Genetic diversity in cocoa germplasm of southern Cameroon revealed by simple sequences repeat (SSRS) markers

The range of polymorphism of about 194 cocoa accessions collected in farms in Southern Cameroon during field surveys and 71 Trinitario and Upper Amazon clones available in genebanks on-station wasassessed using 13 SSR markers. The gene diversity, genetic differentiation and genetic similarities were analysed for the different populations. In total, 282 alleles were detected within all the populations studied. The farm accessions were strongly differentiated based on their geographical origin, with accessions coming from the East province clustering together with local Trinitario accessions from the genebank. Accessions from the Centre-South provinces clustered with Amazon and hybrid accessions, suggesting more uptake of seed garden materials in farms in these provinces. The genetic diversity parameters indicate that the farmers’ planting material is not highly diverse, and is genetically close to parental genotypes available in genebanks. However, some promising Upper Amazon clones (T-clones) that have also been used as parents of released hybrid varieties were genetically distant from the accessions. This result suggests that the progenies of these parents have so far been poorly used in the cocoa farms surveyed. The consequences of these findings for cocoa breeding in Cameroon are discusse

Parentage analysis and outcrossing patterns in cacao (Theobroma cacao L.) farms in Cameroon.

The present study investigates the parentage of farm accessions in Cameroon using data from 12 microsatellite loci. Bayesian analysis suggests that 25.5% of the 400 farm accessions studied is still closely related to the traditional Amelonado variety called 'German Cocoa' by the farmers. Another 46.3% of the farm accessions were found to be direct descendants (20.8% first-generation (F1) hybrids and 25.5% selfed genotypes) from 24 parental clones used in biclonal seed gardens (BSGs) established in the 1970s in southern and western Cameroon. Furthermore, 28.3% of farm accessions appeared to descent from uncontrolled pollination events in cacao farms, which could be related to a common practice of cacao growers to use seeds collected in their own farm for new plantings. All farm accessions descending from BSG could be individually related through parentage analysis to the 24 progenitors of the BSG. Only 25% of progenies distributed from BSG corresponded to F1 hybrids combinations originally planned to be released. Significant biparental inbreeding estimates were observed for all 'traditional' farms and for most 'F1 hybrids' farms due to presence of a high proportion of selfed accessions. Biparental inbreeding occurs when plants receive pollen from genetically related neighbors. High levels of outcrossing observed in 'mixed' farms might be explained by the admixture of traditional varieties and BSG progenies. The implications of our finding for management of seed gardens and for further breeding using farm accessions in Cameroon are discussed

Confocal observations of late-acting self-incompatibility in Theobroma cacao L.

Cocoa (Theobroma cacao) has an idiosyncratic form of late-acting self-incompatibility that operates through the non-fusion of incompatible gametes. Here, we used high-resolution confocal microscopy to define fine level changes to the embryo sac of the strongly selfincompatible cocoa genotype SCA 24 in the absence of pollination, and following compatible and incompatible pollination. All sperm nuclei had fused with the female nuclei by 48 h following compatible pollinations. However, following incompatible pollinations, we observed divergence in the behaviour of sperm nuclei following release into the embryo sac. Incomplete sperm nucleus migration occurred in approximately half of the embryo sacs, where the sperm nuclei had so far failed to reach the female gamete nuclei. Sperm nuclei reached but did not fuse with the female gamete nuclei in the residual cases. We argue that the cellular mechanisms governing sperm nucleus migration to the egg nucleus and those controlling subsequent nuclear fusion are likely to differ and should be considered independently. Accordingly, we recommend that future efforts to characterise the genetic basis of LSI in cocoa should take care to differentiate between these two events, both of which contribute to failed karyogamy. Implications of these results for continuing efforts to gain better understanding of the genetic control of LSI in cocoa are discussed.Caroline S. Ford and Mike J. Wilkinso