Cloning of tri Cluster and Analysis of tri Genes Expressions under Different Trichodermin-producing Conditions in Trichoderma brevicompactum

doi:10.16409/j.cnki.2095-039x.2016.01.014

Abstract

Abstract: Trichodermin is one of trichothecenes compounds, which has strongly antifungal activity on many kinds of plant pathogenic fungi. To preliminary clarify the function of tri cluster on trichodermin biosynthesis by Trichoderma brevicompactum 0248, the full length of tri cluster (24793 bp) was cloned. Sequence analysis of this cluster identified seven orthologs of T. brevicompactum tri genes. The influence of three carbon sources on the trichodermin production as well as the expression levels of tri genes in T. brevicompactum were also investigated. The result showed that the expression levels of tri genes under high trichodermin-producing condition by T. brevicompactum were much higher than that of trichodermin-nonproducing or low trichodermin-producing condition after 40 h incubation. The high expressions of tri genes in the glucose medium resulted in the accumulation of trichodermin after 40 h incubation. The expression level of tri gene was positively correlated with the yield of trichodermin. It provided a basis for further study the functional of tri genes in T. brevicompactum.

Key words: Trichoderma brevicompactum, trichodermin, tri cluster, carbon source, real-time quantitative PCR

CLC Number:

S476

YUAN Xiaofeng, SHENTU Xuping, YU Xiaoping. Cloning of tri Cluster and Analysis of tri Genes Expressions under Different Trichodermin-producing Conditions in Trichoderma brevicompactum[J]. journal1, 2016, 32(1): 93-100.

References

[1] 张广志, 杨合同, 文成敬. 木霉菌形态学分类检索与分子生物学鉴定[J]. 山东农业大学学报(自然科学版), 2011, 42(2):309-316.
[2] Shentu X P, Liu W P, Zhan X H, et al. Transcriptome sequencing and gene expression analysis of Trichoderma brevicompactum under different culture conditions[J]. PLoS ONE, 2014, 9(4):e94203.
[3] 陈捷, 朱杰伟, 张婷, 等. 木霉菌生物防治作用机理与应用研究进展[J]. 中国生物防治学报, 2011, 27(2):145-151.
[4] Shentu X, Zhan X, Ma Z, et al. Antifungal activity of metabolites of the endophytic fungus Trichoderma brevicompactum from garlic[J]. Brazilian Journal of Microbiology:Publication of the Brazilian Society for Microbiology, 2014, 45(1):248-254.
[5] Kimura M, Tokai T, O'Donnell K, et al. The trichothecene biosynthesis gene cluster of Fusarium graminearum F15 contains a limited number of essential pathway genes and expressed non-essential genes[J]. FEBS Letters, 2003, 539(1-3):105-110.
[6] Tijerino A. Overexpression of the trichodiene synthase gene tri5 increases trichodermin production and antimicrobial activity in Trichoderma brevicompactum[J]. Fungal Genetics and Biology, 2011, 48(3):285-296.
[7] Tijerino A, Hermosa R, Cardoza R E, et al. Overexpression of the Trichoderma brevicompactum tri5 gene:effect on the expression of the trichodermin biosynthetic genes and on tomato seedlings[J]. Toxins, 2011, 3(9):1220-1232.
[8] Cardoza R E, Malmierca M G, Hermosa M R, et al. Identification of loci and functional characterization of trichothecene biosynthesis genes in filamentous fungi of the genus Trichoderma[J]. Applied and Environmental Microbiology, 2011, 77(14):4867-4877.
[9] Jiao F, Kawakami A, Nakajima T. Effects of different carbon sources on trichothecene production and Tri gene expression by Fusarium graminearum in liquid culture[J]. FEMS Microbiology Letters, 2008, 285(2):212-219.
[10] Tonukari N J, Scottcraig J S, Waltonb J D. The Cochliobolus carbonum SNF1 gene is required for cell wall-degrading enzyme expression and virulence on maize[J]. The Plant Cell, 2000, 12(2):237-247.
[11] Ospina-Giraldo M D, Mullins E, Kang S. Loss of function of the Fusarium oxysporum SNF1 gene reduces virulence on cabbage and Arabidopsis[J]. Current Genetics, 2003, 44(1):49-57.
[12] Espeso E A, Peñalva M A. Carbon catabolite repression can account for the temporal pattern of expression of a penicillin biosynthetic gene in Aspergillus nidulans[J]. Molecular Microbiology, 1992, 6(11):1457-1465.
[13] Brayner R, Ferrari-Iliou R, Brivois N. Toxicological impact studies based on Escherichia coli bacteria in ultrafine ZnO nanoparticles colloidal medium[J]. Nano Letters, 2006, 6(4):866-870.
[14] 杨勇, 张凤英, 陈岑. PDA培养基改良配方的研究[J]. 酿酒科技, 2012, (4):29-31.
[15] Nakayashiki H, Hanada S, Nguyen B Q, et al. RNA silencing as a tool for exploring gene function in ascomycete fungi[J]. Fungal Genetics and Biology:Fg & B, 2005, 42(4):275-283.
[16] Almeida A J, Carmona J A, Cunha C, et al. Towards a molecular genetic system for the pathogenic fungus Paracoccidioides brasiliensis[J]. Fungal Genetics and Biology, 2007, 44(12):1387-1398.
[17] 申屠旭萍, 石一珺, 俞晓平. GC法测定研究发酵液中木霉素[J]. 中国药学杂志, 2008, 43(22):1755-1756.