水稻RNAi突变体库的创制和筛选

通过转化水稻hpRNA农杆菌文库至粳稻品种台北309,获得143株转基因植株.随机选取36株T0代转基因植株进行检测,其GUS阳性率为77.78％,且其中35份PCR阳性材料的插入片段均与粳稻日本晴的mRNA有较高匹配.对61份T1代突变体进行筛选,获得33份有突变表型的材料.初步获得1份类病斑突变体,序列分析表明其可能干扰基因的编码产物为RAL6-种子过敏蛋白RA5/RA14/RA17的前体,属于淀粉酶/胰蛋白酶抑制剂家族,该类基因在植物自然防御系统中扮演重要较色....

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Published in广东农业科学 Vol. 42; no. 7; pp. 1 - 4
Main Author 谢洪科 韩小霞 李丁 彭选明 邹朝晖 杨振 王磊 李祺
Format Journal Article
LanguageChinese
Published 湖南省核农学与航天育种研究所,湖南长沙,410125%湖南省蔬菜研究所 湖南长沙410125%湖南工业大学生物技术系,湖南株洲,412007%中国农业科学院生物技术研究所,北京,100081 2015
Subjects
RNAi突变体库
水稻
类病斑突变体
表型筛选
表型筛选
水稻
类病斑突变体
RNAi突变体库
Online AccessGet full text
ISSN1004-874X

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Summary:通过转化水稻hpRNA农杆菌文库至粳稻品种台北309,获得143株转基因植株.随机选取36株T0代转基因植株进行检测,其GUS阳性率为77.78％,且其中35份PCR阳性材料的插入片段均与粳稻日本晴的mRNA有较高匹配.对61份T1代突变体进行筛选,获得33份有突变表型的材料.初步获得1份类病斑突变体,序列分析表明其可能干扰基因的编码产物为RAL6-种子过敏蛋白RA5/RA14/RA17的前体,属于淀粉酶/胰蛋白酶抑制剂家族,该类基因在植物自然防御系统中扮演重要较色.
Bibliography:44-1267/S
rice; RNAi mutant library; phenotype screening; lesion mimic mutant
143 transgenic plants were obtained by transforming the hpRNA Agrobacterium library into the callus of japonica variety Taipei 309. The detection of 36 transgenic plants showed that the GUS positive was 77.78%, and the sequencing of 35 PCR positive plants were well matched with japonica rice mRNA sequences. 33 materials with mutant phenotype were screened and one lesion mimic mutant was identified from mutant library with 61 T1 materials. The sequence of insertion showed that the gene production belonging to Alpha-amylase inhibitor protein family was RAL6- Seed allergenic protein RA5/RA14/RA17 precursor, it plays an important role in plant＇s defense system.
XIE Hong-ke;HAN Xiao-xia;LI Ding;PENG Xuan-ming;ZOU Zhao-hui;YANG-Zhen;WANG Lei;LI Qi （ 1. Hunan Nuclear Agronomy and Space Breeding Research Institute, Changsha 410125, China; 2. Hunan Vegetable Research Institute, Changsha 410125, China; 3. Department of Biotechnology, Hunan Unive
ISSN:1004-874X