适于玉米杂交种纯度鉴定的SSR核心引物的确定  

Determination of SSR Core Primers for Maize Hybrid Purity Identification

作　　者：王凤格[1] 赵久然[1] 王璐[1] 易红梅[1] 郭景伦[1] 戴景瑞[2] 原亚萍[3] 卢柏山[1] 杨国航[1] 

机构地区：[1]北京市农林科学院玉米研究中心,北京100097 [2]中国农业大学农学与生物技术学院,北京100094 [3]吉林大学植物科学学院,长春130062

出　　处：《农业生物技术学报》2007年第6期964-969,共6页Journal of Agricultural Biotechnology

基　　金：北京农业育种基础研究创新平台;北京市自然科学基金项目(No.YZPT02-06);北京市科技新星项目(No.2003B23);吉林省科技计划项目(No.20030555)资助。

摘　　要：为了确定一套适于玉米杂交种纯度鉴定的核心SSR引物,利用10个SSR引物对420份已知玉米(Zea mays)杂交种进行DNA指纹分析。综合考虑多态性和杂合率两个指标,确定bnlg161和bnlg1450为玉米杂交种纯度鉴定的首选核心引物,利用这两个引物进行筛选,412个品种(占98%)能够找到具有杂合带型的鉴定引物,确定5个引物bnlg439、bnlg125、umc2105、umc1705和bnlg1792为玉米杂交种纯度鉴定的备选核心引物。研究了纯度鉴定所用特异引物的选择,将特异引物分为两类:第一类是利用单个引物检测,某品种的基因型仅出现1次,将该引物作为该品种在特定品种范围内的特异引物,在420个品种中,累计29个品种具有第一类特异引物,其中umc1705和bnlg1450作为特异引物的次数最多,均为8个,phi072和phi065没有作为特异引物;第二类是根据引物基因频率表,将预测基因型频率低于2.40E-03(即1/420)的基因型作为特异基因型,除了phi072和bnlg161外,其它8个引物均可能成为具有特定基因型品种的第二类特异引物。利用推荐的7个纯度鉴定用核心引物可进一步筛选每个玉米杂交种的双亲互补型引物,建立了已知玉米杂交种的纯度鉴定DNA指纹图谱。In order to determine a set of SSR core primers fitting for maize hybrids＇purity identification, 420 maize （Zea mays） hybrids were analyzed by 10 SSR primers, bnlg439, bnlg125, umc2105, phi072, umcl705, bnlgltl, bnlg1792, bnlg162, phi065 and bnlg1450, respectively, and two primers bnlg161 and bnlg1450 were determined according to their polymorpism and heterogosite rate as preferred core primers for maize hybrid purity identification. Four hundred and twelve hybrids （accounting for 98%） have heterozygosis band type in at least one primer if the two preferred core primers bnlgltl and bnlg1450 were used; five primers bnlg439,bnlg125,umc2105,umc1705 and bnlg1792 were determined as candidate core primers for niaize hybrid purity identification. Specific primers for purity identification were also studied, there were two sorts of specific primer： （1） within specific range of varieties, the genotype of a primer appeared only in one variety, then the primer was used as specific primer of the variety. Twenty-nine varieties had specific primers within the 420 varieties, both umcl705 and bnlg1450 were used as specific primers of 8 varieties, while phi072 and phi065 were not used as specific primers of any variety. （2） the genotype of a primer had a genotype frequency lower than a specific value, then the primer could be used as specific primer of the varieties with such genotype, if genotype frequency lower than 2.40 E-03 （1/420） were treated as specific genotype, eight primers except phi072 and bnlgltl might become the second sort of specific primers of varieties with such genotype. Primers with complementary band types were further screened with the seven recommended core primes, and DNA fingerprint maps for maize purity identification were constructed.

关 键 词：玉米 杂交种 SSR 纯度鉴定 核心引物 

分 类 号：S188[农业科学—农业基础科学]