Functional Analysis of the Isopentenyl Diphosphate Isomerase of Salvia miltiorrhiza via Color Complementation and RNA Interference.
Functional Analysis of the Isopentenyl Diphosphate Isomerase of Salvia miltiorrhiza via Color Complementation and RNA Interference.
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通过颜色互补和 RNA 干扰对丹参异戊烯基二磷酸异构酶进行功能分析。
DOI：
10.3390/molecules201119689
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发表时间：
2015-11-10
期刊：
影响因子：
--
通讯作者：
Huang L
中科院分区：
文献类型：
--
作者：
Zhang X;Guan H;Dai Z;Guo J;Shen Y;Cui G;Gao W;Huang L
异戊烯基二磷酸异构酶（IPI）催化萜类化合物生物合成过程中常见的萜类前体物质异戊烯基二磷酸（IPP）和二甲基烯丙基二磷酸（DMAPP）之间的异构化。在这项研究中，组织表达分析表明，丹参IPI 1基因（SmIPI 1）的表达水平在叶中高于根和茎。进一步采用颜色互补和RNA干扰两种方法从两个方面验证SmIPI 1基因的功能。成功构建了SmIPI 1重组质粒，并将其转入工程菌E.通过与对照组的颜色差异验证SmIPI 1的功能，观察到的颜色差异表明SmIPI 1具有促进番茄红素积累的作用。利用发根农杆菌ACCC 10060介导的方法，成功构建了SmIPI 1 RNA干扰的转化毛状根系。RNA干扰毛状根表现为严重的枯萎、畸形甚至死亡的表型。SmIPI 1在RSi 3根系中的mRNA表达水平仅为野生型的8.4%。此外，丹参酮的含量太低，在RNA干扰线检测不到。这些结果表明SmIPI 1在萜类化合物代谢途径中起着关键作用。外源SmIPI 1基因的加入促进了E. coli中，SmIPI 1对S.丹参毛状根可能导致2-C-甲基-D-β-4-磷酸代谢途径中断。
Isopentenyl diphosphate isomerase (IPI) catalyzes the isomerization between the common terpene precursor substances isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) during the terpenoid biosynthesis process. In this study, tissue expression analysis revealed that the expression level of the Salvia miltiorrhiza IPI1 gene (SmIPI1) was higher in the leaves than in the roots and stems. Furthermore, color complementation and RNA interference methods were used to verify the function of the SmIPI1 gene from two aspects. A recombinant SmIPI1 plasmid was successfully constructed and transferred into engineered E. coli for validating the function of SmIPI1 through the color difference in comparison to the control group; the observed color difference indicated that SmIPI1 served in promoting the accumulation of lycopene. Transformant hairy root lines with RNA interference of SmIPI1 were successfully constructed mediated by Agrobacterium rhizogenes ACCC 10060. RNA interference hairy roots had a severe phenotype characterized by withering, deformity or even death. The mRNA expression level of SmIPI1 in the RSi3 root line was only 8.4% of that of the wild type. Furthermore the tanshinone content was too low to be detected in the RNA interference lines. These results suggest that SmIPI1 plays a critical role in terpenoid metabolic pathways. Addition of an exogenous SmIPI1 gene promoted metabolic flow toward the biosynthesis of carotenoids in E. coli, and SmIPI1 interference in S. miltiorrhiza hairy roots may cause interruption of the 2-C-methyl-D-erythritol-4-phosphate metabolic pathway.
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影响因子：
3.2
作者：
Barkley, SJ;Desai, SB;Poulter, CD
通讯作者：
Poulter, CD
影响因子：
11.6
作者：
Cunningham, FX;Pogson, B;Gantt, E
通讯作者：
Gantt, E
丹参萜类生物合成新基因的全基因组鉴定和表征
影响因子：
6.9
作者：
Ma, Yimian;Yuan, Lichai;Lu, Shanfa
通讯作者：
Lu, Shanfa
影响因子：
56.9
作者：
Jomaa, H;Wiesner, J;Beck, E
通讯作者：
Beck, E
影响因子：
11.6
作者：
Phillips, Michael A.;D'Auria, John C.;Pichersky, Eran
通讯作者：
Pichersky, Eran