Jong-Gyu Woo

Molecular marker information from de novo assembled transcriptomes of chilli pepper ( Capsicum annuum L.) varieties based on next-generation sequencing technology

Next-generation sequencing technique has been known as a useful tool for de novo transcriptome assembly, functional annotation of genes and identification of molecular markers. This study was carried out to mine molecular markers from de novo assembled transcriptomes of four chilli pepper varieties, the highly pungent ‘Saengryeg 211’ and non-pungent ‘Saengryeg 213’ and variably pigmented ‘Mandarin’ and ‘Blackcluster’. Pyrosequencing of the complementary DNA library resulted in 361,671, 274,269, 279,221, and 316,357 raw reads, which were assembled in 23,607, 19,894, 18,340 and 20,357 contigs, for the four varieties, respectively. Detailed sequence variant analysis identified numerous potential single-nucleotide polymorphisms (SNPs) and simple sequence repeats (SSRs) for all the varieties for which the primers were designed. The transcriptome information and SNP/SSR markers generated in this study provide valuable resources for high-density molecular genetic mapping in chilli pepper and Quantitative trait loci analysis related to fruit qualities. These markers for pepper will be highly valuable for marker-assisted breeding and other genetic studies.

Development of gene-based markers for the Bs2 bacterial spot resistance gene for marker-assisted selection in pepper (Capsicum spp.)

The Bs2 gene of pepper (Capsicum spp.) confers resistance to the most commonly detected races of Xanthomonas campestris pv vesicatora (Xcv). Using information from a published database, we developed gene-based markers for selection of Bs2 in pepper. High polymorphism was detected in exon 3, intron 2 and 3’-UTR of the Bs2 gene. The three regions were sequenced and numerous SNPs and In/Dels were identified between resistant and susceptible inbred lines. Seven SNPs were selected to develop SNP markers using the tetra-primer amplification refractory mutation system-PCR methodology (Tetra-primer ARMS-PCR). One SNP (G/C) was used to develop a molecular marker that can perfectly identify the presence of the bacterial spot resistance. The gene-based markers were further validated to test association with resistance in 80 lines and cultivars. The markers successfully identified a bell pepper commercial cultivar 62-2 that harbors the Bs2 gene. The successful development of the markers in this study provides a useful tool for MAS to screen the Bs2 gene in pepper.