软枣猕猴桃ICE1基因克隆与生物信息学分析
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摘要
[目的]为研究软枣猕猴桃ICE1基因的生物学功能及抗寒分子机制奠定基础。[方法]以4℃低温处理24 h的软枣猕猴桃(Actinidia arguta Planch) c DNA为模板,通过反转录PCR技术克隆ICE1基因。[结果]该基因c DNA长1524 bp,可编码507个氨基酸。生物信息学分析显示ICE1基因是编码MYC2型的b HLH转录因子,编码蛋白是定位于细胞核的非跨膜亲水蛋白。推导的氨基酸序列与中华猕猴桃(Actinidia chinensis var. chinensis)和山葡萄(Vitis amurensis) ICE1存在着较高的同源性。[结论]成功克隆到软枣猕猴桃抗寒转录因子命名为AaICE1。
Actinidia arguta Planch cDNA of 24 hours was treated at a low temperature of 4℃ as a template,and ICE1 gene was cloned and named Aa ICE1. Its c DNA is 1 524 bp long and can encode 507 amino acids. Bioinformatics analysis shows that the ICE1 gene is a b HLH transcription factor encoding MYC2 type,and the coding protein is a non-transmembrane hydrophilic protein located in the nucleus. The derived amino acid sequences are homologous with Actinidia chinensis Planch and Vitis amurensis. The cloning of this gene laid the foundation for further research on the cold resistance mechanism of Actinidia arguta Planch.
Actinidia arguta Planch cDNA of 24 hours was treated at a low temperature of 4℃ as a template,and ICE1 gene was cloned and named Aa ICE1. Its c DNA is 1 524 bp long and can encode 507 amino acids. Bioinformatics analysis shows that the ICE1 gene is a b HLH transcription factor encoding MYC2 type,and the coding protein is a non-transmembrane hydrophilic protein located in the nucleus. The derived amino acid sequences are homologous with Actinidia chinensis Planch and Vitis amurensis. The cloning of this gene laid the foundation for further research on the cold resistance mechanism of Actinidia arguta Planch.
引文
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[6]Wang D Z,Jin Y N,Ding X H,et al.Gene regulation and signal transduction in the ICE-CBF-COR signaling pathway during cold stress in plants[J].Biochemistry,2017,82(10):1103-1117.
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[10]崔致学.中国猕猴桃[M].济南:山东科学技术出版社,1993.
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[15]Man L L,Xiang D J,Wang L N,et al.Stress-responsive gene RsICE1 from raphanus sativus increases cold tolerance in rice[J].Protoplasma,2017,254(2):945-956.
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[22]Chinnusamy V,Ohta M,Kanrar S,et al.ICE1:a regulator of coldinduced transcriptome and freezing tolerance in Arabidopsis[J].Genes&Dev,2003,17(8):1043-1054.
[23]Ding Y,Li H,Zhang X,et al.OST1 kinase m odulates freezing tolerance by enhancing ICE1 stability in Arabidopsis[J].Dev.Cell,2015,32(3):278-289.
[24]刘双,陈沼汀,董昭旭,等.拟南芥抗寒基因ICE1的克隆及功能分析[J].西北农林科技大学学报:自然科学版,2017,45(05):175-182.
[25]Zhang T G,Mo J N,Zhou K,et al.Overexpression of brassica campestris BcICE1 gene increases abiotic stress tolerance in tobacco[J].Plant Physiology and Biochemistry,2018,132:515-523.
[26]Feng X M,Zhao Q,Zhao L L,et al.The cold-induced basic helix-loop-helix transcription factor gene MdCIbHLH1 encodes an ICE-like protein in apple[J].BMC Plant Biology,2012,12:22.
[27]Li J T,Wang L N,Zhu W,et al.Characterization of two Vv ICE1genes isolated from‘Muscat Hamburg’grapevine and their effect on the tolerance to abiotic stresses[J].Scientia Horticulturae,2014,165:266-273.
[28]Deng C Y,Ye H Y,Fan M,et al.The rice transcription factors OsICE confer enhanced cold tolerance in transgenic Arabidopsis[J].Plant Signaling&Behavior,2017,12(5).
[29]Lee B H,Henderson D A,Zhu J K.The Arabidopsis cold-responsive transcriptome and its regulation by ICE1[J].The Plant Cell,2005,17:3155-3175.
[2]秦红艳,杨义明,艾军,等.软枣猕猴桃新品种-‘佳绿’的选育[J].果树学报,2015,(04):733-735+520.
[3]陈建权,张梦恬,熊意,等.天山雪莲SiICE2基因克隆及抗寒性分析[J].西北植物学报,2018,38(07):1194-1202.
[4]王意程,王楠,许海峰,等.红肉苹果冷信号基因MdICE1的表达及其蛋白与MdMYB的互作[J].园艺学报,2018,45(05):817-826.
[5]Kim Y S,Lee M,Lee J H,et al.The unified ICE-CBF pathway provides a transcriptional feedback control of freezing tolerance during cold acclimation in Arabidopsis[J].Plant Mol Biol,2015,89(1/2):187-201.
[6]Wang D Z,Jin Y N,Ding X H,et al.Gene regulation and signal transduction in the ICE-CBF-COR signaling pathway during cold stress in plants[J].Biochemistry,2017,82(10):1103-1117.
[7]王艳敏,白卉,张妍妍.小黑杨抗寒转录因子PnsICE1基因克隆与生物信息学分析[J].分子植物育种,2015,13(04):871-876.
[8]Lin M M,Fang J B,Qi X J,et al.i TRAQ-based quantitative proteomic analysis reveals alterations in the metabolism of Actinidia arguta[J].Scientific Reports,2017,7:1-11.
[9]Huang.History of 100 years of domestication and improvement of kiwifruit and gene discovery from genetic introgressed populations in the wild[D].Beijing:Chinese Bulletin of Botany,2009.
[10]崔致学.中国猕猴桃[M].济南:山东科学技术出版社,1993.
[11]赵淑兰,袁富贵,马月申,等.软枣猕猴桃新品种-魁绿[J].园艺学报,1994,21(2):207-208.
[12]黄永红,史修柱,李桂云,等.2016年泰山南麓猕猴桃冻害调查与分析[J].落叶果树,2016,48:17-19.
[13]虞志军,周礼胜,王岚,等.冰冻灾害对庐山植物园猕猴桃生长发育与产量的影响[J].中国南方果树,2012,41:84-88.
[14]张清明.猕猴桃冻害及其预防[J].西北园艺,2008,(10):44-45.
[15]Man L L,Xiang D J,Wang L N,et al.Stress-responsive gene RsICE1 from raphanus sativus increases cold tolerance in rice[J].Protoplasma,2017,254(2):945-956.
[16]Peng P H,Lin C H,Tsai H W,et al.Cold response in Phalaenopsis aphrodite and characterization of PaCBF1 and PaICE1[J].Plant Cell Physiol,2014,55(9):1623-1635.
[17]洪敏,唐月明,潘翠萍,等.枇杷Ej ICE1基因的克隆及其在低温胁迫下的表达分析[J].分子植物育种,2018,16(16):5232-5239.
[18]张倩.蜡梅抗寒相关转录因子CpICE1a、CpICE1b和CpDREB1的克隆及功能初探[D].武汉:华中农业大学,2017.
[19]冯晓明.苹果bHLH基因MdCIbHLH1克隆及功能分析[D].泰安:山东农业大学,2011.
[20]于涛,周桦楠,张海楼,等.甘薯低温胁迫基因Ib ICE1的克隆及表达分析[J].沈阳农业大学学报,2017,48(02):219-225.
[21]全彩虹,李言,田维敏.巴西橡胶树Hb ICE2基因的克隆及表达分析[J].热带生物学报,2017,8(02):133-140.
[22]Chinnusamy V,Ohta M,Kanrar S,et al.ICE1:a regulator of coldinduced transcriptome and freezing tolerance in Arabidopsis[J].Genes&Dev,2003,17(8):1043-1054.
[23]Ding Y,Li H,Zhang X,et al.OST1 kinase m odulates freezing tolerance by enhancing ICE1 stability in Arabidopsis[J].Dev.Cell,2015,32(3):278-289.
[24]刘双,陈沼汀,董昭旭,等.拟南芥抗寒基因ICE1的克隆及功能分析[J].西北农林科技大学学报:自然科学版,2017,45(05):175-182.
[25]Zhang T G,Mo J N,Zhou K,et al.Overexpression of brassica campestris BcICE1 gene increases abiotic stress tolerance in tobacco[J].Plant Physiology and Biochemistry,2018,132:515-523.
[26]Feng X M,Zhao Q,Zhao L L,et al.The cold-induced basic helix-loop-helix transcription factor gene MdCIbHLH1 encodes an ICE-like protein in apple[J].BMC Plant Biology,2012,12:22.
[27]Li J T,Wang L N,Zhu W,et al.Characterization of two Vv ICE1genes isolated from‘Muscat Hamburg’grapevine and their effect on the tolerance to abiotic stresses[J].Scientia Horticulturae,2014,165:266-273.
[28]Deng C Y,Ye H Y,Fan M,et al.The rice transcription factors OsICE confer enhanced cold tolerance in transgenic Arabidopsis[J].Plant Signaling&Behavior,2017,12(5).
[29]Lee B H,Henderson D A,Zhu J K.The Arabidopsis cold-responsive transcriptome and its regulation by ICE1[J].The Plant Cell,2005,17:3155-3175.