Cloning and Expression Analysis of MsJAR1 in Medicago sativa

doi:10.11733/j.issn.1007-0435.2024.05.007

Acta Agrestia Sinica ›› 2024, Vol. 32 ›› Issue (5): 1370-1377.DOI: 10.11733/j.issn.1007-0435.2024.05.007

Cloning and Expression Analysis of MsJAR1 in Medicago sativa

DAI Rui¹, CHEN Qi¹, SHUANG Shuang¹, ZHANG Yan¹, ZHANG Zhi-qiang^1,2, MI Fu-gui^1,2

1. Colleg of Grassland, Resources and Environment, Inner Mongolia Agricultural University, Inner Mongolia Autonomous Region, Hohhot, 010010, China;
2. Technology Engineering Center of Drought and Cold-resistant Grass Breeding in North of the National Forestry and Grassland Administration, Institute of Grassland and Resources Environment, Inner Mongolia Agricultural University, Hohhot, Inner Mongalia 010010, China

Received:2023-12-06 Revised:2024-01-18 Published:2024-06-01

紫花苜蓿MsJAR1基因克隆及表达分析

代蕊¹, 陈崎¹, 爽爽¹, 张岩¹, 张志强^1,2, 米福贵^1,2

1. 内蒙古农业大学草原与资源环境学院, 内蒙古呼和浩特 010010;
2. 内蒙古农业大学草原与资源环境学院, 国家林业草原北方抗旱耐寒草品种育繁工程技术研究中心, 内蒙古呼和浩特 010010

通讯作者: 张志强，E-mail:zhangzq19890102@126.com;米福贵，E-mail:Mfgui@yahoo.com.cn
作者简介:代蕊(1999-)，女，蒙古族，内蒙古赤峰人，硕士研究生，主要从事草育种方面研究，E-mail:18747659997@163.com
基金资助:
内蒙古自治区高等学校“青年科技英才支持项目”(NJYT23009)；国家自然科学基金(32160333；32060388)；苜蓿分子育种体系构建及种质创制(BR22-11-12)；苜蓿新品种选育及制繁种关键技术研发(CCPTZX2023B04);内蒙古自治区研究生科研创新项目(S202311029)资助

Abstract

Abstract: To explore the function of jasmonoyl amino acid conjugate synthase(JAR1) gene in alfalfa,a novel gene, MsJAR1 (OK602801),was cloned,characterized and bioinformatic analyzed from Medicago sativa L.. The results showed that the cDNA sequence of MsJAR1 gene was 1 740 bp in length,and the open reading frame encoded 579 amino acid polypeptide,containing the GH3 conserved domain. Amino acid sequence alignment indicated that the homology of deduced MsJAR1 with Medicago truncatula,Trifolium pratense,Pisum sativum,Cicer arietinum and Abrus precatorius was all more than 80%. Subcellular localization analysis in tobacco showed that the MsJAR1 protein located in the chloroplast. The MsJAR1 gene expression was the highest in old leaves,followed by floral organs. In addition,thrips feeding,salt and drought stress could induce the expression of MsJAR1 gene,which provided a reference for further exploring the function of MsJAR1 gene in alfalfa.

Key words: Alfalfa, MsJAR1, Gene cloning, Expression analysis

摘要： 为探究紫花苜蓿(Medicago sativa L.)茉莉酰氨基酸结合物合成酶(Jasmonoyl amino acid conjugate synthase,JAR1)的基因功能，本试验对'草原4号’紫花苜蓿中克隆到的MsJAR1基因进行了生物信息学和表达模式分析，发现MsJAR1基因cDNA序列长度为1 740 bp，开放阅读框(ORF)编码 579 个氨基酸残基，含有GH3保守结构域。MsJAR1与蒺藜苜蓿(Medicago truncatula)、红三叶(Trifolium pratense)、豌豆(Pisum sativum)、鹰嘴豆(Cicer arietinum)、相思子(Abrus precatorius)的同源性均在80%以上。烟草亚细胞定位显示该蛋白定位于叶绿体。MsJAR1基因在老叶中表达量最高，花器官次之；蓟马取食、盐和干旱胁迫可诱导该基因表达，初步推测其参与紫花苜蓿应答蓟马取食及干旱和盐胁迫防御反应。本研究为进一步探究紫花苜蓿MsJAR1基因功能提供了参考。

关键词: 紫花苜蓿, MsJAR1, 基因克隆, 表达模式

CLC Number:

S892.6

DAI Rui, CHEN Qi, SHUANG Shuang, ZHANG Yan, ZHANG Zhi-qiang, MI Fu-gui. Cloning and Expression Analysis of MsJAR1 in Medicago sativa[J]. Acta Agrestia Sinica, 2024, 32(5): 1370-1377.

代蕊, 陈崎, 爽爽, 张岩, 张志强, 米福贵. 紫花苜蓿MsJAR1基因克隆及表达分析[J]. 草地学报, 2024, 32(5): 1370-1377.

0
/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

URL: https://manu40.magtech.com.cn/Jweb_cdxb/EN/10.11733/j.issn.1007-0435.2024.05.007

https://manu40.magtech.com.cn/Jweb_cdxb/EN/Y2024/V32/I5/1370

References

[1] 王旗旗,解继红,于林清,等. 我国苜蓿育种研究进展及展望[J]. 草学,2023(4):1-7
[2] 刘建宁,胡跃高,王运琦,等. 紫花苜蓿休眠类型与生长特性及生产性能的研究[J]. 草业科学,2003(10):29-32
[3] 杨向黎,韩凤英,秦旭,等. 紫花苜蓿几种主要害虫的化学防治试验[J]. 作物杂志,2014(2):131-136
[4] 爽爽,代蕊,陈崎,等. 基于转录组分析的紫花苜蓿组成型抗蓟马性研究[J]. 江苏农业科学,2023,51(5):44-50
[5] ERB M,MELDAU SANDHOWER G A. Role of phytohormones in insect-specific plant reactions[J]. Trends in plant science,2012,17(5):250-259
[6] DIVEKARIVEKAR P A,RANI V,MAJUMDERAJUMDER S,et al. Protease inhibitors:an induced plant defense mechanism against herbivores[J]. Journal of Plant Growth Regulation,2023;42(10),6057-6073
[7] 林颖,王燕燕,于放. 长春花茉莉酸-异亮氨酸合成酶crjar1生物信息学分析与原核表达[J]. 广西植物,2020,40(8):1181-1187
[8] 杨丽佳,陈欢欢,申龙斌,等. 黄灯笼辣椒茉莉酸氨基合酶基因(Ccjar1)的克隆与表达分析[J]. 分子植物育种,2018,16(22):7241-7247
[9] VALENZUELAalenzuela C E,ACEVEDOCEVEDO-ACEVEDOCEVEDO O,Miranda G S,et al. Salt stress response triggers activation of the jasmonate signaling pathway leading to inhibition of cell elongation in Arabidopsis primary root[J]. Journal of Experimental Botany,2016,67(14):4209-4220
[10] 李晓娜,肖厚贞,万三连,等. 巴西橡胶树生长素响应hbjar1基因克隆与表达分析[J]. 热带作物学报,2017,38(8):1478-1484
[11] WOZNIAK A,KESY J,GLAZINSKA P,et al. The influence of lead and acyrthosiphon pisum (Harris) on generation of pisum sativum defense signaling molecules and expression of genes involved in their biosynthesis[J]. International Journal of Molecular Sciences,2023,24(13):10671-10671
[12] YANG P,ZHANG P,LI B,et al. Effect of nodules on dehydration response in alfalfa (Medicago Sativa L.)[J]. Environmental and Experimental Botany,2013,86:29-34
[13] HE F,ZHANG L,ZHAO G,et al. Genome-wide identification and expression analysis of the nac gene family in alfalfa revealed its potential roles in response to multiple abiotic stresses[J]. International Journal of Molecular Sciences,2022,23(17):10015-10015
[14] 黄馨田,韩慧杰,李宇琛,等. 蒙农杂种冰草acdmyb1基因克隆及表达分析[J]. 草地学报,2023,31(8):2334-2342
[15] LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative pcr and the 2^-Δδct method[J]. Methods,2001,25(4),402-408
[16] PELEG Z, BLUMWALD E. Hormone balance and abiotic stress tolerance in crop plants[J]. Current Opinion in Plant Biology,2011,14(3):290-295
[17] DELFIN J C,KANNO Y,SEO M,et al. Atgh3. 10 is another jasmonic acid-amido synthetase in Arabidopsis thaliana[J]. Plant Journal,2022,110(4):1082-1096
[18] LISCUM E. Faculty opinions recommendation of characterization of an arabidopsis enzyme family that conjugates amino acids to indole-3-acetic acid[J]. Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature,2005;17 (2):616-627
[19] SAMIEE M,KOHNEHROUZ B B, NOROUZI M. Cloning and bioinformatics analysis of accd gene from bell pepper (Capsicum Annuum)[J]. International Journal of Agriculture and Biosciences,2016,5(2):67-72
[20] 张晓峰,焦锋,聂浩,等. 桑树magh3.6基因的克隆与诱导表达及时间特异性[J]. 蚕业科学,2013,39(2):196-203
[21] 常丽丽,吴连成,库丽霞,等. 植物flowering Locus T/Terminal Flower1基因家族的研究进展[J]. 西北植物学报,2008(4):4843-4851
[22] 李春艳,钟华,杜利霞,等. 白羊草bimyb52基因的克隆及转基因拟南芥抗旱性表达分析[J]. 草地学报,2023,31(1):29-39
[23] 黄鑫,夏百成,龙翔宇. 小麦胁迫相关基因Tasf3b2的克隆及表达特性分析[J]. 西北植物学报,2016,36(8):1528-1533
[24] 未丽,刘建利. 植物蛋白质亚细胞定位相关研究概述[J]. 植物科学学报,2021,39(1):93-101
[25] 罗火林,罗丽萍,熊冬金,等. 辣椒gh3基因的电子克隆与生物信息学分析[J]. 华北农学报,2013,28(6):24-29
[26] WASTERNACK CANDAND FEUSSNER I. The oxylipin pathways:biochemistry and function[J]. Annual Review of Plant Biology,2018,69(1):363-386
[27] TSUZUKI H,KITAOKA N,INOUSE S,et al. Airborne methyl jasmonate is metabolized to jasmonic acid and 12-hydroxyjasmonic acid,and induces jasmonate biosynthesis in marchantia polymorpha[J].Chemistry Select,2023,8(21):e202300117
[28] 陈凯,张恒,赵凤佳,等. 番茄茉莉酸-异亮氨酸合成酶基因(Sljar1)的生物信息学鉴定及表达分析[EB/OL]. http://kns.cnkinet/kcms/detail/46.1068.5.20230703.1355.018.html.2023-07-03/2024-01-18
[29] SUZA WANDSTASWICK P E. The role of jar1 in jasmonoyl-L-isoleucine production during arabidopsis wound response[J]. Planta,2008,227(6):1221-1232
[30] ABE H,OHNISHI J,NARUSAKA M,et al. Function of jasmonate in response and tolerance of arabidopsis to thrip feeding[J]. Plant and Cell Physiology,2008,49(1):68-80
[31] CHUNG H S,KOO A J,GAO X,et al. Regulation and function of arabidopsis jasmonate Zim-Domain genes in response to wounding and herbivory[J]. Plant Physiology,2008,146(3):952-964
[32] AI G,HUANG R S,ZHANG D,et al. Slgh3.15,a member of the gh3 gene family,regulates lateral root development and gravitropism. Response by modulating auxin homeostasis in tomato[J]. Plant Science,2023,192(3)：2336-2355
[33] KANG D K,SEO Y J,LEE J D,et al. Jasmonic acid differentially affects growth,ion uptake and abscisic acid concentration in salt-tolerant and salt-sensitive rice cultivars[J]. Journal of Agronomy and Crop Science,2005,191(4):273-282
[34] HEMANTARANJAN A,LALOTRA S, SODANI R. Methyl Jasmonate:a potent player in salinity stress[J]. The Journal of Plant Science Research,2021,37(2):219-224
[35] WU H,YE H,YAO R,et al. Osjaz9 acts as a transcriptional regulator in jasmonate signaling and modulates salt stress tolerance in rice[J]. Plant Science,2015,232:1-12
[36] HAZMAN M,HAUSE B,EICHE E,et al. Increased tolerance to salt stress in opda-deficient rice allene oxide cyclase mutants is linked to an increased ros-scavenging activity[J]. Journal of Experimental Botany,2015,66(11):3339-3352
[37] MAHMUD S,ULLAH C,KORTZ A,et al. Constitutive expression of jasmonate resistant 1 induces molecular changes that prime the plants to better withstand drought[J]. Plant Cell and Environment,2022,45(10):2906-2922

Cloning and Expression Analysis of MsJAR1 in Medicago sativa

紫花苜蓿MsJAR1基因克隆及表达分析

PDF

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	SUO Xin, NIU Qi-Chen, GUO Cheng, GAI Yun-peng, CHEN Ling, YIN Shu-xia. Isolation of a Novel Alfalfa Fusarium Root Rot Pathogen FIESC and Characterization of Strain Biology [J]. Acta Agrestia Sinica, 2024, 32(5): 1327-1338.
[2]	CHEN Qi, DAI Rui, ZHANG Yan, SHUANG Shuang, HUO Xiao-wei, BIAN Lin, GUO Na, ZHANG Zhi-qiang, ZHANG Yu-tong. Genes Expression Analysis of Flavonoid Biosynthesis Induced by Thrips Feeding in Alfalfa [J]. Acta Agrestia Sinica, 2024, 32(4): 1034-1043.
[3]	ZHU Chen-chen, SHI Kun, HE Qin-kun, ZHENG Yu-qin, YUAN Feng, LIU Ya-ling, WANG Zan. Effects of Mixed Saline-Alkali Stress on Physiology and Gene Expression of Alfalfa Seedlings [J]. Acta Agrestia Sinica, 2024, 32(4): 1044-1054.
[4]	ZHOU Zhi-dong, ZHANG Yu, LI Xiang-ru, YANG Shi-jian, YANG Jing, TAN Jing, LIU Ying-cheng, CHEN Yuan-qi. Effects of Waterlogging and Biochar Addition on the Growth and Antioxidant Enzyme Activity of Alfalfa Seedlings [J]. Acta Agrestia Sinica, 2024, 32(4): 1087-1093.
[5]	ZHAO Yao-yao, LIU Mei-jun, ZHANG Zheng, WANG Jie, WANG Yu-xiang, AN Chang-qi, WANG Tiao-xia. Effects of Low Temperature and Drought Stress on Photosystem II of Alfalfa Leaves [J]. Acta Agrestia Sinica, 2024, 32(4): 1120-1130.
[6]	LÜ Yan-zhen, NIE Yu-ting, ZHANG Zhao, CUI Kai-lun, YAN Hui-fang. Comprehensive Evaluation of Seed Storability of Alfalfa and Screening of Key Indicators [J]. Acta Agrestia Sinica, 2024, 32(4): 1131-1141.
[7]	WANG Jie, LIU MEI-jun, ZHAO Yao-yao, ZHANG Zheng, WEI Dan-dan, GONG Liang-zhu, HU Xuan-ming. Effects of Betaine on Respiratory Electron Transport of Alfalfa Seeds During Germination Under Low Temperature [J]. Acta Agrestia Sinica, 2024, 32(3): 763-771.
[8]	JING Fang, SHI Shang-li, NAN Pan, MA Rui-hong, A Yun, LU Bao-fu, GUAN Jian, ZHANG Hui-hui. Comparison of Leaf Characteristics, Photosynthetic Physiological Characteristics, and Yield Traits of Different Alfalfa Varieties [J]. Acta Agrestia Sinica, 2024, 32(2): 369-377.
[9]	LIU Jing-yu, ZHAO Hai-jun, OU Cheng-ming, JIA Zhi-cheng, MAO Pei-sheng. Effects of Melatonin Priming on Alfalfa Seed Germination Characteristics Under Na₂SO₄ Stress [J]. Acta Agrestia Sinica, 2024, 32(2): 378-385.
[10]	YU Jiang-di, LI Yu-zhu, MIAO Jia-min, DING Fei-fei, BAI Xiao-ming, NIU Kui-ju. Cloning and Expression Pattern Analysis of BBML Gene in Poa pratensis(Kentucky Bluegrass) [J]. Acta Agrestia Sinica, 2024, 32(2): 396-408.
[11]	LI Qi-jiao, CUI Dong-yi, DUAN Xin-hui, LIU Li, XU Wen-hua, LI Qing-xi, ZHENG Li-wen, REN Jian, ZHU Li-min, MA Xiang-li. Effects of Melatonin on Seed Germination and Antioxidant Physiology of Alfalfa Seedlings Under Aluminum Stress [J]. Acta Agrestia Sinica, 2024, 32(2): 535-542.
[12]	GONG Ze-lin, EER Demutu, SU Xiao-gang, CHENG Rong-rong, YANG He-ming, ZHANG Xiao-lin, ZHAI Peng-hui. Effects of Water and Fertilizer Addition on Soil Respiration Rate in Grass-field Rotation System [J]. Acta Agrestia Sinica, 2024, 32(2): 562-569.
[13]	LI Xiao-hong, WANG Xiao-tong, MA Xu-xia, CAI Wen-qi, FENG Xue-li, MA Meng-fan, LI Shu-xia. Identification and Analysis of CNGC Gene Family Members in Alfalfa [J]. Acta Agrestia Sinica, 2024, 32(2): 588-598.
[14]	WU Qian-qian, SU Ya, ZENG Xiang-ming, LI Dong-heng, CHENG Wei, HAO Jun. Extraction Method Screening and Process Optimization of Alfalfa and White Clover Selenoprotein [J]. Acta Agrestia Sinica, 2024, 32(2): 654-660.
[15]	LI Xin, WU Jing-ye, CHEN Fei-er, FAN Lu, MA Lin, WANG Xue-min. Effects of Transcriptional Regulation of MsACS2 by MsWRKY33 on Salt Tolerance of Alfalfa [J]. Acta Agrestia Sinica, 2024, 32(1): 54-65.