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RNAi-Mediated Gene Silencing of Trcot1 Induces a Hyperbranching Phenotype in Trichoderma reesei

Journal of Microbiology and Biotechnology 2020년 30권 2호 p.206 ~ 215
 ( Gao Fei ) - Chinese Academy of Agricultural Sciences Feed Research Institute

 ( Li Mengzhu ) - Chinese Academy of Agricultural Sciences Feed Research Institute
 ( Liu Weiquan ) - China Agricultural University College of Biological Sciences
 ( Bai Yingguo ) - Chinese Academy of Agricultural Sciences Feed Research Institute
 ( Tao Tu ) - Chinese Academy of Agricultural Sciences Feed Research Institute
 ( Wang Yuan ) - Chinese Academy of Agricultural Sciences Feed Research Institute
 ( Zhang Jie ) - Chinese Academy of Agricultural Sciences Feed Research Institute
 ( Luo Huiying ) - Chinese Academy of Agricultural Sciences Feed Research Institute
 ( Yao Bin ) - Chinese Academy of Agricultural Sciences Feed Research Institute
 ( Huang Huoqing ) - Chinese Academy of Agricultural Sciences Feed Research Institute
 ( Su Xiaoyun ) - Chinese Academy of Agricultural Sciences Feed Research Institute

Abstract


Trichoderma reesei is the major filamentous fungus used to produce cellulase and there is huge interest in promoting its ability to produce higher titers of cellulase. Among the many factors affecting cellulase production in T. reesei, the mycelial phenotype is important but seldom studied. Herein, a close homolog of the Neurospora crassa COT1 kinase was discovered in T. reesei and designated TrCOT1, which is of 83.3% amino acid sequence identity. Functional disruption of Trcot1 in T. reesei by RNAi-mediated gene silencing resulted in retarded sporulation on potato dextrose agar and dwarfed colonies on minimal medium agar plates containing glucose, xylan, lactose, xylose, or glycerol as the sole carbon source. The representative mutant strain, SUS2/Trcot1i, also displayed reduced mycelia accumulation but hyperbranching in the MM glucose liquid medium, with hyphal growth unit length values decreased to 73.0 μm/tip compared to 239.8 μm/tip for the parent strain SUS2. The hyperbranching phenotype led to slightly but significantly increased cellulase secretion from 24 to 72 h in a batch culture. However, the cellulase production per unit of mycelial biomass was much more profoundly improved from 24 to 96 h.

키워드

Trichoderma reesei; Trcot1; hyperbranching; RNAi; cellulase; biofuel
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