Abstract
The fungal genus Trichoderma has been extensively studied due to its role in the mycoparasitism, and thus developed as biocontrol agent against various plant pathogens. Although the mycoparasitic processes of several Trichoderma species have already been well understood, the information about the mycoparasitic mechanisms of Trichoderma strains resulted from different growth conditions or interacting with different phytopathogens is still limited. In this study, we utilized transcriptome sequencing to identify the differentially expressed genes (DEGs) at 0, 24, 72 and 120 h from T. atroviride strain SS003, growing on an induced-medium with cell walls of Pinus armandii pathogen Cronartium ribicola (CRCW). In total, 86,155,316 reads were obtained with 43,077,658 clean reads. Further, 10,422 genes were identified from four transcriptomes and accounted for 93.89% of annotated genes in T. atroviride IMI 206040 genome, reflecting high-quality sequencing and assembly. In each pairwise comparison, a large number of DEGs were identified with different numbers of genes for up- and down-regulation, respectively. In the presence of CRCW, expression of two main glycoside hydrolase gene families (i.e. chitinase and glucosidase) was induced. Most of 14 secreted enzymes by quantitative real time PCR (qPCR) analysis exhibited a consistent expression pattern with that by RNA-Seq data. This comparative study leads to the identification of phase-specific genes in the interactions of T. atroviride SS003 with C. ribicola, and provides potential molecular targets for improved biocontrol strategies.







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References
Atanasova L, Le Crom S, Gruber S, Coulpier F, Seidl-Seiboth V, Kubicek CP, Druzhinina IS (2013) Comparative transcriptomics reveals different strategies of Trichoderma mycoparasitism. BMC Genom 14:121
Baba Y, Sumitani J-I, Tani S, Kawaguchi T (2015) Characterization of Aspergillus aculeatus β-glucosidase 1 accelerating cellulose hydrolysis with Trichoderma cellulase system. AMB Express 5:3
Baek J-M, Howell RC, Kenerley MC (1999) The role of an extracellular chitinase from Trichoderma virens Gv29-8 in the biocontrol of Rhizoctonia solani. Curr Genet 35:41–50
Bao Z, Ao X, Chen T, Chen Y (2015) Optimizing fermentation conditions for cell wall degrading eymatic production from Trichoderma atroviriade SS003 by response surface method. J South Agric 46:265–270
Benitez T, Rincon AM, Limon MC, Codon AC (2004) Biocontrol mechanisms of Trichoderma strains. Int Microbiol 7:249–260
Cantarel BL, Coutinho PM, Rancurel C, Bernard T, Lombard V, Henrissat B (2009) The Carbohydrate-Active EnZymes database (CAZy): an expert resource for Glycogenomics. Nucleic Acids Res 37:D233–D238
Conesa A, Gotz S, Garcia-Gomez JM, Terol J, Talon M, Robles M (2005) Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research. Bioinformatics 21:3674–3676
de Hoon MJ, Imoto S, Nolan J, Miyano S (2004) Open source clustering software. Bioinformatics 20:1453–1454
Duo-Chuan L (2006) Review of fungal chitinases. Mycopathologia 161:345–360
Free SJ (2013) Fungal cell wall organization and biosynthesis. Adv Genet 81:33–82
Gruber S, Seidl-Seiboth V (2012) Self versus non-self: fungal cell wall degradation in Trichoderma. Microbiology 158:26–34
Gruber S, Vaaje-Kolstad G, Matarese F, Lopez-Mondejar R, Kubicek CP, Seidl-Seiboth V (2011) Analysis of subgroup C of fungal chitinases containing chitin-binding and LysM modules in the mycoparasite Trichoderma atroviride. Glycobiology 21:122–133
Harman GE, Howell CR, Viterbo A, Chet I, Lorito M (2004) Trichoderma species–opportunistic, avirulent plant symbionts. Nat Rev Microbiol 2:43–56
Horton P, Park KJ, Obayashi T, Fujita N, Harada H, Adams-Collier CJ, Nakai K (2007) WoLF PSORT: protein localization predictor. Nucleic Acids Res 35:W585–W587
Howell CR (2003) Mechanisms employed by Trichoderma species in the biological control of plant diseases: the history and evolution of current concepts. Plant Dis 87:4–10
Howell CR, Stipanovic RD (1983) Gliovirin, a new antibiotic from Gliocladium virens, and its role in the biological control of Pythium ultimum. Can J Microbiol 29:321–324
Kanehisa M, Araki M, Goto S, Hattori M, Hirakawa M, Itoh M, Katayama T, Kawashima S, Okuda S, Tokimatsu T et al (2008) KEGG for linking genomes to life and the environment. Nucleic Acids Res 36:D480–D484
Kubicek CP, Herrera-Estrella A, Seidl-Seiboth V, Martinez DA, Druzhinina IS, Thon M, Zeilinger S, Casas-Flores S, Horwitz BA, Mukherjee PK et al (2011) Comparative genome sequence analysis underscores mycoparasitism as the ancestral life style of Trichoderma. Genome Biol 12:R40
Li R, Yu C, Li Y, Lam TW, Yiu SM, Kristiansen K, Wang J (2009) SOAP2: an improved ultrafast tool for short read alignment. Bioinformatics 25:1966–1967
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2–△△CT Method. Methods 25:402–408
Lorito M, Woo SL, Garcia I, Colucci G, Harman GE, Pintor-Toro JA, Filippone E, Muccifora S, Lawrence CB, Zoina A et al (1998) Genes from mycoparasitic fungi as a source for improving plant resistance to fungal pathogens. Proc Natl Acad Sci USA 95:7860–7865
Marra R, Ambrosino P, Carbone V, Vinale F, Woo SL, Ruocco M, Ciliento R, Lanzuise S, Ferraioli S, Soriente I et al (2006) Study of the three-way interaction between Trichoderma atroviride, plant and fungal pathogens by using a proteomic approach. Curr Genet 50:307–321
Martinez D, Berka RM, Henrissat B, Saloheimo M, Arvas M, Baker SE, Chapman J, Chertkov O, Coutinho PM, Cullen D et al (2008) Genome sequencing and analysis of the biomass-degrading fungus Trichoderma reesei (syn. Hypocrea jecorina). Nat Biotechnol 26:553–560
Metcalf DA, Wilson CR (2001) The process of antagonism of Sclerotium cepivorum in white rot affected onion roots by Trichoderma koningii. Plant Pathol 50:249–257
Petersen TN, Brunak S, von Heijne G, Nielsen H (2011) SignalP 4.0: discriminating signal peptides from transmembrane regions. Nat Methods 8:785–786
Ramada MHS, Steindorff AS, Bloch C, Ulhoa CJ (2016) Secretome analysis of the mycoparasitic fungus Trichoderma harzianum ALL 42 cultivated in different media supplemented with Fusarium solani cell wall or glucose. Proteomics 16:477–490
Reithner B, Ibarra-Laclette E, Mach RL, Herrera-Estrella A (2011) Identification of mycoparasitism-related genes in Trichoderma atroviride. Appl Environ Microbiol 77:4361–4370
Saldanha AJ (2004) Java Treeview-extensible visualization of microarray data. Bioinformatics 20:3246–3248
Samolski I, de Luis A, Vizcaíno JA, Monte E, Suárez MB (2009) Gene expression analysis of the biocontrol fungus Trichoderma harzianum in the presence of tomato plants, chitin, or glucose using a high-density oligonucleotide microarray. BMC Microbiol 9:217
Seidl V (2008) Chitinases of filamentous fungi: a large group of diverse proteins with multiple physiological functions. Fungal Biol Rev 22:36–42
Seidl V, Huemer B, Seiboth B, Kubicek CP (2005) A complete survey of Trichoderma chitinases reveals three distinct subgroups of family 18 chitinases. FEBS J 272:5923–5939
Seidl V, Song L, Lindquist E, Gruber S, Koptchinskiy A, Zeilinger S, Schmoll M, Martinez P, Sun J, Grigoriev I et al (2009) Transcriptomic response of the mycoparasitic fungus Trichoderma atroviride to the presence of a fungal prey. BMC Genom 10:567
Singhania RR, Patel AK, Sukumaran RK, Larroche C, Pandey A (2013) Role and significance of beta-glucosidases in the hydrolysis of cellulose for bioethanol production. Bioresour Technol 127:500–507
Steindorff AS, Ramada MHS, Coelho ASG, Miller RNG, Pappas GJ, Ulhoa CJ, Noronha EF (2014) Identification of mycoparasitism-related genes against the phytopathogen Sclerotinia sclerotiorum through transcriptome and expression profile analysis in Trichoderma harzianum. BMC Genom 15:204
Taguri T, Yaginuma K, Yamamoto M, Fujii T, Ando T (2014) Enantiospecific synthesis and filed evaluation of four stereoisomers of 10,14-dimethyloctadec-1-ene, a sex pheromone component secreted by female moths of the apple leafminer. Biosci Biotechnol Biochem 78:761–765
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729
Taylor TN, Hass H, Kerp H, Krings M, Hanlin RT (2005) Perithecial ascomycetes from the 400 million year old Rhynie chert: an example of ancestral polymorphism. Mycologia 97:269–285
Vieira PM, Coelho ASG, Steindorff AS, de Siqueira SJL, Silva RdN, Ulhoa CJ (2013) Identification of differentially expressed genes from Trichoderma harzianum during growth on cell wall of Fusarium solani as a tool for biotechnological application. BMC Genom 14:177
Vinale F, Sivasithamparam K, Ghisalberti EL, Marra R, Woo SL, Lorito M (2008) Trichoderma-plant-pathogen interactions. Soil Biol Biochem 40:1–10
Vizcaino JA, Redondo J, Suarez MB, Cardoza RE, Hermosa R, Gonzalez FJ, Rey M, 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
Wang P, Chen GF, Zhang JS, Xue Q, Zhang JH, Chen C, Zhang QH (2016) Pheromone-trapping the nun moth, Lymantria monacha (Lepidoptera: Lymantriidae) in Inner Mongolia, China. Insect Sci
Webster J, Lomas N (1964) Does Trichoderma viride produce gliotoxin and viridin? Transactions of the British mycological. Society 47:535–540
Weindling R (1934) Studies on a lethal principle effective in the parasitic action of Trichoderma lignorum on Rhizoctonia solani and other soil fungi. Phytopathology 24:1153–1179
Whipps JM, Lumsden RD, 2001. Commercial use of fungi as plant disease biological control agents: status and prospects, In: Butt T, Jackson C, Magan N (eds), Fungal biocontrol agents: progress, problems and potential. CAB, Oxon, pp. 9–22
Witzgall P, Kirsch P, Cork A (2010) Sex Pheromones and Their Impact on Pest Management. J Chem Ecol 36:80–100
Zhou L, Xiao B, Chen Y, Li Y, Wang H (2008) Biological characteristics of the mycelium growth of two Trichoderma isolates. J Nanjing For Univ 32:95–98
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This work was supported by the National Natural Science Foundation of China (31260177) and the Program for Innovative Research Team (in Science and Technology) in the University of Yunnan Province.
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Nai-Yong Liu, Ze-Ran Bao, Jing Li, Xin-Yu Ao, Jia-Ying Zhu and Yu-Hui Chen declare that they have no conflict of interest.
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Liu, NY., Bao, ZR., Li, J. et al. Identification of differentially expressed genes from Trichoderma atroviride strain SS003 in the presence of cell wall of Cronartium ribicola . Genes Genom 39, 473–484 (2017). https://doi.org/10.1007/s13258-016-0512-5
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DOI: https://doi.org/10.1007/s13258-016-0512-5


