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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。
Using Vitis vinifera L. ’Wink’ as the material, a MYB transcription factor gene Vv GAMYB was obtained from grape gibberellin signaling pathway. The open reading frame (ORF) of this gene is 1 680 bp in length and encodes 559 amino acids. The ORF contains the conserved R2R3 DNA binding domain of the GAMYB gene family as well as the Box1, Box2 and Box3 conserved domains. The transient expression of onion epidermal cells showed that VvGAMYB was located in the nucleus. The expression of VvGAMYB and its downstream gene Vv LFY showed a trend of first increasing and then decreasing, whereas GA3 treatment promoted the expression of VvGAMYB and VvLFY, leading to early flowering. VvGAMYB is the target gene of miR159 whose function is regulated by miR159. The negative correlation between miR159 and VvGAMYB expression was observed. Gibberellin treatment down-regulated the expression of miR159 and thus reduced the cleavage of VvGAMYB. From the above results, it can be seen that VvGAMYB participates in the flowering process of grape through the gibberellin flowering pathway. In this process, gibberellin inhibits miR159 cleavage of VvGAMYB, thereby upregulating the gene expression of VvLFY and participating in the regulation of grape flowering.