日本落叶松LoERF017基因的克隆及生物信息学分析

doi:10.7525/j.issn.1673-5102.2020.04.015

摘要/Abstract

摘要： 从日本落叶松中克隆得到的LoERF017基因，该基因全长624 bp，共编码206个氨基酸。预测该基因所编码的蛋白只包括一个AP2结构域，其二级结构包括α-螺旋（30.43%），无规卷曲（54.11%），延伸链（12.56%），β-折叠（2.9%）。对该基因进行亚细胞定位，结果显示LoERF017基因定位在细胞核中。荧光定量PCR结果表明，LoERF017基因在日本落叶松中的表达有一定的组织特异性，该基因在根中的表达量最高，在茎中的表达量最低，在叶中的表达量居中。干旱胁迫下，LoERF017基因的表达量在根中呈上升趋势，复水后，该基因的表达量有下降趋势。可以初步预测该基因主要在日本落叶松根中表达并行使功能，并推测该基因可能与抗旱相关。

关键词: 日本落叶松, LoERF017基因, 生物信息学, 非生物胁迫

Abstract: The LoERF017 gene cloned from Larix kaempferi was sequenced and found to be 624 bp in length, encoding a total of 207 amino acids. The protein encoded by LoERF017 is predicted to include only one AP2 domain, and its secondary structure includes α-helix (30.43%), random coil (54.11%), extended strand 12.56%), and β-sheet (2.9%). Subcellular localization of the gene showed that LoERF017 was located in the nucleus. The results of qRT-PCR showed that the expression level of LoERF017 gene in L.kaempferi had certain tissue specificity. The expression of this gene in root was the highest, the expression in stem was the lowest, and the expression in leaves was middle. Under drought stress, the expression of LoERF017 gene increased in the root, but decreased after rehydration. It was preliminarily predicted that this gene is mainly expressed and functions in root of L.kaempferi, and it was speculated that LoERF017 might be related to drought resistance.

Key words: Larix kaempferi, LoERF017, bioinformatics, abiotic stress

中图分类号:

S722.3+6

马苗苗, 李成浩, 刘晓, 张馨, 杨静莉. 日本落叶松LoERF017基因的克隆及生物信息学分析[J]. 植物研究, 2020, 40(4): 602-612.

MA Miao-Miao, LI Cheng-Hao, LIU Xiao, ZHANG Xin, YANG Jing-Li. Cloning and Bioinformatics Analysis of LoERF017 Gene from Larix kaempferi[J]. Bulletin of Botanical Research, 2020, 40(4): 602-612.

参考文献

[1] 中国科学院中国植物志委员会.中国植物志:第七卷:裸子植物门[M].北京:科学出版社,1978.Chinese Flora Committee of Chinese Academy of Sciences.Flora reipublicae popularis sinicae:tomus 7:gymnosperm[M].Beijing:Science Press,1978.
[2] Guan C Y,Zhang H,Zhang L,et al.Construction of genetic linkage maps of larch (Larix kaempferi×Larix gmelini) by rapd markers and mapping of QTLS for larch[J].Biotechnology & Biotechnological Equipment,2011,25(1):2197-2202.
[3] Zhu J J,Yang K,Yan Q L,et al.Feasibility of implementing thinning in even-aged Larix olgensis plantations to develop uneven-aged larch-broadleaved mixed forests[J].Journal of Forest Research,2010,15(1):71-80.
[4] Lange O L,Kappen L,Schulze E D.Waterand plant life:problems and modern approaches[M].Berlin,Heidelberg:Springer-Verlag,1976.
[5] 马常耕,孙晓梅.我国落叶松遗传改良现状及发展方向[J].世界林业研究,2008,21(3):58-63,doi:10.13348/j.cnki.sjlyyj.2008.03.010.Ma C G,Sun X M.Larch genetic improvement and its future development in China[J].World Forestry Research,2008,21(3):58-63,doi:10.13348/j.cnki.sjlyyj.2008.03.010.
[6] Johnson P R,Ecker J R.The ethylene gas signal transduction pathway:a molecular perspective[J].Annual Review of Genetics,1998,32:227-254.
[7] O'donnell P J,Calvert C,Atzorn R,et al.Ethylene as a signal mediating the wound response of tomato plants[J].Science,1996,274(5294):1914-1917.
[8] Penninckx I A M A,Thomma B P H J,Buchala A,et al.Concomitant activation of jasmonate and ethylene response pathways is required for induction of a plant defensin gene in Arabidopsis[J].The Plant Cell,1998,10(12):2103-2114.
[9] Singh K B,Foley R C,Oñate-Sánchez L.Transcription factors in plant defense and stress responses[J].Current Opinion in Plant Biology,2002,5(5):430-436.
[10] Fujimoto S Y,Ohta M,Usui A,et al.Arabidopsis ethylene-responsive element binding factors act as transcriptional activators or repressors of GCC box-mediated gene expression[J].The Plant Cell,2000,12(3):393-404.
[11] Zhuang J,Cai B,Peng R H,et al.Genome-wide analysis of the AP2/ERF gene family in Populus trichocarpa[J].Biochemical and Biophysical Research Communications,2008,371(3):468-474.
[12] Sakuma Y,Liu Q,Dubouzet J G,et al.DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs,transcription factors involved in dehydration-and cold-inducible gene expression[J].Biochemical and Biophysical Research Communications,2002,290(3):998-1009.
[13] 李聪,郭天麒,梁小红,等.植物ERFs类转录因子在逆境胁迫中的作用[J].生物技术通报,2011,(4):1-6.Li C,Guo T L,Liang X H,et al.Function of ERF transcription factors in plant adverse environmental stresses[J].Biotechnology Bulletin,2011,(4):1-6.
[14] 刘欣,李云.转录因子与植物抗逆性研究进展[J].中国农学通报,2006,22(4):61-65.Liu X,Li Y.Transcription factors re1ated to plant stress-tolerance[J].Chinese Agricultural Science Bulletin,2006,22(4):61-65.
[15] 莫纪波,李大勇,张慧娟,等.ERF转录因子在植物对生物和非生物胁迫反应中的作用[J].植物生理学报,2011,47(12):1145-1154.Mo J B,Li D Y,Zhang H J,et al.Roles of ERF transcription factors in biotic and abiotic stress response in plants[J].Plant Physiology Journal,2011,47(12):1145-1154.
[16] Gao S M,Zhang H W,Tian Y,et al.Expression of TERF1 in rice regulates expression of stress-responsive genes and enhances tolerance to drought and high-salinity[J].Plant Cell Reports,2008,27(11):1787-1795.
[17] Pan Y,Seymour G B,Lu C G,et al.An ethylene response factor(ERF5) promoting adaptation to drought and salt tolerance in tomato[J].Plant Cell Reports,2012,31(2):1-12.
[18] Seo Y J,Park J B,Cho Y J,et al.Overexpression of the ethylene-responsive factor gene BrERF4 from Brassica rapa increases tolerance to salt and drought in Arabidopsis plants[J].Molecules and Cells,2010,30(3):271-277.
[19] 宋佳亮.杨树ERF家族成员基因克隆与表达模式分析[D].南京:南京林业大学,2013.Song J L.Cloning and expression of ERF genes in Populus[D].Nanjing:Nanjing Forestry University,2013.
[20] Trujillo L E,Sotolongo M,Menéndez C,et al.SodERF3,a novel sugarcane ethylene responsive factor(ERF),enhances salt and drought tolerance when overexpressed in tobacco plants[J].Plant and Cell Physiology,2008,49(4):512-525.
[21] Zhang G Y,Chen M,Chen X P,et al.Phylogeny,gene structures,and expression patterns of the ERF gene family in soybean(Glycine max L.)[J].Journal of Experimental Botany,2008,59(15):4095-4107.
[22] 林轶.葡萄ERF基因的克隆及功能分析[D].大连:辽宁师范大学,2009.Lin Y.Isolation and function analysis of grape ERF genes[D].Dalian:Liaoning Normal University,2009.
[23] Qiao Z X,Huang B,Liu J Y.Molecular cloning and functional analysis of an ERF gene from cotton(Gossypium hirsutum)[J].Biochimica et Biophysica Acta(BBA)-Gene Regulatory Mechanisms,2008,1779(2):122-127.
[24] 张金兰,张博,黄东益,等.大薯ERF基因家族的挖掘与特征[J].分子植物育种,2019,17(7):2086-2093.Zhang J L,Zhang B,Huang D Y,et al.Identification and characterization of ERF gene family in Great Yam(Dioscorea alata L.)[J].Molecular Plant Breeding,2019,17(7):2086-2093.
[25] 姜扬.干旱胁迫条件下长春花Str和Pme基因表达变化的研究[D].哈尔滨:东北林业大学,2008.Jiang Y.Str and Pme Gene expression in Catharanthus roseus leaves under drought stress[D].Harbin:Northeast Forestry University,2008.
[26] 胡晓晴.一氧化氮处理后长白落叶松转录组测序及LoERF017基因功能研究[D].哈尔滨:东北林业大学,2016.Hu X Q.Transcriptome sequencing of Larix olgensis Henry induced by nitric oxide and functional characterization of LoERF017 gene[D].Harbin:Northeast Forestry University,2016.