转录因子BvM14-Dof3.4响应盐胁迫的功能研究

doi:10.11924/j.issn.1000-6850.casb2021-0262

中国农学通报 ›› 2021, Vol. 37 ›› Issue (21): 119-125.doi: 10.11924/j.issn.1000-6850.casb2021-0262

所属专题：生物技术

转录因子BvM14-Dof3.4响应盐胁迫的功能研究

杜晓雪¹^,²(), 黄园园¹^,², 马春泉¹^,²(), 李海英¹^,²()

¹黑龙江大学农业微生物技术教育部工程研究中心,哈尔滨 150500
²黑龙江大学生命科学学院,黑龙江省普通高校分子生物学重点实验室,哈尔滨 150080

收稿日期:2021-03-16 修回日期:2021-04-13 出版日期:2021-07-25 发布日期:2021-07-29
通讯作者: 马春泉,李海英
作者简介:杜晓雪,女,1994年出生,内蒙古兴安盟人,硕士,研究方向：寒区植物遗传与功能基因组。通信地址：150080 黑龙江省哈尔滨市学府路74号黑龙江大学生命科学学院320室,E-mail： xuer104@163.com。
基金资助:
国家自然科学基金“转录因子BvGT-1调控乙二醛酶I基因参与甜菜单体附加系耐盐的分子机制”(32072122)

Transcription Factor BvM14-Dof 3.4 in Response to Salt Stress: Functional Study

Du Xiaoxue¹^,²(), Huang Yuanyuan¹^,², Ma Chunquan¹^,²(), Li Haiying¹^,²()

¹Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education, Heilongjiang University, Harbin 150500
²Key Laboratory of Molecular Biology, College of Heilongjiang Province/School of Life Sciences, Heilongjiang University, Harbin 150080

Received:2021-03-16 Revised:2021-04-13 Online:2021-07-25 Published:2021-07-29
Contact: Ma Chunquan,Li Haiying

摘要/Abstract

摘要：

Dof(DNA binding with one finger)家族转录因子是植物特异的转录因子,在胁迫响应、种子发芽、氮同化、光合作用等多种生物学过程中发挥着重要作用。为了探究转录因子BvM14-Dof3.4在响应盐胁迫过程中的生物学功能,本研究以具有强耐盐特性的甜菜M14品系为试验材料,用花序浸染法将BvM14-Dof3.4基因在野生型拟南芥植株中异源表达,经150 mmol/L NaCl处理后发现,BvM14-Dof3.4基因的异源表达促进了盐胁迫下转基因拟南芥植株根的生长,提高了异源表达植株的鲜重和干重,与野生型拟南芥相比异源表达植株中K⁺/Na⁺比值增加1.3倍、甜菜碱含量增加1.1倍以及SOD和POD的酶活性分别上调1.3和1.2倍,从而减少了盐胁迫对异源表达拟南芥植株的损伤。这些结果表明了BvM14-Dof3.4基因响应盐胁迫,并且BvM14-Dof3.4基因的异源表达能够提高拟南芥植株的耐盐能力,该结果对甜菜M14品系优质基因资源的挖掘以及开展栽培甜菜抗逆性的遗传改良工作具有重要意义。

关键词: 甜菜M14品系, 转录因子, BvM14-Dof3.4, 盐胁迫, 异源表达

Abstract:

Dof family transcription factors are plant specific transcription factors, which play an important role in stress response, seed germination, nitrogen assimilation, photosynthesis and other biological processes. In order to explore the biological function of transcription factor BvM14-Dof3.4 in response to salt stress, the sugar beet M14 line with strong salt tolerance was used as the experimental material. The BvM14-Dof3.4 gene was heterologously expressed in wild type Arabidopsis thaliana by inflorescence impregnation. After 150 mmol/L NaCl treatment, it was found that the root growth of transgenic Arabidopsis thaliana under salt stress was promoted, the fresh and dry weight was increased by the heterologous expression of BvM14-Dof3.4 gene; the K⁺/Na⁺ ratio, the content of betaine and the enzyme activities of SOD and POD in heterologous expression plants were increased by 1.3 fold, 1.1 fold and 1.2 fold, respectively compared with the wild type Arabidopsis thaliana, which reduced the damage of heterologous expression Arabidopsis plants caused by salt stress. These results showed that BvM14-Dof3.4 gene was responsive to salt stress and the heterologous expression of BvM14-Dof3.4 gene could improve the salt tolerance of Arabidopsis thaliana. The study is of great significance for the mining of high quality gene resources of sugar beet M14 and the genetic improvement of stress resistance of cultivated sugar beet.

Key words: sugar beet M14 line, transcription factor, BvM14-Dof3.4 gene, salt stress, heterologous expression

中图分类号:

S566.3

杜晓雪, 黄园园, 马春泉, 李海英. 转录因子BvM14-Dof3.4响应盐胁迫的功能研究[J]. 中国农学通报, 2021, 37(21): 119-125.

Du Xiaoxue, Huang Yuanyuan, Ma Chunquan, Li Haiying. Transcription Factor BvM14-Dof 3.4 in Response to Salt Stress: Functional Study[J]. Chinese Agricultural Science Bulletin, 2021, 37(21): 119-125.

图/表 6

引物功能	引物名称	序列
PCR引物	BvM14-Dof3.4-S	5'-ATGGGTGGAGGCGGAGGC-3'
PCR引物	BvM14-Dof3.4-AS	5'-TCACTTGAGACCATTGGCTGATG-3'
荧光定量PCR引物	18S-S	5'-CCCCAATGGATCCTCGTT A-3'
	18S-AS	5'-TGACGGAGAATTAGGGTT CG-3'
	dof -S	5'-GTGGAGGTGGTGGGTTTA-3'
	dof-AS	5'-TCCGTTTTCATTATTATT-3'
构建35S::pCAMBIA1300-BvM14-Dof3.4引物	1300-S	5'-CGAGCTCATGGGTGGAGGCGGAGGCGGAGGT-3'
构建35S::pCAMBIA1300-BvM14-Dof3.4引物	1300-AS	5'-CGAGATCTTCACTTGAGACCATTGGCTGATGT-3'

表1. 引物序列

图1. 35S::pCAMBIA1300-BvM14-Dof3.4重组质粒PCR产物的1%琼脂糖凝胶电泳图 M：DNA Marker DL2000;1~5：35S::pCAMBIA1300-BvM14-Dof3.4重组质粒的PCR产物;6：阴性对照

图2. BvM14-Dof3.4基因异源表达拟南芥植株OX的PCR及RT-PCR检测结果 A：1~4：OX植株的PCR产物;5：WT植株的PCR产物;B：1~3：OX植株的RT-PCR产物;4：WT植株的RT-PCR产物

图3. 150 mmol/L NaCl处理前后WT和OX拟南芥植株的表型及根长、鲜重、干重测定结果 A：表型;B：根长;C：鲜重;D：干重;WT：野生型拟南芥植株;OX：BvM14-Dof3.4基因异源表达拟南芥植株

图4. 150 mmol/L NaCl处理前后WT和OX拟南芥植株的K+/Na+比值和甜菜碱含量测定结果 A：K+/Na+比值;B：甜菜碱含量;WT：野生型拟南芥植株;OX：BvM14-Dof3.4基因异源表达拟南芥植株

图5. 150 mmol/L NaCl处理前后WT和OX拟南芥植株的SOD和POD酶活测定结果 A：SOD酶活;B：POD酶活;WT：野生型拟南芥植株;OX：BvM14-Dof3.4基因异源表达拟南芥植株

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转录因子BvM14-Dof3.4响应盐胁迫的功能研究

Transcription Factor BvM14-Dof 3.4 in Response to Salt Stress: Functional Study

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