title: Gene Overexpression and RNA Silencing Tools for the Genetic Manipulation of the S-(+)-Abscisic Acid Producing Ascomycete Botrytis cinerea.
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.
authors.affiliation: 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. dzhongtaochina@163.com.|University of the Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing 100049, China. dzhongtaochina@163.com.
authors.forename: Zhong-Tao
authors.initials: ZT
authors.lastname: Ding
authors.affiliation: 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. zhangzhi_asia@163.com.|University of the Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing 100049, China. zhangzhi_asia@163.com.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: 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. lddd24@163.com.
authors.forename: Di
authors.initials: D
authors.lastname: Luo
authors.affiliation: 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. zhoujy@cib.ac.cn.
authors.forename: Jin-Yan
authors.initials: JY
authors.lastname: Zhou
authors.affiliation: 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. zhongjuan@cib.ac.cn.
authors.forename: Juan
authors.initials: J
authors.lastname: Zhong
authors.affiliation: 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. yangjie@cib.ac.cn.
authors.forename: Jie
authors.initials: J
authors.lastname: Yang
authors.affiliation: 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. xiaoliang@cib.ac.cn.
authors.forename: Liang
authors.initials: L
authors.lastname: Xiao
authors.affiliation: 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. whosecats@163.com.
authors.forename: Dan
authors.initials: D
authors.lastname: Shu
authors.affiliation: 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. abath@cib.ac.cn.
authors.forename: Hong
authors.initials: H
authors.lastname: Tan
mesh_terms: D000040:Abscisic Acid / Q000096:biosynthesis / Q000235:genetics*; D016960:Agrobacterium tumefaciens / Q000235:genetics; D020171:Botrytis / Q000235:genetics* / Q000378:metabolism; D005800:Genes, Fungal; D005822:Genetic Vectors / Q000235:genetics; D034622:RNA Interference; D059386:Reverse Genetics / Q000379:methods*
chemical_list: D000040:Abscisic Acid