野生稻DNA片段存在于高世代水稻变异系进一步的分子验证  

Molecular Verification of DNA Flow from Wild Rice (O. minuta) to Cultivated Rice

作　　者：王松文[1] 赵炳然 邢全华[3] 阳和华 金德敏[3] 刘霞[1] 王斌[3] 

机构地区：[1]天津农学院,天津300384 [2]中国杂交水稻工程技术研究中心,长沙410125 [3]中国科学院遗传及发育研究所,北京100101

出　　处：《中国农业科学》2006年第11期2170-2177,共8页Scientia Agricultura Sinica

基　　金：湖南省科技厅资助项目;"现代农业生物技术研究与应用"(01NKY1002-01)

摘　　要：【目的】小粒野生稻(O.minuta)DNA导入水稻保持系V20B,第1代(D1)获得变异株,经过连续16代繁育,选出了完全稳定遗传的新不育系及其保持系“野威A”/“野威B”。本试验旨在从分子水平上证明野生稻DNA转移整合进栽培稻“野威B”基因组中,并能稳定遗传。【方法】通过RAPD分析、RAPD扩增特异条带测序分析及AFLP分析等方法揭示了远缘资源基因组DNA导入栽培稻的高世代变异系的基因组整合了远缘基因组片段。【结果】对供体(小粒野生稻)、变异系(野威B)和受体(V20B)进行RAPD分析发现,变异系含有供体存在而受体不存在的“特异带”,在此基础上,对“特异带”,DNA片段进行了核苷酸序列分析,发现RAPD引物OPG-11在变异系与供体中扩增出的1对“特异带”DNA片段的长度均为975 bp,二者间存在97%的同源性,有29个碱基的差异,碱基突变包括转换、颠换、插入及缺失4种类型;同时,AFLP分析表明:高世代(第16代)变异系“野威B”与受体(V20B)存在大量遗传多态性,并含有供体特异AFLP标记。【结论】证明了野生稻DNA向栽培稻的转移整合。[Objective] A new cytoplarnic male sterility line Yewei A and its maintainer line Yewei B, were developed from a maintainer line variant V20B with DNA transformed from wild rice（Oryza. Minuta J.S. Presl. ex C. B. Presl.）. Experiment was carried out to verify if specific DNA fragment from O, minuta was integrated into genomes of the two species （YeweiB and YeweiA）, and to verify if integrated fragment can inherit stably in high generation of YeweiB/YeweiA, [Method] Analysis ofRAPD, DNA sequencing of specific RAPD bands and AFLP in receptor, variant and donor confirmed the integration and its stable inheritance. [ Result ] Results of RAPD analysis of donor, receptor and variant gave that specific bands showed in variant and donor but not in receptor. DNA sequencing of the bands detected in RAPD suggested that primer OPG-11 could be used to amplify a 975 bp product in both variant and donor. A 97% similarity of the products was observed except a 29bp long difference. Mutated base pairs between the two DNA fragments included transition, transversion, insertion and deletion. At the same time, more polymorphisms was displayed in AFLP analysis between variants Yewei B and its receptor V20B with specific DNA fragments from O. minuta integrated [Conclusion] The above results confirmed that genomic DNA transformation from distant rice relatives was an effective approach to create new germplasm.

关 键 词：野生稻 种质资源 AFLP DNA序列 RAPD 

分 类 号：S511.9[农业科学—作物学]