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Soybean transcription factor GmMYBZ2 represses catharanthine biosynthesis in hairy roots of Catharanthus roseus

  • Applied Genetics and Molecular Biotechnology
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Abstract

Catharanthus roseus (L.) G. Don is a plant species known for its production of a variety of terpenoid indole alkaloids, many of which have pharmacological activities. Production of catharanthine in cell cultures or in hairy roots established by transformation with Agrobacterium rhizogenes is of interest because catharanthine can be chemically coupled to the abundant leaf alkaloid vindoline to form the valuable anticancer drug vinblastine. Here, we observed a high amount of catharanthine in hairy roots of C. roseus, established by infecting leaf explants with the A. rhizogenes >agropine-type A4 strain carrying plasmid pRi. T-DNA transfer from plasmid pRi into hairy roots was confirmed by PCR for the essential T-DNA genes rolA and rolB and the agropine synthesis gene ags. The results suggest that integration of T-DNA into the plant DNA plays a positive role on the catharanthine pathway in C. roseus hairy roots. Furthermore, co-transformation with the soybean transcription factor GmMYBZ2 indicated that GmMYBZ2 reduces the catharanthine production by alteration of expression of a number of genes linked to the pathway. Transcription levels of the zinc-finger transcription factor 1 gene ZCT1 were high, and the transcription levels of the anthranilate synthase gene ASα, the strictosidine synthase gene STR, and the key transcription factor gene octadecanoid-responsive Catharanthus APETALA2/ethylene response factor were low. In addition, GmMYBZ2 had a negative effect on the gene expression levels of A-type cyclin CYSA and B-type cyclin CYSB, which was correlated with a reduced growth rate of the hairy roots.

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Acknowledgments

We thank Pang Jun-Feng for technical assistance with RNA extraction, Ma Jiang-Tao for measuring the growth index of hairy roots, Du Hai for work discussions, and Dr. Yang Wen-Jie for providing the plasmid pCAMBIA2301-GmMYBZ2. We are also grateful to Prof. Dr. Johan Memelink, Dr. A. A. N. van Brussel, and Trillian Ross (Plant Cell Physiology, Institute of Biology, Leiden University, The Netherlands) for giving some useful comments on this manuscript. This work was supported by a grant from the National Transgenic Program (2009ZX08005-004B, 2009ZX08009055B, and 2008ZX08005-004).

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  1. Mei-Liang Zhou

    Present address: Plant Cell Physiology, Institute of Biology, Sylvius Laboratory, Sylviusweg 72, Leiden University, P.O. Box 9505, 2300, RA, Leiden, The Netherlands

Authors and Affiliations

  1. School of Life Science, Sichuan Agricultural University, Yaan, Sichuan, 625014, People’s Republic of China

    Mei-Liang Zhou & Ji-Rong Shao

  2. Room 211, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Zhongguancun south street No 12, Haidian district, Beijing, 100081, China

    Mei-Liang Zhou, Yan-Min Wu & Yi-Xiong Tang

  3. Shenzhen Imported Center for Quality Crop Seed, Shenzhen, Guangdong, 518040, People’s Republic of China

    Hong-Li Hou

  4. School of Resources and Environment, Sichuan Agricultural University, Yaan, Sichuan, 625014, People’s Republic of China

    Xue-Mei Zhu

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Zhou, ML., Hou, HL., Zhu, XM. et al. Soybean transcription factor GmMYBZ2 represses catharanthine biosynthesis in hairy roots of Catharanthus roseus . Appl Microbiol Biotechnol 91, 1095–1105 (2011). https://doi.org/10.1007/s00253-011-3288-1

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