Construction Of Fingerprinting And Analysis Of Genetic Diversity With SSR Markers For Nanfeng Tangerine

In this research,22kinds of Nanfeng tangerine germplasm resources, other6kinds of Nanfeng tangerine local cultivars and4related species were used as test materials. Their genomic DNA were extracted. We constructed the Nanfeng tangerine SSR fingerprinting, and analysised the genetic diversity by SSR-PCR amplification and SSR Primers screening, the main research results are as follows:(1) The improved CTAB method and trace extraction method were used to extract selected samples genomic DNA, the high quality DNA was gained for SSR-PCR amplification screening. And the trace extraction method has the character of needing less sample, short time and saving energy.(2)13SSR primers with highly polymorphism were screened from91primer pairs, a total of101alleles, alleles amplified by each pair of primers ranged from2to16, each pair of primers average can detect7.77alleles.The polymorphic information content values(PIC) range of0.375-0.932. with a mean of0.766, the fragment sizes varied from100bp to400bp.(3)13SSR primers with features were screened to constructed Nanfeng tangerine SSR fingerprint, use the13SSR primers combinations can distinguish32Nanfeng tangerine germplasm resources without SS-2,SS-7,SS-12and QB-8.(4) Taking the genetic similarity coefficient0.59as a border, can divide the32selected samples into eight kinds, gather small fruit series ofNanfeng tangerine yangxiao26, SS-1, SS-2, SS-7, SS-9, SS-12, SS-14, SS-28, LX-10, LX-17, LX-21, ST-3, QB-5, QB-8, QW-3,97-1,97-2, guihuadi, precocious series LS-1, precocious series97-1, big fruit series LS-1, big fruit series97-1, miguang for a class, notes for A, indicate miguang and Nanfeng tangerine have close relationships; gather xiaoyeguang, huoguang, huoju for a class, notes for B;gather hongguang, satsuma, ponk, tangerine, bendizao mandarin and kumquat for a class of each, notes for C, D, E, F, G, H,genetic relationship is farther. In the genetic similarity coefficient of0.70, A kind can be divided into two subclasses:a subclass for small fruit series yangxiao26, SS-1, SS-2, SS-7, SS-9, SS-12, SS-14, SS-28, LX-10, LX-17, the LX-21, ST-3, QB-5, QB-8, QW-3,97-1, guihuadi,precocious series LS-1;b subclass for small fruit series97-2, precocious series97-1, big fruit series LS-1, big fruit series97-1and miguang. Nanfeng tangerine strain has not been separately clustering, but small fruit series mainly concentrated in a subclass. The genetic similarity coefficient above0.90are small fruit series SS-1and QW-3; small fruit series SS-9and SS-28; small fruit series SS-2, SS-7, SS-12, SS-14, LX-21and QB-8; precocious series97-1and big fruit series97-1. The genetic similarity coefficient among small fruit series SS-2, SS-7, SS-12and QB-8reached1.0, may be synonyms.