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Int. J. Mol. Sci. 2015, 16(5), 10301-10323; doi:10.3390/ijms160510301

Gene Overexpression and RNA Silencing Tools for the Genetic Manipulation of the S-(+)-Abscisic Acid Producing Ascomycete Botrytis cinerea

1
Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, the Chinese Academy of Sciences, No. 9 Section 4, Renmin Nan Road, Chengdu 610041, China
2
University of the Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing 100049, China
*
Authors to whom correspondence should be addressed.
Academic Editor: Ritva Tikkanen
Received: 15 March 2015 / Revised: 14 April 2015 / Accepted: 17 April 2015 / Published: 6 May 2015
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
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Abstract

The phytopathogenic ascomycete Botrytis cinerea produces several secondary metabolites that have biotechnical significance and has been particularly used for S-(+)-abscisic acid production at the industrial scale. To manipulate the expression levels of specific secondary metabolite biosynthetic genes of B. cinerea with Agrobacterium tumefaciens-mediated transformation system, two expression vectors (pCBh1 and pCBg1 with different selection markers) and one RNA silencing vector, pCBSilent1, were developed with the In-Fusion assembly method. Both expression vectors were highly effective in constitutively expressing eGFP, and pCBSilent1 effectively silenced the eGFP gene in B. cinerea. Bcaba4, a gene suggested to participate in ABA biosynthesis in B. cinerea, was then targeted for gene overexpression and RNA silencing with these reverse genetic tools. The overexpression of bcaba4 dramatically induced ABA formation in the B. cinerea wild type strain Bc-6, and the gene silencing of bcaba4 significantly reduced ABA-production in an ABA-producing B. cinerea strain. View Full-Text
Keywords: Botrytis cinerea; gene overexpression; RNA silencing; ATMT Botrytis cinerea; gene overexpression; RNA silencing; ATMT
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Ding, Z.-T.; Zhang, Z.; Luo, D.; Zhou, J.-Y.; Zhong, J.; Yang, J.; Xiao, L.; Shu, D.; Tan, H. Gene Overexpression and RNA Silencing Tools for the Genetic Manipulation of the S-(+)-Abscisic Acid Producing Ascomycete Botrytis cinerea. Int. J. Mol. Sci. 2015, 16, 10301-10323.

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