以‘赤霞珠’葡萄果实为材料,利用制备好的葡萄查耳酮异构酶(CHI;EC 5.5.1.6)CHI多克隆抗体,采用RT-PCR、蛋白质印记杂交和胶体金免疫电镜技术对葡萄果实发育过程中CHI基因、蛋白表达及亚细胞定位进行了研究。结果表明:CHI基因和蛋白的表达随着果实发育有着明显的变化,在果实发育早期和转色期表达量较高,同果实中总类黄酮含量的积累变化一致;胶体金免疫定位显示在果实发育早期CHI主要分布于葡萄果皮细胞的细胞质中,在液泡及质体中也有少量分布;在转色期主要定位于细胞质、质体和细胞核中,其数量明显增多。在成熟果实中,主要存在于细胞质,少量存在于质体和液泡中。 Polyclonal antibody against CHI chalcone isomerase (CHI; EC 5.5.1.6) was prepared from ’Cabernet Sauvignon’ grapes using RT-PCR, Western blotting and colloidal gold immunoelectron microscopy CHI gene, protein expression and subcellular localization during grape fruit development were studied. The results showed that the expression of CHI gene and protein had obvious changes with the development of fruit, the expression level of CHI gene and protein was higher in the early and middle stages of fruit development, and the accumulation of total flavonoids in fruit was the same. In early development, CHI mainly distributed in the cytoplasm of grape pericarp cells, and there was also a small amount of distribution in vacuoles and plastids. The number of CHI was mainly located in the cytoplasm, plastid and nucleus during the color change. In mature fruits, mainly in the cytoplasm, a small number of plastids and vacuoles.