief (group 47 and 50: half-sibs), Kumbula (group 85: unrelated individuals), Howick (group 400: unrelated individuals) and an unknown area (groups 88, 89, 91: full-sibs). As this investigation was the first of its kind, a DNA isolation method as well as a PCR-RAPD protocol had to be modified. Total genomic DNA was successfully extracted using the CTAB DNA extraction method. This method removed large amounts of tannin present in the cells of the black wattle leaves and extracted high quality DNA to conduct between 50-100 RAPD reactions. The DNA purities ranged from 0.1 to 1.8, with an average of 1.46. A total of fourteen 10-mer RAPD primer sequences were randomly selected from the Operon Technologies primer list A, and tested in this investigation. Of the 14 primers used, only nine primers produced clear, single and repeatable bands. Therefore nine primers were selected for subsequent analyses. Ninety one loci that generated bands ranging from 300-3050 base pairs were produced. Seven to 13 loci per primer were generated. A total of 95.6 % of the loci were polymorphic. The overall expected mean heterozygosity (H = 0.3) obtained in this study was high in comparison to other studies conducted on acacias. The high levels of genetic variation were attributed to mating systems, dissortative mating and geographic distribution. The statistical packages POPGENE and ARLEQUIN were used to analyse the RAPD fingerprints. The genetic measures, Nei's diversity and Shannon's Information Index, showed that there was greater diversity exhibited (Nei's gene diversity = 32.09 % and Shannon's = 48.31 %), in the whole population than in each of the groups (with average of Nei's gene diversity = 20.33 % and Shannon's = 34.64 %). With regards to individual group analyses, low levels of genetic variation was obtained in group 400 (unrelated), from the Howick region, and group 85 (unrelated), from the Kumbula region, (mean 0.14 and 0.17 respectively). The low genetic values were attributed to limited gene exchange occurring in these two areas, bottlenecks and selection pressures. Groups 88, 89 and 91, from the unknown region (full-sib groups), were the most variable in comparison to the other groups, with means of (0.27,0.24 and 0.18 respectively). These high genetic variation values could be due to the fact that gene migration could have occurred between these groups and others in the area. It is thought that most acacias are insect-pollinated and this could have lead to gene migration between groups or populations, thereby explaining the high mean values. The gene flow obtained for the seven groups (FST = 0.174) indicated that great genetic differentiation existed in this population of black wattle studied. This value is higher in comparison to other woody species; however it is similar to other acacia species. UPGMA cluster analysis using Nei's unbiased genetic distance, revealed four distinct clusters of groups corresponding to the distribution areas represented in this study. The Howick (group 400: unrelated) and Kumbula (group 85: unrelated) were more closely related to each other than to the other groups, since both these groups are from Natal. The Piet Retief groups (groups 47 and 50: half-sibs), branched-off together, indicating that they are distinct from the other groups. The pairwise analysis of identity showed that the relationship between the group from Howick (group 400: unrelated) and all the other groups from the other regions was the lowest, ranging from 64 % to 79 %. The relationship between all the groups beside the group from Howick (group 400: unrelated) was reasonably high, ranging from 78 % to 90 %. This distance displayed by group 400 (unrelated) from Howick in relation to the groups, is attributed to the fact that it is frost resistant and the other groups not. Genetic variation was also detected and partitioned, between and within groups, by Analysis of Molecular Variance (AMQVA). Majority of the variation existed within groups (82.65 %) but significant differentiation was recorded between groups (17.44 %). This high level of within group differentiation may be explained by many aspects, such as the species breeding system, genetic drift or genetic isolation of groups or populations. The application of RAPD fingerprinting in black wattle has provided a more in depth understanding of the genetic variation residing in the South African population. The results achieved implementing this technique has shown that significant genetic variation exists within the black wattle population in South Africa. The results obtained in this study are also important since it is contrary to the expectation that the black wattle population in South Africa has low genetic variation. This knowledge is of great value to genetically discriminate between individuals or groups, to improve the selection of superior genotypes and allowing improved quality control in breeding programmes and seed orchard management. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

This thesis reports the cell culture establishment and a somatic cell selection system optimized for the isolation of chlorsulfuron-resistant variants in asparagus (Asparagus officinalis L.). The development of this cell selection system benefited the isolation of chlorsulfuron-resistant variants from an elite asparagus genotype. A cell culture system, suitable for somatic cell selection, was established for asparagus genotype CRD 168. Friable callus was initiated from etiolated shoots in darkness and used to produce a high density of single cells in suspension. Cell density was estimated based on a linear relationship with settled cell volume. A mean plating efficiency of 0.19 % was recorded between 1-4x10⁵ cells/Petri dish. In vitro cell selection techniques were developed to identify mutant asparagus cells with resistance to a sulfonylurea herbicide, chlorsulfuron. A few key aspects were important to achieve this: a cell culture system for cell selection was initially established; a toxic concentration for the complete growth inhibition of the wild type asparagus cells was defined; rare, resistant cell colonies were isolated and characterized; and chlorsulfuron-resistant plants were regenerated. From about 50 million cells, 165 cell colonies were isolated in the presence of 8 nM chlorsulfuron. Characterization of these selected cell colonies yielded 24 escapes, 98 unstable variants, and 43 stable-resistant variants. Callus cultures from 34 of these stable variants retained resistance following 11 months growth in the absence of the selection agent. Plants were regenerated from 36 of these stable herbicide-resistant variants. Six of these chlorsulfuron-resistant variants were screened for their degree of resistance to chlorsulfuron, cross resistance to other acetohydroxyacid synthase (AHAS) inhibiting herbicides and AHAS enzyme activity. Cross resistance to imazamox was evident in four of the resistant variants, while lack of cross resistance to metsulfuron methyl was observed in all six resistant variants. A varying degree of resistance to chlorsulfuron was observed among the resistant variants. Both in the original and secondary callus, an uninhibited AHAS enzyme activity in all six resistant variants was recorded in the presence of high chlorsulfuron concentration (70-140 nM), compared to the total inhibition in the wild type. One chlorsulfuron-resistant variant, R-45, was used to compare the biochemical and physiological basis of resistance with the wild type. The AHAS enzyme activity in the tissue culture and greenhouse foliage of R-45 was significantly higher in the presence of up to 280 nM chlorsulfuron compared with the wild type. Chlorsulfuron retention was considerably higher due to the reduction of epicuticular wax deposits on the foliage of R-45, in comparison with the wild type. Consequently, the resistant line absorbed at least 1.6 fold more chlorsulfuron than the wild type plants. Therefore, foliar application of 15 g a.i./ha Glean (commercial formulation of chlorsulfuron) produced typical symptoms of chlorosis in R-45, similar to the wild type, in the greenhouse plants. Somatic cell selection was carried out using two elite asparagus genotypes, CRD 74 and Clone X. Of the 33 rare cell colonies isolated from Clone X, 22 unstable variants and 6 escapes were discarded. All five remaining resistant variants produced plants. One of the stable-resistant variants (Clone X-24) was evaluated for resistance to chlorsulfuron. Both in vitro shoot cultures and greenhouse-grown plants of Clone X-24 showed increased resistance to chlorsulfuron compared with the wild type. The AHAS enzyme activity in the foliar extracts also showed the presence of higher enzyme activity in Clone X-24.