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花青素已成为植物遗传转化的新型可视化报告基因。为了探索花青素作为报告基因在橡胶树体细胞胚遗传转化中的应用,本研究通过橡胶树体细胞胚花青素HLPC分析,检测橡胶树体细胞胚中是否自身累积花青素及其种类。结果显示,利用3 mol/L的盐酸水溶液沸水浴,从离体培养的橡胶树体细胞胚中分离到花青素,同时,HPLC测定发现橡胶树体细胞胚中花青素主要有两个吸收峰,分别出现在10.571 min和11.763 min,与标准品比对发现保留时间为11.742 min为矢车菊素,其含量为4.613 7μg/g,另一个需要进一步分析。橡胶树体细胞胚在离体培养条件下能够检测到花青素的积累,说明在体细胞胚中存在完整的花青素代谢途径,通过花青素基因调控,有望实现花青素作为橡胶树遗传转化的报告基因。
Anthocyanins have become a new visual reporter for plant genetic transformation. In order to explore the application of anthocyanin as a reporter gene in the genetic transformation of somatic embryos in rubber tree, this study detected the accumulation of anthocyanins and their species in somatic embryos of rubber tree by HLPC analysis of rubber tree somatic embryos. The results showed that anthocyanins were isolated from somatic embryos of rubber tree cells cultured in vitro by using 3 mol / L aqueous hydrochloric acid in boiling water bath. At the same time, anthocyanin in somatic embryos of rubber tree was found to have two absorption peaks, Respectively, appeared at 10.571 min and 11.763 min. Compared with the standard, the retention time was 11.742 min. The content of cyanidin was 4.613 7 μg / g, and the other one needed further analysis. Somatic embryos of rubber tree could detect the accumulation of anthocyanins under in vitro culture conditions, indicating that there is a complete metabolic pathway of anthocyanins in somatic embryos. Through the regulation of anthocyanidin gene, it is expected that anthocyanin can be transformed into rubber tree Of the reporter gene.