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适于大白菜杂交种纯度鉴定的核心引物筛选
Determination a Set of Core Primers for Chinese Cabbage Hybrid Purity Identification
【摘要】 为筛选出一组核心引物用于大白菜杂交种的纯度鉴定,本研究对84份已知大白菜杂交种进行指纹图谱分析,最终确定了8对引物为大白菜品种纯度鉴定的核心引物,分别为Cnu-m477、Cnu-m098、Br ID90113、Nia-m022、Nia-m088、Sau-um044、Cnu-m461、Nia-m051。依照多态性和杂合率两个指标,确定Sau-um044和Nia-m088为本次研究的首选核心引物,并利用Sau-um044和Nia-m088进行鉴定,其中表现为杂合带型的共有81个品种(96.4%)。明确3对备选核心引物分别为:Cnu-m477、Cnu-m098、Br ID90113。根据特异性引物在纯度鉴定时的筛选标准,将特异引物可划分为两种类型:第一种类型是利用某一引物鉴定时,某品种的基因型表现唯一,则该引物可作为该品种的特异引物,在本研究中,表现为第一类特异引物共有13个品种,其中Sau-um044、Cnu-m477、Cnu-m461作为特异引物出现的次数最多,均为3个,而特异引物中没有Br ID90113、Nia-m022、Nia-m088;第二种类型是在特定品种中预测基因型频率低于1.19E-02(即1/84)的引物,作为特异引物。在本研究中,特异引物分别为:Sau-um044、Cnu-m477、Cnu-m461、Nia-m051。
【Abstract】 In order to determine a set of core primers fitting for Chinese cabbage hybrid purity identification. 84 Chinese cabbage varieties were analyzed by 8 primers in our laboratory, Cnu-m477, Cnu-m098, Br ID90113,Nia-m022, Nia-m088, Sau-um044, Cnu-m461 and Nia-m051, respectively. Two primers, Sau-um044 and Nia-m088, were determined according to their polymorphism and heterozgosity rate as preferred core primers for Chinese cabbage hybrid purity identification.. 81 out of 84 hybrids(accounting for 96.4%) could found heterozygostic band type when the two preferred primers were used for PCR amplification. 3 out of 8 primers, Cnu-m477,Cnu-m098 and Br ID90113, were determined as candidate core primers for Chinese cabbage hybrid purity identification. Specific primers for purity identification were also studied. There were two types 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 for the specific variety. 13/84 varieties had specific primers. Sau-um044、Cnu-m477 and Cnu-m461 were used as specific primers for three varieties, while Br ID90113, Nia-m022 and Nai-m088 were not used as specific primers of any variety.(2) the primer could be used as specific primer for the varieties their’s frequency lower than 1.19E-02(1/84) which were treated as specific genotype. Four primers, Sau-um044, Cnu-m477,Cnu-m461 and Nia-m051, were identified as such specific primers
【Key words】 Chinese cabbage; Hybrid; Purity identification; Core primers;
- 【文献出处】 分子植物育种 ,Molecular Plant Breeding , 编辑部邮箱 ,2016年01期
- 【分类号】S634.1
- 【下载频次】135