甘薯盐胁迫响应基因IbNAC14的克隆、生物信息学及表达模式分析

doi:10.16036/j.issn.1000-2650.2019.06.012

摘要/Abstract

摘要： 【目的】探究IbNAC14基因在甘薯胁迫响应过程中的可能功能。【方法】以耐盐性甘薯品种徐薯22与盐敏感性甘薯品种徐薯32为研究材料,从盐胁迫处理后得到的转录组数据中筛选新的NAC基因,命名为IbNAC14。随后对IbNAC14基因进行系统的生物信息学分析,并应用qRT-PCR方法检测其表达模式。【结果】生物信息学分析显示,IbNAC14基因（GenBank登录号：MH660528）包含一个771 bp的开放阅读框,编码着一个具有典型NAC结构域的蛋白,且该蛋白含有多个磷酸化位点。qRT-PCR分析表明,IbNAC14基因的转录水平受盐和干旱胁迫,以及ABA和ACC处理的诱导,暗示其可能在甘薯胁迫过程中发挥重要功能。【结论】IbNAC14基因可能是参与甘薯胁迫响应的重要候选基因。

关键词: 甘薯, IbNAC14, 生物信息学, 非生物胁迫, 表达分析

Abstract: 【Objective】 The aim of the study was to explore the possible functions of IbNAC14 gene in sweet-potato stress response. 【Method】 In this study, the salt tolerant Xushu 22 and the salt sensitive Xushu 32 were used and the IbNAC14 gene was screened from the salt-treated RNA-seq data. A systematic bioinfo-rmatics analysis of the IbNAC14 protein was performed and its expression pattern was detected by qRT-PCR. 【Result】 Bioinformatics analysis showed that IbNAC14 gene contained a 771 bp open reading frame encoding a protein with a typical NAC domain and IbNAC14 protein contained multiple phosphorylation sites. qRT-PCR analysis showed that IbNAC14 expression was induced by salt and drought stresses, as well as ABA and ACC treatments, suggesting that IbNAC14 might play important roles in the stress response of sweetpotato. 【Conclusion】 IbNAC14 gene may be an important candidate gene involved in sweetpotato stress response.

Key words: sweetpotato, IbNAC14, bioinformatics analysis, abiotic stress, expression analysis

中图分类号:

Q812

余静, 孟小庆, 娜菲莎·艾买提江, 李格, 张颖, 李淑清, 朱明库. 甘薯盐胁迫响应基因IbNAC14的克隆、生物信息学及表达模式分析[J]. 四川农业大学学报, 2019, 37(06): 828-835.

YU Jing, MENG Xiaoqing, AMATJIAN Nafisa, LI Ge, ZHANG Ying, LI Shuqing, ZHU Mingku. Cloning,Bioinformatics and Expression Analysis of Salt-Responsive IbNAC14 Gene from Sweetpotato (Ipomoea batatas)[J]. Journal of Sichuan Agricultural University, 2019, 37(06): 828-835.

参考文献

[1] NAKASHIMA K, ITO Y, YAMAGUCHI-SHINOZAKI K.Tra-nscriptional regulatory networks in response to abiotic stresses in Arabidopsis and grasses[J]. Plant Physiology, 2009, 149(1):88-95.
[2] ZHU M, MENG X, CAI J, et al.Basic leucine zipper transcri-ption factor SlbZIP1 mediates salt and drought stress tolerance in tomato[J]. BMC Plant Biology, 2018, 18(1): 83.
[3] ZHU J K.Abiotic stress signaling and responses in plants[J]. Cell, 2016, 167(2): 313-324.
[4] MENG X, LI G, YU J, et al.Isolation,expression analysis,and function evaluation of 12 novel stress-responsive genes of NAC transcription factors in sweetpotato[J]. Crop Science, 2018(58):1328-1341.
[5] NAKASHIMA K, TAKASAKI H, MIZOI J, et al.NAC transc-ription factors in plant abiotic stress responses[J]. Biochimica et Biophysica Acta (BBA)-Gene Regulatory Mechanisms, 2012, 1819(2): 97-103.
[6] PURANIK S, SAHU P P, SRIVASTAVA P S, et al.NAC proteins:regulation and role in stress tolerance[J]. Trends in Plant Science, 2012, 17(6): 369-381.
[7] OLSEN A N, ERNST H A, LEGGIO L L, et al.DNA-binding specificity and molecular functions of NAC transcription factors[J]. Plant science, 2005, 169(4): 785-797.
[8] TRAN L S P, NISHIYAMA R, YAMAGUCHI-SHINOZAKI K,et al. Potential utilization of NAC transcription factors to enhance abiotic stress tolerance in plants by biotechnological approach[J]. GM Crops, 2010, 1(1): 32-39.
[9] XIE Q, FRUGIS G, COLGAN D, et al.Arabidopsis NAC1 tra-nsduces auxin signal downstream of TIR1 to promote lateral root development[J]. Genes & Development, 2000, 14(23): 3024-3036.
[10] PURANIK S, SAHU P P, SRIVASTAVA P S,et al.NAC prot-eins:regulation and role in stress tolerance[J]. Trends in Plant Science,2012(17): 369-381.
[11] 朱明库. 番茄SlNAC4和SlDEAD31基因在果实成熟及非生物胁迫响应中的功能研究[D]. 重庆:重庆大学,2015.
[12] FUJITA M, FUJITA Y, MARUYAMA K,et al.A dehydration-induced NAC protein,RD26,is involved in a novel ABA-depe-ndent stress-signaling pathway[J]. The Plant Journal, 2004, 39(6): 863-876.
[13] HU H, DAI M, YAO J, et al.Overexpressing a NAM,ATAF,and CUC (NAC) transcription factor enhances drought resistance and salt tolerance in rice[J]. Proceedings of the National Acad-emy of Sciences, 2006, 103(35): 12987.
[14] NAKASHIMA K, TRAN L S P, VAN NGUYEN D,et al. Fun-ctional analysis of a NAC-type transcription factor OsNAC6 involved in abiotic and biotic stress-responsive gene expression in rice[J]. The Plant Journal, 2007, 51(4): 617-630.
[15] ZHU M, CHEN G, ZHANG J, et al.The abiotic stress-respo-nsive NAC-type transcription factor SlNAC4 regulates salt and drought tolerance and stress-related genes in tomato (Solanum lycopersicum)[J]. Plant cell reports, 2014, 33(11): 1851-1863.
[16] 马代夫, 李强, 曹清河, 等. 中国甘薯产业及产业技术的发展与展望[J]. 江苏农业学报, 2012, 28(5): 969-973.
[17] CHEN S P, KUO C H, LU H H, et al.The Sweet Potato NAC-Domain Transcription Factor IbNAC1 Is Dynamically Coordinated by the Activator IbbHLH3 and the Repressor IbbHLH4 to Rep-rogram the Defense Mechanism against Wounding[J]. Plos Ge-netics, 2016, 12(10): e1006397.
[18] PARK S C, KIM Y H, CHANG Y J, et al.Stable Internal Reference Genes for the Normalization of Real-Time PCR in Different Sweetpotato Cultivars Subjected to Abiotic Stress Con-ditions[J]. Plos One, 2012, 7(12): e51502.
[19] ZHU M, HU Z, ZHOU S, et al.Molecular characterization of six tissue-specific or stress-inducible genes of NAC transcrip-tion factor family in tomato (Solanum lycopersicum)[J]. Journal of Plant Growth Regulation, 2014, 33(4): 730-744.
[20] JOHN I, HACKETT R, COOPER W, et al.Cloning and char-acterization of tomato leaf senescence-related cDNAs[J]. Plant Molecular Biology, 1997, 33(4): 641-651.
[21] OOKA H, SATOH K, DOI K, et al. Comprehensive analysis of NAC family genes in Oryza sativa and Arabidopsis thaliana[J]. DNA Research, 2003, 10(6)： 239-247.
[22] LU P, CHEN N, AN R, et al.A novel drought-inducible gene,ATAF1,encodes a NAC family protein that negatively regulates the expression of stress-responsive genes in Arabidopsis[J]. Plant Molecular Biology, 2007, 63(2): 289-305.
[23] DELESSERT C, KAZAN K, WILSON I W, et al.The transcr-iption factor ATAF2 represses the expression of pathogenesis-related genes in Arabidopsis[J]. The Plant Journal, 2005, 43(5): 745-757.
[24] TRAN L S P, NAKASHIMA K, SAKUMA Y, et al. Isolation and functional analysis of Arabidopsis stress-inducible NAC transcription factors that bind to a drought-responsive cise-lement in the early responsive to dehydration stress 1 promoter[J]. The Plant Cell, 2004, 16(9): 2481-2498.
[25] HE X, MU R, CAO W, et al.AtNAC2,a transcription factor downstream of ethylene and auxin signaling pathways,is involv-ed in salt stress response and lateral root development[J]. The Plant Journal, 2005, 44(6): 903-916.
[26] TAKADA S, HIBARA K, ISHIDA T, et al.The CUP-SHAPED COTYLEDON1 gene of Arabidopsis regulates shoot apical meri-stem formation[J]. Development, 2001, 128(7): 1127-1135.
[27] AIDA M, ISHIDA T, FUKAKI H, et al.Genes involved in organ separation in Arabidopsis:an analysis of the cup-shaped cotyl-edon mutant[J]. The Plant Cell, 1997, 9(6): 841-857.
[28] GUO Y, GAN S.AtNAP,a NAC family transcription factor, has an important role in leaf senescence[J]. The Plant Journal, 2006, 46(4): 601-612.
[29] CHEN X, LU S, WANG Y, et al.OsNAC2 encoding a NAC transcription factor that affects plant height through mediating the gibberellic acid pathway in rice[J]. Plant Journal for Cell & Molecular Biology, 2015, 82(2): 302-314.
[30] RUSHTON P J, BOKOWIEC M T, HAN S, et al.Tobacco transcription factors:novel insights into transcriptional regulation in the Solanaceae[J]. Plant Physiology, 2008, 147(1): 280-295.
[31] TANG Y, LIU M, GAO S, et al.Molecular characterization of novel TaNAC genes in wheat and overexpression of TaNAC2a confers drought tolerance in tobacco[J]. Physiologia Plantarum,2012, 144(3): 210-224.
[32] FANG Y, YOU J, XIE K, et al.Systematic sequence analysis and identification of tissue-specific or stress-responsive genes of NAC transcription factor family in rice[J]. Molecular Genet-ics and Genomics, 2008, 280(6): 547-563.
[33] SKRIVER K, JENSEN M K, KJAERSGAARD T, et al.The Arabidopsis thaliana NAC transcription factor family:structure-function relationships and determinants of ANAC019 stress sig-naling[J]. Biochem. J.,2010(426):183-196.
[34] ZELLER G, HENZ S R, WIDMER C K, et al.Stress-induced changes in the Arabidopsis thaliana transcriptome analyzed using whole-genome tiling arrays[J]. The Plant Journal, 2009, 58(6):1068-1082.
[35] TRAN L S P, QUACH T N, GUTTIKONDA S K, et al. Molec-ular characterization of stress-inducible GmNAC genes in soybe-an[J]. Molecular Genetics and Genomics, 2009, 281(6): 647-664.
[36] KAZAN K.Diverse roles of jasmonates and ethylene in abiotic stress tolerance[J]. Trends in Plant Science, 2015, 20(4):219-229.
[37] HU H, DAI M, YAO J, et al.Overexpressing a NAM,ATAF,and CUC (NAC) transcription factor enhances drought resistance and salt tolerance in rice[J]. Proceedings of the National Acad-emy of Sciences, 2006, 103(35): 12987-12992.