Functional Analysis of TvGCN5 Gene Encoding Histone Acetylase from Trichoderma viride Tv-1511

doi:10.13560/j.cnki.biotech.bull.1985.2021-0998

Abstract

Abstract:

Trichoderma viride presents a wide range of applications in plant growth promotion and stress resistance,biological control,cellulase production and biomass utilization. Histone acetylation plays an important role in regulating transcriptional expression of gene. GCN5 is the most representative histone acetylase. First,the function of TvGCN5 gene in Trichoderma viride Tv-1511 was analyzed,then the deletion mutants（Tv-1511-△GCN5）was constructed by deleting TvGCN5 gene,and the overexpression mutants（Tv-1511-GCN5-OE）of GCN5 gene in the Tv-1511 was obtained by constructing TvGCN5 gene overexpression vector,for studying the functions of TvGCN5 in the Tv-1511 in growth-promoting and stress resistances as well as enzyme production. The results showed that the over-expression of TvGCN5 resulted in the longer mycelia diameter and more biomass under salt stress and high temperature stress compared with wild-type Tv-1511. Furthermore,the Tv-1511-△GCN5 strains obtained enhanced growth-promoting effect of plant by increasing the synthetic capacity of IAA. Notably,the higher production of cellulase was observed in the Tv-1511-GCN5-OE strain. The deletion mutant（Tv-1511-△GCN5）demonstrated the decrease of all above aspects,i.e.,the effects was weaker. Thus,TvGCN5 gene plays an important role in plant growth promotion,stress tolerance and cellulase production.

Key words: Trichoderma viride, histone acetylase GCN5, gene editing, gene function, physiological characteristics

ZHANG Hao, LI Zhe, GUO Kai, HUANG Yan-hua, HAO Yong-ren. Functional Analysis of TvGCN5 Gene Encoding Histone Acetylase from Trichoderma viride Tv-1511[J]. Biotechnology Bulletin, 2022, 38(5): 136-148.

Figures/Tables 10

Fig. 1 Construction of knockout fragment and overexpression vector of TvGCN5 gene A,B：Fragment construction of knockout TvGCN5 gene. C,D：Construction of overexpression vector of TvGCN5 gene

Fig.2 Identification of overexpressing or deleting TvGCN5 gene in Trichoderma engineering strains by fluores-cence quantitative PCR and Western blot A：TvGCN5 gene transcription level in Trichoderma engineered strains by fluorescence quantitative PCR. B,C：Protein expression level of TvGCN5 in Trichoderma engineered strains by Western blot

Fig.3 Changes in the levels of histone acetylation in T. viride Tv-1511 and its engineered strains A：Quantitative detection of histone H3 acetylation level in T. viride Tv-1511 and its engineered strains by assay kit. B,C：Detection of histone H3 acetylation level in T. viride Tv-1511and its engineered strains by Western blot

Fig.4 Analysis of the salt tolerances of T. viride Tv-1511 original strain and its engineered strains A,B：Petri dish test of the salt tolerance of Trichoderma and quantitative statistics of colony diameter. C：Dry weight of Trichoderma in liquid culture mycelium under salt stress

Fig.5 Analysis of the high temperature tolerance of T. vir-ide Tv-1511 original strain and its engineered strains A,B：Petri dish test of the heat tolerance of Trichoderma and quantitative statistics of colony diameter. C：Dry weight of Trichoderma liquid culture mycelium under high temperature stress

Fig.6 Analysis of the growth-promoting effects of T. viride Tv-1511 original strain and its engineering strains on Arabidopsis thaliana A：Promoting effects of T. viride and its engineering strains on the growth of A. thaliana. B：Quantitative statistics of fresh weight of Arabidopsis treated with T. viride and its engineered strains. C：Quantitative statistics of leaf areas of Arabidopsis treated with T. viride and its engineering strains

Fig.7 Analysis of the growth-promoting effects of T. viride Tv-1511 original strain and its engineering strains on peppermint（Mentha piperita） A：Promoting effects of Trichoderma viride Tv-1511 and its engineering strains on the growth of peppermint. B：Quantitative statistics of fresh weight of peppermint treated with Trichoderma strains. C：Quantitative statistics of leaf area of peppermint treated with Trichoderma strains

Fig.8 Analysis of the promoting effects of T. viride Tv-1511 original strain and its engineering strains on cucum-ber（Cucumis sativus L.） A：Promoting effects of Trichoderma viride Tv-1511 and its engineering strains on the growth of cucumber. B：Quantitative statistics of fresh weight of cucumber treated with Trichoderma strains. C：Quantitative statistics of leaf areas of cucumber treated with Trichoderma strains

Fig.9 Detection of IAA contents in the fermentation broths of T. viride Tv-1511 original strain and its engineer-ing strains A：Chromatogram of IAA standard. B：Quantitative statistics of IAA content in the fermentation broth of T. viride Tv-1511 and its engineering strains

Fig.10 Analysis of cellulase activities of T. viride Tv-1511 original strain and its engineering strains A：Detection of filter paper enzyme activity in the fermentation broth of T. viride Tv-1511 and its engineering strains. B：Detection of hydroxymethyl cellulase activity in the fermentation broth of T. viride Tv-1511 and its engineering strains. C：Detection of exoglucanase activity in the fermentation broth of T. viride Tv-1511 and its engineering strains. D：Detection of β-glucosidase activity in the fermentation broth of T. viride Tv-1511 and its engineering strains

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