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茶树(Camellia sinensis)SSR 标记开发和连锁图谱构建的花转录组测序。

Floral transcriptome sequencing for SSR marker development and linkage map construction in the tea plant (Camellia sinensis).

机构信息

National Center for Tea Improvement, Tea Research Institute of the Chinese Academy of Agricultural Sciences (TRICAAS), Hangzhou, P. R. China ; College of Horticulture, Sichuan Agricultural University, Yaan, P. R. China.

出版信息

PLoS One. 2013 Nov 26;8(11):e81611. doi: 10.1371/journal.pone.0081611. eCollection 2013.

Abstract

Despite the worldwide consumption and high economic importance of tea, the plant (Camellia sinensis) is not well studied in molecular biology. Under the few circumstances in which the plant is studied, C. sinensis flowers, which are important for reproduction and cross-breeding, receive less emphasis than investigation of its leaves or roots. Using high-throughput Illumina RNA sequencing, we analyzed a C. sinensis floral transcriptome, and 26.9 million clean reads were assembled into 75,531 unigenes averaging 402 bp. Among them, 50,792 (67.2%) unigenes were annotated with a BLAST search against the NCBI Non-Redundant (NR) database and 10,290 (16.67%) were detected that contained one or more simple sequence repeats (SSRs). From these SSR-containing sequences, 2,439 candidate SSR markers were developed and 720 were experimentally tested, validating 431 (59.9%) novel polymorphic SSR markers for C. sinensis. Then, a consensus SSR-based linkage map was constructed that covered 1,156.9 cM with 237 SSR markers distributed in 15 linkage groups. Both transcriptome information and the genetic map of C. sinensis presented here offer a valuable foundation for molecular biology investigations such as functional gene isolation, quantitative trait loci mapping, and marker-assisted selection breeding in this important species.

摘要

尽管茶在全球范围内被广泛消费且具有重要的经济价值,但对茶树(Camellia sinensis)的分子生物学研究却相对较少。在对该植物进行研究的少数情况下,茶树的花(对于繁殖和杂交很重要)受到的重视不如对其叶子或根的研究。本研究采用高通量 Illumina RNA 测序技术,分析了茶树的花转录组,共组装出 75531 条 unigenes,平均长度为 402bp,总 clean reads 为 2690 万条。其中,50792 条 unigenes(67.2%)通过与 NCBI 非冗余数据库(NR)的 BLAST 搜索进行注释,10290 条 unigenes(16.67%)包含一个或多个简单序列重复(SSR)。从这些含有 SSR 的序列中开发了 2439 个候选 SSR 标记,其中 720 个经过实验验证,最终确定了 431 个(59.9%)茶树新的多态性 SSR 标记。然后,构建了一个共识 SSR 连锁图谱,覆盖了 1156.9cM,237 个 SSR 标记分布在 15 个连锁群中。本研究提供的茶树转录组信息和遗传图谱为该重要物种的功能基因分离、数量性状位点作图、标记辅助选择育种等分子生物学研究提供了有价值的基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ccce/3841144/98ccfb9f4893/pone.0081611.g001.jpg

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