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

doi:10.11733/j.issn.1007-0435.2023.04.006

Acta Agrestia Sinica ›› 2023, Vol. 31 ›› Issue (4): 984-991.DOI: 10.11733/j.issn.1007-0435.2023.04.006

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

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

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

Abstract

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

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

CLC Number:

Q785

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.

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.

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References

[1] TRIEU A T,HARRISON M J. Rapid transformation of Medicago truncatula:regeneration via shoot organogenesis[J]. Plant Cell Reports,1996,16(1-2):6-11
[2] VIKER K B,STEELE M B,LANKOV I D,et al. Preclinical safety assessment of MV-s-NAP,a novel oncolytic measles virus strain armed with an H. pylori immunostimulatory bacterial transgene[J]. Molecular Therapy - Methods & Clinical Development,2022(26):532-546
[3] ENDRE G,KERESZT A,KEVEI Z,et al. A receptor kinase gene regulating symbiotic nodule development[J]. Nature,2002(417):962-966
[4] 李菲,何小红,张习敏,等. 苜蓿14-3-3基因家族的鉴定与进化和特征分析[J]. 基因组学与应用生物学,2017,36(12):5238-5243
[5] TAO Y Z,ZHENG J,XU Z M,et al. Functional analysis of ZmDWF1,a maize homolog of the Arabidopsis brassinosteroids biosynthetic DWF1/DIM gene[J]. Plant Science,2004,167(4):743-751
[6] 何宇炯,徐如涓,赵毓橘,等. 表油菜素内酯对油菜幼苗生长及其可溶性糖和蛋白质含量的影响[J]. 植物生理学通讯,1995,31(1):37-39
[7] 曲善民,苏一诺,苗阳阳,等. 芸苔素内酯对连作及非连作紫花苜蓿生产调控初探[J]. 草地学报,2019,27(4):874-881
[8] 寇江涛. 外源2,4-表油菜素内酯对NaCl胁迫下燕麦种子萌发和生理的影响[J]. 草地学报,2019,27(6):1562-1568
[9] 邓天福,吴艳兵,李广领,等. 油菜素内酯提高植物抗逆性研究进展[J]. 广东农业科学,2009(11):21-25
[10] WANG Z Y,BAI M Y,OH E,et al. Brassinosteroid signaling network and regulation of photomorphogenesis[J]. Annual Review of Genetics,2012,46(1):701-724
[11] HOSSAIN Z,MCGARVEY B,AMYOT L,et al. Diminuto 1 affects the lignin profile and secondary cell wall formation in Arabidopsis[J]. Planta,2012,235(3):485-498
[12] 张瑾,金乌云,陈莹莹,等. 甜瓜CmDWF1基因的克隆及表达分析[J]. 华北农学报,2019,34(5):8-14
[13] 吴国超. 水稻矮脆突变体dwf1的特性与基因定位[D]. 重庆:西南大学,2014:35-36
[14] 陶亚忠. 玉米与油菜素类固醇生物合成相关的基因ZmDWF1的克隆及其功能分析[D]. 北京:中国农业大学,2004:108
[15] LI Y R Z,WANG M D,GUO T,et al. Overexpression of abscisic acid-insensitive gene ABI4 from Medicago truncatula,which could interact with ABA2,improved plant cold tolerance mediated by ABA signaling[J]. Frontiers in Plant Science,2022,13(13):982715
[16] 舒思晨,王娟,王柏柯,等. 外源MeJA和BR对番茄种子萌发及幼根生长的影响[J]. 农业工程,2022,12(3):138-143
[17] 吴婧,范菠菠,张学峰,等. 蒙古冰草ERF转录因子生物信息学及其表达分析[J]. 草地学报,2022,30(11):2910-2921
[18] CHOE S,DILKES B P,GREGORY B D,et al. The Arabidopsis dwarf1 mutant is defective in the conversion of 24-methylenecholesterol to campesterol in brassinosteroid biosynthesis[J]. Plant Physiology,1999,119(3):897-908
[19] 罗明,周建平,肖月华,等. 棉花油菜素类固醇合成酶基因GhDWF1的克隆和表达分析[J]. 中国农业科学,2007(7):1337-1344
[20] 唐晓,邓孟胜,邹雪,等. 马铃薯StDWF1基因克隆及表达分析[J]. 浙江农业学报,2018,30(6):909-917
[21] SCHULTZ L,HUUB L,KERCKHOFFS J,et al. Molecular characterization of the brassinosteroid-deficient Ikb mutant in pea[J]. Plant Molecular Biology,2001,47(4):491-508
[22] 刚爽. ABA对亚高温强光胁迫下番茄叶片光合作用影响及分子机制研究[D]. 沈阳:沈阳农业大学,2016:9-11
[23] 段方猛,罗秋兰,鲁雪莉,等. 玉米油菜素甾醇生物合成关键酶基因ZmCYP90B1的克隆及其对逆境胁迫的响应[J]. 作物学报,2018,44(3):343-356
[24] VRIET C,RUSSINOVA E,REUZEAU C,et al. From squalene to brassinolide:the steroid metabolic and signaling pathways across the plant kingdom[J]. Molecular Plant,2013,6(6):1738-1757
[25] BEST N B,HARTWIG T,BUDKA J,et al. Nana plant2 encodes a maize ortholog of the Arabidopsis brassinosteroid biosynthesis gene DWARF1,identifying developmental interactions between brassinosteroids and gibberellins[J]. Plant Physiology,2016,171(4):2633-2647

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

蒺藜苜蓿MtDWF1基因克隆、亚细胞定位及表达特征分析

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