葡萄miR159及其靶基因VvGAMYB在花发育过程中的作用分析  

Function Analysis of miR159 and Its Target Gene VvGAMYB in Grape Flower Development

作　　者：王梦琦[1] 解振强[2] 孙欣[1] 李晓鹏[1] 朱旭东[1] 王晨[1] 房经贵[1] 

机构地区：[1]南京农业大学园艺学院,南京210095 [2]江苏农林职业技术学院,江苏句容212400

出　　处：《园艺学报》2017年第6期1061-1072,共12页Acta Horticulturae Sinica

基　　金：中国博士后科学基金项目(802602);江苏省博士后科研资助计划项目(82161015)

摘　　要：以‘魏可’葡萄(Vitis vinifera L.‘Wink’)为试材,克隆得到一个葡萄赤霉素信号途径中的MYB转录因子基因Vv GAMYB。该基因开放阅读框(ORF)长1 680 bp,编码559个氨基酸,该蛋白序列含有GAMYB基因家族保守的R2R3 DNA结合域以及Box1、Box2、Box3保守域。洋葱表皮细胞的瞬时表达结果显示,VvGAMYB定位于细胞核中。VvGAMYB和其下游基因Vv LFY在葡萄花发育过程中的表达都呈现先上升后下降的趋势,而赤霉素处理促进了VvGAMYB和VvLFY的表达,从而使开花提前。VvGAMYB是miR159的靶基因,其作用受到miR159的调控。miR159和VvGAMYB的表达负相关,赤霉素处理下调miR159表达从而也降低了对VvGAMYB的裂解作用。由上述结果可以看出,VvGAMYB通过赤霉素开花途径参与葡萄的开花过程。在这一过程中,赤霉素抑制miR159裂解VvGAMYB的作用,从而上调VvLFY的基因表达,参与了调控葡萄开花过程。VvGAMYB plays an important role in grape flower development. A MYB transcription factor gene in gibberellic acid（GA）signaling pathway,designated as VvGAMYB,was cloned using homology-based cloning and reverse transcription-PCR（RT-PCR） from Vitis vinifera L.＇Wink＇. The open reading frame of VvGAMYB was 1 680 bp,encoding a protein of 559 amino acids. Sequence alignement showed that VvGAMYB protein contained a highly conserved R2R3 domain. In addition,VvGAMYB also contained 3 conserved motifs Box1,Box2 and Box3,which are typical structures in the GAMYB family. The VvGAMYB protein was mainly localized in the nucleus of onion epidermal cell. The expression of VvGAMYB and its downstream gene VvLFY increased and subsequently decreased during the development of grape flower. The GA3 treatment induced the expression of VvGAMYB and VvLFY,which promoted the development of grape flower. VvGAMYB was the target gene of miR159,and its expression was regulated by miR159. The miR159 could negatively regulate expression of VvGAMYB by cleavage. The expression of mi R159 and its target gene VvGAMYB showed a negative correlation,and its expression was decreased by GA3 treatment,which also reduced the cleavage of VvGAMYB. These results suggest that VvGAMYB might paly a crucial role in grape flower development through the gibberellin pathway. In this process,the expression of miR159 was down-regulated while VvGAMYB and VvLFY was up-regulated.

关 键 词：葡萄 GAMYB miR159 赤霉素 花发育 基因表达 

分 类 号：S663.1[农业科学—果树学]