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以酿酒葡萄‘赤霞珠’(VitisviniferaL.‘Cabernet Sauvignon’)为试材,在果实发育过程中定期对植株进行UV-C照射,分别采用分光光度计法、免疫组织定位等方法,对黄烷醇类多酚时空积累及其合成关键酶LAR(leucoanthocyanidin reductase,LAR)活性和定位进行初步研究。结果表明:UV-C诱导果皮和果肉中总酚、黄烷醇类多酚、总黄烷–3–醇的积累,LAR酶活性增强,且这一诱导作用有明显的照射剂量、器官/组织和发育阶段依赖性。UV-C照射并不改变LAR1、LAR2酶蛋白在葡萄果实中的分布,但诱导酶蛋白积累,特别在果皮及果肉维管束中,UV-C照射导致LAR1、LAR2酶蛋白信号明显增强。所有结果表明,UV-C照射诱导果皮和果肉维管束中LAR1、LAR2酶蛋白增加,诱导LAR酶活性增强,最终导致总黄烷–3–醇和黄烷醇类多酚特异性积累。
Taking the wine grape “Cabernet Sauvignon” as the test material, the plants were irradiated with UV-C on a regular basis during the fruit development. By using the methods of spectrophotometer and immunohistochemistry respectively, The spatial and temporal accumulation of alcohol polyphenols and the activity and localization of LAR (key enzyme for the synthesis of LAR) were studied. The results showed that UV-C induced the accumulation of total phenols, flavanols and total flavan-3-ol in pericarp and pulp, and enhanced the activity of LAR. And this induction had significant dose, organ / tissue And developmental stage dependence. UV-C irradiation did not change the distribution of LAR1 and LAR2 enzyme in grape fruit, but induced the accumulation of protein, especially in the peel and pulp vascular bundles, UV-C irradiation led to significantly enhanced LAR1 and LAR2 enzyme signals. All of the results showed that UV-C irradiation induced the increase of LAR1 and LAR2 enzyme proteins in the pericarp and flesh vascular bundles, leading to the enhancement of LAR enzyme activity and finally the specific accumulation of total flavan-3-ol and flavanol polyphenols.