苦荞转录因子基因FtMYC的克隆及其表达与花青素积累的相关性分析
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摘要
【目的】克隆苦荞FtMYC基因,分析非生物胁迫对FtMYC基因表达及花青素含量的影响。【方法】根据苦荞转录组数据克隆FtMYC基因,对其进行生物信息学分析;通过4℃和UV-B胁迫处理芽期苦荞,分析胁迫后苦荞胚轴和子叶中FtMYC基因表达量与花青素积累之间的相关性。【结果】苦荞FtMYC基因DNA全长1 309 bp,由1个外显子和1个内含子构成,符合GT-AG剪切原则;c DNA序列包括1个1 287 bp的开放阅读框,编码428个氨基酸;在进化树上它与参与花青素调控的其他MYC蛋白聚为一簇;UV-B逆境胁迫处理下,苦荞胚轴中FtMYC基因表达量与花青素含量变化显著性相关(r=0.798,P<0.05),在子叶中则极显著相关(r=0.887,P<0.01);在4℃冷胁迫处理下苦荞胚轴中FtMYC的表达量与花青素积累没有明显的相关性(r=0.744),但在子叶中FtMYC的基因表达与花青素积累显著性相关(r=0.814,P<0.05)。此外,对FtMYC基因启动子的序列分析结果显示,pFTMYC序列包含TATA-Box和CAAT-Box等启动子核心元件以及与光、低温和胁迫应答等相关的功能元件。【结论】FtMYC基因可通过参与花青素代谢调控来参与苦荞对UV-B和寒冷等非生物胁迫的应答和适应。
【Objective】Cloning the FtMYC gene and analyzing the effect of abiotic stress on FtMYC expression and anthocyanin content in the tartary buckwheat.【Method】The FtMYC gene was cloned from the data of tartary buckwheat transcriptional group,and analyzed by bioinformatic method;Tartary buckwheat was treated with 4 ℃ and UV-B treatment,and the correlation between FtMYC expression and anthocyanin content was analyzed in the hypocotyl and cotyledons.【Result】The DNA sequence of FtMYC was1 309 bp in length and contained 1 exon and 1 intron,and in line with the principle of GT-AG splicing principle. The cDNA of FtMYC contains a 1 287 bp ORF,which encoded 428 amino acids residues. FtMYC was clustered with other MYC proteins involved in anthocyanin regulation. Under UV-B stress,FtMYC expression was significantly correlated with the anthocyanin content in hypocotyl(r=0.798,P<0.05) and cotyledon(r=0.887,P<0.01). There was no significant correlation between the expression of FtMYC and anthocyanin accumulation in tartary buckwheat under 4 ℃ stress(r=0.744),but the expression of FtMYC in the cotyledon was significant correlation with anthocyanin accumulation(r=0.814,P<0.05). In addition,sequence analysis of the FtMYC gene promoter revealed that pFtMYC contains promoter core elements such as TATA-Box and CAAT-Box as well as functional elements related to light,hypothermia and stress responses.【Conclusion】The FtMYC gene could be involved in the response and adaptation of buckwheat to abiotic stresses such as UV-B and cold by regulating anthocyanin metabolism.
【Objective】Cloning the FtMYC gene and analyzing the effect of abiotic stress on FtMYC expression and anthocyanin content in the tartary buckwheat.【Method】The FtMYC gene was cloned from the data of tartary buckwheat transcriptional group,and analyzed by bioinformatic method;Tartary buckwheat was treated with 4 ℃ and UV-B treatment,and the correlation between FtMYC expression and anthocyanin content was analyzed in the hypocotyl and cotyledons.【Result】The DNA sequence of FtMYC was1 309 bp in length and contained 1 exon and 1 intron,and in line with the principle of GT-AG splicing principle. The cDNA of FtMYC contains a 1 287 bp ORF,which encoded 428 amino acids residues. FtMYC was clustered with other MYC proteins involved in anthocyanin regulation. Under UV-B stress,FtMYC expression was significantly correlated with the anthocyanin content in hypocotyl(r=0.798,P<0.05) and cotyledon(r=0.887,P<0.01). There was no significant correlation between the expression of FtMYC and anthocyanin accumulation in tartary buckwheat under 4 ℃ stress(r=0.744),but the expression of FtMYC in the cotyledon was significant correlation with anthocyanin accumulation(r=0.814,P<0.05). In addition,sequence analysis of the FtMYC gene promoter revealed that pFtMYC contains promoter core elements such as TATA-Box and CAAT-Box as well as functional elements related to light,hypothermia and stress responses.【Conclusion】The FtMYC gene could be involved in the response and adaptation of buckwheat to abiotic stresses such as UV-B and cold by regulating anthocyanin metabolism.
引文
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[15] XIEX, LI S, ZHANG R, et al. The bHLH transcription factor MdbHLH3 promotes anthocyanin accumulation and fruit colouration in response to low temperature in apples[J]. Plant Cell Environment, 2012, 35(11):1884-1897.
[16]张椿雨,龙艳,冯吉,等.植物基因在转录水平上的调控及其生物学意义[J].遗传,2007, 29(7):793-799.
[17]李田,孙景宽,刘京涛.植物启动子研究进展[J].生物技术通报,2015, 31(2):18-25.
[18] YAO P F,LH C L,ZHAO X R,et al. Overexpression of a tartary buckwheatgene,FtbHLH3, enhances drought/oxidative stress tolerance in transgenic Arabidopsis[J]. Frontiers in Plant Science, 2017(8):625.
[2]邹凤莲,寿森炎,叶纨芝,等.类黄酮化合物在植物胁迫反应中作用的研究进展[J].中国细胞生物学学报,2004, 26(1):39-45.
[3] LI S,BAI Y,LI C,et al. Anthocyanins accumulate in tartary buckwheat(Fagopyrum tataricum)sprout in response to cold st-ress[J]. Acta Physiologiae Plantarum, 2015, 37(8):159.
[4] JEON J,KIM J,WU Q,et al. Comparative transcriptomic and metabolomic analyses in response to cold in tartary buckwheat(Fagopyrum tataricum)[J]. Bio Rxiv, 2018:278432.
[5] BOVY A,De V R,KEMPER M,et al. High-flavonol tomatoes resulting from the heterologous expression of the maize transcription factor genes LC and C1[J]. Plant Cell, 2002, 14(10):2509-2526.
[6] ZHANG F, GONZALEZ A, ZHAL M, et al. A network of red-undant bHLH proteins functions in all TTG1-dependent pathw-ays of Arabidopsis[J]. Development, 2003, 130(20):4859-4869.
[7] GONG Z Z, YAMAGISHI E, YAMAZAKI M, et al. A constitutively expressed Myc-like gene involved in anthocyanin biosynthesis from Perilla frutescens:molecular characterization, heterologous expression in transgenic plants and transactivation in yeast cells[J]. Plant Molecular Biology, 1999, 41(1):33-44.
[8]高丽华,刘博欣,李金博,等.陆地棉bHLH转录因子GhMYC4基因的克隆及功能分析[J].中国农业科技导报,2016, 18(5):33-41.
[9] HEIM M, JAKOBY M, WERBER M, et al. The basic HelixLoop-Helix transcription factor family in plants:A genome-wide study of protein structure and functional diversit[J]. Molecular Biology and Evolution,2003,20(5):735-747.
[10] RBOUTORRENT J, ROIGVILLANOVA I, GALSTYAN A, et al.PARI and PAR2 integrate shade and hormone transcriptional networks[J]. Plant Signaling&Behavior,2008,3(7):453-454.
[11]光杨其,宋桂成,张金凤,等.1个新棉花bHLH类基因GhbH LH130的克隆及表达分析[J].棉花学报,2014, 26(4):363-370.
[12] HARBORNE J, WILLIAMS C. Advances in flavonoid research since 1992[J]. Phytochemistry, 2000, 32(7):481-504.
[13] FOYER C, LELANDAIS M, KUNERT K. Photooxidative stress in plants[J]. Physiologia Plantarum, 1994, 92(4):696-717.
[14]罗春丽,王林,李杏,等.紫薯花青素体外抗氧化及对H2O2诱导HepG2细胞氧化损伤的保护作用[J].食品科学,2015,36(3):225-230.
[15] XIEX, LI S, ZHANG R, et al. The bHLH transcription factor MdbHLH3 promotes anthocyanin accumulation and fruit colouration in response to low temperature in apples[J]. Plant Cell Environment, 2012, 35(11):1884-1897.
[16]张椿雨,龙艳,冯吉,等.植物基因在转录水平上的调控及其生物学意义[J].遗传,2007, 29(7):793-799.
[17]李田,孙景宽,刘京涛.植物启动子研究进展[J].生物技术通报,2015, 31(2):18-25.
[18] YAO P F,LH C L,ZHAO X R,et al. Overexpression of a tartary buckwheatgene,FtbHLH3, enhances drought/oxidative stress tolerance in transgenic Arabidopsis[J]. Frontiers in Plant Science, 2017(8):625.