多蘖矮秆基因dwarf-3(D3)是水稻独脚金内酯信号转导过程中的重要节点基因,拟南芥中的MAX2基因与D3 同源,且MAX2 参与拟南芥的抗病防卫反应.本研究以水稻泛素连接酶D3 为诱饵进行酵母双杂筛库,发现水稻抗病相关蛋白维管植物单锌指蛋白VOZ2 与D3 存在潜在的相互作用.通过酵母双杂交试验证实,D3 与VOZ2 存在互作.通过荧光定量PCR证实,接种水稻白叶枯病菌后,VOZ2 基因在转录水平上的表达受到显著诱导.利用水稻原生质体开展的亚细胞共定位实验发现,D3 与VOZ2 共定位于细胞核.双分子荧光互补实验发现,D3 与VOZ2 在烟草叶肉细胞的细胞核和细胞质均产生较强的荧光,进一步证实了D3与VOZ2 的相互作用.研究结果为进一步探究D3 和VOZ2 在水稻抗病防卫反应中的功能与分子机理奠定了基础.
The tiller dwarf gene dwarf-3(D3)is an important node gene in strigolactone signal transduction in rice.It has been reported that the MAX2 gene in Arabidopsis thaliana is homologous to D3,and MAX2 is involved in the defense response of A.thaliana.In this study,the yeast two-hybrid library was screened with D3 protein as bait,and it was found that there was a potential interaction between rice disease resistance-related protein vascular plant one-zinc finger protein 2(VOZ2)and D3.The interaction between D3 and VOZ2 was confirmed by yeast two-hybrid validation.Real-time quantitative PCR experiments showed that the expression of VOZ2 gene was significantly in-duced at the transcriptional level after inoculation with Xanthomonas oryzae pv.oryzae.Subcellular co-localization experiment using rice protoplasts showed that D3 and VOZ2 were co-localized in the nucleus.Bimolecular fluores-cence complementation experiments showed that D3 and VOZ2 produced strong fluorescence in the nucleus and cyto-plasm of tobacco mesophyll cells,further confirming the interaction between D3 and VOZ2.The study result laid a foundation for further exploring of the function and molecular mechanism of D3 and VOZ2 in rice disease resistance and defense response.