Makurazaki Tea Research Station, NARO Institute of Vegetable and Tea Science , 87, Seto, Makurazaki, Kagoshima 898-0087, Japan ; Graduate School of Life and Environmental Sciences, University of Tsukuba , 1-1-1 Tennohdai, Tsukuba, Ibaraki 305-0001, Japan.
Breed Sci. 2012 Jun;62(2):186-95. doi: 10.1270/jsbbs.62.186. Epub 2012 Jun 19.
Tea is one of the most popular beverages in the world and the tea plant, Camellia sinensis (L.) O. Kuntze, is an important crop in many countries. To increase the amount of genomic information available for C. sinensis, we constructed seven cDNA libraries from various organs and used these to generate expressed sequence tags (ESTs). A total of 17,458 ESTs were generated and assembled into 5,262 unigenes. About 50% of the unigenes were assigned annotations by Gene Ontology. Some were homologous to genes involved in important biological processes, such as nitrogen assimilation, aluminum response, and biosynthesis of caffeine and catechins. Digital northern analysis showed that 67 unigenes were expressed differentially among the seven organs. Simple sequence repeat (SSR) motif searches among the unigenes identified 1,835 unigenes (34.9%) harboring SSR motifs of more than six repeat units. A subset of 100 EST-SSR primer sets was tested for amplification and polymorphism in 16 tea accessions. Seventy-one primer sets successfully amplified EST-SSRs and 70 EST-SSR loci were polymorphic. Furthermore, these 70 EST-SSR markers were transferable to 14 other Camellia species. The ESTs and EST-SSR markers will enhance the study of important traits and the molecular genetics of tea plants and other Camellia species.
茶是世界上最受欢迎的饮料之一,茶树(Camellia sinensis(L.)O. Kuntze)是许多国家的重要作物。为了增加茶树的基因组信息量,我们从不同器官构建了七个 cDNA 文库,并利用这些文库生成表达序列标签(ESTs)。总共生成了 17458 个 EST,并组装成 5262 个非冗余基因。大约 50%的非冗余基因被 Gene Ontology 注释。其中一些与涉及重要生物学过程的基因同源,如氮同化、铝响应和咖啡因和儿茶素的生物合成。数字Northern 分析显示,7 个器官中的 67 个基因表达存在差异。在非冗余基因中搜索简单序列重复(SSR)基序,鉴定出 1835 个(34.9%)含有超过六个重复单元的 SSR 基序。100 个 EST-SSR 引物对的一个子集在 16 个茶树品种中进行了扩增和多态性测试。71 个引物对成功扩增了 EST-SSRs,70 个 EST-SSR 位点表现出多态性。此外,这些 70 个 EST-SSR 标记可以转移到其他 14 种山茶花中。这些 ESTs 和 EST-SSR 标记将增强对茶树和其他山茶花重要性状和分子遗传学的研究。