中国50个甘蓝代表品种EST-SSR指纹图谱的构建

doi:10.3864/j.issn.0578-1752.2014.01.012

Abstract

Abstract: 【Objective】In this study, the method of cabbage DNA fingerprint was drawn, and fifty cabbage representative varieties from China were fingerprinted with EST-SSR primers to provide reference for variety distinctness, authenticity, and purity identification. 【Method】First, EST-SSR primers were screened by using the technologies of 6% denaturing polyacrylamide gel electrophoresis and six cabbage varieties that come from different ecogeography. The length of amplified fragments was detected on DNA Analyzer platform using four fluorescent-labels (TAMRA, HEX, ROX and 6-FAM) in 5′end of forward primer, and then defined the reference variety for every alleles. Total twenty core primers were used to establish fifty cabbage varieties SSR fingerprinting, and for ‘Zhonggan 21’ variety identification. 【Result】 Six cabbage varieties of different resources were used to screen 978 EST-SSR primers, out of 128 polymorphic primers were obtained according to the PCR bands stability, high polymorphism information content (PIC), easy discrimination of different alleles and even distribution of molecular markers on each chromosome, and 20 pairs of primers were selected to detect a total of 58 alleles at 20 loci, with 2.22 loci per chromosome and 2.9 alleles per locus on average. The PIC values varied among the primers ranging from 0.34 to 0.76. The length of amplified fragment varied in the range of 143-296 bp. The maximum number of alleles for each primer pairs of BoE607 and BoE723 was five. Fingerprinting database of 50 cabbage representative varieties from China was established with 20 pairs of core primers. The authenticity of ‘Zhonggan 21’ was identified by artificial simulated population and these results were identical with that made from field investigation.【Conclusion】Twenty pairs of core primers were selected and used to establish DNA fingerprint database of 50 cabbage representative varieties from China, and the authenticity of ‘Zhonggan 21’ was identified by artificial simulated population and the accuracy was 100%.

Key words: cabbage , SSR marker , fingerprinting , varieties identification

WANG Qing-Biao, ZHANG Yang-Yong, ZHUANG Mu, YANG Li-Mei, LIU Yu-Mei, 吕Hong-Hao , FANG Zhi-Yuan. EST-SSR Fingerprinting of Fifty Cabbage Representative Varieties from China[J].Scientia Agricultura Sinica, 2014, 47(1): 111-121.

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