Application of Trichoderma and Gliocladium in alginate pellets for control of Rhizoctonia damping-off

Alginate pellets were prepared from wet fermentor biomass of 11 isolates of Trichoderma spp. and Gliocladium virens , with wheat bran as a food base carrier. Pellets with eight of the isolates reduced survival (34-78%) of Rhizoctonia solani in infested beet seed in soil. Pellets containing a T. harzianum isolate (Th-58) and a T. hamatum isolate (TRI-4) were the most effective. All isolates significantly reduced growth of the pathogen from infested beet seed into natural soil. Populations of isolates proliferated in soil to 10⁶−10¹¹ colony-forming units/g (cfu) from propagules within the pellets. Pellets with TRI-4 reduced pathogen survival and growth (>70%) in six different soils and were effective against six R. solani isolates in a natural loamy sand. Survival of R. solani in infested beet seed was not reduced when TRI-4 pellets were added to soil 1-6 weeks before the pathogen; however, saprophytic growth was prevented. Small amounts of biomass (3.0–7.5 g wet weight) in pellets were as effective as a large amount (300 g) in suppressing the pathogen. The storage of pellets for more than 6 weeks at 5 or 25C reduced their effectiveness against R. solani. Pellets prepared with four and three of the 11 isolates prevented damping-off of cotton and sugar beet in the greenhouse, respectively.     

Growth of Coniothyrium minitans,Gliocladium roseum,Trichoderma harzianum and T. viride from alginate pellets and interaction with water availability1

Conidia, chlamydospores and mycelia of Coniothyrium minitans, Gliocladium roseum, Trichoderma harzianum and T. viride were obtained from liquid or solid culture and formulated within alginate pellets. Quantitative assessment of these pellets over a 12-week period showed a decrease in the number of colony-forming units (cfu) from about 10⁷ to 10³-10⁴ g^-1 air-dried pellets. Qualitatively, growth was assessed by direct plating the alginate pellets on rich (potato dextrose agar, PDA) and poor (water agar, WA) substrates. Rate of growth (mm day^-1) from individual pellets was decreased by water potential in the range -0.25 to-2.8 MPa, with growth significantly less on WA than PDA for all species except G. roseum. All species grew from 95 to 100% of direct-plated pellets on PDA over the water-potential range tested. However, on WA, interaction between water stress (-2.8 MPa) and lack of nutrients resulted in a reduction, with less than 30% of pellets of some formulations showing growth. In contrast to the quantitative results, there was little change in the growth rate of test species from pellets over a 12-week period, particularly on PDA. Growth from pellets could not be conclusively demonstrated in unsterile soil. The shelf-life and viability of alginate pellets in soil is assessed.     

Effect of Gliocladium and Trichoderma on damping-off and blight of snapbean caused by Sclerotium rolfsii in the greenhouse

Aqueous suspensions of conidia of 285 wild-type strains and mutants of Gliocladium virens, Trichoderma hamatum, T. harzianum and T. viride were tested against Sclerotium rolfsii in the greenhouse. Ten strains of G. virens and four strains of T. harzianum suppressed damping-off of snapbean by 30-50% and blight by 36-74%. All strains of T. hamatum and T. viride tested as conidia were ineffective. In general, strains of G. virens were more effective in suppressing disease in the greenhouse than strains of T. harzianum. Several strains of G. virens and T. harzianum used alone were equal to or more effective than double and triple mixtures of such strains in disease suppression. Of four formulations of G. virens tested, germlings, alginate-bran-fermenter biomass pellets, and Pyrax-fermenter biomass mixtures reduced disease considerably and all three formulations were more effective than conidia in aqueous suspension. Strain G1-3 of G. virens added to soil as Pyrax-fermenter biomass mixtures in amounts to provide colony-forming units ranging from 1 -5 × 10³ to 1 -2 × 10⁴ per g soil provided statistically significant protection of the host at all concentrations. The extent of biological control with strains G1-3 and G1-21 of G. virens also depended on the strain of the pathogen used. Both strains effectively suppressed disease caused by strain Sr-1 (small sclerotia) of S. rolfsii. They were partially effective against strain Sr-116 (medium size sclerotia) and ineffective against strain Sr-3 (large sclerotia). Although strains G1-3 and G1-21 colonized the sclerotia of all three strains of S. rolfsii in soil effectively, they only reduced germinability of sclerotia of strain Sr-1.     

Biological control of Botrytis cinerea on chickpea seed with Trichoderma spp. and Gliocladium roseum: indigenous versus non-indigenous isolates

Establishment of chickpea seedlings from seed inoculated with Botrytis cinerea was increased under controlled environmental conditions by treatment of seed with Gliocladium roseum or Trichoderma virens . An isolate of G. roseum (GrH) from Horsham in the chickpea-growing region of western Victoria outperformed T. virens in three soil types, including one to which T. virens was indigenous. There was no effect of soil moisture on seedling establishment from seed treated with GrH, but recovery of G. roseum from seed in continuously wet soil was lower than from wet/dry soil. A factorial assay with three isolates of G. roseum and the soils from which they were isolated showed no interaction between isolate and soil type and no difference between isolates for establishment or recovery. In a field trial at Horsham sown in winter (July), seed treatment with GrH increased establishment from 26.0% (control) to 37.5%. Biocontrol by GrH was ineffective in a winter-sown field trial at Bundoora, but all three isolates increased establishment from 47.5% (control) to up to 87.5% in a trial at the same location sown in spring (October). The commercial implications of these observations are discussed.     

Cellulose-decomposing fungi in polder soils and their possible influence on pathogenic fungi

For a study of the colonisation of the IJsselmeerpolders by fungi, attention was fixed on the cellulose-decomposing fungi, since these are considered to represent the more stable part of the mycoflora of soils. If one assumes that in the North East Polder, cultivated for approximately 25 years, the fungal population is approaching stability, the progress of colonisation of the younger polders can be judged by comparison of the number of species found and their frequency.It is shown that the progress of fungal colonisation did not depend only on the time during which a soil has been dry, but also on the use that has been made of it. Of the species known as antagonists of other fungi,Trichoderma spp. provided an increasingly larger proportion of the total population as the soils were further from stability.It is concluded that the increase of fungal antagonists may well be one of the causes of the decline of such parasites asOphiobolus graminis andRhizoctonia solani. Special mention is made ofGliocladium roseum, the parasitism of which towardsR. solani could be demonstrated in paired cultures.     

放牧对短花针茅草原土壤微生物和土壤养分的影响及其季节动态

研究了短花针茅草原不同季节、不同放牧梯度内土壤微生物数量、土壤养分的变化及土壤微生物与土壤养分之间的相关性,结果表明:轻度放牧区(LG)和重度放牧区(HG)内土壤微生物数量春季最高,土壤微生物数量(0-10 cm)在三个放牧区内随着季节的变化均呈下降趋势。而在10-20 cm,对照区内土壤微生物数量的高峰期出现在夏季然后开始下降,轻度放牧区和重度放牧区土壤微生物随着季节的变化而下降。土壤微生物数量到了秋季在三个放牧梯度内均呈现显著性的差异,土壤养分也是在秋季出现了显著性差异(P<0.05)。土壤微生物总数在不同放牧梯度内的变化规律是对照和轻度放牧区均大于重度放牧区。土壤微生物与土壤有机质和土壤全氮呈较强的正相关。     

土壤不同处理对木霉菌定殖及其生防效果的影响

重茬土经施肥改土、高温灭菌后,增施木霉菌的试验结果表明:木霉菌可以在甜瓜根表、根际定殖存活,同时不影响其他有益微生物的正常生长;土壤经高温灭菌后,土壤中的微生物呈真空状态,有利于木霉菌的快速定殖,促进甜瓜生长,降低甜瓜枯萎病的病情指数。甜瓜枯萎病发病率与土壤中镰刀菌孢子的存活量成正相关,与土壤中木霉菌、细菌数量的多少成负相关。     

Effect of benomyl on soil fungi associated with rye. 2. Effect on fungi of culm bases and roots

The mycoflora of culm bases and roots of rye was assessed in field trials, where benomyl was applied at dose rates ranging from 0.24 to 4.80 kg ha^?1. Samples of culm bases were taken three times during the growing season, those of roots only at the harvest date. On culms with various symptoms from untreated plots,Pseudocercosporella herpotrichoides, Alternaria spp. andGerlachia nivalis were prevalent and on those from benomyl-treated plotsAlternaria spp. andFusarium culmorum. In later stages of growth,G. nivalis sharply declined andAlternaria spp. andF. culmorum increased. At the end of the season,Periconia macrospinosa andTyphula incarnata appeared in treated plots. In samples of roots taken more than two months after the last spray, porosporous dematiaceous species (Alternaria, Ulocladium andDendryphion), Mortierella spp. and other resistant fungi were prevalent in plots treated with 1.20 kg ha^?1 or more, but not in those that received 0.24 kg ha^?1, which is recommended for disease control. Lower counts were recorded only for species that are highly sensitive in vitro, e.g.Microdochium bolleyi andTrichoderma spp. Some fusaria were either not affected or tended to be slightly stimulated by the treatment. An attempt was made to attribute the incidence of these moderately sensitive fungi to the effect of the fungicide on non-pathogens.     

The effect of repeated applications of 2,4-D and MCPA on their breakdown in soil

The effect of repeated applications of 2,4-D and MCPA in a long-term field experiment on their subsequent degradation was investigated. The herbicides had been applied annually since 1954, Comparisons were made with plots treated once and then retreated the following year. Repealed applications of the two herbicides resulted in a reduction in degradation lime from 10 weeks for 2,4-D and 20 weeks for MCPA after one application to 4 and 7 weeks respectively after 19 years of annual application. In spite of an increased capability of the soil microflora lo degrade 2,4-D and MCPA 9 months after a previous treatment. no change in numbers of organisms capable of utilizing the substances as carbon sources were found in treated plots as compared with the control. It is obvious that there is a cross adaptation between 2,4-D and MCPA. It seems most likely that two different ‘enrichment’ been induced, each capable of decomposing both herbicides, but with different efficiencies. Effet d'applications répétée de 2,4-D et de MCPA sur leur degradation dans le sol L'effet d'applications répétée de 2.4-D et de MCPA sur leur degradation uléerieure a étéétudié dans une expérience au champ de longue durée. Les herbicides ont été appliqués chaque année depuis 1954. Des comparaisons ont été effectuées avec des parcelles traiées une fois et traitées à nouveau l'année suivante. Les applications répétées des deux herbicides ont provoqué une réduction du temps de dégradation qui est passé de 10 semaines pour le 2,4-D et 20 semaines pour le MCPA après une seule application, à 4 et 7 semaines respectivement, après 19 années d'applications annuelles. En dépit d'une capacité accrue de la microflore du sol à dégrader le 2.4-D et le MCPA, neuf mois après un traitement préalable, aucun changement n'a été constaté dans le nombre des organismes capables d'utiliser les produits comme sources de carbone dans les parcelles traitées, par rapport au témoin. II est évident qu'il y a une adaptation croisée entre le 2,4-D et le MCPA. II apparait très probable que deux ‘enrichissements’ de Hores ont été induits, chacun d'eux étant capable de décomposer les deux herbicides mais avec des efficacités différentes. Die Wirkung wiederholter Behandlungen von 2.4- D und MCPA auf ihren Abbau im Boden Die Wirkung wiederholter Anwendungen von 2,4-D und MCPA auf ihren Abbau wurden in einem Freiland-Versuch untersucht. Die Herbizide wurden jährlich, sell 1954 ange-wendet. Es wurden Vergleiche mil Parzellen angcstellt, die zum erstenmal und im folgenden Jahr nochrnaK bchandelt worden waren. Die widerholien Behandlungen mit den beiden Herbiziden bewirkte elne Verringerung der Ab-bauzeit von 10 Wochen fur 2,4-D und 20 Wochen fiir MCPA nach einer Behandlung, auf 4 bzw. 7 Wochen nach 19 Jahren jahrlicher Anwendung. Trotz der erhohten Ffihigkeit der Bodenmikroflora 2,4-D und MCPA neun Monate nach einer vorhergehenden Behandlung abzubauen, war die Zahl der Organismen, die diese Herbizide als Kohlensloffquelle verwerten konnen, in den behandelten und unbehandelten Parzellen gleich gross. Zwischen 2.4-D und MCPA existiert ofVensichtlich elne Kreuz-Adaplalion. Sehr wahrscheinlich wurden zwei versehiedene ‘enrichment’-Floren induziert, von der jede in der Lage ist beide Herbizide, aber mit unterschiedlicher Effizienz, Hbzubauen.     

钩状木霉菌的生物学特性及对腐皮镰刀菌的抑菌机理研究

从兰花根际中分离出一株木霉菌株LAN2B-1,对其进行形态学鉴定和ITS序列分析,确定菌株为钩状木霉菌Trichoderma hamatum.该菌株菌落生长的最佳条件为PDA培养基、碳源为甘露醇、pH 5、温度25℃～30℃,暗处理有利于菌落生长,但氮源对菌落生长影响不显著.木霉菌发酵液的抑菌活性和作用机理研究表明,钩...     

Effects of annual applications of sodium azide on soil fungal populations with emphasis on Trichoderma species

The effect of a granular formulation of sodium azide, applied annually to pine nursery beds at rates of 0, 67.2, and 134.5 kg a. i. ha^?1 under water seal or plastic seal, on soil fungal populations was determined over a 3-year period. Populations of fungi in the soil decreased following application of the sodium azide each year; the greatest decrease occurred at the highest rate of application. Populations of fungi in soil treated with the azide generally remained lower than in the controls throughout each of the 3 years; however, the population disparity between treated and control plots decreased in magnitude with each succeeding year. Populations of Trichoderma spp., in plots treated with 134.5 kg sodium azide ha^?1, increased 2 weeks after treatment each year, and the population peaks increased in magnitude each year. In addition the effect of sodium azide (technical grade > 99%) at concentrations of 0, 2, 5, 10, 20 and 50 μg ml^?1 in potato dextrose agar and blackstrap molasses agar media was determined in vitro for 14 isolates of Trichoderma harzianum. Growth and sporulation differed among the isolates and between the two media tested. Generally, the azide temporarily inhibited growth of the fungi, but the majority of the isolates were able to grow on either medium containing 50 μg sodium azide ml^?1, although sporulation was more profuse on the molasses than on the potato dextrose agar medium.     

Effect of soil moisture on activity and dynamics of Rhizoctonia solani and Trichoderma harzianum

Journal of Plant Diseases and Protection - The effect of soil moisture on activity and dynamics of the soilborne pathogen Rhizoctonia solani and of its antagonist Trichoderma harzianum were studied...     

地面覆盖的保水增产效应及其机理研究

通过两年的夏玉米微区试验(带遮雨棚)分析麦草覆盖和地膜覆盖的保水增产效果及其作用机理。结果表明:在排除降水影响的条件下,地面覆盖均能增加产量。地膜覆盖增产效果明显,水分利用率高;麦草覆盖保水效果显著,但增产效果较低。休闲时,麦草覆盖和地膜覆盖均明显增加了0~60 cm土层土壤的储水量,分别比不覆盖高10.2 mm和7.4 mm;在种植玉米的条件下,覆草处理的土壤储水量增加16.1 mm;而覆膜处理则减少了13.0 mm。对硝态氮而言,休闲状态下覆草处理的硝态氮在土壤表层的聚集较裸地少,而覆膜处理则相反;种植玉米时,覆草和覆膜处理均可减少硝态氮在表层土壤的聚集,且覆膜的效果大于覆草处理。     

Ochratoxin A-producing fungi in Spanish wine grapes and their relationship with meteorological conditions

Forty vineyards from four wine making regions of Spain were sampled at three different growth stages in 2002 and 2003. The aim was to study the fungi associated with grapes and their ability to produce ochratoxin A (OTA) on synthetic media. Among the total mycoflora, 464 (7.7%) and 648 (10.8%) Aspergillus section Nigri (black aspergilli) strains were isolated in 2002 and 2003, respectively, and were classified into three groups: isolates with uniseriate heads, A. niger aggregate and A. carbonarius. The latter presented the highest percentage of OTA-positive strains (82% in 2002 and 76% in 2003) and produced the highest levels of toxin (2.5–25 μg g⁻¹). The sampling year, sampling date, the region and their interactions presented significant differences in the number of black aspergilli isolated. Most black aspergilli were found in 2003 and at harvest. A positive correlation between the number of black aspergilli found in grapes and the temperature in the field was found. Grapes from 2003, the warmest year, and from Costers del Segre, the warmest region, were significantly the most contaminated. No significant correlation between black aspergilli presence and other meteorological factors such as relative humidity or rainfall could be established. Musts from all the vineyards were also analysed in both years, although no OTA was found in either year.     

Proteomic studies of phytopathogenic fungi,oomycetes and their interactions with hosts

Proteomics, the systematic analysis of the proteome, is a powerful tool in the post-genomic era. Proteomics studies have examined global changes in proteomes of phytopathogenic fungi, oomycetes and their hosts during compatible or incompatible interactions. This article compiles proteomics reports in order to decipher the molecular mechanisms underlying fungal development (infection-related morphogenesis), fungal or oomycete—host plant interactions, and phytopathogenesis.     

Non-target effect of herbicides: I. effect of glyphosate and hexazinone on soil microbial activity. Microbial population,and in-vitro growth of ectomycorrhizal fungi

The effects of two herbicides, glyphosate (as a 359 g litre^?1 SL) and hexazinone (as a 50gkg^?1 granule) on soil microbial population, carbon dioxide evolution, and in-vitro growth of five species of ectomycorrhizal fungi were investigated. Glyphosate at 0–54 and 3.23 kg a.i. ha^?1 and hexazinone at 1. 2 and 8 kg a.i. ha^?1 did not reduce soil microbial population or carbon dioxide evolution in the long term (6 months). However, there was a significant short-term (2 months) effect of glyphosate on both fungal and bacterial counts at the 0.54 kg ha^?1 treatment. In in-vitro tests, Cenococcum graniforme. Hebeloma crustuliniforme and Laccaria laccata were more susceptible to both herbicides than was Suillus tomentosus. which was, in turn, more susceptible than Paxillus involutus. The growth of all five ectomycorrhizal fungi was significantly reduced when subjected to concentrations above 50 μl formulation litre^?1 (glyphosate) or 50 μg formulation litre^?1 (hexazinone).     

Trichoderma species associated with green mould disease of Pleurotus ostreatus and their sensitivity to prochloraz

Green mould disease causes serious economic losses in Pleurotus ostreatus crops worldwide, including in Italy, where prochloraz is the only chemical fungicide allowed to control the disease. The effectiveness of the doses 0.01, 0.05, 0.25 and 1.25 μL L⁻¹ (field dose) of prochloraz (Sponix Flow, 450 g L⁻¹), against colony growth rate and spore germination of Trichoderma pleuroti, T. pleuroticola and T. guizhouense strains on wheat straw extract agar plates were evaluated. Complete inhibition of Trichoderma pleuroti and T. pleuroticola growth was shown by the field dose of prochloraz, and also by the 0.25 μL L⁻¹ dose for T. pleuroti. Complete inhibition of spore germination occurred for all Trichoderma strains at field dose, and at 0.25 μL L⁻¹ for T. pleuroti strains. In in vivo assays, the effect of prochloraz doses 0.05, 0.25 and 1.25 μL L⁻¹ on colonization of straw substrate by T. pleuroti, T. pleuroticola and T. guizhouense inoculated at two spore densities (10² and 10⁵ spores mL⁻¹) immediately after P. ostreatus spawn was studied. Trichoderma pleuroti and T. pleuroticola were both responsible for green mould disease, whereas T. guizhouense was not pathogenic. Trichoderma pleuroti was more aggressive than T. pleuroticola. Prochloraz was effective against T. pleuroti at the field dose, and against T. pleuroticola at 0.25 and 1.25 μL L⁻¹. The study on Trichoderma × Pleurotus interaction type showed that Trichoderma species were active against the mycelial growth of P. ostreatus by competition for space and nutrients, and neither hyphal interaction nor effect by volatile or nonvolatile metabolites occurred.     

Investigation on the transformation of fluchloralin by soil fungi: Identification of some metabolites

Fluchloralin [N-(2-chloroethyl)-α,α,α,-trifluoro-2,6-dinitro-N-propyl-p-toluidine] was transformed by two common soil fungi, Aspergillus flavus Link and Fusarium solani (Mart.) Sacc. in pure culture. The fungi were isolated from enrichment cultures of an analogous herbicidal compound, pendimethalin [N-(1-ethylpropyl)-2,6-dinitro-3,4-xylidine)]. Transformation of fluchloralin by these fungi resulted in the formation of several metabolites, 13 of which were identified. Besides the known mechanisms of microbial transformation of dinitroanilines, methylation of the anilino nitrogen and direct elimination of a nitro group at 2 position from the aromatic ring without any further substitution were also observed.     

The influence of soil temperature and moisture content on the effect of soil fumigants

Laboratory experiments were carried out to test the effect of two levels of soil moisture and two temperatures on the effectivity and persistence of five soil fumigants.Soil moisture had a great influence on the extent of the zone over which the products diffused vertically, as shown by fungicidal and phytotoxic action at various depths in soil columns. In moist soil spread of the gases from the point of application is much poorer than in dry soils. There was little or no temperature effect in the dryer soil, while in the wet soil diffusion was better at 25° than at 10°C.In studying persistence, as measured by assaying inhibition of germination and growth ofLepidium sativum, it appeared that the tested fumigants can be divided into two groups. In the case of chloropicrin and telone persistence is primarily enhanced by high soil moisture content, and in the case of the fumigants having methyl isothiocyanate as the active principle, by a low soil temperature. With chloropicrin and telone, there is a temperature effect only in the moist soil. Persistence of methylisothiocyanate is only slightly affected by soil moisture.Samenvatting In laboratoriumproeven werd de invloed van temperatuur en vochtgehalte van de bodem op de effectiviteit en persistentie van vijf grondontsmettingsmiddelen onderzocht.De bodemvochtigheid heeft een grote invloed op de verspreiding der produkten, zoals bleek bij de toetsing van fungicide en phytotoxische effecten op verschillende diepten in grondkolommen. In vochtige grond met een laag poriënvolume is de verspreiding vanaf het injectiepunt bij deze gasvormige middelen veel geringer dan in droge grond. In vochtige grond was de ontsmette zone groter bij 25°C dan bij 10°C, terwijl in droge grond geen of weinig effect van de temperatuur werd geconstateerd.Bij de bepaling van de persistentie, gemeten als kiem-en groeiremmend effect opLepidium sativum, bleek de getoetste serie middelen in twee groepen uiteen te vallen. De persistentie wordt in het geval van chloorpicrine en telone vooral verlengd door een hoog vochtgehalte, en bij de drie andere, waarvan methylisothiocyanaat het actieve bestanddeel of principe is, door een lage temperatuur. Bij chloorpicrine en telone is er alleen een temperatuureffect in de vochtige grond, terwijl de persistentie van methylisothiocyanaat zeer weinig beïnvloed wordt door het vochtgehalte.Research under the auspices of the IWONL.