标题：Application of T-DNA insertional mutagenesis for improving cellulase production in the filamentous fungus Trichoderma reesei
作者：Zhong, Y.;Wang, X.;Yu, H.;Liang, S.;Wang, T.
作者机构：State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, No.27, Shanda South Road, Jinan 25010
来源：Bioresource Technology: Biomass, Bioenergy, Biowastes, Conversion Technologies, Biotransformations, Production Technologies
关键词：Cellulase;Screening;T-DNA insertional mutagenesis;Trichoderma reesei
摘要：A T-DNA-tagged mutant library created by Agrobacterium-mediated transformation (AMT) was assayed for improvement of cellulase production. After 96-well plate screening for rapid growth on cellulose substrates followed by plate-clearing zone assay, three putative mutants, TA-32, TB-87 and TE-6, with enhanced cellulolytic ability were isolated, exhibiting 38%, 51% and 31% increase in total cellulase activity than the parental strain QM9414, respectively. Endoglucanase, cellobiohydrolase and β-glucosidase activities as well as the hydrolysis efficiencies of the mutants were also improved. Moreover, T-DNA was shown to be integrated at a single site in the genomes of TA-32 and TE-6 while inserted at two copies into the genome of TB-87. Further, the sequences flanking the T-DNA insertion sites were successfully rescued, demonstrating the increased utility of T-DNA insertional mutagenesis for improvement of cellulase production as well as subsequent identification of the tagged genes relevant to cellulolytic ability.