酵母双杂交系统筛选与水稻黑条矮缩病毒P6互作的水稻蛋白

Screening of Rice Proteins Interacting with P6 of Rice Black Streaked Dwarf Virus from Rice cDNA Library by Yeast Two Hybrid System

RBSDV编码的非结构蛋白P6为该病毒的沉默抑制子。为了研究P6与寄主水稻间的互作关系,首先利用RT-PCR扩增,获得水稻黑条矮缩病毒P6基因,并将其构建到酵母表达载体pGBKT7上,自激活验证表明,P6蛋白无自激活活性。利用酵母双杂交系统顺序转化法从水稻cDNA文库中筛选,获得4个与RBSDVP6互作的寄主因子,分别为水稻内囊体抗坏血酸过氧化物酶、醛糖1差向异构酶、immutans蛋白基因同源蛋白和一个功能未知蛋白。分析了3个已知功能的寄主因子在病毒侵染过程中的可能功能。

Citranges transferred with phyB gene were detected at the molecular level and their biological characteristics were analyzed. PCR, RT-PCT and PCR-Southern blot analysis indicated that phyB gene was successfully integrated into citrange genome and was transcribed in citrange. Compared with the non-transgenic plant, the transgenic plant was dwarf, with larger shoot insertion angle and smaller leaf area, and was higher in chlorophyll content, net photosynthesis rate, light saturation point and specific leaf weight. Determination of endogenous hormone by HPLC indicated that the content of GA1, GA4, ABA and Z of the transgenic plant was 32.7%, 50%, 121% and 17.8% higher than that of the non-transgenic plant, respectively, whereas the content of IAA was lower than that of the non-transgenic plant. （GA1＋ GA4）/ABA and Z/ABA ratios of the transgenic plant were 38.4% and 43.2% lower than those of the control, respectively.