玉米赤霉素受体基因ZmGID1a的克隆及功能研究
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摘要
为研究赤霉素(GA)受体GID (gibberellin insensitive dwarf),克隆编码玉米GID的同源基因ZmGID1a,分析其在物种间的保守性和系统进化关系。通过实时定量PCR分析该基因在玉米中的表达特性以及不同激素对该基因表达水平的影响。亚细胞定位确定蛋白在细胞核和膜上,利用酵母双杂技术分析Zm GID1a与GA负调控因子玉米DELLA-8和DELLA-9的互作。还分析了表达Zm GID1a转基因烟草的生理指标。这些结果表明Zm GID1a和其他已报道的同源蛋白功能相似。
In order to know the function of gibberellins receptor GID, the gene encoding maize gibberellin receptor GID1 a was cloned. Based on the comparison of amino acid sequences, phylogenetic relationship of GID1 a proteins among different plant species were analyzed. The expression patterns in maize were analyzed using qPCR, as well as the effect of the hormones on GID1 a expression levels. Subcellular localization showed that the GID1 a was located at nucleus and membrane. Using the yeast two-hybrid system, we identified that the GID1 a was interacted with the DELLA-8 and DELLA-9 as the reported GA negative regulators. The effects of GID1 a in transgenic tobacco plant were analyzed. The current findings present that maize GID1 a is similar to the homologs from other plants.
In order to know the function of gibberellins receptor GID, the gene encoding maize gibberellin receptor GID1 a was cloned. Based on the comparison of amino acid sequences, phylogenetic relationship of GID1 a proteins among different plant species were analyzed. The expression patterns in maize were analyzed using qPCR, as well as the effect of the hormones on GID1 a expression levels. Subcellular localization showed that the GID1 a was located at nucleus and membrane. Using the yeast two-hybrid system, we identified that the GID1 a was interacted with the DELLA-8 and DELLA-9 as the reported GA negative regulators. The effects of GID1 a in transgenic tobacco plant were analyzed. The current findings present that maize GID1 a is similar to the homologs from other plants.
引文
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[15]李针针,刘兆磊,陈发棣,等.荷花R2R3-MYB转录因子NnMYB4对拟南芥木质素合成的影响[J].南京农业大学学报,2016,39(6):932-938.
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[17]时政,高丙德,郭晓恒,等.不同栽培方法对苦荞内源激素及产量和品质的影响[J].安徽农业大学学报,2017,44(4):732-737.
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[20]武健东,洪玲,江海洋,等.玉米抗病相关基因NPR1的同源克隆及表达分析[J].安徽农业大学学报,2015,42(5):651-656.
[21]王西成,王晨,房经贵,等.葡萄VvGA2ox1基因克隆、亚细胞定位及时空表达分析[J].南京农业大学学报,2013,36(1):29-34.
[22]PLACKETT A R G,FERGUSON A C,POWERS S J,et al.DELLA activity is required for successful pollen development in the Columbia ecotype of Arabidopsis[J].New Phytol,2014,201(3):825-836.
[23]SASAKI A.Accumulation of phosphorylated repressor for gibberellin signaling in an F-box mutant[J].Science,2003,299(5614):1896-1898.
[24]董静,尹梦回,杨帆,等.棉花赤霉素不敏感矮化GID1同源基因的克隆和表达分析[J].作物学报,2009,35(10):1822-1830.
[25]葛晖,赵晓,王增,等.葡萄GA受体基因GID1初探及其在果实GA处理条件下的差异表达[J].中国农业大学学报,2011,16(2):58-63.
[26]LI X,YANG W L,LI S J,et al.Molecular characterization of three GIBBERELLIN-INSENSITIVE DWARF1homologous genes in hexaploid wheat[J].J Plant Physiol,2013,170(4):432-443.
[27]WU J,KONG X,WAN J,et al.Dominant and pleiotropic effects of a GAI gene in wheat results from a lack of interaction between DELLA and GID1[J].Plant Physiol,2011,157(4):2120-2130.
[28]王西成,吴伟民,房经贵,等.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):839-848.
[29]EL-SHARKAWY I,SHERIF S,EL KAYAL W,et al.Characterization of gibberellin-signalling elements during plum fruit ontogeny defines the essentiality of gibberellin in fruit development[J].Plant Mol Biol,2014,84(4/5):399-413.
[30]申世辉.纽荷尔脐橙中赤霉素受体基因Cs GID1的功能分析[D].武汉:华中农业大学,2012:9-30.
[31]HUANG D B,WANG S G,ZHANG B C,et al.A gibberellin-mediated DELLA-NAC signaling cascade regulates cellulose synthesis in rice[J].Plant Cell,2015,27(6):1681-1696.
[32]BIEMELT S,TSCHIERSCH H,SONNEWALD U.Impact of altered gibberellin metabolism on biomass accumulation,lignin biosynthesis,and photosynthesis in transgenic tobacco plants[J].Plant Physiol,2004,135(1):254-265.
[2]THOMAS S G,RIEU I,STEBER C M.Gibberellin metabolism and signaling[M]//THOMAS S G,RIEU I,STEBER C M.eds.Plant Hormones","Vitamins&Hormones.Elsevier,2005:289-338
[3]AYA K,UEGUCHI-TANAKA M,KONDO M,et al.Gibberellin modulates anther development in rice via the transcriptional regulation of GAMYB[J].The Plant Cell Online,2009,21(5):1453-1472.
[4]UEGUCHI-TANAKA M,ASHIKARI M,NAKAJIMA M,et al.GIBBERELLIN INSENSITIVE DWARF1 encodes a soluble receptor for gibberellin[J].Nature,2005,437(7059):693-698.
[5]NAKAJIMA M,SHIMADA A,TAKASHI Y,et al.Identification and characterization of Arabidopsis gibberellin receptors[J].Plant J,2006,46(5):880-889.
[6]ALEMAN L,KITAMURA J,ABDEL-MAGEED H,et al.Functional analysis of cotton orthologs of GA signal transduction factors GID1 and SLR1[J].Plant Mol Biol,2008,68(1/2):1-16.
[7]CHANDLER P M,HARDING C A,ASHTON A R,et al.Characterization of gibberellin receptor mutants of barley(Hordeum vulgare L.)[J].Mol Plant,2008,1(2):285-294.
[8]MAURIAT M,MORITZ T.Analyses of GA20ox-and GID1-over-expressing aspen suggest that gibberellins play two distinct roles in wood formation[J].Plant J,2009,58(6):989-1003.
[9]WANG Y J,DENG D X,DING H D,et al.Gibberellin biosynthetic deficiency is responsible for maize dominant dwarf11(d11)mutant phenotype:physiological and transcriptomic evidence[J].PLoS One,2013,8(6):e66466.
[10]CHEN Y,HOU M,LIU L,et al.The maize DWARF1 encodes a gibberellin 3-oxidase and is dual localized to the nucleus and cytosol[J].Plant Physiol,2014,166(4):2028-2039.
[11]LAWIT S J,WYCH,H M,XU D P,et al.Maize della proteins dwarf plant8 and dwarf plant9 as modulators of plant development[J].Plant Cell Physiol,2010,51(11):1854-1868.
[12]ZHAO G,FU J,WANG G,et al.Gibberellin-induced mesocotyl elongation in deep-sowing tolerant maize inbred line 3681-4[J].Plant Breeding,2010,129(1):87-91.
[13]BORLAUG N.Feeding a hungry world[J].Science,2007,318(5849):359.
[14]FU C X,SUNKAR R,ZHOU C,et al.Overexpression of mi R156 in switchgrass(Panicum virgatum L.)results in various morphological alterations and leads to improved biomass production[J].Plant Biotechnol J,2012,10(4):443-452.
[15]李针针,刘兆磊,陈发棣,等.荷花R2R3-MYB转录因子NnMYB4对拟南芥木质素合成的影响[J].南京农业大学学报,2016,39(6):932-938.
[16]李寿田,周健民,朱世东,等.萝卜贮藏期间木质素、纤维素和可溶性糖含量变化及其与糠心的关系[J].安徽农业大学学报,2001,28(3):255-258.
[17]时政,高丙德,郭晓恒,等.不同栽培方法对苦荞内源激素及产量和品质的影响[J].安徽农业大学学报,2017,44(4):732-737.
[18]BAI W Q,XIAO Y H,ZHAO J,et al.Gibberellin overproduction promotes sucrose synthase expression and secondary cell wall deposition in cotton fibers[J].PLoSOne,2014,9(5):e96537.
[19]代丽娟,郑唐春,刘彩霞,等.拟南芥CYCD3;1基因植物表达载体的构建及在烟草中的遗传转化分析[J].安徽农业大学学报,2016,43(6):996-1003.
[20]武健东,洪玲,江海洋,等.玉米抗病相关基因NPR1的同源克隆及表达分析[J].安徽农业大学学报,2015,42(5):651-656.
[21]王西成,王晨,房经贵,等.葡萄VvGA2ox1基因克隆、亚细胞定位及时空表达分析[J].南京农业大学学报,2013,36(1):29-34.
[22]PLACKETT A R G,FERGUSON A C,POWERS S J,et al.DELLA activity is required for successful pollen development in the Columbia ecotype of Arabidopsis[J].New Phytol,2014,201(3):825-836.
[23]SASAKI A.Accumulation of phosphorylated repressor for gibberellin signaling in an F-box mutant[J].Science,2003,299(5614):1896-1898.
[24]董静,尹梦回,杨帆,等.棉花赤霉素不敏感矮化GID1同源基因的克隆和表达分析[J].作物学报,2009,35(10):1822-1830.
[25]葛晖,赵晓,王增,等.葡萄GA受体基因GID1初探及其在果实GA处理条件下的差异表达[J].中国农业大学学报,2011,16(2):58-63.
[26]LI X,YANG W L,LI S J,et al.Molecular characterization of three GIBBERELLIN-INSENSITIVE DWARF1homologous genes in hexaploid wheat[J].J Plant Physiol,2013,170(4):432-443.
[27]WU J,KONG X,WAN J,et al.Dominant and pleiotropic effects of a GAI gene in wheat results from a lack of interaction between DELLA and GID1[J].Plant Physiol,2011,157(4):2120-2130.
[28]王西成,吴伟民,房经贵,等.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):839-848.
[29]EL-SHARKAWY I,SHERIF S,EL KAYAL W,et al.Characterization of gibberellin-signalling elements during plum fruit ontogeny defines the essentiality of gibberellin in fruit development[J].Plant Mol Biol,2014,84(4/5):399-413.
[30]申世辉.纽荷尔脐橙中赤霉素受体基因Cs GID1的功能分析[D].武汉:华中农业大学,2012:9-30.
[31]HUANG D B,WANG S G,ZHANG B C,et al.A gibberellin-mediated DELLA-NAC signaling cascade regulates cellulose synthesis in rice[J].Plant Cell,2015,27(6):1681-1696.
[32]BIEMELT S,TSCHIERSCH H,SONNEWALD U.Impact of altered gibberellin metabolism on biomass accumulation,lignin biosynthesis,and photosynthesis in transgenic tobacco plants[J].Plant Physiol,2004,135(1):254-265.