[1]郭文雅,崔艳梅,喻德跃,等.野生大豆GsAP1基因的克隆及功能分析[J].大豆科学,2016,35(06):919-927.[doi:10.11861/j.issn.1000-9841.2016.06.0919]
　GUO Wen-ya,CUI Yan-mei,YU De-yue,et al.Cloning and Functional Analysis of GsAP1 in Wild Soybean[J].Soybean Science,2016,35(06):919-927.[doi:10.11861/j.issn.1000-9841.2016.06.0919]
点击复制

野生大豆GsAP1基因的克隆及功能分析

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年06期 页码: 919-927 栏目: 出版日期: 2016-11-20
Title:
Cloning and Functional Analysis of GsAP1 in Wild Soybean
作者:
郭文雅; 崔艳梅; 喻德跃; 黄方
南京农业大学 大豆研究所/作物遗传与种质创新国家重点实验室/国家大豆改良中心/农业部大豆生物学与遗传育种实验室（综合性），江苏 南京 210095
Author(s):
GUO Wen-ya;  CUI Yan-mei;  YU De-yue;  HUANG Fang
（Soybean Research Institute, Nanjing Agricultural University/State Key Laboratory of Crop Genetics & Germplasm Enhancement/National Center for Soybean Improvement/Key Laboratory of Biology and Genetic Improvement of Soybean, Ministry of Agriculture, Nanjing 210095, China）
关键词:
野生大豆; 花发育; AP1; 转录因子; 功能研究
Keywords:
Wild soybean;  Floral development;  AP-1;  Transcription factor;  Functional analysis
DOI:
10.11861/j.issn.1000-9841.2016.06.0919
文献标志码:
A
摘要:
AP1转录因子在控制花器官的诱导与发育中起着重要的作用，对植物的成花诱导，促进植物提前开花有重要的意义。从野生大豆中分离克隆得到1个AP1-like 基因，并命名GsAP1，该基因全长711 bp，包含完整的开放阅读框，编码237个氨基酸。利用实时荧光定量PCR技术对GsAP1在野生大豆不同组织中的表达情况进行了分析，结果显示GsAP1只在花中表达，在四轮不同花器官中的表达显示GsAP1在花萼中表达量比较高，在花瓣中表达量较低，在雄蕊和心皮中不表达。洋葱表皮细胞瞬时表达系统表明GsAP1蛋白定位于细胞核中。酵母转录激活试验显示GsAP1具有明显的转录激活活性。在转基因烟草植株中GsAP1的表达量显著提高，并且使转基因植株的开花期比对照大幅度提前。本文探讨GsAP1在大豆成花中的作用机理，为培育大豆新品种提供了分子资源和理论基础。
Abstract:
AP-1 plays an important role in controlling floral organ formation and development. In this report, we isolated an AP1like gene, designated GsAP1, from the wild soybean flowers. The fragment was 711 bp and contained an open reading frame coding a polypeptide of 237 amino acids. Expression of GsAP1 was analyzed by the method of real-time PCR. The result showed that GsAP1 only expressed in flowers; in the four whorls of floral organs, expression of GsAP1 was prominent in sepals while very weak in the petals, not in stamens and capels; and increased during the course of seed development. Subcellular localization assay suggested that GsAP1 was localized in nucleus. The transcriptional activation ability of GsAP1 protein was confirmed by the yeast assay system. Further we found that GsAP1 caused early flowering when ectopically expressed in tobacco. The mechanism of GsAP1 in soybean was discussed in this paper, which provided molecular resources and theoretical basis for the breeding of soybean varieties.

参考文献/References:

［1］Irish V F, Sussex I M. Function of the APETALA1- gene during Arabidopsis- floral development［J］. Plant Cell, 1990(2): 741-754.

［2］Chen Z, Ye M, Su X, et al. Overexpression of AtAP1M3 regulates flowering time and floral development in Arabidopsis and effects key flowering-related genes in poplar［J］. Transgenic Research, 2015, 24: 705-715.
［3］Pandey D K, Chaudhary B. Domestication-driven Gossypium profilin 1(GhPRF1) gene transduces early flowering phenotype in tobacco by spatial alteration of apical/floral-meristem related gene expression［J］. BMC plant biology, 2016, 16(1): 112.
［4］Mandel M A, GustafsonBrown C, Savidge B, et al. Molecular characterization of the Arabidopsis floral homeotic gene APETALA1［J］. Nature, 1992, 360: 273-277.
［5］Bowman J L, Alvarez J, Weigel D, et al. Control of flower development in Arabidopsis thaliana by APETALA1 and interacting genes［J］. Development, 1993, 119: 721-743.
［6］Gustafson-Brown C, Savidge B, Yanofsky M F. Regulation of the Arabidopsis floral homeotic gene APETALA1［J］. Cell, 1994, 76(1): 131-43.
［7］Mandal M A, Yanofsky M F. A gene triggering flower formation in Arabidopsis［J］. Nature, 1995, 377(6549): 522-524.
［8］Thomas J. Molecular and genetic mechanisms of floral control［J］. Plant Cell, 2004,16: 1-17.
［9］Suárez-López P, Wheatley K, Robson F, et al. Constans mediates between the circadian clock and the control of flowering in Arabidopsis［J］. Nature, 2001, 410:1116-1120.
［10］Gocal G F , King R W, Blundell C A, et al. Evolution of floral meristem identity genes. Analysis of Lolium temulentum genes related to APETALA1 and LEAFY of Arabidopsis［J］. Plant Physiology, 2001, 125(4): 1788-1801.
［11］Shimada S, Ogawa T, Kitagawa S, et al. A genetic network of flowering-time genes in wheat leaves, in which an APETALA1/FRUITFULL-like gene, VRN1, is upstream of FLOWERING LOCUS T.［J］. Plant Journal, 2009, 58(4): 668-681.
［12］Chi Y J, Huang F, Liu H C, et al. An APETALA1like gene of soybean regulates flowering time and specifies floral organs［J］. Journal of Plant Physiology, 2011, 168(18): 2251-2259.
［13］Kotoda N, Wada M, Kusaba S, et al. Overexpression of MdM-ADS5, an APETALA1-like gene of apple, causes early flowering in transgenic Arabidopsis［J］. Plant Science, 2002, 162(5): 679-687.
［14］Chen M K, Lin I C, Yang C H. Functional analysis of three lily (Lilium longiflorum).APETALA1-like MADS box genes in regulating floral transition and formation［J］. Plant and Cell Physiology, 2008, 49: 704-717.
［15］Pena L, Martin-Trillo M, Juarez J, et al. Constitutive expression of Arabidopsis LEAFY or APETALA1 genes in citrus reduces their generation time［J］. Nature Biotechnology, 2001, 19(3): 263-267.
［16］Qu G Z, Zheng T, Liu G, et al. Overexpression of a MADS-box gene from birch (Betula platyphylla) promotes flowering and enhances chloroplast development in transgenic tobacco［J］. PLoS One,2013, 8(5): e63398.
［17］Huang H, Wang S, Jiang J, et al. Over expression of BpAP1 induces early flowering and produces dwarfism in Betula platyphylla×Betula pendula［J］ . Physiologia plantarum, 2014, 151(4): 495-506.
［18］杨郁文, 张保龙. 花分生组织决定基因APETALA1转化油菜［J］. 江苏农业学报, 2007, 23(6): 564-567. (Yang Y W, Zhang B L. Flower meristem gene APETALA1 transformed into rape［J］. Jiangsu Agricultural Journal, 2007, 23(6): 564 -567.)
［19］Chen Z, Ye M X, Su X X, et al. Overexpression of AtAP1M3 regulates flowering time and floral development in Arabidopsis and effects key flowering-related genes in poplar［J］.Transgenic Research, 2015, 24(4): 705-715.
［20］Fornara F, Parenicova L, Falasca G, et al. Functional characterization of OsMADS18, a member of the AP1/SQUA subfamily of MADS box genes［J］. Plant Physiology, 2004, 135(4): 2207-2219.
［21］Liu Y X, Song H W, Liu Z L, et al. Molecular characterization of loquat EjAP1 gene in relation to flowering［J］. Plant Growth Regulation, 2013, 70 (3): 287-296.
［22］Kim S, Koh J, Ma H, et al. Sequence and expression studies of A, B, and E.class MADS-box homologues in Eupomatia (Eupomatiaceae): Support for the bracteates origin of the calyptra［J］. International Journal of Plant Sciences, 2005, 166(2): 185-198.
［23］Zhang L, Xu Y, Ma R. Molecular cloning, identification, and chromosomal localization of two MADS box genes in peach (Prunus persica)［J］. Journal of Genetics and Genomics, 2008, 35: 365-372.
［24］刘志雄, 王莹, 吕小蒙, 等. 日本晚樱花器官特征基因-ClAP1的克隆与表达分析［J］. 园艺学报, 2010, 37(4): 649-654. (Liu Z X, Wang Y, Lyu X M, et al. Cloning and expression analysis of a floral organ identity gene ClAP1 from Prunus lannesiana［J］. Acta Horticulturae Sinica, 2010, 37(4): 649 -654.)
［25］Tsaftaris A S, Pasentsis K, Iliopoulos, et al. Isolation of three homologous AP1-like MADS-box genes in crocus (Crocus sativus L.) and characterization of their expression［J］. Plant Science, 2004, 166(5): 1235-1243.
［26］Pillitteri L J, Lovatt C J, Walling L L. Isolation and characterization of LEAFY and APETALA1 homologues from Citrus sinensis L. Osbeck ‘Washington’［J］. Journal of the American Society for Horticultural Science, 2004, 129(6): 846-856.
［27］Kohler C, Hennig L, Spillane C, et al. The Polycomb-guoup protein MEDEA regulates seed development by controlling expression of the MADS-box gene PHERES1［J］. Genes & Development, 2003, 17: 1540-1553.
［28］Buchner P, Boutin J P. A MADS box transcription factor of the AP1/AGL9 subfamily is also expressed in the seed coat of pea(Pisum sativum) during development［J］. Plant Molecular Biology, 1998, 38(6): 1253-1255.
［29］Pabón Mora N, Ambrose B A, Litt A, et al. Poppy APETALA1/FRUITFULL orthologs control flowering time, branching, perianth identity and fruit development［J］. Plant Physiology, 2012, 158(4): 1685-1704.
［30］Tian Y F, Yuan X Y, Jiang S H, et al. Molecular cloning and spatiotemporal expression of an APETALA1/FRUITFULL-like MADS-box gene from the orchid (Cymbidium faberi)［J］. Chinese Journal of Biotechnology, 2013, 29(2): 203-13.
［31］罗聪, 何新华, 陈虎, 等. 芒果AP1同源基因的克隆及其生物信息学分析［J］. 基因组学与应用生物学, 2009, 28(5): 851-858. (Luo C, He X H, Chen H, et al. Cloning and bioinformatics analysis of AP1 homologous gene from mango［J］. Genomics and Applied Biology, 2009, 28(5): 851-858.
［32］安利忻, 刘荣维. 花分生组织决定基因-AP1-转化矮牵牛的研究［J］. 植物学报, 2001, 43(1): 63-66. (An L X, Liu R W. Flower meristem identity gene AP1 transformation of Petunia［J］. Acta Botanica Sinica, 2001, 43(1): 63-66.)
［33］吕山花. 太行花MADS-box基因克隆、表达模式及功能分析［D］. 北京: 中国科学院植物研究所, 2006. (Lyu S H. Taihangia MADS-box gene cloning, expression pattern and functional analysis［D］. Beijing: Academy of Sciences Chinese, 2006.
［34］Fernando D D, Zhang S L. Constitutive expression of the SAP1 gene from willow (Salix discolor) causes early flowering in Arabidopsis thaliana［J］. Development Genes and Evolution, 2006, 216(1): 19-28.
［35］Li Q, Wang Y, Wang F, et al. Functional conservation and diversification of APETALA1/FRUITFULL genes in Brachypodium distachyon［J］. Plant Physiology, 2016.
［36］Zheng T, Li S, Zang L, et al. Overexpression of two PsnAP1 genes from Populus simonii×P. nigra causes early flowering in transgenic tobacco and Arabidopsis［J］. PLoS One, 2013, 9(10): e111725.

相似文献/References:

[1]高越,刘辉,陶波.抗草甘膦野生大豆筛选及其抗性生理机制研究[J].大豆科学,2013,32(01):76.[doi:10.3969/j.issn.1000-9841.2013.01.018]
　GAO Yue,LIU Hui,TAO Bo.Screening and Physiological Mechanisms of Resistance to Glyphosate in Wild Soybeans(Glycine soja)[J].Soybean Science,2013,32(06):76.[doi:10.3969/j.issn.1000-9841.2013.01.018]
[2]王军卫,侯立江,李? 登,等.野生大豆紫色酸性磷酸酶PAP1基因的克隆及分析[J].大豆科学,2013,32(05):596.[doi:10.11861/j.issn.1000-9841.2013.05.0596]
　WANG Jun-wei,HOU Li-jiang,LI Deng,et al.Cloning and Sequence Analysis of Purple Acid Phosphotase PAP1 Gene in Wild Soybean[J].Soybean Science,2013,32(06):596.[doi:10.11861/j.issn.1000-9841.2013.05.0596]
[3]王军卫,侯立江,李 登,等. 野生大豆紫色酸性磷酸酶PAP1基因的克隆及分析[J].大豆科学,2013,32(05):596.
　WANG Jun-wei,HOU Li-jiang,LI Deng,et al. Cloning and Sequence Analysis of Purple Acid Phosphotase PAP1 Gene in Wild Soybean[J].Soybean Science,2013,32(06):596.
[4]王丽燕.硅对野生大豆幼苗耐盐性的影响及其机制研究[J].大豆科学,2013,32(05):659.[doi:10.11861/j.issn.1000-9841.2013.05.0659]
　WANG Li-yan.Effects of Silicon on Salt Tolerance of Glycine soja Seedlings and Its Mechanism[J].Soybean Science,2013,32(06):659.[doi:10.11861/j.issn.1000-9841.2013.05.0659]
[5]陈丽丽,王明玖,何丽君,等.野生大豆ISSR体系的优化及其在远缘杂交后代鉴定中的利用[J].大豆科学,2013,32(04):459.[doi:10.11861/j.issn.1000-9841.2013.04.0459]
　CHEN Li-li,WANG Ming-jiu,HE Li-jun,et al.Optimization for ISSR Reaction System of Wild Soybean and Its Utilization in Distant Hybrid Identification[J].Soybean Science,2013,32(06):459.[doi:10.11861/j.issn.1000-9841.2013.04.0459]
[6]郑世英,萧蓓蕾,金桂芳.NaCl胁迫对野生大豆和栽培大豆叶绿素及光合特性的影响[J].大豆科学,2013,32(04):486.[doi:10.11861/j.issn.1000-9841.2013.04.0486]
　ZHENG Shi-ying,XIAO Bei-lei,JIN Gui-fang.Effect of NaCl Stress on Chlorophyll Content and Photosynthetic Characteristics of Glycine soja and Glycine max[J].Soybean Science,2013,32(06):486.[doi:10.11861/j.issn.1000-9841.2013.04.0486]
[7]徐艳平,胡翠美,张文会,等.干旱胁迫对野生大豆幼苗光合作用相关指标的影响[J].大豆科学,2013,32(03):341.[doi:10.11861/j.issn.1000-9841.2013.03.0341]
　XU Yan-ping,HU Cui-mei,ZHANG Wen-hui,et al.Effect of Simulated Drought Stress on Photosynthesis Related Indexes at Seedling Stage of Wild Soybeans[J].Soybean Science,2013,32(06):341.[doi:10.11861/j.issn.1000-9841.2013.03.0341]
[8]胡卫静,何丽君,何劲莉,等.NaCl胁迫对野生与栽培大豆杂交后代株系生理指标的影响[J].大豆科学,2013,32(03):349.[doi:10.11861/j.issn.1000-9841.2013.03.0349]
　HU Wei-jing,HE Li-jun,HE Jin-li,et al.Effects of NaCl Stress on Physiological Characters of Soybean Hybrids from Glycine max × Glycine soja[J].Soybean Science,2013,32(06):349.[doi:10.11861/j.issn.1000-9841.2013.03.0349]
[9]王 旻,梁 玉,王欣欣,等.即墨野生大豆主要成分及其营养价值分析[J].大豆科学,2013,32(03):355.[doi:10.11861/j.issn.1000-9841.2013.03.0355]
　WANG Min,LIANG Yu,WANG Xin-xin,et al.Assessment on Nutritional Compositions and Value of Jimo Wild Soybean[J].Soybean Science,2013,32(06):355.[doi:10.11861/j.issn.1000-9841.2013.03.0355]
[10]程鹏,徐鹏飞,范素杰,等.野生大豆接种大豆疫霉根腐病菌后过氧化物酶（POD）活性变化[J].大豆科学,2013,32(02):197.[doi:10.3969/j.issn.1000-9841.2013.02.013]
　CHENG Peng,XU Peng-fei,FAN Su-jie,et al.Response of POD Activity in Glycine soja ?Inoculated by Phytophthora sojae[J].Soybean Science,2013,32(06):197.[doi:10.3969/j.issn.1000-9841.2013.02.013]

备注/Memo

基金项目：国家自然科学基金（31571688）。

第一作者简介：郭文雅（1985-），女，硕士，主要从事大豆遗传育种研究。E-mail：cat3.0@163.com。
通讯作者：黄方（1977-），女，博士，教授，主要从事大豆遗传育种研究。E-mail：fhuang@njau.edu.cn。

更新日期/Last Update: 2016-12-09