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Cloning, annotation and expression analysis of mycoparasitism-related genes in Trichoderma harzianum 88

  • Microbial Genetics, Genomics and Molecular Biology
  • Published:
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Abstract

Trichoderma harzianum 88, a filamentous soil fungus, is an effective biocontrol agent against several plant pathogens. High-throughput sequencing was used here to study the mycoparasitism mechanisms of T. harzianum 88. Plate confrontation tests of T. harzianum 88 against plant pathogens were conducted, and a cDNA library was constructed from T. harzianum 88 mycelia in the presence of plant pathogen cell walls. Randomly selected transcripts from the cDNA library were compared with eukaryotic plant and fungal genomes. Of the 1,386 transcripts sequenced, the most abundant Gene Ontology (GO) classification group was “physiological process”. Differential expression of 19 genes was confirmed by real-time RT-PCR at different mycoparasitism stages against plant pathogens. Gene expression analysis revealed the transcription of various genes involved in mycoparasitism of T. harzianum 88. Our study provides helpful insights into the mechanisms of T. harzianum 88-plant pathogen interactions.

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References

  • Adams, M.D., Kelley, J.M., Gocayne, J.D., Dubnick, M., Polymeropoulos, M.H., Xiao, H., Merril, C.R., Wu, A., Olde, B., Moreno, R.F., andet al. 1991. Complementary DNA sequencing: expressed sequence tags and human genome project. Science252, 1651–1656.

    Article  PubMed  CAS  Google Scholar 

  • Alizadeh, F., Abdullah, S.N., Khodavandi, A., Abdullah, F., Yusuf, U.K., and Chong, P.P. 2011. Differential expression of oil palm pathology genes during interactions with Ganoderma boninense and Trichoderma harzianum. J. Plant Physiol. 168, 1106–1113.

    Article  PubMed  CAS  Google Scholar 

  • Almeida, F.B., Cerqueira, F.M., Silva Rdo, N., Ulhoa, C.J., and Lima, A.L. 2007. Mycoparasitism studies of Trichoderma harzianum strains against Rhizoctonia solani: evaluation of coiling and hydrolytic enzyme production. Biotechnol. Lett.29, 1189–1193.

    Article  PubMed  CAS  Google Scholar 

  • Altschul, S.F., Gish, W., Miller, W., Myers, E.W., and Lipman, D.J. 1990. Basic local alignment search tool. J. Mol. Biol.215, 403–410.

    PubMed  CAS  Google Scholar 

  • Bae, H., Roberts, D.P., Lim, H.S., Strem, M.D., Park, S.C., Ryu, C.M., Melnick, R.L., and Bailey, B.A. 2011. Endophytic Trichoderma isolates from tropical environments delay disease onset and induce resistance against Phytophthora capsici in hot pepper using multiple mechanisms. Mol. Plant. Microbe Interact.24, 336–351.

    Article  PubMed  CAS  Google Scholar 

  • Bouck, A. and Vision, T. 2007. The molecular ecologist’s guide to expressed sequence tags. Mol. Ecol.16, 907–924.

    Article  PubMed  CAS  Google Scholar 

  • Buensanteai, N., Mukherjee, P.K., Horwitz, B.A., Cheng, C., Dangott, L.J., and Kenerley, C.M. 2010. Expression and purification of biologically active Trichoderma virens proteinaceous elicitor Sm1 in Pichia pastoris. Protein Expr. Purif. 72, 131–138.

    Article  PubMed  CAS  Google Scholar 

  • Camon, E.B., Barrell, D.G., Dimmer, E.C., Lee, V., Magrane, M., Maslen, J., Binns, D., and Apweiler, R. 2005. An evaluation of GO annotation retrieval for BioCreAtIvE and GOA. BMC Bioinformatics1, S17.

    Article  Google Scholar 

  • Carsolio, C., Gutiérrez, A., Jiménez, B., Van Montagu, M., and Herrera Estrella, A. 1994. Characterization of ech-42, a Trichoderma harzianum endochitinase gene expressed during mycoparasitism. Proc. Natl. Acad. Sci. USA91, 10903–10907.

    Article  PubMed  CAS  Google Scholar 

  • Cortés, C., Gutierrez, A., Olmedo, V., Inbar, J., Chet. I., and Herrera-Estrella, A. 1998. The expression of genes involved in parasitism by Trichoderma harzianum is triggered by a diffusible factor. Mol. Gen. Genet.260, 218–225.

    Article  PubMed  Google Scholar 

  • Druzhinina, I.S., Seidl-Seiboth, V., Herrera-Estrella, A., Horwitz, B.A., Kenerley, C.M., Monte, E., Mukherjee, P.K., Zeilinger, S., and Grigoriev, I.V. 2011. Trichoderma: the genomics of opportunistic success. Nat. Rev. Microbiol.9, 749–759.

    Article  PubMed  CAS  Google Scholar 

  • Ewing, B. and Green, P. 1998. Base-calling of automated sequencer traces using Phred. II. Error probabilities. Genome Res.8, 186–194.

    PubMed  CAS  Google Scholar 

  • Gordon, D. 2003. Viewing and Editing Assembled Sequences Using Consed, pp. 11.2.1–11.2.43. In Baxevanis, A.D. and Davison, D.B. (eds.), Curr. Protoc. Bioinformatics. John Wiley & Co., New York, USA.

    Google Scholar 

  • Götz, S., García-Gómez, J.M., Terol, J., Williams, T.D., Nagaraj, S.H., Nueda, M.J., Robles, M., Talón, M., Dopazo, J., and Conesa, A. 2008. High-throughput functional annotation and data mining with the Blast2GO suite. Nucleic Acids. Res.36, 3420–3435.

    Article  PubMed  Google Scholar 

  • Hermosa, M.R., Grondona, I., Iturriaga, E.A., Diaz Minguez, J.M., Castro, C., Monte, E., and Garcia-Acha, I. 2000. Molecular characterization and identification of biocontrol isolates of Trichoderma spp. Appl. Environ. Microbiol.66, 1890–1898.

    Article  PubMed  CAS  Google Scholar 

  • Kubicek, C.P., Herrera-Estrella, A., Seidl-Seiboth, V., Martinez, D.A., Druzhinina, I.S., Thon, M., Zeilinger, S., Casas-Flores, S., Horwitz, B.A., Mukherjee, P.K., andet al. 2011. Comparative genome sequence analysis underscores mycoparasitism as the ancestral life style of Trichoderma. Genome Biol. 12, R40.

    Article  PubMed  CAS  Google Scholar 

  • Lin, Y.R., Lo, C.T., Liu, S.Y., and Peng, K.C. 2012. Involvement of pachybasin and emodin in self-regulation of Trichoderma harzianum mycoparasitic coiling. J. Agric. Food. Chem.60, 2123–2128.

    Article  PubMed  CAS  Google Scholar 

  • Liu, P.G. and Yang, Q. 2005. Identification of genes with a biocontrol function in Trichoderma harzianum mycelium using the expressed sequence tag approach. Res. Microbiol.156, 416–423.

    Article  PubMed  CAS  Google Scholar 

  • Liu, Z., Yang, X., Sun, D., Song, J., Chen, G., Juba, O., and Yang, Q. 2010. Expressed sequence tags-based identification of genes in a biocontrol strain Trichoderma asperellum. Mol. Biol. Rep.37, 3673–3681.

    Article  PubMed  CAS  Google Scholar 

  • Livak, K. and Schmittgen, T. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods25, 402–408.

    Article  PubMed  CAS  Google Scholar 

  • Morán-Diez, E., Rubio, B., Domínguez, S., Hermosa, R., Monte, E., and Nicolás C. 2012. Transcriptomic response of Arabidopsis thaliana after 24 h incubation with the biocontrol fungus Trichoderma harzianum. J. Plant Physiol. 169, 614–620.

    Article  PubMed  Google Scholar 

  • Petersen, T.N., Brunak, S., Heijne, G.V., and Nielsen, H. 2011. SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat. Methods8, 785–786.

    Article  PubMed  CAS  Google Scholar 

  • Reithner, B., Ibarra-Laclette, E., Mach, R.L., and Herrera-Estrella, A. 2011. Identification of mycoparasitism-related genes in Trichoderma atroviride. Appl. Environ. Microbiol.77, 4361–4370.

    Article  PubMed  CAS  Google Scholar 

  • Restuccia, C., Giusino, F., Licciardello, F., Randazzo, C., Caggia, C., and Muratore, G. 2006. Biological control of peach fungal pathogens by commercial products and indigenous yeasts. J. Food Prot.69, 2465–2470.

    PubMed  Google Scholar 

  • Steiger, M.G., Mach, R.L., and Mach-Aigner, A.R. 2010. An accurate normalization strategy for RT-qPCR in Hypocrea jecorina (Trichoderma reesei). J. Biotechnol.145, 30–37.

    Article  PubMed  CAS  Google Scholar 

  • Vizcaíno, J.A., González, F.J., Suárez, M.B., Redondo, J., Heinrich, J., Delgado-Jarana, J., Hermosa, R., Gutiérrez, S., Monte, E., Llobell, A., and Rey, M. 2006. Generation, annotation and analysis of ESTs from Trichoderma harzianum CECT 2413. BMC. Genomics7, 193.

    Article  PubMed  Google Scholar 

  • Vizcaíno, J.A., Redondo, J., Suárez, M.B., Cardoza, R.E., Hermosa, R., González, F.J., Rey, M., and Monte, E. 2007. Generation, annotation, and analysis of ESTs from four different Trichoderma strains grown under conditions related to biocontrol. Appl. Microbiol. Biotechnol.75, 853–862.

    Article  PubMed  Google Scholar 

  • Yang, C.A., Cheng, C.H., Lee, J.W., Lo, C.T., Liu, S.Y., and Peng, K.C. 2012. Monomeric L-amino acid oxidase-induced mitochondrial dysfunction in Rhizoctonia solani reveals a novel antagonistic mechanism of Trichoderma harzianum ETS 323. J. Agric. Food Chem.60, 2464–2471.

    Article  PubMed  CAS  Google Scholar 

  • Yang, H.H., Yang, S.L., Peng, K.C., Lo, C.T., and Liu, S.Y. 2009. Induced proteome of Trichoderma harzianum by Botrytis cinerea. Mycol. Res.113, 924–932.

    Article  PubMed  CAS  Google Scholar 

  • Zhang, S., Xia, Y.X., Kim, B., and Keyhani, N.O. 2011. Two hydrophobins are involved in fungal spore coat rodlet layer assembly and each play distinct roles in surface interactions, development and pathogenesis in the entomopathogenic fungus, Beauveria bassiana. Mol. Microbiol.80, 811–826.

    Article  PubMed  CAS  Google Scholar 

  • Zhang, H.Y., Yang, Q., Wang, G., and Shang, F.D. 2009. Analysis of expressed sequence tags from Chaetomium cupreum grown under conditions associated with mycoparasitism. Lett. Appl. Microbiol.48, 275–280.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Life Science and Engineering, Institute of Technology, Harbin, 150001, P. R. China

    Lin Yao, Qian Yang, Jinzhu Song, Lianhai Qu & Yun Wang

  2. School of Municipal and Environmental Engineering, Harbin Institute of Technology, Harbin, 150090, P. R. China

    Lin Yao, Chong Tan & Li Wang

  3. Key Laboratory of Molecular and Cytogenetics, Heilongjiang Province, Department of Biology, Harbin Normal University, Harbin, 150025, P. R. China

    Changhong Guo & Li Wang

Authors
  1. Lin Yao
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  2. Qian Yang
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  3. Jinzhu Song
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  4. Chong Tan
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  5. Changhong Guo
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  6. Li Wang
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  7. Lianhai Qu
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  8. Yun Wang
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Correspondence to Qian Yang.

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Yao, L., Yang, Q., Song, J. et al. Cloning, annotation and expression analysis of mycoparasitism-related genes in Trichoderma harzianum 88 . J Microbiol. 51, 174–182 (2013). https://doi.org/10.1007/s12275-013-2545-7

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  • DOI: https://doi.org/10.1007/s12275-013-2545-7

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