甘蓝型油菜转录因子BnaABI5表达特征分析及互作蛋白鉴定

doi:10.16288/j.yczz.24-064

遗传 ›› 2024, Vol. 46 ›› Issue (9): 737-749.doi: 10.16288/j.yczz.24-064

甘蓝型油菜转录因子BnaABI5表达特征分析及互作蛋白鉴定

奥倩倩(), 陆方筱(), 杨柳卿, 李春, 翟增康, 贾东晔, 江元清, 杨博()

西北农林科技大学生命科学学院，杨凌 712100

收稿日期:2024-03-12 修回日期:2024-08-08 出版日期:2024-09-20 发布日期:2024-08-23
通讯作者: 杨博，博士，副教授，研究方向：植物抗逆分子遗传学。E-mail: yangwl@nwafu.edu.cn
作者简介:奥倩倩，硕士研究生，专业方向：植物分子遗传。E-mail: 2628605169@qq.com；
陆方筱，硕士研究生，专业方向：植物分子遗传。E-mail: 392549812@qq.com。
第一联系人：
奥倩倩与陆方筱并列第一作者。
基金资助:
国家自然科学基金项目(32171953)

Analysis of expression characteristics and identification of interaction proteins of transcription factor BnaABI5 in Brassica napus

Qianqian Ao(), Fangxiao Lu(), Liuqing Yang, Chun Li, Zengkang Zhai, Dongye Jia, Yuanqing Jiang, Bo Yang()

College of Life Sciences, Northwest A&F University, Yangling 712100, China

Received:2024-03-12 Revised:2024-08-08 Published:2024-09-20 Online:2024-08-23
Supported by:
National Natural Science Foundation of China(32171953)

摘要/Abstract

摘要：

甘蓝型油菜(Brassica napus L.)是重要的油料作物之一，但是其种植效益在西北地区经常受到干旱等环境逆境的影响。脱落酸(abscisic acid，ABA)信号通路在植物对非生物胁迫的响应与耐受中具有重要作用，而ABFs/AREBs(ABA-responsive element binding factors/ABA-responsive element binding proteins)是调控ABA响应基因表达的核心转录因子。为了解析油菜响应逆境的关键转录因子，本研究首先对甘蓝型油菜转录因子BnaABI5(abscisic acid insensitive 5)进行了亚细胞定位、逆境响应及组织器官表达的检测、转录活性分析以及与激酶BnaMPKs(mitogen-activated protein kinases)的互作筛选。结果显示：BnaABI5-GFP融合蛋白定位于细胞核，该基因响应干旱逆境且主要在油菜的根中表达，并且在酵母体系中呈现出一定的转录激活活性；随后利用烟草瞬时表达体系发现BnaABI5能够激活靶标基因EM6(early methionine-labeled 6)的启动子活性。通过双分子荧光互补(bimolecular fluorescence complementation，BiFC)及酵母双杂交(yeast two-hybrid，Y2H)实验，发现BnaABI5与BnaMPK6及BnaMPK13互作。本研究初步探索了转录因子BnaABI5的表达特征及与BnaMPKs的互作，对于深入理解BnaABI5的功能具有一定的指导意义。

关键词: 甘蓝型油菜, ABI5, MPK, 互作蛋白

Abstract:

Rapeseed is one important oil crop in China. However, its planting benefit is frequently affected by environmental stresses such as drought in the northwest region of China. The abscisic acid(ABA) signaling pathway plays an important role in plant abiotic stress response and tolerance, and ABFs/AREBs(ABA-responsive element binding factors/ABA-responsive element binding proteins) are the core transcription factors that regulate the expression of ABA-responsive genes. To dissect the key transcription factors mediated abiotic stress, we mainly characterized abscisic acid insensitive 5(BnaABI5) in rapeseed, including its subcellular localization, expression pattern in response to various stress and tissue-specific expression analysis, transcriptional activity analysis as well as interaction screening with BnaMPKs(mitogen-activated protein kinases). Our results showed that the BnaABI5-GFP fusion protein was localized in the nucleus, and its transcript level is induced by drought stress and was mainly expressed in the roots of rapeseed. Furthermore, BnaABI5 showed transcriptional activation activity through a yeast transactivation assay and it also activated the promoter activity of EM6 target gene in the transient expression system in tobacco leaves. Moreover, BnaABI5 interacted with BnaMPK6 and BnaMPK13 through BiFC and Y2H analysis. This study preliminarily explored the expression characteristics of transcription factor BnaABI5 and its interaction with BnaMPKs, which might help us for further understanding the function of BnaABI5.

Key words: Brassica napus L., ABI5, MPK, interacting proteins

奥倩倩, 陆方筱, 杨柳卿, 李春, 翟增康, 贾东晔, 江元清, 杨博. 甘蓝型油菜转录因子BnaABI5表达特征分析及互作蛋白鉴定[J]. 遗传, 2024, 46(9): 737-749.

Qianqian Ao, Fangxiao Lu, Liuqing Yang, Chun Li, Zengkang Zhai, Dongye Jia, Yuanqing Jiang, Bo Yang. Analysis of expression characteristics and identification of interaction proteins of transcription factor BnaABI5 in Brassica napus[J]. Hereditas(Beijing), 2024, 46(9): 737-749.

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甘蓝型油菜转录因子BnaABI5表达特征分析及互作蛋白鉴定

Analysis of expression characteristics and identification of interaction proteins of transcription factor BnaABI5 in Brassica napus

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