基于茶树EST-SNP的CAPS标记开发  

Conversion of EST-SNP into CAPS Markers in Tea Plant(Camellia sinensis)

作　　者：张成才[1] 王丽鸳[1] 韦康[1] 成浩[1] 

机构地区：[1]中国农业科学院茶叶研究所,国家茶树改良中心,杭州310008

出　　处：《分子植物育种》2013年第6期817-824,共8页Molecular Plant Breeding

基　　金：现代农业产业技术体系建设专项资金(nycytx-23);浙江省茶产业技术创新战略联盟专项资金共同资助

摘　　要：为了促进SNP技术在茶树遗传育种中的应用,本文探讨了将茶树SNPs转化成CAPS标记进行检测的可行性。首先,对从茶树ESTs中挖掘的候选SNPs进行重测序验证;然后,对确证的SNPs进行限制性内切酶识别位点分析,筛选出引起酶切识别位点改变的SNPs;使用相应的引物从茶树基因组中扩增这些SNPs所在的片段,然后对扩增产物进行酶切和电泳检测,将SNPs转化为CAPS标记;最后,随机选择了14个CAPS标记,在25个茶树品种中进行多态性分析,以验证标记的可用性。结果,经重测序验证,在8个茶树品种中共检测到162个SNPs;将39个SNPs成功转化为CAPS标记;在14个随机选择的CAPS标记中有13个在25个品种中表现出多态性,多态性信息含量(PIC)介于0.043和0.497之间,平均0.311;另外1个标记在25个品种中均为杂合。结果表明,本文构建的将茶树SNPs转化为CAPS标记的方法,可以方便地用于茶树SNPs的检测;该方法能够促进SNP技术在茶树遗传育种研究中的应用;同时,本文报道的39个CAPS标记可以用于茶树遗传多态性检测和茶树遗传图谱构建等方面的研究。CAPS （cleaved amplified polymorphic sequence） has been used as a useful tool for SNP （single nucl- eotide polymorphism） detection. This method was attempted to detect SNPs in tea plant. Fristly, candidate SNPs were mined from tea derived-ESTs and evaluated them in genomic DNA using re-sequencing method. Secondly, the SNPs were analyzed by CodonCode Aligner sot^ware to identified which alter the restrict enzyme recognition sites. Thirdly, restrict enzymes were used to digest the PCR products and checked on agarose gel. Moreover, 14 CAPS markers were randomly selected to validate the usefulness of new markers in genetic study in tea plant. The results showed that 162 SNPs were confirmed in 8 tea cultivars. And 39 SNPs were successfully converted into CAPS markers. Thirteen markers among 14 were shown polymorphism. The PIC value was ranged from 0.043 to 0.497, average 0.311. One marker was homozygous in all accessions. The CAPS method can be used to detect SNPs in tea plant. The new markers reported here, would be useful in tea genetic study. It is also believed that CAPS method will promote the use of SNPs in genetics and breeding research in tea plant.

关 键 词：茶树 EST SNP CAPS 遗传多态性 

分 类 号：S571.1[农业科学—茶叶生产加工]