马铃薯蛋白激酶基因<em>StPki</em>的遗传转化及表达分析

doi:10.7668/hbnxb.2014.01.014

华北农学报 ›› 2014, Vol. 29 ›› Issue (1): 73-77. doi: 10.7668/hbnxb.2014.01.014

所属专题： 薯类作物；生物技术；

马铃薯蛋白激酶基因StPki的遗传转化及表达分析

杨煜¹, 郭晓¹, 郭宝太², 杨晓慧¹, 单伟伟¹, 金黎平³, 马伟清¹, 李广存¹

1. 山东省农业利一学院蔬菜花卉研究所.国家蔬菜改良中心山东分中心.山东省设施蔬菜生物学重点实验室,山东 250100;
2. 青岛农业大学生命利一学院, 山东青岛266109;
3. 中国农业利一学院蔬菜花卉研究所, 北京 100081

收稿日期:2013-10-26 出版日期:2014-02-28
通讯作者: 马伟清(1962-)，女，山东济南人，高级农艺师，主要从事马铃薯育种、组培研究。
作者简介:杨煜(1977-)，男，山东诸城人，副研究员，主要从事马铃薯种质创新研究。杨煜、郭晓为同等贡献作者。
基金资助:
国家“863”项目(2013AA102603-4)；国家科技支撑计划项目(2012BAD02B05-10)；山东省自然科学基金项目(Y2007D24)；山东省良种工程农业生物资源创新利用项目(PTBR2013)

Genetic Transformation of Diploid Potato and Expression Suppression of Its Protein Kinase Gene StPki

YANG Yu¹, GUO Xiao¹, GUO Bao-tai², YANG Xiao-hui¹, SHAN Wei-wei¹, JIN Li-ping³, MA Wei-qing¹, LI Guang-cun¹

1. Institute of Vegetables and Flowers, Shandong Academy of Agriculture Sciences, Nationnal Improvement Center for Vegetable, Shandong Branch,Shandong Key Laboratory for Biology of Greenhouse Vegetables,Ji'nan 250100,China;
2. Life Science College,Qingdao Agricultural University,Qingdao 266109, China;
3. Institute of Vegetables and Flowers,Chinese Academy of Agriculture Sciences,Beijing 100081,China

Received:2013-10-26 Published:2014-02-28

摘要/Abstract

摘要： 以高抗青枯病二倍体马铃薯基因型ED13为材料,克隆了蛋白激酶基因StPki。以StPki基因特异区段为靶标,成功构建了该基因的RNA干扰植物表达载体pCHF1-StPki。利用重组农杆菌株LBA4404(pCHF1-StPki)感染转化ED13茎段外植体,获得了抗庆大霉素的再生植株。利用CaMV35S启动子特异引物对再生植株进行PCR检测,结果表明获得了转基因植株。利用StPki基因的特异引物对转基因植株进行半定量RT-PCR分析,结果显示该基因的转录受到了抑制。马铃薯抗病基因型ED13已被成功转化,且表现出了对StPki基因的RNA干扰活性。

关键词: 马铃薯, 二倍体, 蛋白激酶基因, 遗传转化, 表达抑制

Abstract: Protein kinase gene StPki was cloned from bacterial wilt-resistant diploid potato ED13,and its RNAi plant expression vector was constructed. The expression vector pCHF1-StPki was introduced into Agrobacterium strain LBA4404 and the recombinant strain LBA4404(pCHF1-StPki) was used to transform ED13 stem segment. In the presence of CaMV35S promoter-specific primers,Gen-resistant regeneration plants were detected by PCR and transgenic potato plant producing 500 bp amplification product were obtained. Semi-quantitative RT-PCR analysis indicated that the transcription of StPki gene had beed significantly restrained in the transgenic ED13 plant.

Key words: Potato, Diploid, Protein kinase gene, Genetic transformation, Expression suppression

中图分类号:

杨煜, 郭晓, 郭宝太, 杨晓慧, 单伟伟, 金黎平, 马伟清, 李广存. 马铃薯蛋白激酶基因StPki的遗传转化及表达分析[J]. 华北农学报, 2014, 29(1): 73-77. doi: 10.7668/hbnxb.2014.01.014.

YANG Yu, GUO Xiao, GUO Bao-tai, YANG Xiao-hui, SHAN Wei-wei, JIN Li-ping, MA Wei-qing, LI Guang-cun. Genetic Transformation of Diploid Potato and Expression Suppression of Its Protein Kinase Gene StPki[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2014, 29(1): 73-77. doi: 10.7668/hbnxb.2014.01.014.

http://www.hbnxb.net/CN/Y2014/V29/I1/73

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