[Genetic transformation of the fungus Gliocladium sp. mediated by Agrobacterium tumefaciens].

OBJECTIVE: To construct a transformation system in Gliocladium sp. 6.102, a Cordyceps-colonizing fungus producing a variety of epipolythiodioxopiperazine (ETP) compounds with drug potentials. METHODS: Agrobacterium tumefaciens-mediated transformation (ATMT) was used to transform Gliocladium sp. 6.102. The factors of bacterial cell concentration, co-cultivation time, pH and acetosyringone concentration were optimized. RESULTS; A total of 50 -100 transformants per 10(6) fungal conidia was obtained via the optimal ATMT method. The genes encoding hygromycin B phosphotase and enhanced green fluorescent protein (EGFP) were transferred into Gliocladium sp. by the optimal ATMT method. The marker genes were successfully expressed and stably maintained in the transgenic fungus. CONCLUSION: A transformation system was established for Gliocladium sp. 6.102 and this system may be useful to identify ETP biosynthetic genes in Gliocladium.

High energy biofuel from endophytic fungi?

A paper entitled 'The production of myco-diesel hydrocarbons and their derivatives by the endophytic fungus Gliocladium roseum (NRRL 50072)' (Strobel et al., Microbiology 154, 3319-3328, 2008) demonstrated for the first time that a fungal endophyte can produce volatile hydrocarbons, which are major constituents of diesel fuel. The possible implications of this finding with regards to potential commercial applications and the ecological role of volatiles in endophyte-host relationships are discussed.

Role of zearalenone lactonase in protection of Gliocladium roseum from fungitoxic effects of the mycotoxin zearalenone.

Zearalenone is a mycotoxin with estrogenic effects on mammals that is produced by several species of Fusarium. We found that zearalenone and its derivatives inhibit the growth of filamentous fungi on solid media at concentrations of < or =10 microg/ml. The fungitoxic effect declined in the order zearalenone > alpha-zearalenol > beta-zearalenol. The mycoparasitic fungus Gliocladium roseum produces a zearalenone-specific lactonase which catalyzes the hydrolysis of zearalenone, followed by a spontaneous decarboxylation. The growth of G. roseum was not inhibited by zearalenone, and the lactonase may protect G. roseum from the toxic effects of this mycotoxin. We inactivated zes2, the gene encoding zearalenone lactonase in G. roseum, by inserting a hygromycin resistance cassette into the coding sequence of the gene by means of Agrobacterium tumefaciens-mediated genetic transformation. The zes2 disruption mutants could not hydrolyze the lactone bond of zearalenone and were more sensitive to zearalenone. These data are consistent with a hypothesis that resorcylic acid lactones exemplified by zearalenone act to reduce growth competition by preventing competing fungi from colonizing substrates occupied by zearalenone producers and suggest that they may play a role in fungal defense against mycoparasites.

Enhancing biological control of basal stem rot disease (Ganoderma boninense) in oil palm plantations.

Basal Stem Rot (BSR) disease caused by Ganoderma boninense is the most destructive disease in oil palm, especially in Indonesia and Malaysia. The available control measures for BSR disease such as cultural practices and mechanical and chemical treatment have not proved satisfactory due to the fact that Ganoderma has various resting stages such as melanised mycelium, basidiospores and pseudosclerotia. Alternative control measures to overcome the Ganoderma problem are focused on the use of biological control agents and planting resistant material. Present studies conducted at Indonesian Oil Palm Research Institute (IOPRI) are focused on enhancing the use of biological control agents for Ganoderma. These activities include screening biological agents from the oil palm rhizosphere in order to evaluate their effectiveness as biological agents in glasshouse and field trials, testing their antagonistic activities in large scale experiments and eradicating potential disease inoculum with biological agents. Several promising biological agents have been isolated, mainly Trichoderma harzianum, T. viride, Gliocladium viride, Pseudomonas fluorescens, and Bacillus sp. A glasshouse and field trial for Ganoderma control indicated that treatment with T. harzianum and G. viride was superior to Bacillus sp. A large scale trial showed that the disease incidence was lower in a field treated with biological agents than in untreated fields. In a short term programme, research activities at IOPRI are currently focusing on selecting fungi that can completely degrade plant material in order to eradicate inoculum. Digging holes around the palm bole and adding empty fruit bunches have been investigated as ways to stimulate biological agents.

[Allelopathic effect of root exudates on pathogenic fungi of root rot in continuous cropping soybean].

Allelopathic effect of root exudates on pathogenic fungi of root rot in continuous cropping soybean was studied by sand culture, water culture, and indoor culture experiments. The results showed that allelopathic promotion of root exudates on the growth of Fusarium semitectum, Gliocladium roseum and Fusarium oxysporum, especially Fusarium semitectum reached significant level or especially significant level in continuous cropping soybean compared with the control. Allelopathic promotion of root exudates on the growth of Fusarium semitectum and Gliocladium roseum in continuous cropping soybean was distinctly larger than that in rotation soybean, and the difference reached significant level under their low concentration. Allelopathic promotion of high concentration of root exudates on the growth of Fusarium semitectum was smaller than that of low concentration of root exudates, and the difference reached significant level in continuous cropping soybean. Allelopathic inhibition of high concentration of phthalic acid and propanedioic acid (L5 and B5) on the growth of Fusarium semitectum. Gliocladium roseum and Fusarium oxysporum, especially Fusarium semitectum reached significant level or especially significant level compared with the control. However, allelopathic promotion of low concentration of phthalic acid and propanedioic acid on the growth of Fusarium semitectum, Gliocladium roseum and Fusarium oxysporum partly reached significant level.