转录因子OsMADS25提高水稻对低温的耐受性

doi:10.16288/j.yczz.21-217

遗传 ›› 2021, Vol. 43 ›› Issue (11): 1078-1087.doi: 10.16288/j.yczz.21-217

转录因子OsMADS25提高水稻对低温的耐受性

闫凌月¹(), 张豪健¹, 郑雨晴¹, 丛韫起¹, 刘次桃¹, 樊帆¹, 郑铖¹, 袁贵龙², 潘根³, 袁定阳²(), 段美娟¹()

1. 湖南农业大学农学院,水稻逆境生物学湖南省重点实验室,长沙 410128
2. 湖南杂交水稻研究中心,杂交水稻国家重点实验室,长沙 410125
3. 中国农业科学院麻类研究所,长沙 410205

收稿日期:2021-06-20 修回日期:2021-08-21 出版日期:2021-11-20 发布日期:2021-10-12
通讯作者: 袁定阳,段美娟 E-mail:yanlingyue0203@163.com;yuandingyang@hhrrc.ac.cn;duanmeijuan@163.com
作者简介:闫凌月,在读硕士研究生,专业方向：作物遗传育种。E-mail: yanlingyue0203@163.com
基金资助:
湖南省科技重大专项编号(2018NK1010);湖南省科技人才支持项目编号(2019TJ-Q08);湖南农业大学人才引进启动基金编号(20154/5407419002);杂交水稻国家重点实验室(湖南杂交水稻研究中心)开放课题编号(2020KF05);湖南省自然科学基金青年基金项目编号资助(2019JJ50714)

Transcription factor OsMADS25 improves rice tolerance to cold stress

Lingyue Yan¹(), Haojian Zhang¹, Yuqing Zheng¹, Yunqi Cong¹, Citao Liu¹, Fan Fan¹, Cheng Zheng¹, Guilong Yuan², Gen Pan³, Dingyang Yuan²(), Meijuan Duan¹()

1. Hunan Provincial Key Laboratory of Rice Stress Biology, College of Agriculture, Hunan Agricultural University, Changsha 410128, China
2. State Key Laboratory of Hybrid Rice, Hunan Hybrid Rice Research Center, Changsha 410125, China
3. Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, China

Received:2021-06-20 Revised:2021-08-21 Online:2021-11-20 Published:2021-10-12
Contact: Yuan Dingyang,Duan Meijuan E-mail:yanlingyue0203@163.com;yuandingyang@hhrrc.ac.cn;duanmeijuan@163.com
Supported by:
Supported by the Hunan Science and Technology Major Project No(2018NK1010);the Hunan Science and Technology Talents Support Project No(2019TJ-Q08);the Research Initiation Fund of Hunan Agricultural University No(20154/5407419002);the Open Research Fund of the State Key Laboratory of Hybrid Rice, Hybrid Rice Research Center No(2020KF05);the Hunan Province Natural Science Fund No(2019JJ50714)

摘要/Abstract

摘要：

低温冷害是影响水稻高产的关键环境因素,鉴定和克隆具有重要应用价值的耐低温基因并培育耐低温新品种对于保障粮食安全具有重要意义。MADS转录因子在植物逆境信号途径中扮演着重要的角色。本研究利用qRT-PCR检测,发现OsMADS25受低温和脱落酸(abscisic acid, ABA)诱导表达上调,预示OsMADS25可能参与ABA依赖的逆境信号途径。进一步构建了水稻OsMADS25 的过表达载体pCambia1300-221-OsMADS25-Flag,利用根癌农杆菌介导的遗传转化法将其导入水稻品种中花11 (ZH11),选取两个表达量高的纯合株系进行表型鉴定。结果表明,OsMADS25过表达株系显著提高了水稻苗期对低温的耐受性以及对ABA的敏感性。利用3,3-二氨基联苯胺(diaminobezidine, DAB)和氯化硝基四氮唑蓝(nitrotetrazolium blue chloride, NBT)组织化学染色结果表明：低温处理后,OsMADS25过表达株系比野生型ZH11染色浅,表明过表达株系在低温胁迫下积累的活性氧(reactive oxygen species, ROS)相对较少,增强了对低温的耐受性。综合结果表明,低温逆境下OsMADS25响应ABA信号,通过提高水稻对ROS的清除能力,避免水稻受到低温伤害。

关键词: 水稻, OsMADS25, 脱落酸, 活性氧, 耐低温

Abstract:

Cold stress is the limiting factor of rice growth and production, and it is important to clone cold stress tolerant genes and cultivate cold tolerance rice varieties. The MADS transcription factors play an important role in abiotic stress signaling in rice. This study showed that OsMADS25 was up-regulated by low temperature and abscisic acid (ABA), suggesting that OsMADS25 may be involved in ABA-dependent signaling. The OsMADS25 overexpression vector, pCambia1300-221-OsMADS25-Flag, was constructed and introduced into the rice variety Zhonghua 11 (ZH11) through Agrobacterium tumefacian-mediated genetic transformation. Two homozygous lines with high expression levels were selected for phenotypic identification. OsMADS25 overexpression lines show significantly improved cold stress tolerance and the sensitivity to ABA at the seedling stage of rice. Reactive oxygen species (ROS) was detected by diaminobenzidine (DAB) staining and nitroblue tetrazolium (NBT) staining. After treatment with cold stress, little ROS accumulation was observed in OsMADS25 overexpression lines compared to wild-type ZH11. In conclusion, OsMADS25 plays a role in scavenging reactive oxygen species (ROS) and could improve rice tolerance to cold stress involved in ABA-dependent pathway.

Key words: rice (Oryza sativa L.), OsMADS25, abscisic acid (ABA), reactive oxygen species(ROS), cold tolerance

闫凌月, 张豪健, 郑雨晴, 丛韫起, 刘次桃, 樊帆, 郑铖, 袁贵龙, 潘根, 袁定阳, 段美娟. 转录因子OsMADS25提高水稻对低温的耐受性[J]. 遗传, 2021, 43(11): 1078-1087.

Lingyue Yan, Haojian Zhang, Yuqing Zheng, Yunqi Cong, Citao Liu, Fan Fan, Cheng Zheng, Guilong Yuan, Gen Pan, Dingyang Yuan, Meijuan Duan. Transcription factor OsMADS25 improves rice tolerance to cold stress[J]. Hereditas(Beijing), 2021, 43(11): 1078-1087.

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引物名称	引物序列(5ʹ→3ʹ)
OsMADS25	F: GAGGATCGACAACACGATGAA
OsMADS25	R: GGTGCAGGAGAAGACAATGA
LTG1	F: CGTGCTGAATGGGCTGATAA
LTG1	R: GTTGAGGTTGATGCCAAGGA
OsEREB6	F: TCCGGTTTGTTCCCAGTTTAG
OsEREB6	R: GCCTGGATGTAGTGCATCTG
OsTPP1	F: CCATCTACATTGGAGACGACAG
OsTPP1	R: CCTTGGGAACCTGTGAAACTA
Ubiqitin	F: GCTCCGTGGCGGTATCAT
Ubiqitin	R: CGGCAGTTGACAGCCCTAG

转录因子OsMADS25提高水稻对低温的耐受性

Transcription factor OsMADS25 improves rice tolerance to cold stress

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