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Development of simple sequence repeat markers in Pyropia yezoensis (Bangiales,Rhodophyta) by high‐throughput sequencing technology
Authors:Linbin Huang  Xinghong Yan
Abstract:Pyropia yezoensis is one of the most important economical seaweeds across the world and major efforts are underway to increase the production. However, its genome is relatively unexplored. In this study, genome sequence of wild‐type strain LS was determined through paired‐end sequencing of small‐size fractionated genomic library. In total, 283,606 contigs were assembled and 20,620 SSR‐containing sequences were identified. Most of the SSRs contained tri‐ and di‐nucleotide motifs (95.42% and 2.34% respectively), of which GCC was the most abundant (15.7%). Furthermore, 1,253 pairs of primer sets were designed and 124 of them were selected randomly for validation. The results showed that 120 pairs were successful in PCR, and the remaining four failed to generate PCR product at various conditions. Among the 120 pairs of which the PCR products were subsequently sequenced by Sanger sequencing, 104 pairs amplified SSRs with the same motif and repeat times, three pairs with the same motif but different repeat times and 13 pairs with different motifs. Moreover, 21 primer pairs amplified polymorphic products between LS and an improved strain HT, and tri‐nucleotide SSRs showed higher polymorphism frequency when compared to di‐nucleotide SSRs. These SSR markers may enrich the current molecular resources in Pyezoensis.
Keywords:high‐throughput sequencing  polymorphic marker     Pyropia yezoensis     simple sequence repeat
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