Effects of different temperature treatments on dormancy of sclerotia of ten isolates of Sclerotium cepivorum

The variability of dormancy of sclerotia of ten isolates ofSclerotium cepivorum was investigated. Of all isolates tested, the freshly harvested sclerotia were dormant. After drying for 48 hours the sclerotia of six isolates were able to germinate, two isolates stayed dormant and two isolates were infested by hyperparasitic fungi. After storage in soil at 5°C or 20°C, the sclerotia of the different isolates exhibited considerable differences in respect to germination capability, but all isolates showed highest germination after a treatment of 8 weeks at 20°C followed by 8 weeks at 5°C. The sclerotia of all isolates showed an increased capacity to germinate withoutAllium extracts at 10°C after pretreatment at 30°C for 28 days.     

Inoculum potential of Sclerotinia sclerotiorum sclerotia depends on isolate and host plant

The soilborne fungus Sclerotinia sclerotiorum infects many important crop plants. Central to the success of this pathogen is the production of sclerotia, which enables survival in soil and constitutes the primary inoculum. This study aimed to determine how crop plant type and S. sclerotiorum isolate impact sclerotial production and germination and hence inoculum potential. Three S. sclerotiorum isolates (L6, L17, L44) were used to inoculate plants of bean, carrot, lettuce, oilseed rape (OSR) and potato, and the number and weight of sclerotia per plant quantified. Carpogenic germination of sclerotia collected from different hosts was also assessed for L6. Production of sclerotia was dependent on both crop plant type and S. sclerotiorum isolate, with OSR and lettuce supporting the greatest number (42–122) and weight (1.6–3.0 g) of sclerotia per plant. The largest sclerotia were produced on OSR (33–66 mg). The three S. sclerotiorum isolates exhibited a consistent pattern of sclerotial production irrespective of crop type; L6 produced large numbers of small sclerotia while L44 produced smaller numbers of large sclerotia, with L17 intermediate between the two. Germination rate and percentage was greatest for larger sclerotia (4.0–6.7 mm) and also varied between host plants. Combining sclerotial production data and typical field crop densities suggested that infected carrot and OSR could produce the greatest number (3944 m^?2) and weight (73 g m^?2) of S. sclerotiorum sclerotia, respectively, suggesting these crops potentially contribute a greater increase in inoculum. This information, once further validated in field trials, could be used to inform future crop rotation decisions.     

Effect of Inoculum Rates and Sources of Coniothyrium minitans on Control of Sclerotinia sclerotiorum Disease in Glasshouse Lettuce

Coniothyrium minitans isolate Conio grew on both maizemeal-perlite and ground maizemeal-perlite, producing high numbers (1.6×10⁷ conidiag^–1 inoculum) of germinable conidia. Coniothyrium minitans isolate Conio applied as a preplanting soil incorporation of maizemeal-perlite inoculum at full application rate (0.6lm^–2; 10¹¹ colony forming units (cfu)m^–2) significantly reduced Sclerotinia disease in a sequence of three lettuce crops grown in a glasshouse. No reduction in disease was achieved with any of the reduced rate treatments (10⁸cfum^–2) of a range of C. minitans isolates (Conio ground maizemeal-perlite at reduced rate, Conio and IVT1 spore suspensions derived from maizemeal-perlite, IVT1 spore suspension derived from oats and Contans® WG spore suspension). After harvest of the second and third crops, C. minitans maizemeal-perlite at full rate reduced the number and viability of sclerotia recovered on the soil surface and increased infection by C. minitans compared with spore suspension and reduced rate maizemeal-perlite inocula. Coniothyrium minitans was recovered from the soil throughout the trial, between 10⁵ and 10⁷cfucm^–3 in maizemeal-perlite inoculum full rate treated plots and 10¹–10⁴cfu cm^–3 in all other inoculum treated plots.Pot bioassays were set up corresponding to the inoculum used in the glasshouse, with the addition of Conio ground maizemeal-perlite at a rate corresponding to the full rate maizemeal-perlite. Coniothyrium minitans maizemeal-perlite and ground maizemeal-perlite at full rate significantly decreased carpogenic germination, recovery and viability of sclerotia and increased infection of sclerotia by C. minitans in comparison with spore suspension treatments, reflecting results of the glasshouse trials. Additionally, reduced maizemeal-perlite treatment also decreased apothecial production, recovery and viability of sclerotia compared with the spore suspension treatment, despite being applied at similar rates. Simultaneous infection of sclerotia by several isolates of C. minitans was demonstrated. Inoculum level in terms of colony forming unitscm^–3 of soil appears to be a key factor in both control of Sclerotinia disease and in reducing apothecial production by sclerotia.     

Effect of Combined Treatment of Pasteurisation and <Emphasis Type="Italic">Coniothyrium minitans</Emphasis> on Sclerotia of <Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis> in Soil

Integrated control of soil-borne plant pathogens such as Sclerotinia sclerotiorum is becoming more important as the soil fumigant methyl bromide is being phased out of use. Two alternative methods of control that have been found to reduce viability of sclerotia are steam sterilisation (pasteurisation) of soil or the application of the mycoparasite Coniothyrium minitans. This work investigated the possibility of integrating these two control measures. Soil was pasteurised in an autoclave, using a temperature of 80 °C for 3 min to simulate the possible temperatures reached by soil steaming machines for field use. Coniothyrium minitans was subsequently applied to the pasteurised soil to assess the effects of the combination of control measures in reducing sclerotial viability of S. sclerotiorum. Similar results were found in two soil types. Either method used individually was effective in decreasing the number of viable sclerotia, but no further reduction in sclerotial viability was seen when the two methods were combined. Coniothyrium minitans was found to colonise pasteurised sclerotia significantly quicker than untreated sclerotia, and it was seen that there was an increase in number of C. minitans in pasteurised soil in the presence of sclerotia. Experiments were also conducted to investigate the effect of application timing of the biocontrol agent to soil following pasteurisation, in relation to sclerotial infection. Here, two different isolates of S. sclerotiorum were used, with similar results. Application of C. minitans to soil immediately following pasteurisation resulted in sclerotial infection by the mycoparasite, but application 7 days or more after soil pasteurisation resulted in low recovery of the biocontrol agent from sclerotia, possibly due to the mycoparasite being masked by the presence of other fungi which colonised the sclerotia first.     

Quantitative Aspects of Infection of Sclerotinia sclerotiorum Sclerotia by Coniothyrium minitans – Timing of Application, Concentration and Quality of Conidial Suspension of the Mycoparasite

White mould disease leads to production of sclerotia, which subsequently survive in soil and may be responsible for future epidemics. The effect of the mycoparasite Coniothyrium minitans in decreasing survival of sclerotia of Sclerotinia sclerotiorum was studied. Infection of sclerotia of S. sclerotiorum by C. minitans can be achieved by a single conidium. Under optimal conditions, 2 conidia per sclerotium produced 63% of the maximum infection (ca. 90%) of sclerotia produced by up to 1000 conidia. Similar results were observed on the infection of stem pieces infected by S. sclerotiorum. In field trials, application of conidial suspensions of C. minitans to a bean crop soon after white mould outbreak led to a higher percentage of sclerotial infection than later applications. Ninety per cent infection of sclerotia was obtained within 3 weeks of application by C. minitans suspensions in the range of 5 × 10⁵ and 5 × 10⁶ conidia ml^–1 at 1000 l ha^–1. The concentration of the conidial suspensions and the isolate used were of less importance. The result was marginally affected by the germinability of the conidia (75% against 61% infected sclerotia at 91% and 16% viability of isolate IVT1, respectively). Less apothecia of S. sclerotiorum developed in soil samples collected after 2 months from plots sprayed immediately after disease outbreak than from those treated 11–18 days later. It is concluded that a suspension of 10⁶ conidia ml^–1 in 1000 l ha^–1 (= 10¹² conidia ha^–1) sprayed immediately after the first symptoms of disease are observed, results in > 90% infection of sclerotia of S. sclerotiorum. The infection of sclerotia, which prevents their carry-over, occurs within a broad range of inoculum quality.     

Effect of inoculum type and timing of application of Coniothyrium minitans on Sclerotinia sclerotiorum: influence on apothecial production

The effects of different inocula of the mycoparasite Coniothyrium minitans on carpogenic germination of sclerotia of Sclerotinia sclerotiorum at different times of year were assessed. A series of three glasshouse box bioassays was used to compare the effect of five spore-suspension inocula of C. minitans , including three different isolates (Conio, IVT1 and Contans), with a standard maizemeal–perlite inoculum. Apothecial production, as well as viability and C. minitans infection of S. sclerotiorum sclerotia buried in treated soil, were assessed. Maizemeal–perlite inoculum at 10⁷ CFU per cm³ soil reduced sclerotial germination and apothecial production in all three box bioassays, decreasing sclerotial recovery and viability in the second bioassay and increasing C. minitans infection of sclerotia in the first bioassay. Spore-suspension inocula applied at a lower concentration (10⁴ CFU per cm³ soil) were inconsistent in their effects on sclerotial germination in the three box bioassays. Temperature was an important factor influencing apothecial production. Sclerotial germination was delayed or inhibited when bioassays were made in the summer. High temperatures also inhibited infection of sclerotia by C. minitans . Coniothyrium minitans survived these high temperatures, however, and infected the sclerotia once the temperature decreased to a lower level. Inoculum level of C. minitans was an important factor in reducing apothecial production by sclerotia. The effects of temperature on both carpogenic germination of sclerotia and parasitism of sclerotia by C. minitans are discussed.     

Production,survival and evaluation of solid-substrate inocula ofConiothyrium minitans againstSclerotinia sclerotiorum

Coniothyrium minitans grew on all ten solid-substrates (barley, barley-rye-sunflower, bran-vermiculite, bran-sand, maizemed-perlite, millet, oats, peat-bran, rice and wheat) tested, producing high numbers of germinable pycnidiospores (1.9–9.3×10⁸ g^–1 air dry inocula). All solid substrate inocula survived better in the laboratory at 5 and 15 °C than at 30 °C for at least 64 weeks.In pot bioassays carried out in the glasshouse and field, soil incorporations of each inoculum almost completely inhibited carpogenic germination ofS. sclerotiorum. In the field bioassay, no sclerotia were recovered after 38 weeks fromC. minitans-treated pots compared to 56% from control pots. In the glasshouse bioassay, 9–30% of sclerotia were recovered after 20 weeks fromC. minitans-treated pots, but 88–100% of these were infected by the antagonist. The antagonist also spread to infect sclerotia in control pots.In larger scale glasshouse trials, single preplanting soil-incorporations of five inocula (barley-ryesunflower, maizemeal-perlite, peat-bran, rice and wheat) controlled Sclerotinia disease in a sequence of lettuce crops, with only small differences between the types of inocula tested. At harvest,C. minitans reduced sclerotial populations on the soil surface and over 74% of sclerotia recovered fromC. minitans-treated plots were infected by the antagonist.C. minitans survived in soil in all solid-substrate inocula-treated plots for at least 39 weeks at levels of 10⁴–10⁵ colony forming units cm^–3 soil and spread to infect over 36% of sclerotia recovered from control plots.     

Bioassay-led isolation of Myrothecium verrucaria and verrucarin A as germination inhibitors of Orobanche crenata

A total of 188 fungal isolates was obtained from the rhizosphere of Vicia faba grown in an Egyptian soil heavily infested with Orobanche species. Agar cultures of 58 isolates inhibited the germination of conditioned seed of Orobanche crenata exposed to the germination stimulant, GR24. Filtrates of inhibitory fungi grown in liquid medium for 9–15 days were also assayed and those of five isolates, which were morphologically similar, inhibited germination even when diluted 16-fold. The fungus was identified as Myrothecium verrucaria (Alb. & Schwein.) Ditmar by its morphology and the nucleotide sequence of the ITS1 and ITS2 regions of the ribosomal repeat unit. Purification of the inhibitor to homogeneity was accomplished by solvent partitioning, flash chromatography on silica gel, semi-preparative HPLC on a reversed phase C18 column, solid phase extraction and tlc on silica gel. The inhibitor was identified as verrucarin A by nuclear magnetic resonance spectroscopy and comparison of the spectra with those of an authentic sample of the compound. A preliminary experiment demonstrated that infection of V. faba by O. crenata could be prevented by addition of spores of the fungus to soil infested by the parasite.     

Single and combined colonization of Sclerotinia sclerotiorum sclerotia by the fungal mycoparasites Coniothyrium minitans and Microsphaeropsis ochracea

Potential enhancement of mycoparasitic efficacy of Coniothyrium minitans and Microsphaeropsis ochracea through concomitant colonization of Sclerotinia sclerotiorum sclerotia was investigated, following observation that the two mycoparasites did not exhibit any mutual antagonism in dual culture assays. Simultaneous application of both mycoparasites increased sclerotia mortality in a temperature range from 16 to 26°C compared to single application, indicating a predominantly additive interaction. With increasing temperature the efficacy of M. ochracea decreased, but C. minitans was unaffected. Degradation of sclerotia by C. minitans proceeded slightly faster than with M. ochracea. Simultaneous colonization of sclerotia was studied at the histopathological level with mycoparasite strains transformed via Agrobacterium tumefaciens‐mediated transformation (ATMT) with reporter genes encoding for DsRed and GFP. Sclerotia colonization followed by fluorescence microscopy revealed effective penetration of the sclerotial rind, growth and formation of pycnidia in the cortex and medulla by both antagonists, resulting in complete degradation of sclerotia within 25 days after single inoculation. Upon simultaneous inoculation, both antagonists concomitantly colonized the sclerotial tissue and independently formed pycnidia in the sclerotial medulla and on the sclerotial rind, demonstrating their ability to co‐colonize the same host fungus. Although the individual growth of the two mycoparasites in dual inoculations was slightly delayed, the sclerotia degrading effects were additive, suggesting a complementary antagonistic interaction. The combined application of two different species of mycoparasites cooperating on the same host fungus and differing in temperature requirements may be advantageous for making biocontrol applications in the field less sensitive to varying environmental and host conditions.     

Opportunities for the microbial control of Allium white rot1

The microbial control of Allium white rot (Sclerotium cepivorum) has shown promise experimentally but has yet to be used in commerce. Allium crops are grown from dry or fluiddrilled seed at a range of seed rates, from bare root and module-grown transplants and from sets and cloves. The period of susceptibility to infection may be prolonged or short in cool and hot climates, respectively. Primary infection of the Allium host results from the growth of hyphae from germinating sclerotia to the roots; secondary infection results from hyphal growth between plants. Sclerotial populations in the soil are often low, hence sclerotia are widely dispersed and therefore inaccessible to microbial control. New sclerotia form on host stem tissue, rarely on roots, rendering them accessible to microbial control. Hyphal parasites may therefore be effective at reducing primary and secondary infection and the formation of new sclerotia. Parasites of sclerotia may restrict the survival of new sclerotia and hence reduce primary infection in future, but not the current season's crops. The opportunities and problems that the variety of growing techniques and conditions of Allium crops present for microbial control of white rot will be reviewed.     

Dumontinia root rot of liver leaf caused by <Emphasis Type="Italic">Dumontinia tuberosa</Emphasis>

Foliar wilt as well as crown and root rot with sclerotia formation has affected potted liver leaf (Hepatica nobilis var. japonica f. magna) in Ojiya, Niigata Prefecture, Japan, since 2006. Apothecia developed from the sclerotia on soil surface of pots with the diseased plants in March. A fungus forming the apothecia was identified as Dumontinia tuberosa (Sclerotiniaceae) based on its morphology and demonstrated to cause the disease. We coined the name “Dumontinia root rot (Dumontinia-negusare-byo in Japanese) of liver leaf” for the new disease.     

Germination of Sclerotinia minor and S. sclerotiorum Sclerotia Under Various Soil Moisture and Temperature Combinations

ABSTRACT Sclerotial germination of three isolates each of Sclerotinia minor and S. sclerotiorum was compared under various soil moisture and temperature combinations in soils from Huron and Salinas, CA. Sclerotia from each isolate in soil disks equilibrated at 0, -0.03, -0.07, -0.1, -0.15, and -0.3 MPa were transferred into petri plates and incubated at 5, 10, 15, 20, 25, and 30 degrees C. Types and levels of germination in the two species were recorded. Petri plates in which apothecia were observed were transferred into a growth chamber at 15 degrees C with a 12-h light-dark regime. All retrievable sclerotia were recovered 3 months later and tested for viability. Soil type did not affect either the type or level of germination of sclerotia. Mycelial germination was the predominant mode in sclerotia of S. minor, and it occurred between -0.03 and -0.3 MPa and 5 and 25 degrees C, with an optimum at -0.1 MPa and 15 degrees C. No germination occurred at 30 degrees C or 0 MPa. Soil temperature, moisture, or soil type did not affect the viability of sclerotia of either species. Carpogenic germination of S. sclerotiorum sclerotia, measured as the number of sclerotia producing stipes and apothecia, was the predominant mode that was affected significantly by soil moisture and temperature. Myceliogenic germination in this species under the experimental conditions was infrequent. The optimum conditions for carpogenic germination were 15 degrees C and -0.03 or -0.07 MPa. To study the effect of sclerotial size on carpogenic germination in both S. minor and S. sclerotiorum, sclerotia of three distinct size classes for each species were placed in soil disks equilibrated at -0.03 MPa and incubated at 15 degrees C. After 6 weeks, number of stipes and apothecia produced by sclerotia were counted. Solitary S. minor sclerotia did not form apothecia, but aggregates of attached sclerotia readily formed apothecia. The number of stipes produced by both S. minor and S. sclerotiorum was highly correlated with sclerotial size. These results suggest there is a threshold of sclerotial size below which apothecia are not produced, and explains, in part, why production of apothecia in S. minor seldom occurs in nature.     

Relationships between aflatoxin production and sclerotia formation among isolates of <Emphasis Type="Italic">Aspergillus</Emphasis> section <Emphasis Type="Italic">Flavi</Emphasis> from the Mississippi Delta

Aspergillus section Flavi isolates, predominately A. flavus, from different crops and soils differed significantly in production of aflatoxin and sclerotia. About 50% of the isolates from corn, soil and peanut produced large sclerotia, while only 20% of the rice isolates produced large sclerotia. There was a higher frequency of small sclerotia-producing isolates from rice compared to the other sources and isolates that did not produce sclerotia were significantly less likely to be toxigenic than strains that produced large sclerotia.     

The effect of temperature and water potential on the production of conidia by sclerotia of Botrytis squamosa

The rate of conidiogenic germination of Botrytis squamosa was highest at 16°C and the greatest numbers of conidia per sclerotium (up to 5 × 10⁴) were produced at temperatures of 5–10°C. At temperatures above 20°C, the percentage of sclerotia producing conidia declined rapidly. Decreasing water potential reduced the rate at which conidia were produced and also resulted in fewer conidia produced per sclerotium. However, conidia were produced at water potentials as low as −2 MPa, at which sclerotial germination was at least 60%. A simulation model that included effects of both temperature and water potential was developed from laboratory and field data obtained for conidial production in sclerotia exposed for periods of 1, 2, 3 or 4 weeks during an entire year. There was good agreement between conidiogenic germination predicted by the model and conidial production observed in onion plots artificially inoculated with sclerotia. Temperature and water potential were therefore considered to be the principal microclimatic factors affecting conidial production by B. squamosa. The role of sclerotia in the context of UK onion production is discussed.     

Interactions Between the Mycoparasite Pythium oligandrum and Sclerotia of the Plant Pathogen Sclerotinia sclerotiorum

Pythium oligandrum Drechsler is a mycoparasite which parasitizes hyphae of many fungal species. A detailed study of the interactions between P. oligandrum and the sclerotia of the plant pathogen Sclerotinia sclerotiorum (Lib.) de Bary is presented. Pythium oligandrum was present in Danish soils at concentrations between 4 and 26 cfu g^–1 soil. An increase in the natural population of P. oligandrum by addition of P. oligandrum zoospores to a soil reduced the ability of sclerotia of S. sclerotiorum to germinate myceliogenically and the sclerotia were colonized internally by P. oligandrum. This colonization and reduction of germination of sclerotia were also seen when sclerotia and P. oligandrum were incubated together in water. Small sclerotia were significantly more susceptible to parasitism by P. oligandrum than large sclerotia, and increasing the incubation time caused a further reduction in the germination ability of the sclerotia. P. oligandrum was able to pass through its entire life-cycle from zoospores to oogonia both with sclerotia as sole nutrient-source and in water containing exudates from the sclerotia. The cell wall degrading enzymes N-acetyl--D-glucosaminidase (NAGase), endo-chitinase, protease, -glucanase, -glucosidase and cellobiohydrolase were detected in culture filtrates of P. oligandrum cultivated with S. sclerotiorum. These findings suggest that P. oligandrum has a potential to reduce the survival of S. sclerotiorum sclerotia present naturally in soils, through mycoparasitic activity.     

Interaction betweenSclerotinia sclerotiorum andConiothyrium minitans strains with different aggressiveness

The effects ofSclerotinia sclerotiorum live and autoclaved sclerotia, and sclerotial exudates, and commercial oxalic acid were testedin vitro on sevenConiothyrium minitans strains differing in aggressiveness towardsS. sclerotiorum. Only sclerotial exudates and autoclaved sclerotia affected the mycelial growth rate of almost all the strains tested, whereas a change in theC. minitans mycelial growth pattern was observed in the presence of autoclaved sclerotia and live sclerotia germinating by the myceliogenic eruptive germination. In addition, sclerotial exudates had a stimulatory effect on spore germination. These findings indicate that the various treatments could influence theC. minitans strains regardless of their aggressiveness.     

Forecasting Sclerotinia Disease on Lettuce: A Predictive Model for Carpogenic Germination of Sclerotinia sclerotiorum Sclerotia

ABSTRACT A predictive model for production of apothecia by carpogenic germination of sclerotia is presented for Sclerotinia sclerotiorum. The model is based on the assumption that a conditioning phase must be completed before a subsequent germination phase can occur. Experiments involving transfer of sclerotia from one temperature regime to another allowed temperature-dependent rates to be derived for conditioning and germination for two S. sclerotiorum isolates. Although the response of each isolate to temperature was slightly different, sclerotia were fully conditioned after 2 to 6 days at 5 degrees C in soil but took up to 80 days at 15 degrees C. Subsequent germination took more than 200 days at 5 degrees C and 33 to 52 days at 20 degrees C. Upper temperature thresholds for conditioning and germination were 20 and 25 degrees C, respectively. A predictive model for production of apothecia derived from these data was successful in simulating the germination of multiple burials of sclerotia in the field when a soil water potential threshold of between -4.0 and -12.25 kilopascals (kPa) was imposed. The use of a germination model as part of a disease forecasting system for Sclerotinia disease in lettuce is discussed.     

盾壳霉对油菜长效专用配方肥料的敏感性评价

油菜菌核病是油菜生产中的重要病害,盾壳霉是核盘菌的重寄生真菌,在菌核病防治方面具有重要的生防潜力。为了明确盾壳霉与油菜长效专用配方肥料混合施用的可行性,本文研究了盾壳霉对油菜长效专用配方肥的敏感性。结果发现,低浓度的油菜长效专用配方肥（1.5和7.5 mg/mL）对盾壳霉菌丝生长、菌落及菌丝尖端形态和分生孢子萌发等无明显影响,高浓度的油菜长效专用配方肥对盾壳霉生长和孢子萌发有一定影响,在饱和浓度（560 mg/mL）条件下的油菜长效专用配方肥,24 h盾壳霉孢子的萌发率为0.83%,然而在96 h时萌发率可达到95%;油菜长效专用配方肥对盾壳霉产孢和寄生致腐菌核的能力无明显影响,寄生菌核30 d后,致腐指数均在50以上。研究结果表明,在田间生产中,盾壳霉可以与油菜长效专用配方肥料混合施用,达到轻简化栽培的目的。     

Forecasting sclerotinia disease on lettuce: toward developing a prediction model for carpogenic germination of sclerotia

ABSTRACT The feasibility of developing a forecasting system for carpogenic germination of Sclerotinia sclerotiorum sclerotia was investigated in the laboratory by determining key relationships among temperature, soil water potential, and carpogenic germination for sclerotia of two S. sclerotiorum isolates. Germination of multiple burials of sclerotia to produce apothecia also was assessed in the field with concurrent recording of environmental data to examine patterns of germination under different fluctuating conditions. Carpogenic germination of sclerotia occurred between 5 and 25 degrees C but only for soil water potentials of >/=-100 kPa for both S. sclerotiorum isolates. Little or no germination occurred at 26 or 29 degrees C. At optimum temperatures of 15 to 20 degrees C, sclerotia buried in soil and placed in illuminated growth cabinets produced stipes after 20 to 27 days and apothecia after 27 to 34 days. Temperature, therefore, had a significant effect on both the rate of germination of sclerotia and the final number germinated. Rate of germination was correlated positively with temperature and final number of sclerotia germinated was related to temperature according to a probit model. Thermal time analysis of field data with constraints for temperature and water potential showed that the mean degree days to 10% germination of sclerotia in 2000 and 2001 was 285 and 279, respecttively, and generally was a good predictor of the observed appearance of apothecia. Neither thermal time nor relationships established in the laboratory could account for a decline in final percentage of germination for sclerotia buried from mid-May compared with earlier burials. Exposure to high temperatures may explain this effect. This, and other factors, require investigation before relationships derived in the laboratory or thermal time can be incorporated into a forecasting system for carpogenic germination.     

An Experiment to Assess Losses Caused by Frit-fly (Oscinella frit L) Shoot Attack and the Application of Phorate in a Crop of Sweet Corn (Zea mays L)

Wilt caused by the fungus Fusarium oxysporum f. sp. ciceris adversely affects the productivity of cultivated chickpea. For the management of this disease, seed and soil application formulations developed from another fungus, Trichoderma species, were evaluated. In pot experiments, T. harzianum-based formulations Pusa 5SD for seed dressing and Pusa Biopellet (PBP) 10G and Pusa Biogranule (PBG) 5 for soil application, and T. viride-based formulations Pusa 5SD for seed dressing and PBP 4G and PBG 4 for soil application, were found to be highly effective against the disease. A combination of PBP 4G (T. viride) for soil application and Pusa 5SD (T. harzianum) for seed treatment together with a fungicide, carboxin, provided the highest seed germination, shoot and root lengths and grain yield with the lowest incidence of wilt in chickpea under field conditions. Individually, soil application of PBP 4G, and seed treatment with Pusa 5SD were effective in reducing the incidence of wilt and increasing the grain yield of chickpea, but their effectiveness was greater when applied as a combination. Thus, combined application of the formulations of two different species of Trichoderma in two modes of application is recommended for the management of chickpea wilt.