谷子SiRLK35基因克隆及功能分析

doi:10.16288/j.yczz.17-027

遗传 ›› 2017, Vol. 39 ›› Issue (5): 413-422.doi: 10.16288/j.yczz.17-027

谷子SiRLK35基因克隆及功能分析

王一帆^1,²(),李臻²,潘教文²,李颖秀²,王庆国²,管延安³,刘炜²()

1. 青岛农业大学生命科学学院,青岛 266000
2. 山东省农业科学院生物技术研究中心,山东省作物遗传改良与生理生态重点实验室,济南 250100
3. 山东省农业科学院作物研究所,济南 250100

收稿日期:2017-01-20 修回日期:2017-04-01 出版日期:2017-04-24 发布日期:2017-12-25
作者简介:王一帆,硕士研究生,专业方向：植物分子生物学。E-mail: 1356978348@qq.com|刘炜,博士,研究员,研究方向：作物分子育种。E-mail: wheiliu@163.com
基金资助:
山东省重大科技专项(2015ZDGS03001-2);山东省农业科学院重大科技成果培育计划项目(2015CGPY10);山东省自然科学基金面上项目(2016ZRC02178);山东农业大学作物生物学国家重点实验室开放课题(2015KF15)

Cloning and functional analysis of the SiRLK35 gene in Setaria italic L.

Wang Yifan^1,²(),Li Zhen²,Pan Jiaowen²,Li Yingxiu²,Wang Qingguo²,Guan Yan'an³,Liu Wei²()

1. College of Life Science, Qingdao Agricultural University, Qingdao 266000, China
2. Biotechnology Research Center, Shandong Academy of Agricultural Sciences /Shandong Provincial Key Laboratory of Crop Genetic Improvement, Ecology and Physiology, Ji'nan 250100, China;
3. Crop Research Institute, Shandong Academy of Agricultural Sciences, Ji'nan 250100, China;

Received:2017-01-20 Revised:2017-04-01 Online:2017-04-24 Published:2017-12-25
Supported by:
the Major in Shandong Province Science and Technology Projects(2015ZDGS03001-2);Shandong Academy of Agricultural Sciences(2015CGPY10);the Natural Science Foundation of Shandong Province(2016ZRC02178);the State Key Laboratory of Crop Biology of Shandong Aguricultural University(2015KF15)

摘要/Abstract

摘要：

为探讨谷子(Setaria italica L.)耐旱抗逆机制,解析类受体蛋白激酶(receptor like protein kinase, RLKs)基因功能,进而为培育谷子抗逆新品种提供依据,本文以干旱处理的谷子“豫谷1号”为材料,通过iTRAQ技术筛选到1个干旱响应的类受体蛋白激酶基因,命名为SiRLK35。以谷子RNA反转录的单链cDNA为模板,经PCR扩增获取SiRLK35基因全长序列。应用qRT-PCR方法,对SiRLK35在NaCl、PEG、ABA、GA、MeJA等不同处理下的表达模式进行分析。进一步构建基因原核表达载体pET28a-SiRLK35,结合斑点法对SiRLK35的抗盐能力进行初步评价。同时构建过表达载体pCAMBIA1301P-SiRLK35转化水稻,并对转基因植株抗盐能力进行检测。结果显示：胁迫及激素处理均可不同程度诱导SiRLK35基因的表达;斑点法研究结果显示,在相同NaCl浓度的LB平板上,含有SiRLK35基因的原核表达载体的大肠杆菌菌株生长状态较阴性对照好,SiRLK35具有一定的抗盐能力;获得的转SiRLK35基因水稻植株对盐胁迫的耐受性高于对照。SiRLK35基因对不同胁迫均可以产生响应,但对盐胁迫的响应较为明显,推测该基因可能在谷子的抗盐及抗逆过程中发挥作用。

关键词: 谷子, 类受体蛋白激酶, 表达模式, 抗旱及抗逆, 品种培育

Abstract:

Receptor like protein kinases (RLKs) play vital roles in both plant development and stress conditions. Using drought-treated "Yugu 1" as materials, a drought-responsive RLK gene, SiRLK35, was isolated through iTRAQ analysis. In this study, the further analyses of the gene functions were carried out. First, full-length SiRLK35 was amplified by PCR using the cDNA of foxtail millet seedlings as a template. The expression patterns of SiRLK35 under NaCl, PEG, ABA, GA and MeJA treatment were analyzed by quantitative real-time PCR (qRT-PCR), and we found that the expression of SiRLK35 could be induced under different treatments, especially under NaCl treatment. Second, the prokaryotic expression plasmid of SiRLK35 was constructed, and the salt resistance of SiRLK35 was detected by the bacterial plaque growth method. And we uncovered that the growth and tolerance of SiRLK35-containing Escherichia coli strains were in better conditions than control under the NaCl stress. Lastly, pCambia1301P-SiRLK35 was constructed and transformed into rice to obtain transgenic plants. The tolerance of transgenic rice plants to salt stress was higher than that of controls through physiological analysis. We propose that SiRLK35 may participate in salt and stress resistance processes, which could provide potential theoretical foundation for the stress resistance varieties cultivation and breeding of foxtail millet.

Key words: Setaria italica L., receptor like protein kinase, expression pattern, salt and stress resistance, varieties cultivation and breeding

王一帆,李臻,潘教文,李颖秀,王庆国,管延安,刘炜. 谷子SiRLK35基因克隆及功能分析[J]. 遗传, 2017, 39(5): 413-422.

Wang Yifan,Li Zhen,Pan Jiaowen,Li Yingxiu,Wang Qingguo,Guan Yan'an,Liu Wei. Cloning and functional analysis of the SiRLK35 gene in Setaria italic L.[J]. Hereditas(Beijing), 2017, 39(5): 413-422.

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谷子SiRLK35基因克隆及功能分析

Cloning and functional analysis of the SiRLK35 gene in Setaria italic L.

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