Development of a set of genomic microsatellite markers in tea (Camellia L.) (Camelliaceae)

Abstract

Tea (Camellia L.) is the world’s most consumed health drink and is also important economically. Due to its self-incompatible and outcrossing nature, tea is composed of highly heterogeneous germplasm. It is a perennial, slow-growing crop and hence the successful release of new improved cultivars following conventional breeding methods takes years. In this context, a DNA marker-based molecular breeding approach holds great promise in accelerating genetic improvement programs in tea. Here we describe the isolation of a set of highly polymorphic genomic microsatellite markers using the enrichment approach, which may be useful for phylogenetic and marker-assisted breeding programs in tea. The enriched library comprising 3,205 clones was screened for the presence of microsatellites using a three-primer-based colony PCR method. Four hundred positive clones were selected and sequenced, to reveal 153 sequences containing simple sequence repeats. Seventy-eight primer pairs were designed from repeat-positive sequences, out of which 40 primer pairs produced successful amplifications. Twenty-two of these primer pairs, when tested on a panel of 21 diverse tea clones and accessions, were found to be highly polymorphic, resulting in 137 alleles with an average of 6.76 alleles per primer pair. The polymorphic information content (PIC), expected heterozygosity (H _e) and observed heterozygosity (H _o) of the polymorphic markers ranged from 0.1 to 0.9, 0.1–0.9 and 0.0–0.8, with average values of 0.6 ± 0.18, 0.7 ± 0.17 and 0.5 ± 0.22, respectively. These markers can be applied for various diversity analyses, mapping programs and genotyping of tea crop.