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花青素是一种简单、安全和肉眼直接可辨转基因筛选标记。本研究为了建立橡胶树体细胞胚花青素调控基因发掘的实验体系,探讨了茉莉酸处理方式和时间对橡胶树体细胞胚累积花青素的影响,以及分析了花青素合成关键结构基因在茉莉酸处理前后表达。结果显示140μmol/L茉莉酸甲酯液体培养4 h后,再转移到无茉莉酸甲酯的固体培养基上继续培养,不但能够诱导花青素累积,而且较直接固体培养基添加茉莉酸甲酯,提早1~2 d观察到花青素的累积,同时,累积花青素主要为矢车菊,处理后矢车菊含量提高了5倍以上。再者,花青素合成相关13个结构基因在茉莉酸甲酯处理后均上调表达,尤其Hb LDOX上调表达13.29倍。本研究证实了茉莉酸是诱导橡胶树体细胞胚花青素累积的有效诱导剂,而且Hb LDOX可能是橡胶树体细胞胚花青素累积关键限速步骤。
Anthocyanins are a simple, safe and visually distinguishable direct gene screening marker. In this study, in order to establish an experimental system for the excision of anthocyanin regulated genes of rubber tree somatic embryos, the effects of jasmonic acid treatment and time on the accumulation of anthocyanins in somatic embryos of rubber trees were discussed. And the key structural genes of anthocyanin synthesis in jasmine Acid treatment before and after expression. The results showed that 140μmol / L methyl jasmonate liquid cultured 4 h, then transferred to non-methyl jasmonate solid medium to continue the culture, not only to induce the accumulation of anthocyanins, but also more direct solid medium added jasmonic acid methyl ester , The accumulation of anthocyanins was observed 1 ~ 2 days earlier, meanwhile, the accumulation of anthocyanin was mainly cornflower, and the content of cornflower increased more than 5 times. In addition, 13 structural genes related to anthocyanin synthesis were upregulated after methyl jasmonate treatment, especially 13.29 times higher than that of Hb LDOX. This study confirmed that jasmonic acid is an effective inducer of ECG accumulation in rubbery somatic embryos, and that Hb LDOX may be the key rate-limiting step in the accumulation of ECs in rubber tree.