Abstract
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.
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Acknowledgment
The authors are grateful to Professors J. Memelink (Leiden University, Netherlands) for providing the vectors containing G10H or ORCA3 genes, and Dr. Wang Yan-zhang (Shanghai Institutes for Biological Sciences, China) for providing A. rhizogenes MSU440, and Chen Jun (Shanghai Institutes for Biological Sciences, China) for HPLC analysis. This work was supported by the following grants: the Knowledge Innovation Program of Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Grant No. 2007KIP304); the National Basic Research Program of China (Grant No. 2006CB100106; 2010CB126600); the National Natural Science Foundation of China (Grant No. 30571196; 0933ZF11C1; 0933Z411C1); The Ministry of Science and Technology of China (Grant No. 2007AA10Z187); Shanghai Science and Technology Commission (Grant No. 08DZ2270800).
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Communicated by K. Chong.
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Wang, CT., Liu, H., Gao, XS. et al. Overexpression of G10H and ORCA3 in the hairy roots of Catharanthus roseus improves catharanthine production. Plant Cell Rep 29, 887–894 (2010). https://doi.org/10.1007/s00299-010-0874-0
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DOI: https://doi.org/10.1007/s00299-010-0874-0