[1]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):59-64.[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(4):59-64.[doi:10.7685/j.issn.1000-2030.2012.04.011]
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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()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
35卷
期数:
2012年4期
页码:
59-64
栏目:
出版日期:
2012-06-15

文章信息/Info

Title:
Spatiotemporal expression of microRNA156b and microRNA172c and their target genes during flower development of winter buds growing on cut-back treated shoots of grapevine
作者:
王晨1 张演义2 房经贵1 宋长年1 刘洪1 王西成1
1. 南京农业大学园艺学院,江苏 南京 210095;
2. 聊城大学农学院,山东 聊城 252059
Author(s):
WANG Chen1 ZHANG Yan-yi2 FANG Jing-gui1 SONG Chang-nian1 LIU Hong1 WANG Xi-cheng1
1. College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China;
2. College of Agriculture,Liaocheng University,Liaocheng 252059,China
关键词:
葡萄microRNAs靶基因冬芽实时荧光定量RT-PCRRLM-5’-RACE
Keywords:
grapevinemicroRNAstarget geneswinter budsquantitative real-time RT-PCRRLM-5’-RACE
分类号:
S663.1
DOI:
10.7685/j.issn.1000-2030.2012.04.011
摘要:
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-SPL9Vv-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miRNAs 5’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。
Abstract:
The expression of two important regulatory factors(microRNA156b(Vv-miR156b)and microRNA172c(Vv-miR172c)),their target genes and their interaction during the second flower development in winter buds growing on shoots of grapevine ‘Fujimori’subjected to cut-back treatments at various times were studied.Semi-quantitative and quantitative real-time RT-PCR were employed in analyzing variations in the expression levels of the two miRNAs,while RLM-5’-RACE was used to validate the modes by which these miRNAs regulate their target genes.Observations showed that grapevine winters only in the third topping treatment could have flower bud differentiation and form inflorescences while no flower bud differentiation occurred in the other treatments and control winters.Two RT-PCR results with identity showed that both miRNAs exhibited distinctly decreased expression and were least expressed in the inflorescence,while the corresponding target genes for these miRNAs exhibited inverse expression trends with those of their miRNAs.The RLM-5’-RACE results showed that Vv-miR156b and Vv-miR172c could negatively regulate expression of their target genes by cleavage,and their target cleavage sites were between the tenth and eleventh nucleotide at the 5’-end of the miRNA.All results suggested that microRNA156b and microRNA172c were involved in the process of the second flower development of grape winters through regulating expression levels of target genes by cleavage.

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备注/Memo

备注/Memo:
收稿日期:2010-10-31;改回日期:。
基金项目:中央高校基本科研业务费专项(KYJ200909)
作者简介:王晨,博士研究生。
通讯作者:房经贵,教授,博导,研究方向为果树分子生物学,E-mail:fanggg@njau.edu.cn。
更新日期/Last Update: 1900-01-01