基于NYT 1433-2014中48对SSR引物的94份杂交稻亲本DNA分子数字指纹库研究
收稿日期: 2015-12-29
修回日期: 2016-03-12
网络出版日期: 2016-11-10
基金资助
国家自然科学基金资助项目（3147430）； 生态学重点学科资助项目（0713）。
Research on DNA Molecular Digital Fingerprint Database Based on 48 Pairs of SSR Primers for 94 Hybrid Rice Parents in NYT 1433-2014
Received date: 2015-12-29
Revised date: 2016-03-12
Online published: 2016-11-10
建立方法简便、分辨率高的水稻品种遗传多态性和真实性鉴定的分子指纹技术对于指导水稻育种和规范种子市场都具有重要意义。农业部颁布的水稻品种鉴定技术规程行业新标准是基于35个不同遗传特点的代表性水稻品种建立的SSR分子标记技术规程。本研究根据该标准方法，对94份杂交水稻亲本材料的遗传多态性和特异性进行了比较分析，结果表明，供试品种间至少具有3对以上引物扩增的DNA片段差异，即利用该标准能很好地区分供试杂交水稻亲本的遗传差异。对新标准中48对推荐引物的比较与分析表明，46对引物扩增的DNA片段多态性较高，而RM176和RM551两对引物扩增带多态性较低，因此在其染色体的其他位点可进一步研究多态性更高的分子标记。与标准中35个水稻品种的指纹库进行比较，发现了16个新的等位变异，这些位点可作为标准指纹库的信息补充，丰富标准库中的遗传信息。对94个杂交水稻亲本的分子指纹比较分析，发现23个亲本材料具有特异性分子标记，这些特异分子标记可应用于杂交组合的真实性以及杂交种子纯度的分子鉴定。根据供试亲本的数字分子指纹，构建了87个不育系与7个父本杂交的虚拟组合数字分子指纹库以及虚拟组合的真实性和纯度快速鉴定的特异数字分子标记。
林亦霞,王梓辛,刘欢,王征,梁满中,戴小军*,陈良碧* . 基于NYT 1433-2014中48对SSR引物的94份杂交稻亲本DNA分子数字指纹库研究[J]. 中国水稻科学, 2016 , 30(6) : 593 -602 . DOI: 10.16819/j.1001-7216.2016.6028
It is of great significance to establish the simple molecular fingerprinting technique with high resolution for identification of the genetic polymorphism and the authenticity of different rice varieties, so as to guide rice breeding and regulate its seed market. The new standards of technical regulation on identification of the rice varieties with SSR markers, formulated by the Ministry of Agriculture of P.R. China, recommended the use of 35 control standard reference samples with different genetic characteristics for identification of rice varieties. The genetic polymorphism and specificity of 94 hybrid rice parents were compared based on the standard method. The results indicated that the tested varieties differed at least three pairs of mutated loci or the genetic differences between the parents of hybrid rice could been well distinguished. By comparing the polymorphism of the 48 recommended primers of the new standard, 46 primers showed higher polymorphism except RM176 and RM551. Thus, higher polymorphic alternative molecular markers would be identified in other loci of the same chromosome. 16 new allelic variation sites were found and could be used as supplementary of standard fingerprint database and enrich genetic variation sites information. By analyzing molecular fingerprint of 94 hybrid parental materials, 23 have specific molecular markers and can be used in authenticity analysis of hybrid combination and purity identification of hybrid seeds. According to the digital molecular fingerprint of the tested rice parent, we constructed a virtual digital molecular fingerprint database including 87 female sterile lines and 7 male parents, and specific digital molecular marker of virtual hybridized combination authenticity and rapid seed purity identification.
Key words： hybrid rice; SSR primer; digital fingerprint
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