Browsing by Author "Bitalo, Daphne Nyachaki"

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    Implementation of molecular markers for triticale cultivar identification and marker-assisted selection
    (Stellenbosch : Stellenbosch University, 2012-12) Bitalo, Daphne Nyachaki; Botes, Willem; Stellenbosch University. Faculty of AgriSciences. Dept. of Genetics.
    Triticale is an amphidiploid that consists of wheat (A and B) and rye (R) genomes. This cereal is fast becoming important on a commercial basis and warrants further assessment for the better management and breeding of the hybrid. The assessment of the genetic diversity among the wheat and rye genomes within triticale can be obtained by using molecular markers developed in both donor genomes. Simple sequence repeats markers (SSRs) and amplified fragment length markers (AFLPs) have been previously used to assess the genetic diversity among triticale lines. SSRs are highly polymorphic markers that are abundant and which have been shown to be highly transferable between species in previous studies while AFLP markers are known to generate plenty of data as they cover so many loci. Thus, the aim of this study was to develop a marker system suitable to assess the genetic diversity and relationships of advanced breeding material (and cultivars) of the Stellenbosch University’s Plant Breeding Laboratory (SU-PBL). Therefore, both AFLP and SSR markers were initially analysed using eight triticale cultivars (with known pedigrees) to facilitate cultivar identification. Fourty-two AFLP primer combinations and 86 SSR markers were used to assess the genetic diversity among the Elite triticale cultivars. The AFLP primer combinations generated under average polymorphism information content (PIC) values. Furthermore, these markers generated neighbour-joining (NJ) and unweighted pair group method with arithmetic average (UPGMA) dendograms that displayed relationships that did not correspond with the available pedigree information. Therefore, this marker system was found not to be suitable. A set of 86 SSRs previously identified in both wheat and rye, was used to test the genetic diversity among the eight cultivars. The markers developed in wheat achieved 84% transferability while those developed in rye achieved 79.3% transferability. A subset of SSR markers was able to distinguish the cultivars, and correctly identify them by generating NJ and UPGMA dendograms that exhibited relationships that corroborated the available pedigree data. This panel of markers was therefore chosen as the most suitable for the assessment of the advanced breeding material. The panel of seven SSR markers was optimised for semi-automated analysis and was used to screen and detect the genetic diversity among 306 triticale entries in the F6, Senior and Elite phases of the SU-PBL triticale breeding programme. An average PIC value of 0.65 was detected and moderate genetic variation was observed. NJ and UPGMA dendograms generated showed no clear groupings. However, the panel of markers managed to accurately identify all cultivars within the breeding program. The marker panel developed in this study is being used to routinely distinguish among the advanced breeding material within the SU-PBL triticale breeding programme and as a tool in molecular-assisted backcross.
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    Population genetics of Galeorhinus galeus, Carcharhinus brachyurus and Rhinobatos annulatus- implications for regional fisheries and elasmobranch conservation
    (Stellenbosch : Stellenbosch University, 2016-03) Bitalo, Daphne Nyachaki; Bester-van der Merwe, Aletta Elizabeth; Roodt-Wilding, Rouvay; Stellenbosch University. Faculty of AgriSciences. Dept. of Genetics.
    ENGLISH ABSTRACT: Elasmobranchs (sharks, skates and rays) are highly exploited world-wide and more vulnerable than most teleosts due to their life history traits (e.g. late age at maturity, low fecundity and slow growth). Most elasmobranchs are either targeted by commercial fisheries or unintentionally taken as bycatch in mixed-species fisheries. Among these, the tope shark Galeorhinus galeus, the copper shark Carcharhinus brachyurus and the southern African endemic lesser sandshark Rhinobatos annulatus, are targeted globally and locally in demersal, pelagic and recreational fisheries. Across the Southern Hemisphere, the International Union for the Conservation of Nature (IUCN) categorizes both the tope and copper sharks as “vulnerable” while the lesser sandshark as “data deficient” within its region of endemism. Information is urgently needed on their regional genetic structure and diversity to help delineate management units (MUs) for better fisheries monitoring and conserving local biodiversity. Regional and local population genetic structure of these species was assessed using previously optimised cross-species microsatellite panels and/or the mitochondrial NADH2 and NADH4 genes. Patterns of evolutionary and demographic history were inferred using coalescent and Bayesian statistical methods. For G. galeus, the data showed a lack of contemporary gene flow and deep historical divergence across the Southern Hemisphere. Two geographically distinct mitochondrial clades were recovered, one including the Atlantic and Indo-Pacific collections (ARG, SA and AUS) and one comprising the Pacific samples (NZ and CHI) as well as single divergent haplotype restricted to South Africa. Nuclear data also revealed large population subdivisions (FST = 0.050 to 0.333, P < 0.05) indicating very limited gene flow for tope sharks across ocean basins. On a local scale, F-statistics, multivariate and clustering analyses supported gene flow with substantial admixture along the South African coastline (FST = 0.016 to 0.048, P > 0.05), with some degree of genetic structure between the Atlantic and Indian Ocean samples. The east coast samples of Port Elizabeth were significantly differentiated from the rest (FST = 0.023 to 0.091, P > 0.05). For C. brachyurus, estimates of pairwise population differentiation were significant (average FST = 0.031, P = 0.000) indicating some degree of gene flow between sampling sites while the sub-structuring observed at Strandfontein indicated the existence of a possible distinct, more admixed group of individuals. Neither AMOVA (FCT = -0.011, P = 1.000) nor Bayesian clustering analyses indicated genetic discontinuity or significant population structure across the Atlantic/Indian boundary. Although the ND4 results also alluded to historical dispersal across this boundary, the population of Mossel Bay harboured four highly divergent haplotypes, indicating that this region might be a potential nursery site for C. brachyurus. The genetic diversity and genetic connectivity of R. annulatus was inferred using cross-amplified polymorphic microsatellite loci across the Agulhas bioregion that coincides with the warm temperate biogeographical province of South Africa. Significant genetic differentiation was observed over a small sampling range (FST = 0.016 to 0.094, P < 0.050) implying that the species might be highly structured throughout its entire geographical range. Overall effective population size for R. annulatus was very low (Ne = 106) and not in accordance to the abundance proposed for the species. As this is the first regional assessment for all three of these species, the findings of this study could have immediate implications for the regional management and conservation of commercial and recreational sharks.