Journal
BIORESOURCE TECHNOLOGY
Volume 100, Issue 23, Pages 5941-5946Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.biortech.2009.06.031
Keywords
ATMT; Trichoderma atroviride; Dichlorvos; Microbial degradation
Funding
- Shanghai Rising-star Program [07QA14035]
- National High Technology Research and Development Program of China [2006AA10Z409, 2006AA10A211]
- Foundation of Anhui Educational Committee [2003kj282]
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A protocol for efficient Agrobacterium tumefaciens-mediated transformation (ATMT) of biocontrol fungus Trichoderma atroviride strain T23 was developed to construct mutants with improved dichlorvos-degradation ability. A transformation frequency of 5 x 10(-6) was achieved. Among 110 genetically stable T-DNA transformants of T. atroviride T23, two transformants, AMT-12 and AMT-28, confirmed by Southern blot analysis to have single-copy inserts of T-DNA, showed an increase in dichlorvos-degradation ability of more than 10% compared to that of the wild type, exhibited similar tolerance to the pesticide, but lower spore formation ability. Five transformants exhibited a reduction in degradation of more than 70%, exhibited wild-type spore formation, and tolerated up to 800 mu g/ml. of dichlorvos. The left-flanking sequence of the insertion site in AMT-12 was cloned as a 1845-bp fragment and shown to have 89% identity to the DNA from T. atroviride IMI 206040; however, the involvement of this DNA in dichlorvos degradation remains still to be determined. This study can promote both a more efficient isolation of DNA sequence flanking T-DNA integration site in T. atroviride mutants and a more rational utilization of these transformants in dichlorvos degradation. (C) 2009 Elsevier Ltd. All rights reserved.
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