Expression of ethylene-forming enzyme (EFE) of Pseudomonas syringae pv. glycinea in Trichoderma viride

  • Li Tao
  • Hong-Jun Dong
  • Xi Chen
  • San-Feng Chen
  • Tian-Hong Wang
Biotechnological Products and Process Engineering


The efe gene encoding an ethylene-forming enzyme from Pseudomonas syringae pv. glycinea has been expressed for the first time under the control of Trichoderma reesei cbh1 promoter in Trichoderma viride. Reverse transcription polymerase chain reaction analysis showed that transformant Y2 produced mRNA of the efe gene. Southern blot analysis showed that there was one copy of efe gene which was integrated into the chromosomal DNA of T. viride. Ethylene production by transformant Y2 was efficiently induced by cellulose, while very low level of ethylene was produced when sodium carboxymethyl cellulose or lactose was used as carbon source. Peptone exerted a much greater stimulatory effect on ethylene production. A high level of ethylene was produced when transformant Y2 was cultured in solid fermentation medium containing wheat straw, indicating that plant wastes could be directly converted to ethylene by the recombinant filamentous fungus.


Ethylene Ethylene-forming enzyme (EFE) cbh1 promoter Trichoderma viride Pseudomonas syringae 



We wish to thank Dr. Qun He for discussions. This work was supported by China high Technology (863) Project (Grant No. 2006AA10A213).


  1. Calo L, García I, Gotor C, Romero LC (2006) Leaf hairs influence phytopathogenic fungus infection and confer an increased resistance when expressing a Trichoderma α-1,3-glucanase. J Exp Bot 57:3911–3920CrossRefGoogle Scholar
  2. Chague V, Elad Y, Barakat R, Tudzynski P, Sharon A (2002) Ethylene biosynthesis in Botrytis cinerea. FEMS Microbiol Ecol 40:143–149Google Scholar
  3. Chalutz E, Lieberman M (1977) Methionine-induced ethylene production by Penicillium digitatum. Plant Physiol 60:402–406Google Scholar
  4. Hall MA, Smith AR (1995) Ethylene and the responses of plants to stress. Bulg J Plant Physiol 21:71–79Google Scholar
  5. Henrique-Silva F, El-Gogary S, Carle-Urioste JC, Matheucci EJ, Crivellaro O, El-Dorry H (1996) Two regulatory regions controlling basal and cellulose-Induced expression of the gene encoding cellobiohydrolase I of Trichoderma reesei are adjacent to Its TATA Box. Biochem Biophys Res Commun 228:229–237CrossRefGoogle Scholar
  6. Ilmen M, Saloheimo A, Onnela ML, Penttila ME (1997) Regulation of cellulase gene expression in the filamentous fungus Trichoderma reesei. Appl Environ Microbiol 63:1298–1306Google Scholar
  7. Ishihara K, Matsuoka M, Inoue Y, Tanase S, Ogawa T, Fukuda H (1995) Overexpression and in vitro reconstitution of the ethylene-forming enzyme from Pseudomonas syringae. J Ferment Bioeng 79:205–211CrossRefGoogle Scholar
  8. Jiao XZ, Philosoph-Hadas S, Su LY, Yang SF (1986) The Conversion of 1-(malonylamino)cyclopropane- l-carboxylic acid to 1-aminocyclopropane-1-carboxylic acid in plant tissues. Plant Physiol 81:637–641Google Scholar
  9. Joutsjoki V, Torkkeli T, Nevalainen H (1993) Transformation of Trichoderma reesei with the Hormoconis resinae glucoamylase P (gamP) gene: production of a heterologous glucoamylase by Trichoderma reesei. Curr Genet 24:223–229CrossRefGoogle Scholar
  10. Margolles-Clark E, Hayes CK, Harman GE, Penttila M (1996a) Improved production of Trichoderma harzianum endochitinase by expression in Trichoderma reesei. Appl Environ Microbiol 62:2145–2151Google Scholar
  11. Margolles-Clark E, Harman GE, Penttila M (1996b) Enhanced expression of endochitinase in Trichoderma harzianum with the cbh1 Promoter of Trichoderma reesei. Appl Environ Microbiol 62:2152–2155Google Scholar
  12. Miettinen-Oinonen A, Torkkeli T, Paloheimo M, Nevalainen H (1997) Overexpression of the Aspergillus niger pH 2.5 acid phosphatase gene in a heterologous host Trichoderma reesei. J Biotechnol 58:13–20CrossRefGoogle Scholar
  13. Nagahama K, Ogawa T, Fujii T, Tazaki M, Tanase S, Morino Y, Fukuda H (1991) Purification and properties of ethylene-forming enzyme from Pseudomonas syringae pv. phaseolicola PK2. J Gen Microbiol 137:2281–2286Google Scholar
  14. Nagahama K, Ogawa T, Fujii T, Fukuda H (1992) Classification of ethylene producing bacteria in terms of biosynthetic pathways to ethylene. J Ferment Bioeng 73:1–5CrossRefGoogle Scholar
  15. Penttila M, Nevalainen H, Ratto M, Salminen E, Knowles J (1987) A versatile transformation system for the cellulolytic filamentous fungus Trichoderma reesei. Gene 61:155–164CrossRefGoogle Scholar
  16. Sakai M, Ogawa T, Matsuoka M, Fukuda H (1997) Photosynthetic conversion of carbon dioxide to ethylene by the recombinant cyanobacterium, Synechococcus sp. PCC 7942, which harbours a gene for the ethylene-forming enzyme of Pseudomonas syringae. J Ferment Bioeng 84:434–443CrossRefGoogle Scholar
  17. Sato M, Watanabe K, Yazawa M, Takikawa Y, Nishiyama K (1997) Detection of new ethylene-producing bacteria, Pseudomonas syringae pvs. cannabina and sesami, by PCR amplification of genes for the ethylene-forming enzyme. Phytopathology 87:1192–1196CrossRefGoogle Scholar
  18. Spalding DH, Lieberman M (1964) Factors affecting the production of ethylene by Penicillium digitatum. Plant Physiol 40:645–648CrossRefGoogle Scholar
  19. Sun T, Liu BH, Liu DM, Li ZH (1997) Effect of elevated temperature on Trichoderma viride SL-1 in solid state fermentations. Biotechnol Lett 19:171–174CrossRefGoogle Scholar
  20. Takahama K, Matsuoka M, Nagahama K, Ogawa T (2003) Construction and analysis of a recombinant cyanobacterium expressing a chromosomally inserted gene for an ethylene-forming enzyme at the psbA1 locus. J Ferment Bioeng 95:302–305Google Scholar
  21. Tel-Zur N, Abbo S, Myslabodski D, Mizrahi Y (1999) Modified CTAB procedure for DNA isolation from epiphytic cacti of the genera Hylocereus and Selenicereus (cactaceae). Plant Mol Biol Report 17:249–254CrossRefGoogle Scholar
  22. Umikalsom MS, Ariff AB, Zulkifli HS, Tong CC, Hassan MA, Karim MIA (1997) Production of cellulase by a wild strain of Chaetomium globosum using delignified oil palm empty-fruit-bunch fibre as substrate. Appl Microbiol Biotechnol 47:590–595CrossRefGoogle Scholar
  23. Van Wyk JPH, Mohulatsi M (2003) Biodegradation of wastepaper by cellulase from Trichoderma viride. Bioresource Technol 86:21–23CrossRefGoogle Scholar
  24. Völksh B, Weingart H (1997) Comparison of ethylene-producing Pseudomonas syringae strains isolated from kudzu (Pueraria lobata) with Pseudomonas syringae pv. phaseolicola and Pseudomonas syringae pv. glycinea. Eur J Plant Pathol 103:795–802CrossRefGoogle Scholar
  25. Wang TH, Liu L, Wu ZH, Liu SL, Qu YB (2004) Novel cellulase profile of Trichoderma reesei strains constructed by cbh1 gene replacement with eg3 gene expression cassette. Acta Biochim Biophys 36:667–672Google Scholar
  26. Weingart H, Völksch B (1997) Ethylene production by Pseudomonas syringae pathovars in vitro and in planta. Appl Environ Microbiol 63:156–161Google Scholar
  27. Weingart H, Völksch B, Ullrich MS (1999) Comparison of ethylene production by Pseudomonas syringae and Ralstonia solanacearum. Phytopathology 88:360–365CrossRefGoogle Scholar
  28. Weingart H, Ullrich MS, Geider K, Völksch B (2001) The role of ethylene production in virulence of Pseudomonas syringae pvs. glycinea and phaseolicola. Phytopathology 91:511–518CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Li Tao
    • 1
  • Hong-Jun Dong
    • 1
  • Xi Chen
    • 1
  • San-Feng Chen
    • 1
  • Tian-Hong Wang
    • 2
  1. 1.National Key Laboratory for Agrobiotechnology, College of Biological Sciences and Center for Biomass EngineeringChina Agricultural UniversityBeijingPeople’s Republic of China
  2. 2.The State Key Laboratory of Microbial TechnologyShandong UniversityJinanChina

Personalised recommendations