Functional analysis of the class II hydrophobin gene HFB2-6 from the biocontrol agent Trichoderma asperellum ACCC30536

• Published in: Microbiological research 2015-02, Vol.171, p.8-20
• Main Authors: Huang, Ying, Mijiti, Gulijimila, Wang, Zhiying, Yu, Wenjing, Fan, Haijuan, Zhang, Rongshu, Liu, Zhihua
• Format: Article
• Language: English
• Subjects: Amino Acid Motifs; Amino Acid Sequence; Binding Sites; Biocontrol; Cloning, Molecular; Computational Biology; Fungal Proteins - chemistry; Fungal Proteins - genetics; Fungal Proteins - metabolism; Gene Expression Regulation, Fungal; Genome, Fungal; Hydrophobin; Models, Molecular; Molecular Sequence Data; Multigene Family; Phylogeny; Populus - genetics; Populus - growth & development; Populus - metabolism; Prokaryotic expression; Promoter Regions, Genetic; Protein Binding; Protein Conformation; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; RT-qPCR; Sequence Alignment; Sequence Analysis, DNA; Trichoderma - classification; Trichoderma - genetics; Trichoderma - metabolism; Trichoderma asperellum ACCC30536
• ISSN: 0944-5013; 1618-0623
• DOI: 10.1016/j.micres.2014.12.004

A class II hydrophobin gene, HFB2-6, was cloned from Trichoderma asperellum ACCC30536 and its biocontrol function was studied. According to our previous transcriptome data, six of the eight class II hydrophobin genes were obviously differential expression in four inducing conditions, especially the gene HFB2-6. Moreover, HFB2-6 proven to be differentially transcribed under eight different treatments. HFB2-6 transcripts were up-regulated under 1% Alternaria alternata cell wall and 5% A. alternata fermentation liquid treatments, and by nutritional stress conditions, suggesting that HFB2-6 plays roles in interactions with both biotic and abiotic environmental conditions. HFB2-6 expression was down-regulated under 1% poplar leaf powder culture conditions, but its expression was up-regulated under 1% poplar root powder, indicating that HFB2-6 has a function in root colonization. Furthermore, the recombinant hydrophobin rHFB2-6 was successfully expressed in Escherichia coli BL21-HFB2-6 and purified from the recombinant strain. Genes related to both the jasmonic acid and salicylic acid signal transduction pathways were up-regulated by interaction with renatured rHFB2-6. The ORCA3 (octadecanoid-derivative responsive Catharanthus AP2-domain) gene of the poplar jasmonic acid signal transduction pathway showed a peak expression of 4.48 times at 2h, and the peak expression of PR1 (pathogenesis-related protein gene) in the salicylic acid signal transduction pathway was 4.58 times at 72h. Two genes, MP (monopteros) and GH3.17 (auxin original response gene), in the auxin signal transduction pathway were also up-regulated after induction with rHFB2-6, indicating that rHFB2-6 can promote poplar growth and confer broad-spectrum resistance to pathogens.