2个杂交籼稻和2个粳稻品种SSR指纹图谱的构建及双重PCR技术的初步研究

Construction of Molecular Fingerprinting Database and Dual PCR Assay for Two Indica Rice Hybrids and Two Japonica Rice Varieties by Using SSR Markers

用460对水稻SSR引物对5个杂交籼稻及其双亲和16个粳稻品种(系)进行了PCR扩增和聚丙烯酰胺凝胶电泳,筛选出12对稳定性好、多态性高、杂带少的SSR引物作为核心引物,构建了国稻6号和两优培九的SSR指纹图谱。另外,还筛选出9对核心引物,构建了粳稻品种盐稻8号和徐稻4号的分子指纹图谱。其中,筛选出国稻6号的特异性标记5个,两优培九的特异性标记3个,盐稻8号的特异性标记1个,徐稻4号的特异性标记2个。这些特异标记可用于种子纯度的快速鉴定。另外,挑选扩增条带稳定、特异性较好的引物组合,进行了双重PCR分析。

A total of 460 SSR primers were used for amplification of five indica hybrid rice combinations and their parents,and 16 japonica rice varieties（or elite lines）.By using the identified co-dominant specific primers,molecular fingerprinting databases for Guodao 6,Liangyoupeijiu,Yandao 8 and Xudao 4 were constructed.A total of 5,3,1 and 2 SSR primers were identified to be specific for Guodao 6,Liangyoupeijiu,Yandao 8 and Xudao 4,respectively.These fingerprints can be used to examine the seed purity of these rice varieties.In addition,to develop dual PCR assay in examination of seed purity and authenticity,SSR primers which could stably amplify clear bands and have good specificity were selected and used in multiplex popymerase chain reaction amplification.