National Key Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai, 200032, China.
Plant Cell Rep. 2010 Aug;29(8):887-94. doi: 10.1007/s00299-010-0874-0. Epub 2010 Jun 10.
A number of genes that function in the terpenoid indole alkaloids (TIAs) biosynthesis pathway have been identified in Catharanthus roseus. Except for the geraniol 10-hydroxylase (G10H) gene, which encodes a cytochrome P450 monooxygenase, several of these genes are up-regulated by ORCA3, a jasmonate-responsive APETALA2 (AP2)-domain transcript factor. In this study, the G10H gene was transformed independently, or co-transformed with ORCA3 into C. roseus, using Agrobacterium rhizogenes MSU440. Hairy root clones expressing the G10H gene alone, or both the G10H and ORCA3 genes, were obtained. Alkaloid accumulation level analyses showed that all transgenic clones accumulated more catharanthine, with the highest accumulation level in the transgenic clone OG12 (6.5-fold higher than that of the non-expression clone). Following treatment with ABA, accumulation of catharanthine reached 1.96 mg/g DW in the transgenic clone OG12. The expression levels of TIAs biosynthesis genes in transgenic and non-transgenic clones were also investigated.
在长春花中鉴定出了许多在萜类吲哚生物碱 (TIAs) 生物合成途径中起作用的基因。除了编码细胞色素 P450 单加氧酶的香叶醇 10-羟化酶 (G10H) 基因外，这些基因中的几个都受到茉莉酸响应 APETALA2 (AP2) 结构域转录因子 ORCA3 的上调。在这项研究中，G10H 基因被独立地或与 ORCA3 一起通过发根农杆菌 MSU440 转化到长春花中。获得了单独表达 G10H 基因的毛状根克隆，或同时表达 G10H 和 ORCA3 基因的毛状根克隆。生物碱积累水平分析表明，所有转基因克隆都积累了更多的长春质碱，其中转基因克隆 OG12 的积累水平最高（比非表达克隆高 6.5 倍）。在用 ABA 处理后，转基因克隆 OG12 中的长春质碱积累达到 1.96 mg/g DW。还研究了转基因和非转基因克隆中 TIAs 生物合成基因的表达水平。
