紫化茶树UDP-糖基转移酶启动子的克隆及其生物信息学分析
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摘要
以紫化茶树武夷奇种C18的叶片为材料,采用染色体步移技术获得该材料的UDP-糖基转移酶(UDPG)基因5′端上游启动子序列大小为902 bp,利用软件进行生物信息学分析.结果表明,该调控序列含有启动子核心元件TATA-box、CAAT-motif以及多个光响应元件(GT1-motif、GATA-motif等)、胚乳特异性表达元件(GCN4-motif、Skn-1-motif)、水杨酸响应元件(TCA-element)等特殊顺式作用元件.
Genomewalking method was used to clone the promoter sequence of UDP-glycosyltransferases(UDPG) gene from the purple bud and leaf of tea tree sinensis Wuyiqizhong C18, and finally a 902 bp fragment flanking 5′-upstream of UDPG was obtained. Bioinformatics analysis by software showed that the promoter of UDPG contains core promoter elements of TATA-box and CAAT-motif, several light responsive elements(CT1-motif, GATA-motif, et al), endosperm expression elements(GCN4-motif, Skn-1-motif, et al), and a salicylic acid responsiveness element of TCA-element.
Genomewalking method was used to clone the promoter sequence of UDP-glycosyltransferases(UDPG) gene from the purple bud and leaf of tea tree sinensis Wuyiqizhong C18, and finally a 902 bp fragment flanking 5′-upstream of UDPG was obtained. Bioinformatics analysis by software showed that the promoter of UDPG contains core promoter elements of TATA-box and CAAT-motif, several light responsive elements(CT1-motif, GATA-motif, et al), endosperm expression elements(GCN4-motif, Skn-1-motif, et al), and a salicylic acid responsiveness element of TCA-element.
引文
[1] 宛晓春.茶叶生物化学[M].北京:中国农业出版社,2003.
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[3] 唐秀华,周喆,唐琴,等.茶树CsPAL3基因cDNA全长克隆及其表达分析[J].茶叶科学,2018,38(1):33-42.
[4] 王惠聪,黄旭明,胡桂兵,等.荔枝果皮花青苷合成与相关酶的关系研究[J].中国农业科学,2004,37(12):2 028-2 032.
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[13] 郝迪萩,赵琳,陈李淼,等.克隆启动子方法的比较及应用[J].东北农业大学学报,2010,41(3):154-160.
[14] LIN Y L,LAI Z X.Superoxide dismutase multigene family in longan somatic embryos:a comparison of Cu Zn-SOD,Fe-SOD,and Mn-SOD gene structure,splicing,phylogeny,and expression [J].Molecular Breeding,2013,32(3):595-615.
[15] 练从龙,卢秉国,赖钟雄,等.荔枝古树胚性愈伤组织LcCu/Zn-SOD3基因启动子的克隆及功能验证[J].西北植物学报,2015,35(1):16-22.
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[17] 金琦芳,陈志丹,孙威江,等.茶树CsANS基因及其启动子的克隆与生物信息学分析[J].茶叶科学,2016,36(2):219-228.
[18] XU G,MA H,NEI M,et al.Evolution of F-box genes in plants:different modes of sequence divergence and their relationships with functional diversification [J].Proc Natl Acad Sci USA,2009,106(3):835-840.
[19] JONES P,MESSNER B,NAKAJIMA J I,et al.UGT73C6 and UGT78D1,glycosyltransferases involved in flavonol glycoside biosynthesis in Arabidopsis thaliana [J].Journal of Biological Chemistry,2003,278:43 910-43 918.
[20] ONO E,YU H,HORIKAWA M,et al.Functional differentiation of the glycosyltransferases that contribute to the chemical diversity of bioactive flavonol glycosides in grapevines (Vitis vinifera) [J].The Plant Cell,2010,22(8):2 856-2 871.
[21] HALL D,YUAN X X,MURATA J,et al.Molecular cloning and biochemical characterization of the UDP-glucose:flavonoid 3-O-glucosyltransferase from Concord grape (Vitis labrusca) [J].Phytochemistry,2012,74(3):90-99.
[22] HALL D,KIM K H,LUCA V D.Molecular cloning and biochemical characterization of three concord grape (Vitis labrusca) flavonol 7-O-glucosyltransferases [J].Planta,2011,234:1 201-1 214.
[23] GRUBB C D,ZIPP B J,LUDWIGMüLLER J,et al.Arabidopsis glucosyltransferase UGT74B1 functions inglucosinolate biosynthesis and auxin homeostasis [J].The Plant Journal,2004,40:893-908.
[24] JACKSON R G,KOWALCZYK M,LI Y,et al.Over-expression of an Arabidopsis gene encoding a glucosyltransferase of indole-3-acetic acid:phenotypic characterisation of transgenic lines [J].The Plant Journal,2002,32(4):573-583.
[25] 郑晓瑜,郭晋艳,张毅,等.植物非生物胁迫诱导启动子顺式作用元件的研究方法[J].植物生理学报,2011,47(2):129-135.
[26] 王婧,李冰,刘翠翠,等.启动子结构和功能研究进展[J].生物技术通报,2014,8(1):40-45.
[27] 王志新,赵琳,李文滨.植物诱导型启动子的研究进展[J].大豆科技,2011(3):5-9.
[28] 周云.水稻胚乳特异强表达基因的筛选及其启动子功能的初步分析[D].福州:福建农林大学,2011.
[2] 唐秀华,孙威江,唐琴.紫化茶树生理生化及其花青素调控机理研究进展[J].天然产物研究与开发,2017,29(6):1 077-1 083.
[3] 唐秀华,周喆,唐琴,等.茶树CsPAL3基因cDNA全长克隆及其表达分析[J].茶叶科学,2018,38(1):33-42.
[4] 王惠聪,黄旭明,胡桂兵,等.荔枝果皮花青苷合成与相关酶的关系研究[J].中国农业科学,2004,37(12):2 028-2 032.
[5] 田艳维.茶树L组UDPG-糖基转移酶基因的克隆及原核表达[D].合肥:安徽农业大学,2012.
[6] VOGT T,JONES P.Glycosyltransferases in plant natural product synthesis:characterization of a supergene family [J].Trends in Plant Science,2000,5(9):380-386.
[7] BUTLER J E,KADONAGA J T.The RNA polymerase Ⅱ core promoter:a key component in the regulation of gene expression [J].Genes & Development,2002,16(20):2 583-2 592.
[8] 聂丽娜,夏兰琴,徐兆师.植物基因启动子的克隆及其功能研究进展[J].植物遗传资源学报,2008,9(3):385-391.
[9] 王颖,麦维军,梁承邺,等.高等植物启动子的研究进展[J].西北植物学报,2003,23(11):2 040-2 048.
[10] 文添龙,刘雪梅,冀亚萍,等.高等植物胁迫诱导型启动子的研究进展[J].西北植物学报,2014,34(1):206-214.
[11] PELLEGRINESCHI A,REYNOLDS M,PACHECO M,et al.Stress-induced expression in wheat of the Arabidopsis thaliana DREB1A gene delays water stress symptoms under greenhouse conditions [J].Genome,2004,47(3):493-500.
[12] 张建军,文国琴,刘震.拟南芥糖基转移酶基因UGT71C5启动子的克隆和功能分析[J].四川大学学报(自然科学版),2011,48(4):955-960.
[13] 郝迪萩,赵琳,陈李淼,等.克隆启动子方法的比较及应用[J].东北农业大学学报,2010,41(3):154-160.
[14] LIN Y L,LAI Z X.Superoxide dismutase multigene family in longan somatic embryos:a comparison of Cu Zn-SOD,Fe-SOD,and Mn-SOD gene structure,splicing,phylogeny,and expression [J].Molecular Breeding,2013,32(3):595-615.
[15] 练从龙,卢秉国,赖钟雄,等.荔枝古树胚性愈伤组织LcCu/Zn-SOD3基因启动子的克隆及功能验证[J].西北植物学报,2015,35(1):16-22.
[16] 杨晓娜,赵昶灵,李云,等.启动子序列克隆和功能分析方法的研究进展[J].云南农业大学学报(自然科学版),2010,25(2):283-290.
[17] 金琦芳,陈志丹,孙威江,等.茶树CsANS基因及其启动子的克隆与生物信息学分析[J].茶叶科学,2016,36(2):219-228.
[18] XU G,MA H,NEI M,et al.Evolution of F-box genes in plants:different modes of sequence divergence and their relationships with functional diversification [J].Proc Natl Acad Sci USA,2009,106(3):835-840.
[19] JONES P,MESSNER B,NAKAJIMA J I,et al.UGT73C6 and UGT78D1,glycosyltransferases involved in flavonol glycoside biosynthesis in Arabidopsis thaliana [J].Journal of Biological Chemistry,2003,278:43 910-43 918.
[20] ONO E,YU H,HORIKAWA M,et al.Functional differentiation of the glycosyltransferases that contribute to the chemical diversity of bioactive flavonol glycosides in grapevines (Vitis vinifera) [J].The Plant Cell,2010,22(8):2 856-2 871.
[21] HALL D,YUAN X X,MURATA J,et al.Molecular cloning and biochemical characterization of the UDP-glucose:flavonoid 3-O-glucosyltransferase from Concord grape (Vitis labrusca) [J].Phytochemistry,2012,74(3):90-99.
[22] HALL D,KIM K H,LUCA V D.Molecular cloning and biochemical characterization of three concord grape (Vitis labrusca) flavonol 7-O-glucosyltransferases [J].Planta,2011,234:1 201-1 214.
[23] GRUBB C D,ZIPP B J,LUDWIGMüLLER J,et al.Arabidopsis glucosyltransferase UGT74B1 functions inglucosinolate biosynthesis and auxin homeostasis [J].The Plant Journal,2004,40:893-908.
[24] JACKSON R G,KOWALCZYK M,LI Y,et al.Over-expression of an Arabidopsis gene encoding a glucosyltransferase of indole-3-acetic acid:phenotypic characterisation of transgenic lines [J].The Plant Journal,2002,32(4):573-583.
[25] 郑晓瑜,郭晋艳,张毅,等.植物非生物胁迫诱导启动子顺式作用元件的研究方法[J].植物生理学报,2011,47(2):129-135.
[26] 王婧,李冰,刘翠翠,等.启动子结构和功能研究进展[J].生物技术通报,2014,8(1):40-45.
[27] 王志新,赵琳,李文滨.植物诱导型启动子的研究进展[J].大豆科技,2011(3):5-9.
[28] 周云.水稻胚乳特异强表达基因的筛选及其启动子功能的初步分析[D].福州:福建农林大学,2011.