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Use of Trichoderma spp. for biological control of the livestock feed contaminant fungus Fusarium proliferatum

浙江大学学报（农业与生命科学版） 2004年第4期30卷 414-414页

作者： Ruocco M Pane F Ritieni A lanzuise s Ambrosino P Marra R Woo s L Ciliento R soriente I Ferraioli s scala F Lorito M Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 ItalyDip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 ItalyDip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 ItalyDip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 ItalyDip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 ItalyDip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Uni

Fusarium spp. are pathogens of many important agricultural crops, and are often strong mycotoxin producers. Fusarium proliferatum, in particular, causes disease in cereals and secretes the toxin Beauvaricin that contaminates livestock feed and cereals, producing a variety of toxicity symptoms ranging from poor weight gain to mortality. Beauvaricin is a cyclodepsipeptide and acts as a potent mycotoxin known to have insecticidal properties. This compound is highly toxic to human cell lines, where it induces apoptosis and specifically inhibits cholesterol acetyltransferase. Nothing is known about the role of this mycotoxin during the interaction of F. proliferatum with other microorganisms, including the fungal antagonists Trichoderma spp. In vitro tests have demonstrated that the antagonistic and mycoparasitic activity of Trichoderma is not inhibited by the presence of Beauvaricin at concentrations up to 10 mg/kg in the substrate. In vivo biocontrol assays on barley and wheat with Trichoderma against F. proliferatum isolates, producing and non-producing Beauvaricin, confirmed the ability of the antagonist to control this pathogen in all cases. Also Trichoderma culture filtrates obtained in conditions that promote Cell Wall Degrading Enzyme (CWDE) secretion, were able to inhibit. spore. germination. of. different. F.. proliferatum. isolates.. These. results. suggest. the. possibility. of. using. Trichoderma. and/or. its. metabolites. to. control. contaminants. of. livestock. feed. by. mycotoxin-producing. Fusarium..

关键词：家畜霉菌饲料污染物镰刀霉木霉属生物防治

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Cloning of first abc transporter encoding gene from Trichoderma spp. and its expression during stress and mycoparasitism

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浙江大学学报（农业与生命科学版） 2004年第4期30卷 450-450页

作者： lanzuise s Ruocco M scala V Catapano L Woo s Ciliento R Ferraioli s soriente I Vinale F scala F Del sorbo G Lorito M Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 ItalyDip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 ItalyDip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 ItalyDip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 ItalyDip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Un

Trichoderma in its natural environment competes for nutrient uptake and is required to protect itself from adverse natural toxic compounds, such as those produced by plants and other microbes in the soil community, or synthetic toxic compounds released human activity. One of the most important metabolic pathways for drug resistance and substrate uptake, both in prokaryotes and eukaryotes, is ATP dependent. The role of ABC transporter proteins in the biology of Trichoderma is still not known. We present the cloning of the first four ABC transporter genes (TABC1, TABC2, TABC3, TABC4 ) in Trichoderma, and in particular T. atroviride P1, and the characterization of TABC2 The complete sequence of this gene is 6535 bp, which includes a promoter of 1624 bp, a terminator of 642 bp and a coding region of 4264 bp. The promoter contains many of the potential transcription factor binding sites found in the 5’ upstream region of the ech42 gene of T. atroviride P1. These included: heat shock factors (HsF), a nitrogen-regulating factor (Nit-2), a stress-response element (sTRE), a GCR1 elements, and a Cre BP1 motif. Northern analysis and RT-PCR demonstrated that TABC2 is highly expressed when Trichoderma is subjected to nitrogen starvation, grown in the presence of culture filtrates of Botrytis cinerea, Rhizoctonia solani, and Pythium ultimum, or when N-acetylglucosamine is added to the substrate. TABC2 appears to be co-regulated with some CWDE-encoding genes, suggesting that this is the first ABC transporter encoding gene involved in mycoparasitic events. It’s role in the interaction of Trichoderma with fungal hosts or plants is being investigated by targeted gene disruption and overexpression.

关键词：真菌寄生现象应激反应木霉属真菌克隆病原体

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The Trichoderma-plant interaction is mediated by avirulence proteins produced by this fungus

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浙江大学学报（农业与生命科学版） 2004年第4期30卷 422-422页

作者： Ruocco M lanzuise s Woo s L Ambrosino P Marra R Turrà D Gigante s Formisano E scala F Kip N P J G M de Wit Lorito M Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 ItalyDip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica Università di Napoli "Federico II" and CNR-IPP Via Università 100 Portici (NA) 80055 Italy Dip. Arboricoltura Botanica e Patologia Vegetale-Sez. Patologia Vegetale Lab. di Lotta Biologica

The molecular basis of Trichoderma -plant interaction is very complex and still not completely understood. The colonization of the root system by rhizosphere competent strains of Trichoderma results in increased development of root/aerial systems, in improved yields and in plant disease control. Other beneficial effects, such as the induction of plant systemic resistance, have also been described. To understand the mechanisms involved we are using different approaches, including the making of transformants expressing genes that encode for compounds able to affect plant response to pathogens. Trichoderma transformants carrying the avirulence gene Avr4 from Cladosporium fulvum under the control of constitutive and inducible promoters were obtained and tested on tomato plants having the Cf4 resistance gene. Necrosis and suberification zones, similar to the symptoms appearing during Cladosporium-tomato interaction, were found when the roots of the Cf4 plants were treated with Avr4-Trichoderma. This demonstrates that selected Trichoderma strains are able to transfer to the plant molecules that may deeply affect metabolism, disease resistance etc. Therefore, these beneficial fungi can be regarded as biotechnological tools to provide a variety of crops with useful compounds. Moreover, in in vitro competition assays the transformants were found to be more effective as antagonists against Alternaria alternata than the wild type. Trichoderma sends a variety of biochemical signals to the plants including avirulence molecules; therefore the presence of avr-like proteins in the fungus proteome was investigated. Proteome analysis has permitted us to isolate and sequence many proteins potentially having this function. From the extracellular protein extracts, we have purified and sequenced a protein with structural characteristics similar to Avr4 of C. fulvum. The protein, Hytra1, was found to be a hydrophobin with chitin binding activity, the typical 8 cysteine residues, and 4 dis

关键词：无毒力蛋白质真菌木霉属生产植物

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