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本文利用发根农杆菌A4诱导甜叶菊毛状根再生,建立毛状根生产绿原酸类物质的培养体系,并研究茉莉酸甲酯和水杨酸对毛状根中绿原酸类物质积累的影响。结果表明,经发根农杆菌A4侵染的甜叶菊叶片外植体在共培养14 d后诱导出了毛状根;PCR检测结果表明,发根农杆菌Ri质粒中rol B和rol C基因均已整合到毛状根中;毛状根在MS液体培养基中,培养到第14 d时分别加入不同浓度(50、100、200μmol/L)的水杨酸和茉莉酸甲酯进行诱导,处理第1、3、6 d后均抑制了毛状根的生长,但茉莉酸甲酯促进了毛状根中绿原酸类物质的积累,而水杨酸却抑制了毛状根中绿原酸类物质的积累。由此说明,发根农杆菌A4侵染甜叶菊可诱导出毛状根,该毛状根可用于绿原酸类物质的生产,茉莉酸甲酯可提高绿原酸类物质的含量。
In this paper, Agrobacterium rhizogenes A4 was used to induce the regeneration of hairy roots and the culture system of hairy roots for the production of chlorogenic acids was established. The effects of methyl jasmonate and salicylic acid on the accumulation of chlorogenic acids in hairy roots Impact. The results showed that the hairy roots were induced after 14 days of co-cultivation by the leaf explants of Agrobacterium rhizogenes A4 infected with Agrobacterium rhizogenes. PCR results showed that both rol B and rol C genes of Agrobacterium rhizogenes Ri plasmid Hairy roots were integrated into hairy roots. Hairy roots were cultured in liquid MS medium and cultured on the 14th day. Salicylic acid and methyl jasmonate with different concentrations (50,100 and 200μmol / L) Hairy root growth was inhibited on the 1st, 3rd and 6th day, but methyl jasmonate promoted the accumulation of chlorogenic acids in the hairy roots, whereas salicylic acid inhibited the chlorogenic acid Accumulation of substances. Thus, Agrobacterium rhizogenes A4 could infect hairy roots by inoculation with Stevia rebaudiana, which could be used for the production of chlorogenic acids. Methyl jasmonate could increase the content of chlorogenic acids.