基于SSR分子标记构建19个杧果品种(系)的DNA指纹图谱

Construction of DNA Fingerprint of 19 Mango Cultivars(lines)Based on SSR Molecular Markers

杧果(Mangifera indica Linn.)是一种热带亚热带常绿乔木果树,果实香甜、营养丰富、肉质细滑多汁,富有“热带果王”之美誉。为了加速杧果品种(系)的分子鉴定,本研究根据19份试验材料的表型特征,选择其差异较大的5份杧果材料(红象牙杧、金煌杧、红杧6号、红玉杧、南逗迈4号杧)为模板,进行SSR标记引物筛选,PAGE电泳检测筛选得到多态性较好的引物;将得到的引物再采用HEX、FAM两个荧光标记进行5’端修饰,再通过毛细管电泳技术检测,其扩增片段大小通过数字和字母对每对引物的多态性位点进行编码,构建杧果的DNA指纹图谱和DNA分子身份证。结果表明:经PCR扩增以及PAGE电泳检测,从已开发得到115对SSR标记引物中筛选得到9对稳定性较好、多态性好的引物,引物名称分别为M9、M15、M35、M40、M53、M59、M101、M102、M103。经毛细管电泳检测,结合“数字+字母”处理获得19个杧果品种(系)的指纹图谱代码,如金煌杧的指纹图谱代码为9EE9F8933,黔山杧2号的指纹图谱代码为H6F7564IC,黔山杧4号的指纹图谱代码为3DGE47279。以19份材料的指纹图谱代码和基本信息为载体,通过在线条形码生成程序、二维码生成程序生成其条形码DNA分子身份证、二维码DNA分子身份证。该研究结果为标准化构建杧果DNA分子身份证库奠定基础,为杧果遗传育种、优异基因挖掘提供数据参考。

Mangifera indica Linn.is a tropical and subtropical evergreen tree fruit,sweet fruit,rich nutrition,meat smooth and juicy,which is rich“tropical fruit king”reputation.In order to speed up the molecular identification of mango cultivars(lines),five mango cultivars(Red Ivory Mango,Jinhuang Mango,Red Mango No.6,Red Jade Mango,Nan Doc Mai No.4 Mango)with different phenotypic characteristics were selected from 19 test materials to screen SSR marker primers.The primers with better polymorphism were obtained by PAGE electrophoresis.Then,the primers were labeled with 5'-end modification with HEX and FAM fluorescent labels.The size of the amplified fragments were de-tected by capillary electrophoresis,and the polymorphic sites of each pair of primers were coded by numbers and letters.Finally,DNA fingerprint and DNA molecular ID card of mango were constructed by capillary electrophoresis.The re-sults show that nine pairs of primers with good polymorphism were screened from the developed SSR primers from 115 pair primers through PCR amplification and PAGE electrophoresis.The primers were named M9,M15,M35,M40,M53,M59,M101,M102,and M103.After capillary electrophoresis detection and data analysis,bar code DNA molecular identification and quick response code DNA molecular identification of the mango cultivars(lines)were obtained com-bined with"number+letter"processing.For example,the fingerprint code of Jinhuang Mango is 9EE9F8933,the fin-gerprint code of Qianshan Mango No.2 is H6F7564IC,and the fingerprint code of Qianshan Mango No.4 is 3DGE47279.Finally,with the fingerprint code and basic information of 19 materials as the carrier,barcode DNA mo-lecular ID and two-dimensional code DNA molecular ID were generated by online barcode generation program and two-dimensional code generation program.This study laid a foundation for standardizing the construction of molecular identification database of DNA of mango,and could provide data reference for mango genetics and breeding and excel-lent gene excavating.