不同生态条件下棉花赤霉素合成代谢途径关键酶基因差异表达分析
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摘要
为揭示棉花植株体内赤霉素合成途径关键酶基因表达量与不同组织及环境条件的关系,以"中棉所49"(CCRI49)和"高脚棉"(396289)为材料,利用实时荧光定量PCR方法中的相对定量,以UBQ7为内参基因,对新疆维吾尔自治区阿拉尔和河南省安阳,2种不同生态环境下的CCRI49和396289不同组织内赤霉素合成途径相关基因的表达量进行检测。结果显示,在阿拉尔地区生长条件下,CCRI49果枝始生节位相比于安阳地区平均降低2.0,高脚棉平均降低2.6。在安阳地区,CCRI49和396289中赤霉素合成途径各关键酶基因在不同组织内的表达趋势基本一致,茎组织中各相关基因表达量高,为植株生长提供充足的活性赤霉素;在新疆阿拉尔地区,CCRI49和396289主要表现为赤霉素合成早期阶段CPS1、KS和KO等基因的表达量下调,GA2ox1表达量上调,以减少活性赤霉素的合成。结果表明,棉花植株赤霉素合成代谢途径关键酶基因的表达量发生改变,从而调控植株的生长发育来适应新疆干旱和盐渍化的生长环境。
To explore the relationship between the key enzyme gene expression of gibberellin synthesis pathway and different tissues or environmental condition,cotton CCRI49 and 396289 were selected as material,and UBQ7 was selected as reference gene,Relative quantification real-time PCR was developed to determine the gibberellin key gene expression of different cotton organizations under two kinds of ecological environment.The results showed that:compared with Anyang,the average node of first fruiting/sympodial branch of CCRI49 and 396289 were repectively decreased by 2.0 and 2.6 under the growth condition of Alar.In Anyang region,the expression trends of gibberellin related gene were basically identical in tissues,but the expression level in stem was higher,which was beneficial for the growth of plants.In Alar region,the CPS1,KS,and KO gene expressions were significantly down-regulated in CCRI49 and 396289,while GA2 ox1 gene expression was up-regulated.The down-regulation of CPS1,KS and KO reduced the synthesis rate of gibberellin,and the up-regulation of GA2 ox1 decomposed the active gibberellin.To adapt to the arid and salinized growth environment of Xinjiang,the cotton plant regulate its growth and development by changing the expression of the key enzyme gene of gibberellin in the plant.
To explore the relationship between the key enzyme gene expression of gibberellin synthesis pathway and different tissues or environmental condition,cotton CCRI49 and 396289 were selected as material,and UBQ7 was selected as reference gene,Relative quantification real-time PCR was developed to determine the gibberellin key gene expression of different cotton organizations under two kinds of ecological environment.The results showed that:compared with Anyang,the average node of first fruiting/sympodial branch of CCRI49 and 396289 were repectively decreased by 2.0 and 2.6 under the growth condition of Alar.In Anyang region,the expression trends of gibberellin related gene were basically identical in tissues,but the expression level in stem was higher,which was beneficial for the growth of plants.In Alar region,the CPS1,KS,and KO gene expressions were significantly down-regulated in CCRI49 and 396289,while GA2 ox1 gene expression was up-regulated.The down-regulation of CPS1,KS and KO reduced the synthesis rate of gibberellin,and the up-regulation of GA2 ox1 decomposed the active gibberellin.To adapt to the arid and salinized growth environment of Xinjiang,the cotton plant regulate its growth and development by changing the expression of the key enzyme gene of gibberellin in the plant.
引文
[1]李四游.赤霉素对莲株型影响及其相关基因克隆与时空表达分析[D].武汉:华中农业大学,2015Li S Y.Gibberellin impact on lotus plant height and cloning and expression analysis of its related genes[D]. Wuhan:Huazhong Agricultural University,2015(in Chinese)
[2] Carrera E,Bou J,Garcia-Martinez J L,Prat S.Changes in GA20-oxidase gene expression strongly affect stem length,tuber induction and tuber yield of potato plants[J].Plant Journal,2000,22(3):247-256
[3] Sakamoto T,Miura K,Itoh H,Tatsumi T,Ueguchi-Tanaka M,Ishiyama K,Kobayashi M,Agrawal G K,Takeda S,Abe K,Miyao A,Hirochika H,Kitano H,Ashikari M,Matsuoka M.An overview of gibberellin metabolism enzyme genes and their related mutants in rice[J].Plant Physiology,2004,134(4):1642-1653
[4] Hedden P,Stephen GT.Gibberellin biosynthesis and its regulation[J].Biochemical Journal,2012,444(1):11-25
[5]虞慧芳,曹家树,王永勤.植物矮化突变体的激素调控[J].生命科学,2002,14(2):85-88,76Yu H F,Cao J S,Wang Y Q.Hormones regulation in plant dwarfing mutants[J].Chinese Bulletin of Life Sciences,2002,14(2):85-88,76(in Chinese)
[6]刘洁,李润植.作物矮化基因与GA信号转导途径[J].中国农学通报,2005,21(1):37-40Liu J,Li R Z.Dwarfing genes and GA signal transduction pathway in crops[J].Chinese Agricultural Science Bulletin,2005,21(1):37-40(in Chinese)
[7] Toyomasu T,Kawaide H,Sekimoto H,Numers C,Phillips A L,Hedden P,Kamiya Y.Cloning and characterization of a cDNA encoding gibberellin 20-oxidase from rice(Oryza sativa)seedlings[J].Physiologia Plantarum,1997,99(1):111-118
[8]岳川,曾建明,曹红利,郝心愿,章志芳,王新超,杨亚军.茶树赤霉素受体基因CsGID1a的克隆与表达分析[J].作物学报,2013,39(4):599-608Yue C,Zeng J M,Cao H L,Hao X Y,Zhang Z F,Wang X C,Yang Y J.Cloning and expression analysis of gibberellin receptor gene CsGID1ain tea plant(Camellia sinensis)[J].ACTA Agronomica Sinica,2013,39(4):599-608(in Chinese)
[9]王秀莉,姚颖垠,彭惠茹,张一,逯腊虎,倪中福,孙其信.赤霉素代谢及其调控相关基因的差异表达与小麦株高杂种优势的关系研究[J].科学通报,2009,54(16):2320-2324Wang X L,Yao Y Y,Peng H R,Zhang Y,Lu L H,Ni Z F,Sun Q X.The relationship of differential expression of genes in GA biosynthesis and response pathways with heterosis of plant height in a wheat diallel cross[J].Chinese Science Bulletin,54(16):2320-2324(in Chinese)
[10]刘斌.黄瓜赤霉素受体基因CsGID1a的功能验证与表达谱分析[D].北京:中国农业大学,2017Liu B. Functional verification and expression spectrum analysis of CsGID1aof cucumber gibberellin receptor gene[D].Beijing:China Agricultural University,2017(in Chinese)
[11]李强,吴建明,梁和,黄杏,丘立杭.高等植物赤霉素生物合成及其信号转导途径[J].生物技术通报,2014(10):16-22Li Q,Wu J M,Liang H,Huang X,Qiu L H.Gibberellins biosynthesis and signaling transduction pathway in higher plant[J].Biotechnology Bulletin,2014,(10):16-22(in Chinese)
[12]鲁井云,卢龙,邢佳毅,舒晓娟,唐丁,胡建芳.外施赤霉素对‘峰后’葡萄坐果及VvAG基因表达的影响[J].中国农业大学学报,2015,20(1):53-59Lu J Y,Lu L,Xing J Y,Shu X J,Tang D,Hu J F.Improvement of fruit set and expression of VvAG in‘Fenghou’grapevine with exogenous gibberellin[J].Journal of China Agricultural University,2015,20(1):53-59(in Chinese)
[13]谈心,马欣荣.赤霉素生物合成途径及其相关研究进展[J].应用与环境生物学报,2008,14(4):571-577Tan X, Ma X R. Advance in research of gibberellin biosynthesis pathway[J].Chinese Journal of Applied&Environmental Biology,2008,14(4):571-577(in Chinese)
[14]张驰宇,徐顺高,黄新祥.一种新颖简便的荧光实时RTPCR相对定量方法的建立[J].生物化学与生物物理进展,2005,32(9):883-888Zhang C Y,Xu S G,Huang X X.A Novel and convenient relative quantitative method of fluorescence real time RTPCR Assay based on slope of standard curve[J].Progress in Biochemistry and Biophysics,2005,32(9):883-888(in Chinese)
[15]倪中福,孙其信,吴利民,解超杰,孟凡荣.小麦种间与品种间杂交种及其亲本之间基因差异表达比较研究[J].农业生物技术学报,2001,9(4):366-370Ni Z F,Sun Q X,Wu L M,Xie C J,Meng F R.Alteration of gene expression in wheat hybrid F1 and their parental seedling leaves[J].Journal of Agricultural Biotechnology,2001,9(4):366-370(in Chinese)
[16]王西成,任国慧,房经贵,李阿英,刘洪,吴伟民,赵密珍.葡萄赤霉素合成相关基因克隆、亚细胞定位和表达分析[J].中国农业科学,2012,45(11):2224-2231Wang X C,Ren G H,Fang J G,Li A Y,Liu H,Wu W M,Zhao M Z.Cloning,subcellular localization and expression analysis of genes related to the synthesis of gibberellin from grapevine[J].Scientia Agricultural Sinica,2012,45(11):2224-2231(in Chinese)
[17]王西成,吴伟民,房经贵,钱亚明,王晨,宋长年,赵密珍.葡萄赤霉素受体基因VvGID1A的分离、亚细胞定位及表达分析[J].园艺学报,2013,40(5):839-848Wang X C,Wu W M,Fang J G,Qian Y M,Wang C,Song C N,Zhao M Z.Isolation,subcellular localization and expression analysis of gibberellin receptor gene VvGID1A from grapevine[J].Acta Horticulturae Sinica,2013,40(5):839-848(in Chinese)
[18]钟希琼,王惠珍.高等植物赤霉素生物合成及其调节研究进展[J].植物学通报,2001,18(3):303-307Zhong X Q,Wang H Z.Progress on GA biosynthesis and regulation in higher plants[J].Chinese Bulletin of Botany,2001,18(3):303-307(in Chinese)
[19]王丽.赤霉素合成抑制剂DPC调控棉花营养生长的分子机制研究[D].北京:中国农业大学,2014Wang L.Molecular mechanism of vegetative growth control by a gibberellin biosynthesis inhibitor DPC in cotton(Gossypium hirsutum L)[D].Beijing:China Agricultural University,2014(in Chinese)
[20]范术丽,喻树迅,张朝军,原日红,宋美珍.短季棉常用亲本早熟性状的遗传及配合力研究[J].棉花学报,2004,16(4):211-215Fan S L,Yu S X,Zhang C J,Yuan R H,Song M Z.Study on heredity and combining ability of earliness of short season cotton[J].Cotton Science,2004,16(4):211-215(in Chinese)
[21]李晨晨,侯雷,尹亮,赵金凤,袁守江,张文会,李学勇.水稻极矮突变体s2-47对赤霉素的响应及基因定位研究[J].作物学报,2013,39(10):1766-1774Li C C,Hou L,Yin L,Zhao J F,Yuan S J,Zhang W H,Li X Y.Gibberellin responsiveness and gene mapping of rice extreme dwarf mutant s2-47[J].Acta Agronomica Sinica,2013,39(10):1766-1774(in Chinese)
[22]郭献平,李兴亮,段续伟,邓舒,曹庆芹,秦岭.板栗野生和芽变雄花序赤霉素合成关键酶基因比较分析[J].中国农业大学学报,2012,17(4):91-95Guo X P,Li X L,Duan X W,Deng S,Cao Q Q,Qin L.Comparation of key enzymes of gibberellin biosynthesis between normal catkin and short catkin in Castanea mollissima BI[J]. Journal of China Agricultural University,2012,17(4):91-95(in Chinese)
[23]王成祥,李长生,侯蕾,刘学英,安静,万书波,毕玉平,王兴军.花生赤霉素2-氧化酶基因的克隆和表达研究[J].山东农业科学,2013,45(1):14-18,24Wang C X,Li C S,Hou L,Liu X Y,An J,Wan S B,Bi Y P,Wang X J.Cloning and expression analysis of Gibberellin2-Oxidase gene from peanut(Arachis hypogaea L)[J].Shandong Agricultural Sciences,2013,45(1):14-18,24(in Chinese)
[24] Lee D J,Zeevaart J A.Differential regulation of RNA levels of gibberellin dioxygenases by photoperiod in spinach[J].Plant Physiology,2002,130(4):2085-2094
[2] Carrera E,Bou J,Garcia-Martinez J L,Prat S.Changes in GA20-oxidase gene expression strongly affect stem length,tuber induction and tuber yield of potato plants[J].Plant Journal,2000,22(3):247-256
[3] Sakamoto T,Miura K,Itoh H,Tatsumi T,Ueguchi-Tanaka M,Ishiyama K,Kobayashi M,Agrawal G K,Takeda S,Abe K,Miyao A,Hirochika H,Kitano H,Ashikari M,Matsuoka M.An overview of gibberellin metabolism enzyme genes and their related mutants in rice[J].Plant Physiology,2004,134(4):1642-1653
[4] Hedden P,Stephen GT.Gibberellin biosynthesis and its regulation[J].Biochemical Journal,2012,444(1):11-25
[5]虞慧芳,曹家树,王永勤.植物矮化突变体的激素调控[J].生命科学,2002,14(2):85-88,76Yu H F,Cao J S,Wang Y Q.Hormones regulation in plant dwarfing mutants[J].Chinese Bulletin of Life Sciences,2002,14(2):85-88,76(in Chinese)
[6]刘洁,李润植.作物矮化基因与GA信号转导途径[J].中国农学通报,2005,21(1):37-40Liu J,Li R Z.Dwarfing genes and GA signal transduction pathway in crops[J].Chinese Agricultural Science Bulletin,2005,21(1):37-40(in Chinese)
[7] Toyomasu T,Kawaide H,Sekimoto H,Numers C,Phillips A L,Hedden P,Kamiya Y.Cloning and characterization of a cDNA encoding gibberellin 20-oxidase from rice(Oryza sativa)seedlings[J].Physiologia Plantarum,1997,99(1):111-118
[8]岳川,曾建明,曹红利,郝心愿,章志芳,王新超,杨亚军.茶树赤霉素受体基因CsGID1a的克隆与表达分析[J].作物学报,2013,39(4):599-608Yue C,Zeng J M,Cao H L,Hao X Y,Zhang Z F,Wang X C,Yang Y J.Cloning and expression analysis of gibberellin receptor gene CsGID1ain tea plant(Camellia sinensis)[J].ACTA Agronomica Sinica,2013,39(4):599-608(in Chinese)
[9]王秀莉,姚颖垠,彭惠茹,张一,逯腊虎,倪中福,孙其信.赤霉素代谢及其调控相关基因的差异表达与小麦株高杂种优势的关系研究[J].科学通报,2009,54(16):2320-2324Wang X L,Yao Y Y,Peng H R,Zhang Y,Lu L H,Ni Z F,Sun Q X.The relationship of differential expression of genes in GA biosynthesis and response pathways with heterosis of plant height in a wheat diallel cross[J].Chinese Science Bulletin,54(16):2320-2324(in Chinese)
[10]刘斌.黄瓜赤霉素受体基因CsGID1a的功能验证与表达谱分析[D].北京:中国农业大学,2017Liu B. Functional verification and expression spectrum analysis of CsGID1aof cucumber gibberellin receptor gene[D].Beijing:China Agricultural University,2017(in Chinese)
[11]李强,吴建明,梁和,黄杏,丘立杭.高等植物赤霉素生物合成及其信号转导途径[J].生物技术通报,2014(10):16-22Li Q,Wu J M,Liang H,Huang X,Qiu L H.Gibberellins biosynthesis and signaling transduction pathway in higher plant[J].Biotechnology Bulletin,2014,(10):16-22(in Chinese)
[12]鲁井云,卢龙,邢佳毅,舒晓娟,唐丁,胡建芳.外施赤霉素对‘峰后’葡萄坐果及VvAG基因表达的影响[J].中国农业大学学报,2015,20(1):53-59Lu J Y,Lu L,Xing J Y,Shu X J,Tang D,Hu J F.Improvement of fruit set and expression of VvAG in‘Fenghou’grapevine with exogenous gibberellin[J].Journal of China Agricultural University,2015,20(1):53-59(in Chinese)
[13]谈心,马欣荣.赤霉素生物合成途径及其相关研究进展[J].应用与环境生物学报,2008,14(4):571-577Tan X, Ma X R. Advance in research of gibberellin biosynthesis pathway[J].Chinese Journal of Applied&Environmental Biology,2008,14(4):571-577(in Chinese)
[14]张驰宇,徐顺高,黄新祥.一种新颖简便的荧光实时RTPCR相对定量方法的建立[J].生物化学与生物物理进展,2005,32(9):883-888Zhang C Y,Xu S G,Huang X X.A Novel and convenient relative quantitative method of fluorescence real time RTPCR Assay based on slope of standard curve[J].Progress in Biochemistry and Biophysics,2005,32(9):883-888(in Chinese)
[15]倪中福,孙其信,吴利民,解超杰,孟凡荣.小麦种间与品种间杂交种及其亲本之间基因差异表达比较研究[J].农业生物技术学报,2001,9(4):366-370Ni Z F,Sun Q X,Wu L M,Xie C J,Meng F R.Alteration of gene expression in wheat hybrid F1 and their parental seedling leaves[J].Journal of Agricultural Biotechnology,2001,9(4):366-370(in Chinese)
[16]王西成,任国慧,房经贵,李阿英,刘洪,吴伟民,赵密珍.葡萄赤霉素合成相关基因克隆、亚细胞定位和表达分析[J].中国农业科学,2012,45(11):2224-2231Wang X C,Ren G H,Fang J G,Li A Y,Liu H,Wu W M,Zhao M Z.Cloning,subcellular localization and expression analysis of genes related to the synthesis of gibberellin from grapevine[J].Scientia Agricultural Sinica,2012,45(11):2224-2231(in Chinese)
[17]王西成,吴伟民,房经贵,钱亚明,王晨,宋长年,赵密珍.葡萄赤霉素受体基因VvGID1A的分离、亚细胞定位及表达分析[J].园艺学报,2013,40(5):839-848Wang X C,Wu W M,Fang J G,Qian Y M,Wang C,Song C N,Zhao M Z.Isolation,subcellular localization and expression analysis of gibberellin receptor gene VvGID1A from grapevine[J].Acta Horticulturae Sinica,2013,40(5):839-848(in Chinese)
[18]钟希琼,王惠珍.高等植物赤霉素生物合成及其调节研究进展[J].植物学通报,2001,18(3):303-307Zhong X Q,Wang H Z.Progress on GA biosynthesis and regulation in higher plants[J].Chinese Bulletin of Botany,2001,18(3):303-307(in Chinese)
[19]王丽.赤霉素合成抑制剂DPC调控棉花营养生长的分子机制研究[D].北京:中国农业大学,2014Wang L.Molecular mechanism of vegetative growth control by a gibberellin biosynthesis inhibitor DPC in cotton(Gossypium hirsutum L)[D].Beijing:China Agricultural University,2014(in Chinese)
[20]范术丽,喻树迅,张朝军,原日红,宋美珍.短季棉常用亲本早熟性状的遗传及配合力研究[J].棉花学报,2004,16(4):211-215Fan S L,Yu S X,Zhang C J,Yuan R H,Song M Z.Study on heredity and combining ability of earliness of short season cotton[J].Cotton Science,2004,16(4):211-215(in Chinese)
[21]李晨晨,侯雷,尹亮,赵金凤,袁守江,张文会,李学勇.水稻极矮突变体s2-47对赤霉素的响应及基因定位研究[J].作物学报,2013,39(10):1766-1774Li C C,Hou L,Yin L,Zhao J F,Yuan S J,Zhang W H,Li X Y.Gibberellin responsiveness and gene mapping of rice extreme dwarf mutant s2-47[J].Acta Agronomica Sinica,2013,39(10):1766-1774(in Chinese)
[22]郭献平,李兴亮,段续伟,邓舒,曹庆芹,秦岭.板栗野生和芽变雄花序赤霉素合成关键酶基因比较分析[J].中国农业大学学报,2012,17(4):91-95Guo X P,Li X L,Duan X W,Deng S,Cao Q Q,Qin L.Comparation of key enzymes of gibberellin biosynthesis between normal catkin and short catkin in Castanea mollissima BI[J]. Journal of China Agricultural University,2012,17(4):91-95(in Chinese)
[23]王成祥,李长生,侯蕾,刘学英,安静,万书波,毕玉平,王兴军.花生赤霉素2-氧化酶基因的克隆和表达研究[J].山东农业科学,2013,45(1):14-18,24Wang C X,Li C S,Hou L,Liu X Y,An J,Wan S B,Bi Y P,Wang X J.Cloning and expression analysis of Gibberellin2-Oxidase gene from peanut(Arachis hypogaea L)[J].Shandong Agricultural Sciences,2013,45(1):14-18,24(in Chinese)
[24] Lee D J,Zeevaart J A.Differential regulation of RNA levels of gibberellin dioxygenases by photoperiod in spinach[J].Plant Physiology,2002,130(4):2085-2094