基于荧光标记SSR的CID法快速鉴定福建茶树品种

茶树品种真实性鉴定是茶树种质资源研究的重要组成部分,是茶树品种知识产权保护的前提。在茶树基因组上按照均匀分布的原则选择45个SSR标记合成荧光标记引物,利用这些标记引物对54个福建无性系茶树品种进行PCR扩增,标记基因型分型后进一步筛选出9个SSR标记核心引物。这批核心引物的多态性信息含量(Polymorphism information content,PIC)均大于0.5,等位位点数在3~5之间,基因型数在5~10之间,属于高度多态性引物且方便进行基因型数据统计,适宜用来进行无性系茶树品种的真实性鉴定。利用其中的6个标记核心引物建立了54个福建无性系茶树品种的品种鉴别图(Cultivar identification diagram,CID),该CID能直观地提供对其中任意品种进行鉴定所需要的标记引物以及相应的标记基因型,且能够方便地进行扩容以用来对更多的无性系茶树品种进行真实性鉴定。利用该方法对无性系茶树品种进行真实性鉴定,操作方便、快速准确、稳定性高、实用性强。本实验所获得的茶树品种鉴定图(CID)对茶树品种知识产权保护,以及促进茶树品种选育工作健康可持续发展具有重要的意义。

An Efficient Identification of Tea Cultivars in Fujian with A Strategy of Cultivar Identification Diagram(CID) Based on Fluorescent Labeled SSR Markers

The authenticity identification of tea cultivars is an important part of the research on tea germplasm resources, and the precondition of intellectual property protection of tea cultivars, 45 SSR markers were selected to synthesize fluorescent labeled SSR primer pairs based on the principle of uniform distribution in the genome of the tea plant. Using these markers, 54 clonal tea cultivars in FUJIAN province were amplified by PCR and 9 SSR markers were selected as the core primer pairs after genotyping. The polymorphism information content (PIC) of the core primer pairs was more than 0.5, and the number of all alleles was varied from 3 to 5, the number of genotypes was varied from 5 to 10. These core primer pairs were suitable for identifying the authenticity of tea cultivars for 54 Fujian clonal tea cultivars due to high polymorphism and convenient to genotype. The cultivar identification diagram (CID) was then established using 6 of these core primer pairs. The CID provided primer pairs and its corresponding genotype, by which any cultivar in the CID could be identified. The capacity of the CID can be expanded so as to facilitate the authenticity identification of more clonal tea cultivars. The method was considered to be easy, fast, accurate, stable and practical for the authenticity identification of tea cultivars. The CID is important for the protection of intellectual property of tea cultivars, and will be helpful for the healthy and sustainable development of tea breeding.