柑橘栽培品种（系）DNA指纹图谱库的构建

 【目的】构建柑橘栽培品种（系）的DNA指纹图谱数据库,为建立柑橘种苗纯度及真实性鉴定技术体系和技术规范奠定基础。【方法】利用SSR标记和ISSR标记对102个柑橘栽培品种（系）进行DNA指纹分析,筛选适合的特征引物,构建柑橘栽培品种（系）的指纹图谱数据库。【结果】从200对SSR引物中筛选到重复性好、多态性丰富的12对引物作为柑橘品种（系）鉴定的特征引物,12对SSR特征引物组合可鉴别42个品种（系）;在此基础上,对未出现SSR特征指纹的60个品种（系）进行ISSR指纹分析,从40个ISSR引物中筛选到2个可用于品种鉴定的特征引物,结合SSR标记,可快速、准确地鉴别70个柑橘的品种（系）。并利用这12对SSR特征引物和2个ISSR特征引物构建了70个柑橘栽培品种（系）的DNA特征指纹图谱数据库。【结论】SSR和ISSR标记适于构建柑橘栽培品种DNA指纹图谱库;指纹图谱库的建立为柑橘种苗纯度及真实性鉴定奠定了基础。

Construction of DNA Fingerprinting Database of Citrus Cultivars(Lines)

【Objective】 The aims of this study were to construct a DNA fingerprinting database of major citrus cultivars, which could provide a possibility of establishment of a technical standard system for purity and authenticity identification of citrus nursery trees. 【Method】 Simple sequence repeat (SSR) and inter-simple sequence repeat (ISSR) markers were used to analyze the DNA fingerprinting of 102 citrus cultivars(lines) and then to choose suitable primers for the construction of DNA fingerprinting database of citrus cultivars (lines). 【Result】 12 SSR primer pairs, with high polymorphisms and good repeatability, regarded as specific primers, which could identify 42 cultivars (lines), were selected from 200 SSR primer pairs. Meanwhile, 40 ISSR primers were used to analyze the DNA fingerprinting of 60 cultivars (lines) that can not be distinguished by SSR markers and 2 ISSR specific primers were chosen. As a result, a total of 70 citrus cultivars (lines) could be identified by the 12 SSR primers and 2 ISSR primers. Finally, a DNA fingerprinting database containing 70 major citrus cultivars (lines) was constructed by using those 12 SSR and 2 ISSR primers. 【Conclusion】 The results indicated that SSR and ISSR markers are suitable for the construction of DNA fingerprinting database of citrus cultivars (lines). Moreover, the fingerprinting database will provide a basis for genetic purity and authenticity fast evaluation of citrus nursery trees.