Effects of organomercury treatment of wheat seed on Fusarium seedling disease in inoculated soil

In growth room experiments, treatment of wheat seed with phenyl mercuric acetate significantly reduced seedling disease caused by soil-borne Fusarium culmorum, F. nivale and F. avenaceum in inoculated soil. Seed treatment improved the low germination caused by F. culmorum, but did not affect the reduced seedling vigour caused by F. nivale. Seed treatment was normally as effective in conditions favouring disease as it was in less favourable conditions. Conditions favouring disease in soil inoculated with F. culmorum were high inoculum level, deep sowing and dry soil.     

Isolates of Microdochium nivale and M. majus Differentiated by Pathogenicity on Perennial Ryegrass (Lolium perenne L.) and in vitro Growth at Low Temperature

Pink snow mould is a serious disease on grasses and winter cereals in cold and temperate zones during winter. To better understand the basis for the variation in pathogenicity between different isolates of Microdochium nivale and M. majus and to simplify selection of highly pathogenic isolates to use when screening for resistance to pink snow mould in perennial ryegrass, we sought traits correlated with pathogenicity. Isolates of M. nivale were more pathogenic on perennial ryegrass than isolates of M. majus, as measured by survival and regrowth of perennial ryegrass after infection and incubation under simulated snow cover. Pathogenicity as measured by relative regrowth was highly correlated with fungal growth rate on potato dextrose agar (PDA) at 2°C. Measuring fungal growth on PDA therefore seems to be a relatively simple method of screening for potentially highly pathogenic isolates. In a study of a limited number of isolates, highly pathogenic isolates showed an earlier increase and a higher total specific activity of β‐glucosidase, a cell wall‐degrading enzyme, compared with less pathogenic isolates. None of the M. majus isolates was highly pathogenic on perennial ryegrass. Our results indicate biological differences between M. nivale and M. majus and thus strengthen the recently published sequence‐based evidence for the elevation of these former varieties to species status.     

Fungi pathogenic to ryegrass seedlings

Summary Fungi borne on or in ryegrass (Lolium spp.) seeds or invading ryegrass seedlings grown on field soils were isolated and identified. Selected isolates were tested to determine their pathogenicity to ryegrass seedlings. Seed-borne fungi were generally weakly virulent or non-pathogenic to ryegrass seedlings. Pathogenic seed-borne fungi includedChaetomium globosum Kunze: Fr.,Curvularia trifolii (Kauffm.) Boedijn, and species ofPenicillium Link andAspergillus Mich. ex Link. Species of fungi isolated from seedlings grown on field soils de pended on soil and temperature. Soil-borne fungi pathogenic to seedlings includedFusarium avenaceum (Fr.) Sacc.,F. culmorum (W. G. Smith) Sacc.,F. equiseti (Corda) Sacc.,F. oxysporum Schlecht.: Fr.,F. solani (Mart.) Sacc.,Pythium afertile Kanouse and Humphrey,P. debaryanum auct. non Hesse,P. irregulare Buisman,P. ultimum Trow, a sphaerosporangiatePythium sp.,Chaetomium globosum, Thanatephorus cucumeris (Frank) Donk,Trichoderma koningii Oudem., and aPhomopsis sp. Individual isolates of fungi differed in virulence to ryegrass seedlings, and ryegrass cultivars differed in susceptibility to seedling pathogens.     

Glasshouse studies on the effect of low temperature on infection of perennial ryegrass seedlings by Fusarium nivale

In glasshouse experiments, isolates of Fusarium nivale, applied as mycelial macerates, caused marked reductions in the number and length of roots, and in the height and fresh weight of S. 321 perennial ryegrass seedlings. Spore inocula failed to cause infection. Damage by F. nivale was increased when inoculation was immediately followed by four alternate 12 h periods in a growth cabinet at 0°C and in a warm glasshouse at c. 17°C, succeeded by a further 11 days in the same glasshouse. Although inoculated seedlings receiving no cold treatments initially showed damage, regrowth of roots and some recovery of the shoots occurred in a fortnight. Exposure of the plants after that time to four intermittent 12 h periods at 0°C, however, arrested their recovery and caused further root damage. The implication of these results in the importance of low temperature in the winterkill syndrome is discussed.     

Temperature and seedling age affect susceptibility of perennial ryegrass seedlings to pathogenic fungi

Summary Effects of temperature and seedling age on survival of perennial ryegrass (Lolium perenne L.) seedlings grown on sand-wheat wholemeal cultures of different isolates ofFusarium spp. (9 isolates),Pythium spp. (9 isolates), andChaetomium spp. (1 isolate) are reported. Some isolates were virulent over the whole range of temperatures tested (7.5–27.5°C). The virulence of others depended on temperature. Most isolates were less virulent at intermediate temperatures (12.5–22.5°C) than at higher or lower temperatures. At 25°C ryegrass seedlings were susceptible to fungal attack for only a limited period after germination commenced. This period differed for different fungi, but for most isolates tested, seedlings were resistant after 2–3 days.     

The effect of water potential on soil-borne diseases of wheat seedlings

Under glasshouse conditions, Fusarium culmorum caused more injury to wheat seedlings in soil at 33% saturation (approximately – 1 bar) than at 66% (–0·1 bar). The same effect occurred with inoculated seeds raised on soil moisture tension tables at –0·1 and –0·5 bar, at 20° and at 15°C. In controls, spontaneous infection by soil-borne F. culmorum was unaffected by soil water potential. Inoculated F. nivale, at 127deg; and at 15°C, showed a similar, but smaller, effect to that with inoculated F. culmorum. Water potential in this range did not affect disease incidence with inoculated Gaeumannomyces graminis, but disease was more severe in the drier soil. On tension tables, seedlings did not consistently develop faster under wetter conditions, and factors other than host growth rate probably played a part in limiting fungal attack.     

Identity and Pathogenicity of Fungi Associated with Root and Crown Rot of Soft Red Winter Wheat Grown on the Upper Coastal Plain Land Resource Area of Mississippi

Seedling stand, disease severity and fungal incidence were determined from untreated ‘Wakefield’ soft red winter wheat planted on a Leeper silty clay loam in field tests conducted at the Mississippi Agricultural and Forestry Experiment Station, Plant Science Research Center, Mississippi State University, Starkville, Mississippi during the 1996–97 and 1997–98 growing seasons. Seedling stand was reduced by 40% each year in plots established with untreated seed. Cochliobolus sativus was the most frequently isolated fungus. Fusarium acuminatum, Fusarium equiseti and Fusarium solani were the most prevalent Fusarium spp. Seven other Fusarium spp. and 23 species of other fungal genera were isolated. Pathogenicity tests with three isolates each of C. sativus, Cochliobolus spicifer, F. acuminatum, F. solani, F. equiseti, Fusarium compactum, Embellisia chlamydospora and Microdochium bolleyi were performed in test tube culture and two isolates each of C. sativus, C. spicifer, F. acuminatum, E. chlamydospora and M. bolleyi under greenhouse conditions. In test tubes and in the greenhouse, seedlings infected with isolates of C. sativus developed seedling blight, discoloration and necrosis, primarily in seminal roots and crowns. In the greenhouse, C. sativus induced lesions on the lower leaf sheath and reduced seedling height, seedling emergence, dry and fresh weight of roots and shoots. Isolates of F. acuminatum, F. solani, F. equiseti, F. compactum, E. chlamydospora and M. bolleyi induced slight to moderate orange to light‐brown discoloration of crown and seminal roots in test tubes. Cochliobolus spicifer isolates had the most pre‐emergence activity, inducing black root discoloration and root pruning of wheat seedlings and reducing seedling emergence, root fresh weight and shoot dry weight. In the greenhouse, F. acuminatum reduced seedling height, seedling emergence and root and shoot dry weights. Microdochium bolleyi and E. chlamydospora reduced fresh and dry weight of roots, plant emergence and shoot dry weight. Fusarium acuminatum and C. spicifer reduced the growth rate of wheat seedlings. All fungi evaluated showed increased disease severity compared to the untreated control. The high frequency of isolation of C. sativus from crown and root tissues can be partially explained by the dry, warm conditions during the early stages of wheat seedling development in the Upper Coastal Plain Land Resource Area of Mississippi.     

The Effect of Chitosan and Bion on Resistance to Pink Snow Mould in Perennial Ryegrass and Winter Wheat

The effects of chitosan on resistance to pink snow mould (Microdochium nivale) were studied in young winter wheat (Triticum aestivum L.) and perennial ryegrass (Lolium perenne L.) under controlled environmental conditions. In perennial ryegrass, the putative defence activator Bion was also tested. Resistance was measured as regrowth of plants after inoculation with M. nivale and incubation in darkness at 2°C. In winter wheat, pre‐treatment with chitosan at 1000 μg per plant increased resistance to subsequent infection by M. nivale, but this effect was less significant in a replicate experiment. Chitosan‐treated winter wheat plants expressed the gene for the pathogenesis‐related protein chitinase at higher levels than non‐treated plants. Chitinase gene expression was also stimulated by M. nivale infection in winter wheat. Perennial ryegrass pre‐treated with Bion or chitosan and inoculated with M. nivale did not display better regrowth after incubation than non‐treated, inoculated plants. Rather, regrowth was reduced in some of the Bion‐treated plants after incubation. We speculate that the cost or the mechanism of induced resistance makes Bion non‐effective in plants that are not actively growing. Bion at concentrations of 10, 100 and 1000 μg active ingredient per ml, and the highest concentration of chitosan used (2000 μg per ml) reduced in vitro growth of the pathogen, suggesting that both defence activators possess antifungal activity.     

Effects of infection time and moisture on development of ear blight and deoxynivalenol production by Fusarium spp. in wheat

Wheat ears were inoculated with conidia of Fusarium spp. at different growth stages between ear emergence and harvest and moist conditions were maintained for up to 7 days subsequently by mist irrigation. Of the fungi tested (Fusarium culmorum, F. avenaceum, F. tricinctum, F. sporotrichioides and Microdochium nivale), only F. culmorum produced ear blight symptoms and grain samples were found subsequently to contain deoxynivalenol. Most ear infection and deoxynivalenol formation occurred following inoculation at about mid-anthesis. Small amounts of deoxynivalenol were formed and some F. culmorum was isolated even in the absence of ear blight symptoms. An overnight wet period was sufficient to initiate infection and deoxynivalenol formation but both were increased by extending the wet period up to at least 3 days. Recovery of Fusarium spp. from harvested grain was usually possible whether or not symptoms developed. F. culmorum usually persisted and often increased to moderately high levels after storage for 7 wk in a range of moisture conditions.     

Suppression of gray leaf spot (blast) of perennial ryegrass turf by Pseudomonas aeruginosa from spent mushroom substrate

Bacteria isolated from spent mushroom substrate (SMS) were evaluated for the suppression of Pyricularia grisea, the causal agent of gray leaf spot of perennial ryegrass (Lolium perenne) turf. Thirty-two of 849 bacterial isolates (3.8%) from SMS significantly inhibited the mycelial growth of P. grisea in vitro. Six bacterial isolates that afforded maximum inhibition of P. grisea were also refractory to Rhizoctonia solani, Rhizoctonia cerealis, Sclerotinia homoeocarpa, and Fusarium culmorum. Each of the six isolates was identified as Pseudomonas aeruginosa by fatty acid profile analysis. The biocontrol activity of the bacterial isolates was not compromised by a prolonged exposure to UV radiation in vitro. In controlled-environment chamber experiments, all 32 bacterial isolates were tested for suppression of gray leaf spot on Pennfine perennial ryegrass when applied as seed treatment or foliar sprays. Foliar applications of the bacteria (10⁸ cfu/ml 0.1% carboxymethylcellulose), but not the seed treatment, significantly reduced disease severity and incidence. The three most efficient isolates from foliar application treatments, which were among the six bacterial isolates identified as P. aeruginosa, were further evaluated for suppression of gray leaf spot as a function of timing of application. The three isolates of P. aeruginosa suppressed gray leaf spot in perennial ryegrass in Cone-tainers when applied at 1, 3, and 7 days prior to inoculation with P. grisea both in controlled-environment chamber experiments, and in potted ryegrass plants maintained in the field. All application intervals, regardless of the bacterial isolate, provided significant reduction of gray leaf spot severity. Suppression of gray leaf spot by isolates of P. aeruginosa under controlled-environment chamber conditions was not different from that observed in potted ryegrass plants maintained in the field. In field experiments, an isolate of P. aeruginosa provided significant suppression of gray leaf spot when applied at 1, 7, and 14 days prior to inoculation with P. grisea. The bacterium proved effective against gray leaf spot of perennial ryegrass maintained as fairway and rough heights. These results indicate that P. aeruginosa may be a potential biocontrol agent for gray leaf spot of perennial ryegrass turf.     

Zearalenone-producing species of Fusarium on barley seed

Isolates of Fusarium were obtained from samples of barley grain at harvest in 1971, 1973 and 1974. The ability of these isolates to produce the myco-toxin zearalenone was examined, the grain also being tested for its presence in 1974. The most common isolate was F. culmorum, 60% of which in 1974 produced zearalenone. Other species isolated which produced this myco-toxin were F. monilijorme, F. graminearum, F. avenaceum, F. nivale and F. sambucinum var coeruleum, the last three species have not previously been recorded as producing zearalenone.     

Old-field seedling responses to insecticide,seed addition,and competition

The controls of seedling emergence and survival determine the potential distribution of adult plants and, thereby, plant community structure. Seed availability, competition from established neighbors, and seedling predation may all limit seedling recruitment. In this field experiment, we followed the emergence and survival of seedlings of three perennial forbs, Achillea millefolium, Hypericum perforatum, and Monarda fistulosa, in old-fields in southeastern Michigan, USA. As adults, all three have aromatic foliage that may deter herbivory, but seedlings may be more susceptible than adults. To establish the relative importance of potential controls on seedling numbers, we manipulated seed availability through seed additions, the influence of competitors by neighbor-removals, and the influence of insect herbivores with insecticide in a fully factorial field experiment. Seed addition and insecticide never affected seedling emergence for any species. Competition from established neighbors controlled seedling emergence for all three species and decreased Achillea survival. Insecticide significantly increased Monarda seedling survival in competition plots, significantly increased Hypericum survival in open plots, and had no effect on Achillea. Notably, insecticide increased survival of the native Monarda fistulosa more than the two introduced species. While neighbors strongly reduced emergence and survival of all three species, herbivores acted on a species-specific basis. These results suggest the differential effects of insects may contribute more to the seedling species composition and abundance patterns than the less-selective influence of competition.     

Phytotoxity and pathogenicity of Fusarium nivale towards cereal seelings

Strong phytotoxic activity towards whew seedling has been derected in culatural extracts of 12 Fusarium nivale strains. The exmined isolates have been found strong pathogens of rye. triticale and wheat seedlings. However, their phytotoxity was not corrclued with their pathogenicity.     

Auswinterungsschäden in Raigras-Reinbeständen, die Bedeutung pilzlicher Krankheitserreger und ihre mögliche Eindämmung

Winterkilling in puresown ryegrass, the importance of fungal pathogens and possibilities for their control I. Results from field trials with natural infection Rotten plants, incomplete stands, a reduced number of shoots and a decline in dry matter yield were observed after hibernations of perennial and Italian ryegrass. Isolations from damaged plants showed that the main pathogen was the causal agent of snow mould Monographella nivalis (Schaffn.) E. Müll., asexual stage Gerlachia nivalis (Ces. ex Sacc.) W. Gams et E. Müll., synonym Fusarium nivale Ces. ex Sacc. The following fungi were also isolated: Laetisaria fuciformis(Mc Alp.) Burds., synonym Corticium fuciforme (Berk.) Wakef., Gibberella avenacea Cook, asexual stage Fusarium avenaceum (Fr.) Sacc. and Fusarium culmorum (W. G. Sm.) Sacc. The field trials were located at 440 and 630 m a.s.l. These elevations correspond to plain and mid-mountain climates, respectively. The tetraploid varieties ‘Lipo’, ‘Tetila’, ‘Citadel’ and ‘Bastion’ showed a higher level of resistance to snow mould compared to the diploid varieties ‘Ursus’, ‘Lemtal’, ‘Melino’ and ‘Pablo’. Tetraploid ryegrasses contaminated the soil with Gerlachia nivalis to a lesser extend than diploid ryegrasses. The influence of the date of cutting and of nitrogen fertilization in autumn on snow mould was also tested. Lowest values at both locations were observed when the last cutting occurred at the end of October. Severity was higher when the last cutting, took place earlier (mid September, end of September, mid October). Since parts of stands of the last cutting were strongly damaged by the cold, last cutting should take place in the middle of October, although infection with snow mould will not be as much reduced as if the last cutting was done at the end of October. The plant rotting caused by Gerlachia nivalis was increased with 80 kg N/ha in autumn compared with 40 kg N/ha.     

Pathogenicity of fungi associated with wheat and barley seedling emergence and fungicide efficacy of seed treatment

Presented study focused on the influence of Cochliobolus sativus isolates origin on pathogenicity towards wheat and barley seedlings in comparison with pathogenicity of certain Fusarium species and Microdochium nivale. The efficacy of fungicide seed treatment against C. sativus was estimated. The C. sativus isolates were collected from different locations and were isolated from wheat, barley and sunflower seeds. The pathogenicity of C. sativus, Fusarium species and M. nivale towards germinating seedlings were expressed as germination (GA) retardation and coleoptile growth rate retardation (CGR). Of wheat only, the CGR was significantly influenced by the isolate origin. The C. sativus isolates obtained from sunflower seeds were the most aggressive. Of the barley seeds, the barley isolates were the most aggressive. Barley was significantly more susceptible to damage by C. sativus isolates than wheat. The pathogenicity of tested fungal species declined in the order: F. culmorum, F. graminearum, C. sativus, F. avenaceum, M. nivale, F. poae for both barley and wheat. The results highlighted high pathogenicity potential of C. sativus equal to that of F. avenaceum and M. nivale. The symptoms of C. sativus on coleoptile and roots were very similar or the same as the symptoms caused by Fusarium species and M. nivale, except of white, pink or red colours. Of wheat sprouts, the fungicide efficacy (FE) against C. sativus declined in the order: tebuconazole + thiram, carboxin + thiram, quazatine, difenoconazole, iprodione + triticonazole (in term of GA) and carboxin + thiram, iprodione + triticonazole, tebuconazole + thiram, difenoconazole, quazatine (in term of CGR). In barley, the FE declined in the order: carboxin + thiram, iprodione + triticonazole, tebuconazole + thiram, difenoconazole, quazatine (in term of GA) and carboxin + thiram, tebuconazole + thiram, difenoconazole, iprodione + triticonazole, quazatine (in term of CGR).     

Effect of Microdochium nivale and Low Temperature on Winter Survival of Perennial Ryegrass

Winter survival of perennial ryegrass (Lolium perenne L.) depends mainly on the plant resistance to low freezing temperature and to snow mould fungi (Microdochium nivale). Field evaluation of these plant characters gave irreproducible results. A comparison of field trials with laboratory tests pointed to the close correlation between the yield level of examined varieties and strains and their resistance to Microdochium nivale and frost. A lack of correlation between snow mould resistance and frost tolerance was shown.     

Pathogenicity of Fusarium avenaceum Isolates from Cereals and their Ability to Produce Substance with Yellow Fluorescence

Fusarium avenaceum isolates from wheat, rye, barley, triticale, corn and potato formed substance with yellow fluorescence with properties similar to citrinin. Pathogenicity of 17 isolates tested against cereals seedlings was weaker than F. culmorum isolates; one isolate only was strong, two medium and the remaining 82 % were very weak or non pathogenic.     

Establishment and growth of seedlings of Eucalyptus obliqua: Interactive effects of litter,water, and pathogens

We studied how leaf litter, water and fungal pathogens affect the establishment, survivorship, and growth of Eucalyptus obliqua using glasshouse and field experiments. In a glasshouse experiment, the presence of leaf litter of E. obliqua increased the establishment of seedlings when the seeds were placed beneath the litter, but dense litter layers reduced seedling emergence when seeds were placed on top. Results from a complementary experiment suggest that the effect of litter may be mediated by the high humidity microenvironment it creates, which may enhance imbibition of unburied seeds. In another glasshouse experiment, litter in combination with excessive water applications reduced seedling emergence and increased seedling mortality, but the addition of fungicide reverted these effects. Litter with lower water application or fungicide added increased seedling emergence. In all cases higher biomass in seedlings growing in litter treatments was associated with earlier emergence. In two recently burnt sites, the density of seedlings of E. obliqua was positively correlated to the presence of litter. Experimental addition of litter in the field reduced soil temperature during the summer, and together with watering, increased soil water content, and the water potential of established seedlings. However, neither treatment had any positive effect on their growth or survivorship. On the contrary, litter addition reduced the height and number of leaves, probably as a result of increased herbivory. We conclude that in Mediterranean-type systems litter may produce various effects on seedling establishment depending on their developmental stage, and the season.     

Influence of fungal isolates on germination and growth of perennial ryegrass

Summary The influence of fungi isolated from perennial ryegrass roots on the germination and development of seedlings of perennial ryegrass was investigated. The basic procedure employed was to sterilise the seed surface and then inoculate with fungi and plant in non-sterile soil. It was realised that the fungal isolate inoculated on to the sterile seed surface would not remain dominant in the root region of the host and would have an influence on the host which would decline with time from when the seed germinated. This was because it would have to face antagonism from the normal components of the root microflora present in the non-sterile soil.Trichoderma viride delayed the emergence of the seedlings and reduced the production of herbage, an observation consistent with results of some other investigators. A sterile hyaline fungus stimulated the emergence of the seedlings, but subsequent tests showed that the presence of the microflora of the seed coat, or the soil microflora, or the sterile hyaline fungus was effective in promoting the rate of germination of seed that had been surface sterilised. Leaching seed in water brought about an increase in the rate of seed germination, and it is suggested that there might be a germination inhibitor soluble in water present in the seed coat, which might be inactivated by saprophytic micro-organisms.     

Preservation of fungi in water (Castellani): 20 years

Sixty-two isolates of Fusarium were obtained from pasture grass and soil from various areas of New Zealand and identified as F. anthophilum [2], F. avenaceum [17], F. crookwellense [8], F. culmorum [4], F. graminearum [1], F. nivale [3], F. oxysporum [3], F. sambucinum [17], F. semitectum [1], F. tricinctum [1] and an unidentified Fusarium spp. [5]. These isolates were grown on autoclaved rice and tested for toxicity to rats in feeding tests. Eighty two percent of the isolates were toxic, of which twenty-four percent were severely toxic and caused hemorrhages of stomach and intestine, hematuria, and finally death. Cultures of the most toxic isolates contained 0.1 to 104 ppm of deoxynivalenol, 0.7 and 7 ppm of 15- and 3-acetyldeoxynivalenol respectively, 0.2 to 4 ppm of fusarenon- X, 11 to 1021 ppm zearalenone, 40 to 272 ppm of the hemorrhagic factor (wortmannin), 2,100 to 7,200 ppm of moniliformin, 565 ppm of the cytotoxic factor (HM-8) and enniatin in substantial concentrations. F. sambucinum is reported as a moniliformin producer for the first time.