Research Article | Volume : 1 Issue : 8, October 2011

Enhancement of camptothecin production in Camptotheca acuminata hairy roots by overexpressing ORCA3 gene

Xiaoning Ni,  Shiqiu Wen,  Wei Wang,  Xiaoyun Wang,  Hui Xu,  Guoyin Kai   
Xiaoning Ni2, Shiqiu Wen3, Wei Wang1, Xiaoyun Wang3, Hui Xu1, Guoyin Kai1

1Laboratory of Plant Biotechnology, College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, P. R. China.

2Department of General Surgery, Zhongshan Hospital, Fudan University, Shanghai 200032, PR China. Laboratory of Plant Biotechnology, College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, P. R. China.

3Department of Gynaecology and Obstetrics, Kunshan Hospital of Traditional Chinese Medicine, Kunshan, Jiangsu Province 215300, People’s Republic of China.

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Abstract

Being a good anti-tumor drug, camptothecin (CPT) is a kind of modified monoterpene indole alkaloid firstly isolated from the deciduous Chinese happy tree with rapidly increasing clinical demand. Due to the great importance and low content of this compound, it is very important to improve CPT production by modern biotechnology. ORCA3 is a jasmonateresponsive APETALA2-domain transcript factor isolated from Catharanthus roseus, with strong ability to up-regulate expression of serveral key genes involved in TIA biosynthetic pathway. To investigate physiological function of ORCA3 gene in Camptotheca acuminata, the ORCA3 gene was transformed using Agrobacterium-mediated gene transfer technology. PCR analysis confirmed that the ORCA3 gene was integrated into the plant genome. HPLC showed that overexpression of ORCA3 in transgenic hairy root lines can effectively enhance the production of camptothecin with 1.5-fold compared with the control (1.12 mg/g dw). The results revealed that ORCA3 is an effective regulatory gene for improving metabolic flux in camptothecin biosynthetic pathway at the first time.


Keyword:     Camptotheca acuminata ORCA3 camptothecin transformation hairy root cultures

Copyright:The Author(s). This is an open access article distributed under the Creative Commons Attribution Non-Commercial License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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