黄枸杞中调控花青素生物合成候选基因AN2相关功能验证

Related function verification of anthocyanin biosynthesis candidate gene AN2 from lycium barbarum L

花青素是一种存在于植物中的显色水溶性类黄酮化合物,具有较高的营养价值.黄枸杞果实中色素的大量积累与花青素生物合成代谢密切相关.前人研究发现MYB转录因子LrAN2主要参与调控黑枸杞黑色果实中花青素生物合成代谢途径,而黄枸杞中花青素合成代谢的分子机理尚不明确.本研究利用同源克隆的方式分离黄枸杞AN2基因,通过Gateway技术构建植物中过量表达AN2载体PJAM1502:AN2.利用农杆菌侵染的方式将AN2基因转染到模式植物烟草中,得到紫色烟草表型.花青素含量测定发现烟草叶片中花青素含量最高,同时qPCR验证检测出转基因烟草中花青素合成代谢途径中的结构基因都显著上调,其中结构基因ANS最为明显.本研究发现黄枸杞中AN2基因与黄枸杞黄色表型的形成密切相关,为黄枸杞的分子育种研究提供可靠的理论基础.

Anthocyanin is a colored water-soluble flavonoid existing in plants,which has high nutritional value.The accumulation of pigments in Lycium barbarum L fruit is closely related to anthocyanin biosynthesis and metabolism.Previous studies have found that MYB transcription factor LrAN2 is mainly involved in regulating the pathway of anthocyanin biosynthesis and metabolism in black fruits of Lycium ruthenicum Murr,while the molecular mechanism of anthocyanin biosynthesis and metabolism in Lycium barbarum L is still unclear.In this study,the AN2 gene of Lycium barbarum L was isolated by homologous cloning,and the overexpression vector PJAM1502:AN2 in plants was constructed by Gateway technique.The AN2 gene was transfected into model tobacco by Agrobacterium tumefaciens,and the purple phenotype was obtained.The content of anthocyanin in tobacco leaves was the highest,and the structural genes in anthocyanin biosynthesis and metabolism pathway in transgenic tobacco were significantly up-regulated by qPCR,among which structural genes ANS was the most obvious.The study found that AN2 gene in Lycium barbarum L was closely related to the formation of yellow phenotype of Lycium barbarum L and provided a reliable theoretical basis for molecular breeding of Lycium barbarum L.