2-氨基磷酸茚处理对新疆紫草悬浮细胞次生代谢物积累的影响

Secondary metabolic responses to treatment of 2-aminoindan-2-phosphonic acid in cell lines from Arnebia euchroma

用不同浓度2-氨基磷酸茚(AIP),并结合茉莉酸甲酯(Me JA)处理新疆紫草悬浮细胞,测定不同处理组、不同处理时间细胞中迷迭香酸及乙酰紫草素、脱氧紫草素、β,β'-二甲基丙烯酰紫草素、异戊酰紫草素等次生代谢物的含量,考察AIP抑制苯丙氨酸通路后对紫草悬浮细胞中次生代谢物合成、积累的影响。结果显示Me JA处理能够明显促进新疆紫草细胞中次生代谢物的合成积累;AIP处理能够抑制新疆紫草细胞中上述次生代谢物的积累,并在二者联合处理时能一定程度上抵消Me JA的促进作用,且抑制作用的强弱与处理剂量、处理时间存在正相关性。表明苯丙氨酸途径在紫草素类化合物生物合成中具有重要作用,可为通过细胞培养方式生产紫草素类化合物的代谢调控及紫草素类化合物生物合成等进一步研究提供参考。

The accumulation of rosmarinic acid, acetylshikonin, deoxyshikonin, β, β′-dimethylacrylshikonin and isovalerylshikonin was investigated in cell suspension cultures of Arnebia euchroma （Royle） Johnst under the influence of 2-aminoindan-2-phosphonic acid （AIP）, an inhibitor of phenylalanine ammonia lyase （PAL）, and of the effector methyl jasmonate （MeJA）.The results showed that methyl jasmonate promoted the accumulation of rosmarinic acid and shikonin derivatives.Conversely, 2-aminoindan-2-phosphonic acid suppressed the formation of rosmarinic acid, which indicated that AIP, indeed, was able to inhibit the phenylpropanoid pathway in A.euchroma.Meanwhile, the content of total shinkonins and other four kinds of shikonin derivatives, though varied in degrees, was also inhibited.And the inhibition was dose-dependent and time-dependent.Acetylshikonin responsed most rapidly to the treatment of AIP, the content reduced after 24 h of treatment and decreased to only half of those untreated control 48 h after teratment.β, β′-Dimethylacrylshikonin, difffer from acetylshikonin, responded much slowly to the treatment, inhibition could only be observed 96 h later.These suggest that phenylpropanoid pathway plays an important role in the shikoninsbiosynthesis, and this study provides a reference for the further research in metabolic regulation of producing shikonins by cell culture technology and biosynthesis pathways of shikonin derivatives.Still, shikonins biosynthesis pathways is complicated, the exact dose-and time-effect relationship of AIP and interaction between AIP and other effectors like MeJA need further research.