Antifungal activity of Bacillus subtilis filtrate to control Fusarium oxysporum f.sp. lentis, the causal organism of lentil vascular wilt

In dual cultures, the supernatant filtrate of the biological control agent Bacillus subtilis was evaluated against (Fusarium oxysporum f.sp. lentis) the causal organism of lentil vascular wilt. The antagonistic activity was evaluated as percent reduction of fungal growth (certainly due, in part, to the antifungal metabolites produced by the antagonistic bacterium). The in-vitro experiments showed that B. subtilis filtrate, whether solid or liquid media, had a strong inhibiting activity on the spore germination and mycelial growth of F. oxysporum f. sp. lentis. In a glasshouse experiment, soil was drenched with B. subtilis filtrate at 30 ml/kg (vol/wt) around seedlings of a susceptible lentil line (ILL 4605). In this treatment there was only 31% mortality compared with 100% kill of plants in the control treatment (P≤0.05).

Antagonistic activities of selected fungal isolates against Armillaria mellea

The antagonistic activities of six selected fungal isolates against Armilloria mellea were studied on two different concentrations of three media, on fungicides-amended malt extract agar (MEA) medium, and in glasshouse pots filled with John Innes No.2 compost and natural field soil. Trichoderma hamatum isolate Tham1 was found the most effective in reducing Armillaria growths on both the low and high concentrations of malt extract, potato dextrose and V-8 juice in MEA, potato dextrose agar (PDA) and V-8 juice agar (VJA), respectively, followed by T. harzianum isolate Th2 and T. viride isolate Tv3. Neither dose rate (200 or 2000 mg l(-1)) of fenpropidin allowed any growth of Armillaria on MEA, while that of the antagonists was also completely inhibited or greatly restricted. However, both dose rates of fosetyl-A1 allowed the growth of Armillaria and almost all the antagonists. Data on colony diameters of Armillaria showed Tham1 as the most effective antagonist along with Th2, Th23 and Tv3. Tham1 was also found the most effective in protecting hazel billets from colonization by Armillaria, followed by Th2 and Th23. Compared with 7.1 colonized billets in the inoculated controls, only 1.3, 2.6 and 2.7 billets (out of ten) were colonized, respectively, when protected with these antagonists. The results indicate that the Trichoderma isolates are able to maintain their antagonistic effects on A. mellea under a variety of nutritional, chemical and edaphic regimes. More investigations are needed to develop a system of control for the disease with these potential antagonists.

Cultural techniques for improvement in biocontrol potential of fungal antagonists for biological control of Armillaria root rot of strawberry plants under glasshouse conditions

Several in vitro and in vivo experiments were conducted to develop an effective technique for culturing potential fungal antagonists (isolates of Trichoderma harzianum, Dactylium dendroides, Chaetomium olivaceum and one unidentified fungus) selected for activity against Armillaria mellea. The antagonists were inoculated onto (1) live spawn of the oyster mu shroom (Pleurotus ostreatus), (2) extra-moistened or sucrose-enriched mushroom composts containing living or autoclaved mycelia of P. ostreatus or Agaricus bisporus (button mushroom), (3) pasteurized compost with or without A. bisporus mycelium, wheat bran, wheat germ and (4) spent mushroom composts with living mycelia of A. bisporus, P. ostreatus or Lentinus edodes (the Shiitake mushroom). In one experiment, a representative antagonist (isolate Th2 of T. harzianum) was grown together with the A. bisporus mycelium, while in another one, the antagonist was first grown on wheat germ or wheat bran and then on mushroom compost with living mycelium of A. bisporus. Some of the carrier substrates were then added to the roots of potted strawberry plants in the glasshouse to evaluate their effectiveness against the disease. The antagonists failed to grow on the spawn of P. ostreatus even after reinoculations and prolonged incubation. Providing extra moisture or sucrose enrichment also did not improve the growth of Th2 on mushroom composts in the presence of living mycelia of A. bisporus or P. ostreatus. The antagonist, however, grew rapidly and extensively on mushroom compost with autoclaved mycelia, and also on wheat germ and wheat bran. Colonization of the substrates by the antagonist was positively correlated with its effectiveness in the glasshouse studies. Whereas only 33.3% of the inoculated control plants survived in one experiment monitored for 560 days, 100% survival was achieved when Th2 was applied on wheat germ or wheat bran. Growth of the antagonist alone on pasteurized or sterilized compost (without A. bisporus mycelia) and simultaneous growth of the antagonist and mushroom on pasteurized compost did not improve survival over the inoculated controls, but growth over mushroom compost with the living mycelium resulted in 50% survival rate. C. olivaceum isolate Co was the most effective, resulting in overall survival rate of 83.3% compared with only 8.3% for the inoculated and 100% for the uninoculated (healthy) controls. This antagonist gave the highest survival rate of 100% on spent mushroom compost with L. edodes. T harzianum isolate Th23, with 75% survival rate, was the most effective on spent mushroom compost with P. ostreatus, while D. dendroides isolate SP resulted in equal survival rates of 50% on all the three mushroom composts.

Comparisons between the in vitro and in vivo efficacies of potential fungal antagonists of Armillaria mellea

Seventeen fungal isolates were tested in vitro as potential antagonists of two isolates of the root rot pathogen, Armillaria mellea. Some of the isolates were also added on mushroom composts with living mycelia to the roots of Armillaria-inoculated potted strawberry plants in the glasshouse to find out if they had the same degree of efficacy against the disease. Dactylium dendroides isolate SP was the most effective in reducing mycelial growth of A. mellea isolate 1 (Am1), followed by Trichoderma harzianum isolate Th2 and T. viride isolate Tv4. Th2, Th22, Tv3 and SP grew extensively over Am1 colonies, disintegrating the rhizomorphs. Isolate Tham1 of T hamatum was the most effective in reducing mycelial growth of A. mellea isolate 2 (Am2), followed by Tv3. Th12, Th22, Tv1, Tv3 and SP inhibited the initiation and growth of rhizomorphs of Am2. Regeneration tests showed that both Am1 and Am2 attacked by Trichoderma isolates and SP were no longer viable. Th23 and SP were almost as effective in vivo as in vitro. But isolate Co of Chaetomium olivaceum, which was ineffective in vitro, was found effective in vivo. Conversely, Th2, which exhibited good antagonistic activity in vitro, performed poorly in vivo. These results show that the in vitro and in vivo efficacies of potential antagonists may not necessarily be closely correlated. Hence, there is a danger that potentially effective isolates may be discarded if decisions are made only on the basis of preliminary screening tests carried out under laboratory conditions.

Genotype and temperature influence pea aphid resistance to a fungal entomopathogen

The influence of temperature on life history traits of four Acyrthosiphon pisum clones was investigated, together with their resistance to one genotype of the fungal entomopathogen Erynia neoaphidis . There was no difference among aphid clones in development rate, but they did differ in fecundity. Both development rate and fecundity were influenced by temperature, but all clones showed similar responses to the changes in temperature (i.e. the interaction term was nonsignificant). However, there were significant differences among clones in susceptibility to the pathogen, and this was influenced by temperature. Furthermore, the clones differed in how temperature influenced susceptibility, with susceptibility rankings changing with temperature. Two clones showed changes in susceptibility which mirrored changes in the in vitro vegetative growth rate of E. neoaphidis at different temperatures, whereas two other clones differed considerably from this expected response. Such interactions between genotype and temperature may help maintain heritable variation in aphid susceptibility to fungal pathogen attack and have implications for our understanding of disease dynamics in natural populations. This study also highlights the difficulties of drawing conclusions about the efficacy of a biological control agent when only a restricted range of pest genotypes or environmental conditions are considered.

Modelling the uptake and growth kinetics of Penicillium glabrum in a tannic acid-containing solid-state fermentation for tannase production

A mathematical growth model for the batch solid-state fermentation process for fungal tannase production was developed and tested experimentally. The unstructured model describes the uptake and growth kinetics of Penicillium glabrum in an impregnated polyurethane foam substrate system. In general, good agreement between the experimental data and model simulations was obtained. Biomass, tannase and spore production are described by logistic kinetics with a time delay between biomass production and tannase and spore formation. Possible induction mechanisms for the latter are proposed. Hydrolysis of tannic acid, the main carbon source in the substrate system, is reasonably well described with Michaelis-Menten kinetics with time-varying enzyme concentration but a more complex reaction mechanism is suspected. The metabolism of gallic acid, a tannase-hydrolysis product of tannic acid, was shown to be growth limiting during the main growth phase. (c) 2004 Elsevier Ltd. All rights reserved.

Long-term conditioning of soil by plantation eucalypts and pines does not affect growth of the native jarrah tree

Plant species can condition the physico-chemical and biological properties of soil in ways that modify plant growth via plant–soil feedback (PSF). Plant growth can be positively affected, negatively affected or neutrally affected by soil conditioning by the same or other plant species. Soil conditioning by other plant species has particular relevance to ecological restoration of historic ecosystems because sites set aside for restoration are often conditioned by other, potentially non-native, plant species. We investigated changes in properties of jarrah forest soils after long-term (35 years) conditioning by pines (Pinus radiata), Sydney blue gums (Eucalyptus saligna), both non-native, plantation trees, and jarrah (Eucalyptus marginata; dominant native tree). Then, we tested the influence of the conditioned soils on the growth of jarrah seedlings. Blue gums and pines similarly conditioned the physico-chemical properties of soils, which differed from soil conditioning caused by jarrah. Especially important were the differences in conditioning of the properties C:N ratio, pH, and available K. The two eucalypt species similarly conditioned the biological properties of soil (i.e. community level physiological profile, numbers of fungal-feeding nematodes, omnivorous nematodes, and nematode channel ratio), and these differed from conditioning caused by pines. Species-specific conditioning of soil did not translate into differences in the amounts of biomass produced by jarrah seedlings and a neutral PSF was observed. In summary, we found that decades of soil conditioning by non-native plantation trees did not influence the growth of jarrah seedlings and will therefore not limit restoration of jarrah following the removal of the plantation trees.

Host and habitat filtering in seedling root-associated fungal communities: taxonomic and functional diversity are altered in ‘novel’ soils

Climatic and land use changes have significant consequences for the distribution of tree species, both through natural dispersal processes and following management prescriptions. Responses to these changes will be expressed most strongly in seedlings near current species range boundaries. In northern temperate forest ecosystems, where changes are already being observed, ectomycorrhizal fungi contribute significantly to successful tree establishment. We hypothesised that communities of fungal symbionts might therefore play a role in facilitating, or limiting, host seedling range expansion. To test this hypothesis, ectomycorrhizal communities of interior Douglas-fir and interior lodgepole pine seedlings were analysed in a common greenhouse environment following growth in five soils collected along an ecosystem gradient. Currently, Douglas-fir’s natural distribution encompasses three of the five soils, whereas lodgepole pine’s extends much further north. Host filtering was evident amongst the 29 fungal species encountered: 7 were shared, 9 exclusive to Douglas-fir and 13 exclusive to lodgepole pine. Seedlings of both host species formed symbioses with each soil fungal community, thus Douglas-fir did so even where those soils came from outside its current distribution. However, these latter communities displayed significant taxonomic and functional differences to those found within the host distribution, indicative of habitat filtering. In contrast, lodgepole pine fungal communities displayed high functional similarity across the soil gradient. Taxonomic and/or functional shifts in Douglas-fir fungal communities may prove ecologically significant during the predicted northward migration of this species; especially in combination with changes in climate and management operations, such as seed transfer across geographical regions for forestry purposes.