大叶落地生根KdSRG1基因的克隆、表达分析及自激活检测

doi:10.11733/j.issn.1007-0435.2023.05.009

草地学报 ›› 2023, Vol. 31 ›› Issue (5): 1349-1358.DOI: 10.11733/j.issn.1007-0435.2023.05.009

大叶落地生根KdSRG1基因的克隆、表达分析及自激活检测

栾雅琳, 潘佳妮, 王梦迪, 李殷睿智, 曾会明

北京林业大学草业与草原学院, 北京 100083

收稿日期:2022-11-02 修回日期:2023-01-18 出版日期:2023-05-15 发布日期:2023-05-31
通讯作者: 曾会明,E-mail:sciinfo@bjfu.edu.cn
作者简介:栾雅琳(1995-)，女，汉族，山东泰安人，硕士研究生，主要从事草地植物分子生物学研究，E-mail:yalinluan@163.com

Cloning,Expression Analysis and Self-activation Detection of KdSRG1 Gene from Kalanchoe daigremontiana

LUAN Ya-lin, PAN Jia-ni, WANG Meng-di, LI Yin-rui-zhi, ZENG Hui-ming

School of Grassland Science, Beijing Forestry University, Beijing 100083, China

Received:2022-11-02 Revised:2023-01-18 Online:2023-05-15 Published:2023-05-31

摘要/Abstract

摘要： 为探究大叶落地生根(Kalanchoe daigremontiana)中衰老相关基因1(Senescence-related gene 1,SRG1)的功能,以野生型大叶落地生根为材料,克隆KdSRG1基因,对其进行生物信息学分析、亚细胞定位和基因表达分析。结果表明:KdSRG1基因开放阅读框为219 bp,编码72个氨基酸;KdSRG1蛋白是一个亲水性的不稳定酸性蛋白,定位于细胞核和细胞质,不含跨膜结构和信号肽,并且具有物种上的特异性;该基因在大叶落地生根的不定芽和叶中的相对表达量较高,说明其参与不定芽和叶的生长调控;通过分析其启动子顺式作用元件,推测KdSRG1基因参与植物分生组织表达和栅栏细胞分化过程,并且其表达可能受光信号、脱落酸(Abscisic acid,ABA)和茉莉酸甲酯(Methyl jasmonate,MeJA)调控,同时可能在植物逆境胁迫调控方面发挥作用;自激活检测表明KdSRG1蛋白没有自激活活性,可用于后续的酵母双杂实验。本研究为进一步研究KdSRG1基因的作用机制及功能奠定了基础。

关键词: 大叶落地生根, KdSRG1基因, 基因克隆, 亚细胞定位, 表达分析

Abstract: In order to explore the functions of the senescence-related gene (SRG1) in Kalanchoe daigremontiana,the gene of KdSRG1 was cloned from the wild-type K daigremontiana,and then analyzed for its bioinformatics,subcellular localization,and gene expression analysis. The results showed that the open reading frame of KdSRG1 was 219 bp in length,encoding a peptide of 72 amino acids. The KdSRG1 protein is a hydrophilic unstable acidic protein,which was localized in the nucleus and the cell membrane,and had no transmembrane structures or signal peptides and species specific in property. The relative expression of KdSRG1 gene in K. daigremontiana was higher in its plantlets and leave than that in the roots and stems,which demonstrated that gene may be involved in the regulation of adventitious bud and leaf growth. By analyzing the cis-acting elements of KdSRG1 promoter,it was speculated that KdSRG1 gene would take a role in that plant meristem formation and palisade mesophyll cells differentiation,and the expression of this gene may be regulated by light signal,abscisic acid (ABA),and methyl jasmonate (MeJA). Moreover,the KdSRG1 gene may play a role in that plant stress regulation. The self-activation detection showed that KdSRG1 protein had no self-activation activity and could be used to the subsequent yeast two-hybrid experiments. This study could be used for further research on the functional mechanisms of KdSRG1 gene.

Key words: Kalanchoe daigremontiana, KdSRG1 gene, Gene cloning, Subcellular localization, Expression analysis

中图分类号:

Q781

栾雅琳, 潘佳妮, 王梦迪, 李殷睿智, 曾会明. 大叶落地生根KdSRG1基因的克隆、表达分析及自激活检测[J]. 草地学报, 2023, 31(5): 1349-1358.

LUAN Ya-lin, PAN Jia-ni, WANG Meng-di, LI Yin-rui-zhi, ZENG Hui-ming. Cloning,Expression Analysis and Self-activation Detection of KdSRG1 Gene from Kalanchoe daigremontiana[J]. Acta Agrestia Sinica, 2023, 31(5): 1349-1358.

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参考文献

[1] KAWAI Y S K,ONO E,MIZUTANI M S H R. Evolution and diversity of the 2-oxoglutarate-dependent dioxygenase superfamily in plants[J]. The Plant Journal,2014,78(2):328-343
[2] SCHOFIELD C J,ZHANG Z H. Structural and mechanistic studies on 2-oxoglutarate-dependent oxygenases and related enzymes[J]. Current Opinion in Structural Biology,1999,9(6):722-731
[3] DE CAROLIS E,DE LUCA V. 2-oxoglutarate-dependent dioxygenase and related enzymes:biochemical characterization[J]. Phytochemistry,1994,36(5):1093-1107
[4] ISIAM M S,LEISSING T M,CHOWDHURY R,et al. 2-Oxoglutarate-dependent oxygenases[M]. Annual Review of Biochemistry,2018:87,585-620
[5] WILMOUTH R C,TURNBULL J J,WELFORD R W,et al. Structure and mechanism of anthocyanidin synthase from Arabidopsis thaliana[J]. Structure,2002,10(1):93-103
[6] 刘浩. 拟南芥一个2OG-Fe(II)氧化还原酶在GA合成及其与ABA互作的功能分析[D]. 开封:河南大学,2011:83
[7] WANG L,YU B,ZHAO Y,et al. A putative 2OG-Fe(II) oxygenase's response to gibberellin deficiency is related to the internodal growth of columnar apples[J]. Acta Physiologiae Plantarum,2021,43(5):1-11
[8] PARK S,KIM D H,PARK B R,et al. Molecular and functional characterization of Oryza sativa flavonol synthase (OsFLS),a bifunctional dioxygenase[J]. Journal of Agricultural and Food Chemistry,2019,67(26):7399-7409
[9] SUN F L,CHEN Q J,CHEN Q,et al. Genome-wide identification and expression analysis of the 2OG-Fe(II) oxygenase gene family in upland cotton (Gossypium hirsutum L.)[J]. Physiology and Molecular Biology of Plants,2021,27(9):1969-1977
[10] LI J J,LI Y,YIN Z,et al. OsASR5 enhances drought tolerance through a stomatal closure pathway associated with ABA and H₂O₂ signalling in rice[J]. Plant Biotechnology Journal,2017,15(2):183-196
[11] VAN DAMME M,HUIBERS R P,ELBERSE J,et al. Arabidopsis DMR6 encodes a putative 2OG-Fe(II) oxygenase that is defense-associated but required for susceptibility to downy mildew[J]. Plant Journal,2008,54(5):785-793
[12] FANG L K,ZHAO F M,CONG Y F,et al. Rolling-leaf14 is a 2OG-Fe (II) oxygenase family protein that modulates rice leaf rolling by affecting secondary cell wall formation in leaves[J]. Plant Biotechnology Journal,2012,10(5):524-532
[13] 王熙昂,王彪,李海燕,等. 植物中α-酮戊二酸/Fe(Ⅱ)依赖双加氧酶研究进展[J]. 植物生理学报,2021,57(11):2077-2090
[14] 中国科学院中国植物志编辑委员会.中国植物志[M]. 北京:科学出版社,1984,34(1):11-17
[15] SUPRATMAN U,FUJITA T,AKIYAMA K,et al. Anti-tumor promoting activity of bufadienolides from Kalanchoe pinnata and K. daigremontiana x tubiflora[J]. Bioscience,Biotechnology,and Biochemistry,2001,65(4):947-949
[16] SUPRATMAN U,FUJITA T,AKIYAMA K,et al. New insecticidal bufadienolide,bryophyllin C,from Kalanchoe pinnata[J]. Bioscience,Biotechnology,and Biochemistry,2000,64(6):1310-1312
[17] GARCES H,SINHA N. The 'mother of thousands' (Kalanchoe daigremontiana):a plant model for asexual reproduction and CAM studies[J]. Cold Spring Harbor protocols,2009(10):pdb-emo133
[18] 朱晨. 干旱条件下差异表达基因KdSOC1和KdN41调控开花及不定芽发生机制的研究[D]. 北京:北京林业大学,2017:13
[19] 黄伟伟,杨曦,张常娥,等. 烟草黄酮醇合成酶基因的克隆及其序列分析[J]. 植物生理学通讯,2006(6):1059-1062
[20] 贾会丽,石永红,王学敏,等. 紫花苜蓿MsLEA4启动子的克隆及表达分析[J]. 草地学报,2019,27(4):789-796
[21] LAU O S,DENG X W. Plant hormone signaling lightens up:integrators of light and hormones[J]. Current Opinion in Plant Biology,2010,13(5):571-577
[22] 崔泽田,商贺阳,张金旭,等. 甘蔗2OG-Fe(II)加氧酶基因家族的全基因组鉴定及表达模式分析[J/OL]. 分子植物育种,2022:1-19
[23] 李颖,鱼小军,赵一珊,等. 水杨酸和脱落酸浸种对低温下扁蓿豆种子萌发和幼苗生长的影响[J]. 草地学报,2021,29(1):174-181
[24] WASTERNACK C,HAUSE B. Jasmonates and octadecanoids:signals in plant stress responses and development[J]. Progress in Nucleic Acid Research and Molecular Biology,2002,72:165-221
[25] FERRO E,TRABALZINI L. The yeast two-hybrid and related methods as powerful tools to study plant cell signalling[J]. Plant Molecular Biology,2013,83(4-5):287-301
[26] 朱玉贤,李毅,郑晓峰,等. 现代分子生物学[M]. 第四版. 北京:高等教育出版社,2013:220
[27] CAUSIER B,DAVIES B. Analysing protein-protein interactions with the yeast two-hybrid system[J]. Plant Molecular Biology,2002,50(6):855-870

大叶落地生根KdSRG1基因的克隆、表达分析及自激活检测

Cloning,Expression Analysis and Self-activation Detection of KdSRG1 Gene from Kalanchoe daigremontiana

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