蒺藜苜蓿MtDWF1基因克隆、亚细胞定位及表达特征分析

doi:10.11733/j.issn.1007-0435.2023.04.006

草地学报 ›› 2023, Vol. 31 ›› Issue (4): 984-991.DOI: 10.11733/j.issn.1007-0435.2023.04.006

• 研究论文 • 上一篇

蒺藜苜蓿MtDWF1基因克隆、亚细胞定位及表达特征分析

谢宏¹, 李舒文¹, 董迪¹, 王梦迪¹, 贾薇¹, 晁跃辉¹, 赵彦²

1. 北京林业大学草业与草原学院, 北京 100083;
2. 内蒙古农业大学草原与资源环境学院, 内蒙古呼和浩特 010018

收稿日期:2022-12-02 修回日期:2023-01-13 发布日期:2023-04-28
通讯作者: 晁跃辉,E-mail:chaoyuehui@bjfu.edu.cn
作者简介:谢宏(1998-),女,汉族,内蒙古包头人,硕士研究生,主要从事草地植物遗传育种研究,E-mail：xiehong20211014@163.com;
基金资助:
教育部草地资源教育部重点实验室开放课题资助

Cloning, Subcellular Localization and Expression Pattern of MtDWF1 Gene in Medicago truncatula

XIE Hong¹, LI Shu-wen¹, DONG Di¹, WANG Meng-di¹, JIA Wei¹, CHAO Yue-hui¹, ZHAO Yan²

1. School of Grassland Science, Beijing Forestry University, Beijing 100083, China;
2. College of Grassland, Resources Environment, Inner Mongolia Agricultural University, Hottot, Inner Mongolia 010018, China

Received:2022-12-02 Revised:2023-01-13 Published:2023-04-28

摘要/Abstract

摘要： Delta24-甾醇还原酶(Delta24-sterol reductase，DWF1)是油菜素内酯(Brassinosteroide，BR)合成途径中的关键酶，在植物生长发育中起着关键性的作用。为了研究DWF1基因在蒺藜苜蓿(Medicago truncatula)中的功能，本试验从蒺藜苜蓿R108中克隆MtDWF1基因全长序列，该基因开放阅读框(ORF)1 704 bp，编码567个氨基酸，分子量65.911 kD，理论等电点为8.53。亚细胞定位显示，该蛋白定位于细胞质内。进化分析表明，蒺藜苜蓿MtDWF1蛋白与鹰嘴豆(Cicer arietinum)、红三叶(Trifolium pratense)及豌豆(Pisum sativum)亲缘关系较为接近。MtDWF1基因在蒺藜苜蓿根、茎、叶中均有表达，在茎和叶中的表达水平较高。外源激素脱落酸(Abscisic acid，ABA)能够明显提高MtDWF1表达，水杨酸(Salicylic acid，SA)能够显著降低MtDWF1表达水平。同时，经油菜素内酯诱导后，MtDWF1表达水平整体呈现增加趋势。本研究为进一步探索MtDWF1基因在蒺藜苜蓿发育过程中的作用提供理论研究基础。

关键词: MtDWF1, 蒺藜苜蓿, 基因克隆, 表达分析, 亚细胞定位

Abstract: Delta24-sterol reductase (DWF1) is a crucial enzyme in the biosynthesis of brassinosteroids and has a significant regulation on plant growth and development. To examine the function of the DWF1 gene in Medicago truncatula,the full-length sequence of the MtDWF1 gene was cloned from M. truncatula (R108). The open reading frame of MtDWF1 was found to be 1 704 bp in length and encoded a peptide of 567 amino acids with a molecular weight of 65.911 kDa and a predicted isoelectric point of 8.53. The protein of the MtDWF1 was localized in the cytoplasm as determined by subcellular localization analysis. Evolutionary analysis indicated that the MtDWF1 protein of M. truncatula is closely related to those of Cicer arietinum,Trifolium pratense,and Pisum sativum. The MtDWF1 gene was expressed in the roots,stems,and leaves of M. truncatula,with higher expression levels observed in stems and leaves than that in roots. Exogenous application of abscisic acid (ABA) caused the MtDWF1 expression significantly increased,while salicylic acid (SA) significantly reduced. Brassinosterol (BR) treatment resulted in an overall increase of MtDWF1 expression. This study provides a foundation for further investigating on the role of the MtDWF1 gene in M. truncatula development.

Key words: MtDWF1, Medicago truncatula, Gene clone, Expression analysis, Subcellular localization

中图分类号:

Q785

谢宏, 李舒文, 董迪, 王梦迪, 贾薇, 晁跃辉, 赵彦. 蒺藜苜蓿MtDWF1基因克隆、亚细胞定位及表达特征分析[J]. 草地学报, 2023, 31(4): 984-991.

XIE Hong, LI Shu-wen, DONG Di, WANG Meng-di, JIA Wei, CHAO Yue-hui, ZHAO Yan. Cloning, Subcellular Localization and Expression Pattern of MtDWF1 Gene in Medicago truncatula[J]. Acta Agrestia Sinica, 2023, 31(4): 984-991.

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蒺藜苜蓿MtDWF1基因克隆、亚细胞定位及表达特征分析

Cloning, Subcellular Localization and Expression Pattern of MtDWF1 Gene in Medicago truncatula

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