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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

来源：花匠小妙招时间：2025-10-29 09:05

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DOI: 10.7685/j.issn.1000-2030.2012.04.011, PP. 59-64

Keywords: 葡萄,microRNAs,靶基因,冬芽,实时荧光定量RT-PCR,RLM-5’-RACE

Abstract:

利用半定量RT-PCR与实时荧光定量RT-PCR(qRT-PCR)技术检测‘藤稔’葡萄microRNA156b(Vv-miR156b)和microRNA172c(Vv-miR172c)及其靶基因(Vv-SPL9和Vv-AP2)在摘心处理(5月14日、5月20日、5月28日)后不同发育时期冬芽中的表达情况,并利用RLM-5’-RACE验证了miRNAs作用其靶基因的方式。结果表明仅5月28日摘心处理的冬芽能够进行花芽分化并形成花序,而其他处理及对照未能花芽分化。2种RT-PCR检测结果基本一致,5月28日摘心处理的2条miRNAs表达水平明显下降,且花序中有最低表达。相应的靶基因在冬芽二次成花过程中的表达水平呈现出与其miRNAs相反的变化趋势。RLM-5’-RACE结果显示,2条miRNAs介导靶基因裂解,裂解位点均在miRNAs5’端的第10和11位碱基之间。表明microRNA156b和microRNA172c通过介导相应靶基因的裂解调控其靶基因的表达,从而影响葡萄冬芽的二次成花。

References[1]  王晨,刘洪,房经贵,等.基于EST库的葡萄(Vitis vinifera)APETAAL2基因cDNA克隆及其表达分析[J].果树学报,2010,27(2):207-212
[2]  Levy Y Y,Deng C.The transition to flowering[J].The Plant Cell,1998,10:1973-1990
[3]  Glazinska P,Zienkiewicz A,Wojciechowski W,et al.The putative miR172 target gene APETALA2-like is involved in the photoperiodic flower induction of Ipomoea nil[J].Plant Physiology,2009,166(16):1801-1813
[4]  Wang J W,Czech B,Weigel D.miR156-regulated SPL transcription factors define an endogenous flowering pathway in Arabidopsis thaliana[J].Cell,2009,138(4):738-749
[5]  Lauter N,Kampani A,Carlson S,et al.MicroRNA172 down regulates glossy15 to promote vegetative phase change in maize[J].Proceedings of the National Academy of Sciences USA,2005,102:9412-9417
[6]  Chen X.A microRNA as a translational repressor of APETALA2 in Arabidopsis flower development[J].Science,2004,303:2022-2025
[7]  陶磅,贾克功,舒群,等.葡萄一、二次果的花器官及果实发育比较[J].西南农业学报,2002,15(2):100-105
[8]  房玉林,李华,宋建伟,等.葡萄产期调节的研究进展[J].西北农业学报,2005,14(3):98-101
[9]  郭磊,王晨,曹雪,等.葡萄夏芽成花过程中相关基因的cDNA-RAPD分析[J].华北农学报,2011,26(2):43-48
[10]  邹秀华.利用葡萄冬芽二次结果的原理与技术讨论[J].潍坊高等职业教育,2010,6(1):64-66
[11]  Jaillon O,Aury J M,Noel B,et al.The grapevine genome sequence suggests ancestral hexaploidization in major angiosperm phyla[J].Nature,2007,449:463-467
[12]  Wang C,Wang X,Kibet K N,et al.Deep sequencing of grapevine flower and berry short RNA library for discovery of novel microRNAs and validation of precise sequences of grapevine microRNAs deposited in miRBase[J].Physiologia Plantarum,2011,143:64-81
[13]  曹雪,王晨,房经贵,等.葡萄SPL9和SPL10基因全长cDNA克隆、亚细胞定位和表达分析[J].园艺学报,2011,38(4):240-250
[14]  Wang C,Shangguan L F,Kibet K N,et al.Characterization of microRNAs identified in a table grapevine cultivar with validation of computationally predicted grapevine miRNAs by miR-RACE[J].PLoS One,2011,6(7):e21259
[15]  Song C N,Fang J G,Wang C,et al.MiR-RACE,a New efficient approach to determine the precise sequences of computationally identified trifoliate orange(Poncirus trifoliate)microRNAs[J].PLoS One,2010,5:e10861
[16]  Chang S,Purvear J,Caimey J.A simple and efficient method for isolating RNA from pine trees[J].Plant Molecular Biology Reporter,1993,11:113-116
[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
[18]  Song C,Fang J,Li X,et al.Identification and characterization of 27 conserved microRNAs in citrus[J].Planta,2009,230:671-685
[19]  Song C N,Wang C,Zhang C Q,et al.Deep sequencing discovery of novel and conserved microRNAs in trifoliate orange(Citrus trifoliate)[J].BMC Genomics,2010,11:431
[20]  Morrison J C.Bud development in Vitis vinifera L.[J].Botanical-Gazette-Chicago,1991,152:304-315
[21]  Jung J H,Seo P J,Kang S K,et al.miR172 signals are incorporated into the miR156 signaling pathway at the SPL3/4/5 genes in Arabidopsis developmental transitions[J].Plant Molecular Biology,2011,76:35-45

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