Scientia Agricultura Sinica ›› 2015, Vol. 48 ›› Issue (5): 882-888.doi: 10.3864/j.issn.0578-1752.2015.05.06

• PLANT PROTECTION • Previous Articles     Next Articles

Isolation and Purification of Active Compound from Trichoderma viridescens and Its Inhibitory Activities Against Phytopathogens

ZHANG Liang, ZHANG Jing-ze   

  1. Institute of Biotechnology, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058
  • Received:2014-09-09 Online:2015-03-01 Published:2015-03-01

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Abstract: 【Objective】An antagonistic compound against Phytophthora spp. was obtained from Trichoderma spp. and its biocontrol potential was evaluated for controlling phytopathogens, thus providing a scientific basis for utilization of biocontrol strains and their antagonistic compounds. 【Method】 Trichoderma strains producing antagonistic compounds were screened by the cellophane method and grew on PDA as inoculants, and then were transfered into rice media for isolation of antagonistic compounds. The cultures of the Trichoderma strains from the rice media were extracted, filtered and concentrated, and initial crude extract was obtained. The crude extract was further purified by column and thin-layer chromatography and the active fractions were ascertained by the biological activity determination. Based on the chemical property of the active fraction obtained, their chemical structure was identified by analysis of gas chromatography-mass spectrometry (GC-MS). The biological activity of different types of plant pathogens was determined with the antagonistic compound obtained, including the oomycete P. capsici and P. melonis, ascomycete Fusarium oxysporum and basidiomycete Rhizoctonia solani. 【Result】 The initial screening assays showed that the strain TS0404 of T. viridescens produced an antagonistic compound with strong inhibitory activity against P. capsici. The results of isolation, purification and bioactivity determination displayed that the active fraction obtained was yellow oily liquid. Mass spectra revealed the biggest ion peak of active fraction was 166, which was identified as 6-pentyl-2H-pyran-2-one (6-PP). The bioactive determination demonstrated that the 6-PP had significant inhibitory activity against hyphal growth of P. capsici, P. melonis, R. solani and F. oxysporum (EC50 115.26, 99.58, 126.46 and 315.75 μg?mL-1, respectively) but the inhibitory effect on P. melonis was the best among them and hyphal growth was completely inhibited when its concentration reached 300 μg?mL-1. Similarly, the 6-PP had a conspicuous inhibitory activity against zoosporangial germination of P. capsici and P. melonis but the inhibitory effect on P. melonis was the best and zoosporangial germination of P. melonis was completely inhibited at 400 μg?mL-1 level. In addition, the 6-PP had significant inhibitory effect on conidial germination of F. oxysporum (EC50 151.81 μg?mL-1) and sclerotial germination of R. solani (300 μg?mL-1 for complete inhibitory concentration). 【Conclusion】 The 6-PP is an antagonistic compound isolated from T. viridescens, which showed strong inhibitory activities against the fruiting bodies of P. melonis, P. capsici, F. oxysporum and R. solani. It was concluded that the 6-PP is a type of broad-band antagonistic compound, which has great potential values of application in biocontrol.

Key words: Trichoderma viridescens, biocontrol fungal screening, antagonistic compound, structural identification, 6-pentyl- 2H-pyran-2-one

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