In common bean (Phaseolus vulgaris), the main seed storage pests are the bruchid beetles. Damage done to the seed by the larvae has a large impact on seed quality and yield. Arcelin (ARC), phytohaemagglutinin (PHA), and α-amylase inhibitor (α-AI) are linked seed storage proteins that form the APA locus on chromosome Pv04 and are associated with resistance. A major breeding objective is to introduce bruchid resistance into common bean from a resistant tepary genotype, G40199, by introgressing the resistant APA locus into susceptible common bean backgrounds. Here we developed a molecular marker that tracks the introgression. A set of PCR primers to the α-amylase inhibitor locus amplified a DNA fragment that showed a 45 base pair insertion in the middle of a lectin Leg_b domain. This enhanced locus characterization and insertion/deletion marker may preclude the need for bruchid resistance screening early in the breeding. / United States. Agency for International Development / United States. Global Hunger and Food Security Initiative (Cooperative Agreement No. EDH-A-00-07-00005-00)

Biological control of pest Bruchidae may provide an important management strategy against infestation of stored grain legumes, a key source of dietary protein in developing countries. Previous related research has focused on the potential of parasitoids to control bruchids; the role of generalist predators in this application has not yet been extensively explored. / The anthocorid true bug Xylocoris flavipes (Reuter) exhibited a Type II density dependent functional response to five species of adult bruchids. The rate of kill of these large prey was quite low but fairly consistent and female predators were generally more effective. Of the species examined, only the eggs and neonate larvae of A. obtectus were accessible and predation on these stages was high. / Population interaction studies evaluating the effects of predator density and of time elapsed between infestation of commodity and predator addition indicated that adding the predator simultaneously with the pests significantly reduced the number of F$ sb1$, bruchid progeny for all species. Predator density contributed less to bruchid suppression than time of predator addition and bruchid progeny suppression was much greater than anticipated given the rate of kill observed in the functional response experiments. Reproduction by A. obtectus was almost entirely inhibited by the predator. / The high levels of suppression achieved with the predator indicated a significant biological control potential; however, the more fecund bruchid species with inaccessible immature stages continued to produce a large number of progeny. The predator was then combined with larval parasitoids capable of utilizing the internally-developing stages of the bruchids; bruchid suppression was considerably enhanced over the predator alone, and for the most fecund pests, suppression was greater than for the parasitoids alone.

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<p>Multiple mating by females is common in nature. Yet, the evolution and maintenance of polyandry remains a bit of an evolutionary puzzle. It was my aim in this thesis to reach a greater understanding of this phenomenon as well as to investigate the consequences of polyandry on the evolution of reproductive divergence in insects. In an extensive meta analysis addressing the direct effects of multiple mating on female fitness in insects, I found that insects gain from multiple matings in terms of increased lifetime offspring production. In species without nuptial feeding, increased mating rate leads to decreased female lifespan and my results strongly support the existence of an intermediate optimal female mating rate. However, results from an experimental study where I examined the relationship between female fitness and mating rate in the bean weevil (Callosobruchus maculatus) showed that female fitness was maximized at two alternative mating rates, indicating that some species may exhibit a more complex relationship between the costs and benefits of mating. In the meta analysis on species with nuptial feeding, I found only positive effects of increased mating rate and the puzzle is rather what constrains the actual mating rates of females in these groups.</p><p>Sexual selection is a very potent driver of rapid evolutionary change in reproductive characters. Most research has focussed on precopulatory sexual selection, but in promiscuous species sexual selection continues after copulation and variance in male fertilization success gives rise to postcopulatory sexual selection. In this thesis I found that three allopatric populations of the red flour beetle (Tribolium castaneum) have diverged in traits related to reproduction. Male genotype affected all aspects of female reproduction, but more interestingly, males and females interacted in their effect on offspring production and reproductive rate, showing that the divergence was due to the evolution of both male and female reproductive traits.</p><p>When studying postcopulatory sexual selection, sperm competition has been put forward as the main source of variance in fertilization success. The results from a set of double-mating experiments, using the same populations of flour beetles, provided strong evidence that cryptic female choice is also important in generating variance in male fertilization success. I found not only main effects of female genotype on male fertilization success but also male-female interactions which provide more unambiguous evidence for cryptic female choice. Finally, I attempted to uncover which male signals-female receptors are involved in the reproductive divergence observed in the Tribolium populations. In a double-mating experiment I manipulated female perception of two male reproductive signals, copulatory courtship and cuticular hydrocarbons, and the results indicate that, within populations, both signals are sexually selected. However, only male cuticular hydrocarbons seem to be involved in the reproductive divergence between the populations. </p><p>In conclusion, multiple mating by female insects can be understood solely in terms of direct fitness benefits resulting from increased offspring production. I have shown that postcopulatory sexual selection can lead to rapid divergence in reproductive traits related to mating and that cryptic female choice plays an important role in this divergence.</p>

Multiple mating by females is common in nature. Yet, the evolution and maintenance of polyandry remains a bit of an evolutionary puzzle. It was my aim in this thesis to reach a greater understanding of this phenomenon as well as to investigate the consequences of polyandry on the evolution of reproductive divergence in insects. In an extensive meta analysis addressing the direct effects of multiple mating on female fitness in insects, I found that insects gain from multiple matings in terms of increased lifetime offspring production. In species without nuptial feeding, increased mating rate leads to decreased female lifespan and my results strongly support the existence of an intermediate optimal female mating rate. However, results from an experimental study where I examined the relationship between female fitness and mating rate in the bean weevil (Callosobruchus maculatus) showed that female fitness was maximized at two alternative mating rates, indicating that some species may exhibit a more complex relationship between the costs and benefits of mating. In the meta analysis on species with nuptial feeding, I found only positive effects of increased mating rate and the puzzle is rather what constrains the actual mating rates of females in these groups. Sexual selection is a very potent driver of rapid evolutionary change in reproductive characters. Most research has focussed on precopulatory sexual selection, but in promiscuous species sexual selection continues after copulation and variance in male fertilization success gives rise to postcopulatory sexual selection. In this thesis I found that three allopatric populations of the red flour beetle (Tribolium castaneum) have diverged in traits related to reproduction. Male genotype affected all aspects of female reproduction, but more interestingly, males and females interacted in their effect on offspring production and reproductive rate, showing that the divergence was due to the evolution of both male and female reproductive traits. When studying postcopulatory sexual selection, sperm competition has been put forward as the main source of variance in fertilization success. The results from a set of double-mating experiments, using the same populations of flour beetles, provided strong evidence that cryptic female choice is also important in generating variance in male fertilization success. I found not only main effects of female genotype on male fertilization success but also male-female interactions which provide more unambiguous evidence for cryptic female choice. Finally, I attempted to uncover which male signals-female receptors are involved in the reproductive divergence observed in the Tribolium populations. In a double-mating experiment I manipulated female perception of two male reproductive signals, copulatory courtship and cuticular hydrocarbons, and the results indicate that, within populations, both signals are sexually selected. However, only male cuticular hydrocarbons seem to be involved in the reproductive divergence between the populations. In conclusion, multiple mating by female insects can be understood solely in terms of direct fitness benefits resulting from increased offspring production. I have shown that postcopulatory sexual selection can lead to rapid divergence in reproductive traits related to mating and that cryptic female choice plays an important role in this divergence.

Dry bean (Phaseolus vulgaris L.) is economically and nutritionally an important legume, not only in Malawi, but in many parts of Africa and Latin America. Unfortunately, two bruchid species (Acanthoscelides obtectus Say, and Zabrotes subfasciatus Boheman) are known to cause extensive damage in storage, reducing the economic importance, food value and planting value of the crop. The aim of this study was to: i) ascertain farmers’ perceptions of the importance of bruchids as storage pests, and to identify their preferred varietal traits in dry beans; ii) screen Malawian dry bean landraces for effective and adaptable sources of resistance to the two bruchid species; iii) determine the gene action and inheritance of bruchid resistance. Farmers’ perceptions on the importance of the two bruchid species to beans both in the field and in storage were established using a participatory rural appraisal (PRA) in three extension planning areas (EPAs) in Lilongwe agricultural development division (ADD). Results confirmed that the two bruchid species are important storage pests, causing serious storage losses among smallholder farmers. In the absence of any control measures, farmers indicated that more than 50% of their stored beans could be lost to bruchids. Indigenous bruchid control measures are not very effective, making it necessary to search for other control methods. It was also clear from the PRA results that breeders need to consider both agronomic and culinary traits in bean cultivar development. This would enhance uptake of newly developed varieties. To address the problem of bruchid damage experienced by smallholder farmers, a total of 135 dry bean genotypes, comprising 77 landraces and 58 improved varieties (obtained from collaborating partners) were tested under laboratory infestation (nochoice test methods) and field infestation (free-choice test methods). The objective of this study was to identify effective sources of resistance to the two bruchid species. Results of the study showed that there was a wide variation among the genotypes for resistance to the two bruchid species. Overall results showed that 88% of the genotypes ranged from susceptible to highly susceptible to Z. subfasciatus and only 12% of the genotypes were moderately resistant to resistant. Genotype screening for resistance to A. obtectus showed that only 12.5% were resistant, whereas 87.5% were moderately to highly susceptible. All of the improved genotypes were 100% susceptible to A. obtectus in storage. One landrace, KK35, consistently showed a high level of resistance to both bruchids under laboratory infestation, with results similar to the resistant checks (SMARC 2 and SMARC 4), while another landrace, KK90, displayed stable resistance under both laboratory and field infestation. However, performance of most genotypes was not consistent with field and laboratory screenings, suggesting that mechanisms of bruchid resistance in the field are different from that in the laboratory and field screening should always be used to validate laboratory screening. Resistance in the field was not influenced by morphological traits. The seed coat played a significant role in conferring resistance to both bruchid species in the laboratory, whereas arcelin did not play any significant role in conferring resistance in the landraces. The inheritance of resistance to A. obtectus was studied in a 6 x 6 complete diallel mating design, involving crosses of selected Malawian dry bean landraces. The F1 crosses, their reciprocals, and six parents were infested with seven F1 generation (1 to 3 d old) insects of A. obtectus in a laboratory, no-choice test. There were significant differences among genotypes for general combining ability (GCA) and specific combining ability (SCA). However, SCA accounted for 81% of the sum of squares for the crosses, indicating predominance of the non-additive gene action contributing to bruchid resistance. A chi-square test for a single gene model showed that 5 of the 13 F2 populations fitted the 1:2:1 segregation ratio of resistant, intermediate and susceptible classes, respectively indicating partial dominance. The eight F2 populations did not conform to the two gene model of 1:4:6:4:1 segregation ratio of resistant, moderately resistant, moderately susceptible, susceptible and highly susceptible classes, respectively. Average degree of dominance was in the partial dominance range in five F3 populations, but in general resistance was controlled by over-dominance gene action in the F2 populations. The additive-dominance model was adequate to explain the variation among genotypes indicating that epistatic effects were not important in controlling the bruchid resistance. The frequency distribution of the 13 F3 populations for resistance to A. obtectus provided evidence for transgressive segregation, suggesting that resistance is conditioned by more than one gene. Reciprocal differences were not significant in the F2 generation seed; but were significant in four crosses in the F3 generation seed for adult bruchid emergence, suggesting that maternal effects or cytoplasmic gene effects also played a role in the inheritance of resistance to the common bean weevil. Through this study, important sources of bruchid resistance in dry bean have been identified in Malawian landraces (KK35, KK90 and KK73). These resistant sources will be used in a breeding programme to develop bruchid resistant bean cultivars, as well as improve resistance in susceptible commercial bean cultivars currently grown by farmers in Malawi. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2007.