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以黄色金鱼草(Antirrhinum majus)、“雪球”海棠(Malus‘Snow drift’)为试材,采用农杆菌介导法,将构建的金鱼草素合酶基因(aureusidin synthase,AmAS1)和查尔酮4’-O-葡萄糖苷转移酶基因(chalcone 4’-O-glucosyltransferase,Am4’CGT)双价表达载体在“雪球”活体花瓣上进行瞬时表达,并通过高效液相色谱分析和基因表达分析对瞬时表达效果进行鉴定。结果表明:成功构建双价表达载体CGT-AS-pCambia 1302,农杆菌侵染“雪球”花苞后,花苞呈现淡黄色,表明目的基因已在花瓣内转录并表达。高效液相色谱分析发现,侵染后的花苞出现新的吸收峰,进一步确定花瓣内有橙酮物质的合成。
Using the methods of Agrobacterium-mediated transformation, the antifungal activities of aureusidin synthase (AmAS1) and anti-rhodopseudanidin synthase (AmAS1) were tested using the primers Antirrhinum majus and Malus’ Snow drift. The chalcone 4’-O-glucosyltransferase (Am4’CGT) bivalent expression vector was transiently expressed on “Snowball” living petals and analyzed by high performance liquid chromatography Analysis and Gene Expression Analysis Identify transient expression effects. The results showed that the CGT-AS-pCambia 1302 was successfully constructed, and the flower buds were yellowish after the Agrobacterium tumefaciens infected “Snowball” buds, indicating that the target gene has been transcribed and expressed in the petals. High performance liquid chromatography analysis showed that there was a new absorption peak in the buds after infection, which further confirmed the synthesis of the orange ketone material in the petals.