Establishment of Agrobacterium tumefaciens-mediated Transformation System of Aspergillus ochraceus

Abstract

Abstract: Filamentous fungus Aspergillus ochraceus TCCC41060is an industrial strain used for microbial steroid C11-α hydroxylation. In order to create genetic tools for A. ochraceus, the transformation system of A. ochraceus was establishedusing Agrobacterium tumefaciens- mediated system（ATMT）. A. tumefaciens LBA4404 was used as the infective strain and hygromycin B gene as the selection marker. The major factors affecting the transformation efficiency including the concentrations of A. tumefaciens celland A. ochraceus spores, co-culture time, and co-culture temperature were investigated. Under the best conditions, the transformation efficiency of 57 transformants/10⁷ spores was obtained. A. ochraceus transformants were confirmed by PCR amplification . Randomly selected 8 transformants were shown to be genetically stable after 10 rounds of successive cultivation. Results indicated this ATMT transformation system would provide important means for the directed genetic manipulation of this important industrial strain A. ochraceus TCCC41060.

Key words: Aspergillus ochraceus, Agrobacterium tumefaciens, Transformantion system

Cui Qian, Li Jie, Liu Xiaoguang. Establishment of Agrobacterium tumefaciens-mediated Transformation System of Aspergillus ochraceus[J]. Biotechnology Bulletin, 2014, 0(6): 199-204.

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https://biotech.aiijournal.com/EN/Y2014/V0/I6/199

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