生物技术通报 ›› 2018, Vol. 34 ›› Issue (9): 195-201.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0155

• 研究报告 • 上一篇    下一篇

利用SSR标记构建枸杞品种分子身份证

尹跃, 赵建华, 安巍, 李彦龙, 何军, 曹有龙   

  1. 宁夏农林科学院国家枸杞工程技术研究中心,银川 750002
  • 收稿日期:2018-02-23 出版日期:2018-09-26 发布日期:2018-10-10
  • 作者简介:尹跃,男,硕士,研究实习员,研究方向:枸杞分子标记辅助育种;E-mail:yueyin0112@aliyun.com
  • 基金资助:
    国家自然科学基金项目(31760218),一二三产业融合发展农业技术推广示范项目(YES-16-0405),宁夏农林科学院科技创新先导资金(NKYZ-16-0405)

Establishment of Molecular Identity for Wolfberry Cultivars Based on SSR Markers

YIN Yue, ZHAO Jian-hua, AN Wei, Li Yan-long, HE Jun, CAO You-long   

  1. National Wolfberry Engineering Research Center,Ningxia Academy of Agriculture and Forestry Sciences,Yinchuan 750002
  • Received:2018-02-23 Published:2018-09-26 Online:2018-10-10

摘要: 以16个枸杞品种为材料,利用SSR标记构建枸杞分子身份证,为枸杞品种鉴定和保护提供参考。从600对SSR引物中筛选出多态性高、稳定性好、均匀分布在枸杞12条染色体上的24对引物,对16个枸杞品种进行扩增,共检测到等位基因155个,每对引物检测到等位基因数在3-10,平均为6.5个;共检测到基因型208个,每对引物检测到基因型在3-14个,平均为8.7个;多态信息含量(PIC)变幅在0.461-0.848,平均为0.682;Shannon’s信息指数变幅在0.939-2.055,平均为1.487。基于最少引物鉴定最多种质的原则,筛选出等位基因数、基因型数和PIC值均大于平均值,且Shannon’s信息指数>1.7引物,最终确定出引物LBSSR0052和LBSSR0423组合可区分全部供试品种。根据这两对引物对所有品种扩增获得等位基因按照由小到大排序,然后将每个品种在这两个SSR位点的赋值依次组合,构建16个枸杞品种的分子身份证。结果表明SSR标记技术可以作为枸杞品种鉴定和分子身份证构建的有效技术手段。

关键词: 枸杞, 品种, SSR, 分子身份证

Abstract: Using 16 cultivars as experimental materials,wolfberry molecular ID was constructed by SSR marker,aiming at providing references for identification and protection for wolfberry cultivars. Twenty-four primer pairs with high polymorphism,repeatability and distribution on the 12 chromosomes of wolfberry were screened from 600 pairs of SSR primers,and amplified 16 wolfberry cultivars. A total of 155 alleles were detected,the number of alleles per pair of primers ranged in 3-10,averagely 6.5 alleles per locus. Two hundred and eight genotypes were obtained ranging from 3 to 14 with average of 8.7 genotypes per locus. Polymorphic information content(PIC)ranged from 0.461 to 0.848 with an average of 0.682 per locus. Shannon’s information index ranged from 0.939 to 2.055 with an average of 1.487. Based on the principle of identifying the most cultivars using the minimum pairs of primers,the number of screened alleles,the number of genotypes,and PIC values were great than the average,and the value of Shannon’s information index was great than 1.7. Finally,the combination of primers LBSSR0052 and LBSSR0423 distinguished all tested cultivars. Based on these two pairs of primers,the alleles of all the cultivars were sequenced in descending order,and then the values of each cultivar in these two SSR loci were sequentially combined,thus the molecular ID of 16 wolfberry cultivars were established. The results indicate that SSR marker can be used as an effective technique for the variety identification and molecular ID construction of wolfberry.

Key words: wolfberry, cultivars, SSR, molecular ID