[1]崔梦杰,郭凤菲,王晨,等.葡萄VvAGL11和VvAGL15基因的鉴定及其在赤霉素诱导葡萄无核果实发育过程中的作用[J].南京农业大学学报,2019,42(2):261-269.[doi:10.7685/jnau.201803054]
 CUI Mengjie,GUO Fengfei,WANG Chen,et al.Identification and roles of VvAGL11 and VvAGL15 gene in the development process of seedless grape berry induced by gibberellin[J].Journal of Nanjing Agricultural University,2019,42(2):261-269.[doi:10.7685/jnau.201803054]
点击复制

葡萄VvAGL11VvAGL15基因的鉴定及其在赤霉素诱导葡萄无核果实发育过程中的作用()
分享到:

《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
42卷
期数:
2019年2期
页码:
261-269
栏目:
植物科学
出版日期:
2019-03-19

文章信息/Info

Title:
Identification and roles of VvAGL11 and VvAGL15 gene in the development process of seedless grape berry induced by gibberellin
作者:
崔梦杰1 郭凤菲2 王晨1 纠松涛1 朱旭东1 房经贵1
1. 南京农业大学园艺学院, 江苏 南京 210095;
2. 南京大学生命科学学院, 江苏 南京 210023
Author(s):
CUI Mengjie1 GUO Fengfei2 WANG Chen1 JIU Songtao1 ZHU Xudong1 FANG Jinggui1
1. College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China;
2. College of Life Science, Nanjing University, Nanjing 210023, China
关键词:
葡萄VvAGL11VvAGL15克隆种胚发育
Keywords:
Vitis vinifera L.VvAGL11VvAGL15cloneseed development
分类号:
S663.1
DOI:
10.7685/jnau.201803054
摘要:
[目的] 本文旨在研究VvAGL11VvAGL15在应答赤霉素(GA3)信号诱导葡萄无核果实发育过程中的作用。[方法] 以‘巨峰’葡萄为试材,鉴定了VvAGL11VvAGL15的cDNA全长序列,并通过分析基因的启动子顺式作用元件预测其潜在功能,同时运用RT-qPCR方法检测对照和GA3处理组葡萄VvAGL11VvAGL15的时空表达水平。[结果] GA3处理能够诱导葡萄果实无核化,使葡萄果粒和果穗显著伸长,穗轴增粗。同时,鉴定到VvAGL11(MG581423)和VvAGL15(MG581424)的cDNA全长序列。VvAGL11和VvAGL15蛋白序列与可可和棉花的亲缘关系较近;二者均含有MADS-MEF2-like和K-box家族保守结构域,属于MADS-box转录因子家族;其主要二级结构元件为α-螺旋和无规则卷曲,且主要定位于细胞核中,蛋白结构具有保守性。启动子顺式作用元件分析显示:二者均含有响应赤霉素和胚乳发育相关的motifs。RT-qPCR分析显示,在果实硬核期和转色期的种子中,VvAGL11VvAGL15的表达量较高,而GA3处理能够显著抑制其在种子中的表达。[结论] 鉴定到MADS-box基因家族中的2个成员,分别为VvAGL11VvAGL15。GA3处理可抑制VvAGL11VvAGL15在种子区的表达,影响种胚的正常发育,从而形成无核果实。
Abstract:
[Objectives] The paper aimed to study the role of VvAGL11 and VvAGL15 in development process of seedless grape berry induced by gibberellin.[Methods] Full-length of cDNA of VvAGL11 and VvAGL15 was obtained from ‘Kyoho’. Potential function of VvAGL11 and VvAGL15 was predicted by analyzing its promoter motif elements. Spatio-temporal expression level of VvAGL11 and VvAGL15 in grape berry from control and GA3 treatment groups was detected by RT-qPCR.[Results] Seedless grape berries were induced by GA3 treatment which can significantly elongate grape berry and clusters and thickening spike stalk. At the same time,cDNA full-length of VvAGL11(MG581423) and VvAGL15(MG581424) was identified. Both VvAGL11 and VvAGL15 contained conserved domains of MADS-MEF2-like and K-box,belonging to the MADS-box transcription factor family. The main secondary structural elements were α-helix and random coil,and these two genes were mainly located in the nucleus. The protein structure was conservative. Promoter cis-acting element analysis showed that both of them contained the gibberellin-responsive elements and the elements required for endosperm expression. RT-qPCR results showed that VvAGL11 and VvAGL15 expressed highly in grape stone hardening stage and veraison stage. GA3 treatment could significantly down-regulate the expression of them only in the seed region.[Conclusions] VvAGL11 and VvAGL15 were identified as members of the MADS-box gene family. GA3 can inhibit the development of seed by down-regulating the expression of VvAGL11 and VvAGL15.

参考文献/References:

[1] 陈俊伟,谢鸣,吴江. 遗传工程无籽果实原理及其研究进展[J]. 浙江农业学报,2003,15(6):365-371. Chen J W,Xie M,Wu J. The principle of breeding of seedless fruit by genetic engineering and its study advance[J]. Acta Agriculturae Zhejiangensis,2003,15(6):365-371(in Chinese with English abstract).
[2] Yao J L,Dong Y H,Bret A M. Parthenocarpic apple fruit production conferred by transposon insertion mutations in a MADS-box transcription factor[J]. Proc Natl Acad Sci USA,2001,98(3):1306-1311.
[3] Singh D P,Jermakow A W,Swain S M. Gibberellins are required for seed development and pollen tube growth in Arabidopsis[J]. Plant Cell,2002,14(12):3133-3147.
[4] 崔梦杰,王晨,张文颖,等. 无核葡萄研究进展[J]. 植物生理学报,2017,53(3):317-330. Cui M J,Wang C,Zhang W Y,et al. Progress in the study of seedless grapes[J]. Plant Physiology Journal,2017,53(3):317-330(in Chinese with English abstract).
[5] 孙其宝,俞飞飞,孙俊. 葡萄无核化研究进展[J]. 安徽农业科学,2004,32(2):360-362. Sun Q B,Yu F F,Sun J. Progress in the study of grapevine berry seedless[J]. Journal of Anhui Agricultural Sciences,2004,32(2):360-362(in Chinese with English abstract).
[6] 肖祥希,李明,邱栋梁. 果实无核机理研究进展[J]. 经济林研究,2009,27(2):104-110. Xiao X X,Li M,Qiu D L. Research progress of seedless mechanism of fruits[J]. Nonwood Forest Research,2009,27(2):104-110(in Chinese with English abstract).
[7] Alvarezbuylla E R,Liljegren S J,Pelaz S,et al. MADS-box gene evolution beyond flowers:expression in pollen,endosperm,guard cells,roots and trichomes[J]. Plant Journal for Cell & Molecular Biology,2010,24(4):457-466.
[8] Battaglia R,Brambilla V,Colombo L,et al. Functional analysis of MADS-box genes controlling ovule development in Arabidopsis using the ethanol-inducible alc gene-expression system[J]. Mechanisms of Development,2006,123(4):267-276.
[9] Becker A,Theissen G. The major clades of MADS-box genes and their role in the development and evolution of flowering plants[J]. Molecular Phylogenetics & Evolution,2003,29(3):464-489.
[10] Huang B,Routaboul J M,Liu M,et al. Overexpression of the class D MADS-box gene Sl-AGL11 impacts fleshy tissue differentiation and structure in tomato fruits[J]. Journal of Experimental Botany,2017,68(17):4869-4884.
[11] Ocarez N,Mejía N. Suppression of the D-class MADS-box AGL11 gene triggers seedlessness in fleshy fruits[J]. Plant Cell Reports,2016,35(1):239.
[12] Wang H,Caruso L V,Downie A B,et al. The Embryo MADS domain protein AGAMOUS-Like 15 directly regulates expression of a gene encoding an enzyme involved in gibberellin metabolism[J]. Plant Cell,2004,16(5):1206-1219.
[13] Zheng Y,Perry S E. Global identification of targets of the Arabidopsis MADS domain protein AGAMOUS-Like 15[J]. The Plant Cell,2009,21(9):2563.
[14] 王文举,马治国. 赤霉素和链霉素及促生灵促进巨峰葡萄果粒增大和无核的试验[J]. 落叶果树,2003,35(4):43-44. Wang W J,Ma Z G. Test of gibberellin,streptomycin and tomatotone on promoting Kyoho grape berry enlargement and seedless[J]. Deciduous Fruits,2003,35(4):43-44(in Chinese with English abstract).
[15] 刘佳,刘晓,陈建. 四川地区赤霉素和链霉素处理对巨峰葡萄无核化及果实发育的影响[J]. 西南农业学报,2011,24(1):220-224. Liu J,Liu X,Chen J. Effect of GA3 and SM treatment on inducing seedless and berry growth of Kyoho grape in Sichuan area[J]. Southwest China Journal of Agricultural Sciences,2011,24(1):220-224(in Chinese with English abstract).
[16] 陈发河,蔡慧农,冯作山,等. 葡萄浆果发育过程中激素水平的变化[J]. 植物生理与分子生物学学报,2002,28(5):391-395. Chen F H,Cai H N,Feng Z S,et al. Changes of hormone levels during the development of grape berries[J]. Journal of Plant Physiology and Molecular Biology,2002,28(5):391-395(in Chinese with English abstract).
[17] 张彦苹,王晨,于华平,等. 适于葡萄不同组织RNA提取方法的筛选[J]. 西北农业学报,2010,19(11):135-140. Zhang Y P,Wang C,Yu H P,et al. Screening of RNA extraction methods for various grapevine organs and tissues[J]. Acta Agriculturae Boreali-occidentalis Sinica,2010,19(11):135-140(in Chinese with English abstract).
[18] Livak K J,Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CT method[J]. Methods,2001,25(4):402-408.
[19] Riechmann J L,Meyerowitz E M. Determination of floral organ identity by Arabidopsis MADS domain homeotic proteins AP1,AP3,PI,and AG is independent of their DNA-binding specificity[J]. Molecular Biology of the Cell,1997,8(7):1243-1259.
[20] Folter S,Shchennikova A V,Franken J,et al. A Bsister MADS-box gene involved in ovule and seed development in petunia and Arabidopsis[J]. The Plant Journal,2006,47(6):934-946.
[21] Deng W,Chen G,Peng F,et al. Transparent testa16 plays multiple roles in plant development and is involved in lipid synthesis and embryo development in canola[J]. Plant Physiology,2012,160(2):978-989.
[22] Xu W,Fiume E,Coen O,et al. Endosperm and nucellus develop antagonistically in Arabidopsis seeds[J]. The Plant Cell,2016,28(6):1343-1360.
[23] Yang Z,Li C,Wang Y,et al. GhAGL15s,preferentially expressed during somatic embryogenesis,promote embryogenic callus formation in cotton(Gossypium hirsutum L.)[J]. Molecular Genetics & Genomics,2014,289(5):873-883.
[24] 董庆龙,冀志蕊,迟福梅,等. 苹果MADS-box转录因子的生物信息学及其在不同组织中的表达[J]. 中国农业科学,2014,47(6):1151-1161. Dong Q L,Ji Z R,Chi F M,et al. Bioinformatics of the MADS-Box transcription factor and their expression in different apple tissues[J]. Scientia Agricultura Sinica,2014,47(6):1151-1161(in Chinese with English abstract).
[25] 蔡英卿. 龙眼体胚发生过程中SERK等胚性相关基因的克隆与表达分析[D]. 福州:福建农林大学,2011. Cai Y Q. Cloning and epression of SERK and sme other somatic embryogenesis-related genes during somatic embryogenesis in Dimocarpus longan Lour[D]. Fuzhou:Fujian Agriculture and Forestry University,2011(in Chinese with English abstract).
[26] Parenicová L,De S F,Kieffer M,et al. Molecular and phylogenetic analyses of the complete MADS-box transcription factor family in Arabidopsis:new openings to the MADS world[J]. The Plant Cell,2003,15(7):1538-1551.
[27] 王飞. 葡萄无核品种及其杂种胚败育机理与胚挽救技术研究[D]. 杨凌:西北农林科技大学,2002. Wang F. Mechanism of embryo abortion in stonespermic seedless grape and embryo rescue with seedless grape[D]. Yangling:Northwest A&F University,2002(in Chinese with English abstract).

相似文献/References:

[1]宗成文,章镇,房经贵,等.葡萄LEAFY基因启动子的克隆与序列分析[J].南京农业大学学报,2007,30(4):20.[doi:10.7685/j.issn.1000-2030.2007.04.005]
 ZONG Cheng-wen,ZHANG Zhen,FANG Jing-gui,et al.Cloning and sequence analysis of LEAFY gene promoter from grape(Vitis vinifera×V.labrusca)[J].Journal of Nanjing Agricultural University,2007,30(2):20.[doi:10.7685/j.issn.1000-2030.2007.04.005]
[2]黄非,盛炳成.宿晓红葡萄与葡萄属有关种亲缘关系初探[J].南京农业大学学报,1993,16(4):49.[doi:10.7685/j.issn.1000-2030.1993.04.009]
 Huang Fei Sheng Bingcheng.STUDY ON THE RELATIONSHIP BETWEEN SUXIAOHONG GRAPE AND SOME SPECIES OF VITIS[J].Journal of Nanjing Agricultural University,1993,16(2):49.[doi:10.7685/j.issn.1000-2030.1993.04.009]
[3]周培根,戚晓玉,P.A.Braell,等.葡萄中β-damascenone前体的初步纯化[J].南京农业大学学报,1989,12(3):79.[doi:10.7685/j.issn.1000-2030.1989.03.017]
[4]韩浩章,姜卫兵,费宪进,等.葡萄和油桃自然休眠解除过程中H2O2含量和抗氧化酶活性的变化[J].南京农业大学学报,2007,30(1):50.[doi:10.7685/j.issn.1000-2030.2007.01.010]
 HAN Hao-zhang,JIANG Wei-bing,FEI Xianjin,et al.Changes in H_2O_2 content and activities of antioxidant enzymes of grape and nectarine during natural dormancy-release[J].Journal of Nanjing Agricultural University,2007,30(2):50.[doi:10.7685/j.issn.1000-2030.2007.01.010]
[5]郭磊,上官凌飞,房经贵*,等.葡萄EST鄄SSR 标记的开发及其应用[J].南京农业大学学报,2011,34(4):23.[doi:10.7685/j.issn.1000-2030.2011.04.005]
 GUO Lei,SHANGGUAN Ling-fei,FANG Jing-gui *,et al.Development of SSR markers from grape ESTs and its application[J].Journal of Nanjing Agricultural University,2011,34(2):23.[doi:10.7685/j.issn.1000-2030.2011.04.005]
[6]孙兴民,余智莹,张萌,等.葡萄未成熟胚诱导体细胞胚发生和植株再生与遗传鉴定[J].南京农业大学学报,2012,35(3):13.[doi:10.7685/j.issn.1000-2030.2012.03.003]
 SUN Xing-min,YU Zhi-ying,ZHANG Meng,et al.Somatic embryogenesis from immature zygotic embryos and genetic fidelity identificating of regenerated plants of grapevine[J].Journal of Nanjing Agricultural University,2012,35(2):13.[doi:10.7685/j.issn.1000-2030.2012.03.003]
[7]王晨,张演义,房经贵,等.葡萄microRNA156b和microRNA172c及其靶基因在冬芽二次成花过程中的表达特性研究[J].南京农业大学学报,2012,35(4):59.[doi:10.7685/j.issn.1000-2030.2012.04.011]
 WANG Chen,ZHANG Yan-yi,FANG Jing-gui,et al.Spatiotemporal expression of microRNA156b and microRNA172c and their target genes during flower development of winter buds growing on cut-back treated shoots of grapevine[J].Journal of Nanjing Agricultural University,2012,35(2):59.[doi:10.7685/j.issn.1000-2030.2012.04.011]
[8]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.[doi:10.7685/j.issn.1000-2030.2013.01.006]
 WANG Xicheng,WANG Chen,FANG Jinggui,et al.Cloning,subcellular localization and spatiotemporal expression of VvGA2ox1 gene from grapevine[J].Journal of Nanjing Agricultural University,2013,36(2):29.[doi:10.7685/j.issn.1000-2030.2013.01.006]
[9]谢荔,成学慧,冯新新,等.氨基酸肥料对‘夏黑’葡萄叶片光合特性与果实品质的影响[J].南京农业大学学报,2013,36(2):31.[doi:10.7685/j.issn.1000-2030.2013.02.006]
 XIE Li,CHENG Xuehui,FENG Xinxin,et al.Effects of an amino acid fertilizer on the leaf photosynthesis and fruit quality of ‘Summer Black’grape[J].Journal of Nanjing Agricultural University,2013,36(2):31.[doi:10.7685/j.issn.1000-2030.2013.02.006]
[10]任国慧,上官凌飞,房经贵,等.葡萄DELLA家族成员VvRGA和VvRGL1的预测、验证及生物信息学分析[J].南京农业大学学报,2013,36(3):15.[doi:10.7685/j.issn.1000-2030.2013.03.003]
 REN Guohui,SHANGGUAN Lingfei,FANG Jinggui,et al.The prediction,validation and bioinformatics analysis of the grape DELLA family members about VvRGA and VvRGL1[J].Journal of Nanjing Agricultural University,2013,36(2):15.[doi:10.7685/j.issn.1000-2030.2013.03.003]

备注/Memo

备注/Memo:
收稿日期:2018-3-27。
基金项目:江苏省农业科技自主创新资金[CX(16)1013];中央高校基本科研业务费专项资金(KYTZ201602)
作者简介:崔梦杰,硕士研究生。
通信作者:王晨,硕导,研究方向为葡萄遗传育种与基因组学,E-mail:wangchen@njau.edu.cn。
更新日期/Last Update: 1900-01-01