[1]高世敏,董阳,王武,等.葡萄赤霉素合成关键基因VvGA20ox2的克隆、亚细胞定位和表达分析[J].江苏农业学报,2018,(06):1331-1338.[doi:doi:10.3969/j.issn.1000-4440.2018.06.019]
　GAO Shi-min,DONG Yang,WANG Wu,et al.Cloning, subcellular localization and expression analysis of the key gene VvGA20ox2 in gibberellin synthesis of grapevine[J].,2018,(06):1331-1338.[doi:doi:10.3969/j.issn.1000-4440.2018.06.019]
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葡萄赤霉素合成关键基因VvGA20ox2的克隆、亚细胞定位和表达分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2018年06期
页码:
1331-1338
栏目:
园艺
出版日期:
2018-12-25

文章信息/Info

Title:
Cloning, subcellular localization and expression analysis of the key gene VvGA20ox2 in gibberellin synthesis of grapevine
作者:
高世敏; 董阳; 王武; 陶建敏
（南京农业大学园艺学院，江苏南京210095）
Author(s):
GAO Shi-min; DONG Yang; WANG Wu; TAO Jian-min
(College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China)
关键词:
葡萄; 赤霉素; VvGA20ox2; 亚细胞定位; 表达分析
Keywords:
Vitis vinifera; gibberellin(GA3); VvGA20ox2; subcellular localization; expression analysis
分类号:
S663.103.53
DOI:
doi:10.3969/j.issn.1000-4440.2018.06.019
文献标志码:
A
摘要:
本研究建立在对金手指葡萄果形转录组分析的基础上，通过分析样品间表达差异发现，与对照相比，GA20ox2基因的转录水平在GA3处理后有显著差异。为了进一步从分子水平阐述GA20ox2在赤霉素合成途径中的作用机理，本研究以金手指葡萄幼果为材料，克隆了GA20ox2基因，分析了此基因碱基序列特征，并通过构建融合表达载体对其进行亚细胞定位分析。研究结果表明，克隆了葡萄赤霉素合成途径中的VvGA20ox2基因，测序得CDS全长为1 134 bp，编码 377个氨基酸。VvGA20ox2编码的氨基酸序列与NCBI上其他物种GA20ox序列相似性在 51%～67%，其中该基因与可可（EOX92603.1）和杨树（PNT28192.1）的亲缘关系较近。该基因定位在细胞核和细胞膜中。外源GA3处理后的成熟葡萄果实与对照相比纵径显著伸长，且在幼果中该基因表达量在一段时间内明显上升，推测VvGA20ox2基因可能参与葡萄果形拉长的调控。
Abstract:
In this study, based on the transcriptome analysis of grape berry shape change, it was found that there was a significant expression difference among samples in the transcriptional level of the GA20ox2 gene after gibberellin(GA3) treatment compared with the control. In order to study molecular mechanism of GA20ox2 in GA biosynthesis pathway, VvGA20ox2 gene was cloned from Golden Finger grape. The base sequence characteristics of GA20ox2 gene were analyzed, and subcellular localization analysis was performed by constructing a fusion expression vector. The results showed that the VvGA20ox2 gene related to gibberellin synthesis pathway was successfully cloned, and the total length of CDS was 1 134 bp, and 377 amino acids were encoded. The sequence similarity of VvGA20ox2 to other nine species GA20ox protein sequences in NCBI was 51%-67%,it had closer relationship with cacao (EOX92603.1) and poplar (PNT28192.1). The VvGA20ox2 gene was located in both the plasma membrane and nucleus. The longitudinal diameter of mature grape berries treated with GA3 was significantly longer than that of the control, and the expression level of VvGA20ox2 gene in young fruit increased significantly. It is speculated that the VvGA20ox2 gene may be involved in the regulation of grape shape.

参考文献/References:

[1]HEDDEN P, KAMIYA Y. Gibberellin biosynthesis：enzymes, genes and their regulation[J]. Annual Review of Plant Physiology Plant Molecular Biology,1997,48:431-460.
[2]葛长军，闫良，徐丽荣．赤霉素处理对黄瓜雌性系的诱雄效果[J]．江苏农业科学，2017,45(5):111-114.
[3]ERIKSSON S, BHLENIUS H, MORITZ T, et al. GA4 is the active gibberellin in the regulation of LEAFY transcription and Arabidopsis floral initiation[J]. Plant Cell,2006,18 (9):2172-2181.
[4]林玲，白先进，张瑛，等. 赤霉素对巨峰葡萄冬果花序拉长效果及果实品质的影响[J]．南方农业学报，2017，48（6）：1031-1035.
[5]HARTWECK L M, OLSZWSKI N E. Rice GIBBERELLIN INSENSITIVE DWARF1 is a gibberellins receptor that illuminates and raises questions about GA signaling[J]. The Plant Cell,2006,18 (2):278-282.
[6]黄桃鹏，李媚娟，王睿，等.赤霉素生物合成及信号转导途径研究进展[J]. 植物生理学报,2005,51(8):1241-1247.
[7]陈峰源，王先俱，陈亚君，等．赤霉素对杂交粳稻组合孕穗期生育进程的影响[J]．江苏农业科学，2016,44(2):92-94.
[8]张晓莹，宋长年，房经贵，等. 赤霉素的生物合成及其在葡萄栽培上的应用[J]. 浙江农业科学, 2011(5): 1015-1018.
[9]熊书. 赤霉素的生物合成和信号转导研究进展[J]. 教育教学论坛，2015（16）：95-96.
[10]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.
[11]HEDDEN P. Recent advances in gibberellin biosynthesis[J]. Journal of Experimental Botany，1999, 50（334）:553-563.
[12]黄先忠，蒋才富，廖立力, 等. 赤霉素作用机理的分子基础与调控模式研究进展[J]. 植物学通报, 2006, 23(5): 499-510.
[13]石琰璟，沙广利，束怀瑞. 赤霉素生物合成及其分子机理研究进展[J].西北植物学报, 2006, 26(7): 1482-1489.
[14]KHALILURREHMAN M, SUN L,LI C X, et al. Comparative RNA-seq based transcriptomic analysis of bud dormancy in grape[J].BMC Plant Biology,2017,17(1):18.
[15]柴丹丹. 黄瓜果实圆形性状的SRAP分子标记[D]. 西北农林科技大学, 2008.
[16]WANG Z, ZHAO F, ZHAO X, et al. Proteomic analysis of berry-sizing effect of GA3 on seedless Vitis vinifera L.[J]. Proteomics, 2012,12 (1):86-94.
[17]郑焕，季晨飞，陶建敏. GA3和CPPU对钟山红葡萄果实品质的影响[J].中国南方果树, 2013,42(1): 22-24.
[18]WANG S, CHANG Y, GUO J, et al. Arabidopsis ovate family protein 1 is a transcriptional repressor that suppresses cell elongation[J]. Plant Journal, 2007, 50(5): 858-872.
[19]TOYOOKA K, GOTO Y, ASATSUMA S, et al. A mobile secretory vesicle cluster involved in mass transport from the Golgi to plant cell exterior[J]. Plant Cell， 2009,21(4):1212-1229.
[20]LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta delta C(T)) method[J]. Methods, 2001, 25(4): 402-408.
[21]王西成，任国慧，房经贵，等. 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):2224-2231.
[22]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.
[23]刘炳臣，王茜，王跃进，等.葡萄活性赤霉素合成关键基因VvGA3ox家族的克隆和表达分析[J].园艺学报,2016,43(12):2293-2303.
[24]JUNG C J, HUR Y Y, JUNG S M, et al. Transcriptional changes of gibberellin oxidase genes in grapevines with or without gibberellin application during inflorescence development[J]. Plant Res,2014,127(2):359-371
[25]GIACIMELLI L, ROTA-STABELLI O, MASUERO D, et al. Gibberellin metabolism in Vitis vinifera L. during bloom and fruit-set：funcational characterization and evolution of grapevine gibberellin oxidase[J]. Journal of Experimental Botany,2013,64(14):4403-4419.
[26]ATIAKO K, ACHEAMPONG,CHUANLIN ZHENG, et al. Abnormal endogenous repression of GA signaling in a seedless table grape cultivar with high berry growth response to GA application[J].Frontier in Plant Science,2017, 8:850.
[27]邢浩然，刘丽娟，刘国振.植物蛋白质的亚细胞定位研究进展[J].华北农学报,2006,21(增刊):1-6.
[28]KING K E, MORITZ T, HARBERD N P. Gibberellins are not required for normal stem growth in Arabidopsis thaliana in the absence of GAI and RGA[J].Genetics,2001,159(2):767-776.
[29]RIEU I, RUIZ-RIVERO O, FERNANDEZ-GARCIA N, et al. The gibberellin biosynthetic genes AtGA20ox1 and AtGA20ox2 act, partially redundantly, to promote growth and development throughout the Arabidopsis life cycle[J]. The plant journal, 2008, 53(3):488-504.
[30]陶建敏,韩传光,章镇,等. GA3在葡萄生产上的应用[J].中外葡萄与葡萄酒, 2003(6):33-35.

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备注/Memo

备注/Memo:
收稿日期：2018-03-18 基金项目：国家现代农业葡萄产业技术体系项目（CARS-29）；国家“948”重点项目（2016-X19）；江苏省农业重大新品种创制项目（PZCZ201723） 作者简介：高世敏，（1992-）男，山西临汾人，硕士研究生，主要从事果树生理与分子技术研究。（E-mail）547579531@qq.com 通讯作者：陶建敏，（E-mail）tjm266@sina.com
更新日期/Last Update: 2018-12-28