大豆GmRLP19基因克隆及胁迫应答模式分析
|
摘要
以大豆品种Williams82为试验材料,克隆GmRLP19基因,通过生物信息学分析qPCR检测探讨该基因不同生育期品种、不同组织、不同非生物胁迫下表达模式。结果表明,该基因编码区全长3 138 bp,编码1 045个氨基酸,编码蛋白具有胞外信号肽,LRRNT域,富含亮氨酸重复结构域,跨膜结构域和一段胞内短肽,属于受体类蛋白家族,定位于质膜。该基因在不同生育期品种间差异表达,在豆和豆荚中特异性表达,且在5种非生物胁迫中均有响应。研究为探究WRKY20基因功能及挖掘和完善基因调控通路奠定基础。
In this paper, the cloning and bioinformatics analysis of the gene was carried out using the soybean(Glycine max) variety Williams82. The expression pattern of the gene at different growth stages, different tissues and different abiotic stresses was further investigated by qPCR. The results showed that the coding region of this gene was 3 138 bp in length and encoded 1 045 amino acids. The encoded protein had an extracellular signal peptide, LRRNT domain, leucine-rich repeat domain,transmembrane domain and a short intracellular peptide. It belonged to the family of RLP and was localized to the plasma membrane. The gene was differentially expressed at different growth stages,specifically expressed at beans and pods, and responded in five abiotic stresses. The research laid the foundation for further exploring the function of the gene and excavating and perfecting the WRKY20 gene regulatory pathway.
In this paper, the cloning and bioinformatics analysis of the gene was carried out using the soybean(Glycine max) variety Williams82. The expression pattern of the gene at different growth stages, different tissues and different abiotic stresses was further investigated by qPCR. The results showed that the coding region of this gene was 3 138 bp in length and encoded 1 045 amino acids. The encoded protein had an extracellular signal peptide, LRRNT domain, leucine-rich repeat domain,transmembrane domain and a short intracellular peptide. It belonged to the family of RLP and was localized to the plasma membrane. The gene was differentially expressed at different growth stages,specifically expressed at beans and pods, and responded in five abiotic stresses. The research laid the foundation for further exploring the function of the gene and excavating and perfecting the WRKY20 gene regulatory pathway.
引文
[1]梁建秋,张明荣,吴海英.大豆抗旱性研究进展[J].大豆科学,2010,29(2):341-346.
[2]Benjamin P,Stéphane H,Sébastien D,et al.Genome analysis of poplar LRR-RLP gene clusters reveals RISP,a defense-related gene coding a candidate endogenous peptide elicitor[J].Frontiers in Plant Science,2014,5:111.
[3]吴锦斌,宋银,徐姗姗,等.植物类受体蛋白参与植物生长与发育的研究进展[J].植物生理学报,2012,48(12):1121-1127.
[4]Wang G D,Fiers M,Ellendorff U,et al.The diverse roles of extracellular leucine-rich repeat-containing receptor-like proteins in plants[J].Critical Reviews in Plant Sciences,2010,29(5):285-299.
[5]Takken F L,Thomas C M,Joosten M H,et al.A second gene at the tomato Cf-4 locus confers resistance to Cladosporium fulvum through recognition of a novel avirulence determinant[J].Plant Journal for Cell&Molecular Biology,1999,20(3):279-288.
[6]Wang G,Zhang Z,Angenent G C,et al.New aspects of CLAVATA2,a versatile gene in the regulation of Arabidopsis development[J].Journal of Plant Physiology,2011,168(4):403-407.
[7]Taguchi-Shiobara F,Yuan Z,Hake S,et al.The fasciated ear2gene encodes a leucine-rich repeat receptor-like protein that regulates shoot meristem proliferation in maize[J].Genes&Development,2001,15(20):2755-2766.
[8]Bhave N S,Veley K M,Nadeau J A,et al.TOO MANY MOUTHSpromotes cell fate progression in stomatal development of Arabidopsis stems[J].Planta,2009,229(2):357-367.
[9]Wu J,Liu Z,Zhao Z,et al.Transcriptional regulation of receptorlike protein genes by environmental stresses and hormones and their overexpression activities in Arabidopsis thaliana[J].Journal of Experimental Botany,2016,67(11):3339-3351.
[10]Luo X,Bai X,Sun X,et al.Expression of wild soybean WRKY20in Arabidopsis enhances drought tolerance and regulates ABAsignalling[J].Journal of Experimental Botany,2013,64(8):2155-2169.
[11]Tang L,Cai H,Zhai H,et al.Overexpression of Glycine soja WRKY20 enhances both drought and salt tolerance in transgenic alfalfa(Medicago sativa L.)[J].Plant Cell,Tissue and Organ Culture,2014,118(1):77-86.
[12]Wang G,Ursula E,Ben K,et al.A genome-wide functional investigation into the roles of receptor-like proteins in Arabidopsis[J].Plant Physiol,2008,147(2):503-517.
[13]潘君飞,彭丽云,赵春丽,等.苋菜amaARF6基因克隆及其外源激素处理下在幼苗中响应分析[J].东北农业大学学报,2018,49(5):27-35.
[14]Fritz-Laylin L K,Nandini K,Mahmut T,et al.Phylogenomic analysis of the receptor-like proteins of rice and Arabidopsis[J].Plant Physiology,2005,138(2):611-623.
[15]Stergiopoulos I,Wit P J G M De.Fungal effector proteins[J].Annual Review of Phytopathology,2009,47(1):233-263.
[16]Pan L X,Lv S,Yang N,et al.The multifunction of CLAVATA2 in plant development and immunity[J].Frontiers in Plant Science,2016,7:1-7.
[17]宁文峰,庞添,杨艳玲,等.大豆GmWRKY20基因表达特性研究[J].大豆科学,2016,35(5):748-753.
[18]Magali L,Patrice D,Gert T,et al.PlantCARE,a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences[J].Nucleic Acids Research,2002,30(1):325-327.
[19]薛仁镐,金圣爱.茉莉酸甲酯:一种重要的植物信号转导分子[J].生物技术通讯,2006,17(6):985-988.
[20]Yamamoto S,Nakano T,Suzuki K,et al.Elicitor-induced activation of transcription via W box-related cis-acting elements from a basic chitinase gene by WRKY transcription factors in tobacco[J].Biochimica Et Biophysica Acta Gene Structure&Expression,2004,1679(3):279-287.
[2]Benjamin P,Stéphane H,Sébastien D,et al.Genome analysis of poplar LRR-RLP gene clusters reveals RISP,a defense-related gene coding a candidate endogenous peptide elicitor[J].Frontiers in Plant Science,2014,5:111.
[3]吴锦斌,宋银,徐姗姗,等.植物类受体蛋白参与植物生长与发育的研究进展[J].植物生理学报,2012,48(12):1121-1127.
[4]Wang G D,Fiers M,Ellendorff U,et al.The diverse roles of extracellular leucine-rich repeat-containing receptor-like proteins in plants[J].Critical Reviews in Plant Sciences,2010,29(5):285-299.
[5]Takken F L,Thomas C M,Joosten M H,et al.A second gene at the tomato Cf-4 locus confers resistance to Cladosporium fulvum through recognition of a novel avirulence determinant[J].Plant Journal for Cell&Molecular Biology,1999,20(3):279-288.
[6]Wang G,Zhang Z,Angenent G C,et al.New aspects of CLAVATA2,a versatile gene in the regulation of Arabidopsis development[J].Journal of Plant Physiology,2011,168(4):403-407.
[7]Taguchi-Shiobara F,Yuan Z,Hake S,et al.The fasciated ear2gene encodes a leucine-rich repeat receptor-like protein that regulates shoot meristem proliferation in maize[J].Genes&Development,2001,15(20):2755-2766.
[8]Bhave N S,Veley K M,Nadeau J A,et al.TOO MANY MOUTHSpromotes cell fate progression in stomatal development of Arabidopsis stems[J].Planta,2009,229(2):357-367.
[9]Wu J,Liu Z,Zhao Z,et al.Transcriptional regulation of receptorlike protein genes by environmental stresses and hormones and their overexpression activities in Arabidopsis thaliana[J].Journal of Experimental Botany,2016,67(11):3339-3351.
[10]Luo X,Bai X,Sun X,et al.Expression of wild soybean WRKY20in Arabidopsis enhances drought tolerance and regulates ABAsignalling[J].Journal of Experimental Botany,2013,64(8):2155-2169.
[11]Tang L,Cai H,Zhai H,et al.Overexpression of Glycine soja WRKY20 enhances both drought and salt tolerance in transgenic alfalfa(Medicago sativa L.)[J].Plant Cell,Tissue and Organ Culture,2014,118(1):77-86.
[12]Wang G,Ursula E,Ben K,et al.A genome-wide functional investigation into the roles of receptor-like proteins in Arabidopsis[J].Plant Physiol,2008,147(2):503-517.
[13]潘君飞,彭丽云,赵春丽,等.苋菜amaARF6基因克隆及其外源激素处理下在幼苗中响应分析[J].东北农业大学学报,2018,49(5):27-35.
[14]Fritz-Laylin L K,Nandini K,Mahmut T,et al.Phylogenomic analysis of the receptor-like proteins of rice and Arabidopsis[J].Plant Physiology,2005,138(2):611-623.
[15]Stergiopoulos I,Wit P J G M De.Fungal effector proteins[J].Annual Review of Phytopathology,2009,47(1):233-263.
[16]Pan L X,Lv S,Yang N,et al.The multifunction of CLAVATA2 in plant development and immunity[J].Frontiers in Plant Science,2016,7:1-7.
[17]宁文峰,庞添,杨艳玲,等.大豆GmWRKY20基因表达特性研究[J].大豆科学,2016,35(5):748-753.
[18]Magali L,Patrice D,Gert T,et al.PlantCARE,a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences[J].Nucleic Acids Research,2002,30(1):325-327.
[19]薛仁镐,金圣爱.茉莉酸甲酯:一种重要的植物信号转导分子[J].生物技术通讯,2006,17(6):985-988.
[20]Yamamoto S,Nakano T,Suzuki K,et al.Elicitor-induced activation of transcription via W box-related cis-acting elements from a basic chitinase gene by WRKY transcription factors in tobacco[J].Biochimica Et Biophysica Acta Gene Structure&Expression,2004,1679(3):279-287.