Germination and production ofMacrophomina phaseolina microsclerotia as affected by oxygen and carbon dioxide concentration

Summary Germination of microsclerotia ofMacrophomina phaseolina was observed at O₂ concentrations of 16% or higher in autoclaved soil. Germination was delayed but otherwise unaffected as O₂ decreased from 21 to 16% and was in all cases complete in 32 hours. Laboratory-produced microsclerotia consistently germinated more rapidly and seemed more independent of O₂ concentrations within the range that permitted germination than naturallyproduced microsclerotia.Population changes in soil as measured by microsclerotial counts were inversely correlated with depth of interment and reduced O₂ concentration. Our inability to detect significantly growth responses ofM. phaseolina in non autoclaved soil was apparently related to limited O₂ although other possibilities are discussed.Contribution of the Missouri Agricultural Experiment Station Scientific Journal Series No. 9124.     

Factors affecting germination, mycoparasitism, and survival of Sporidesmium sclerotivorum.

Macroconidia of Sporidesmium sclerotivorum, a mycoparasite of Sclerotinia spp., germinated after 3 days in soil adjacent to sclerotia of S. minor and on membrane filters placed on soil containing sclerotia. Germination increased with time up to 18 days and with concentration of sclerotia. Conidia as distant as 9 mm from single sclerotia germinated. Germination of conidia was maximum on a sclerotial agar medium in the range of pH 5 to pH 7. Cultivation of S. sclerotivorum parasitically on living sclerotia proceeded optimally in moist, fine quartz sand amended with 1 to 2% (w/w) sclerotia and 0.07% (w/w) CaCO3, at 25 degrees C. Infection of sclerotia in sand reached 100% by 5 weeks. Conidia production paralled infection resulting in logarithmic increase in numbers; a maximum of 3 x 10(5) to 4 x 10(5) conidia/g was reached in 6 to 12 weeks. Viability of air-dried sand-sclerotial cultures of S. sclerotivorum was reduced after 1 and 6 days, but viability was undiminished in air-dried soil. Sporidesmium sclerotivorum survived in moist and air-dried soils stored at room temperature for 15 months.     

Epidemiological studies on jute diseases

Summary Hyphae of M. phaseoli failed to grow on unsterilized natural soil and were completely lysed within 4 days of exposure. Germination of sclerotia in natural soil was inhibited indicating soil fungistasis. Lysis of mycelium and inhibition of germination of sclerotia could be annulled by addition of various organic nutrients and fertilizers to natural soil or by autoclaving the soil. Germination of dormant sclerotia in natural soil was stimulated by root-exudates of host and non-host plants. Population of sclerotia buried in unsterilized natural soil gradually declined and after 15 months only 35 per cent of the initial number could be recovered; more than 80 per cent of these germinated when nutrients were added. Data suggest poor saprophytic ability of M. phaseoli in mycelial form and the involvement of dormant sclerotia in the survival of the organism in soil.     

The effect of species of Allium on soil bacteria in relation to germination of sclerotia of Sclerotium cepivorum Berk

Extracts made from Allium species were shown to exhibit marked antibiotic properties in a variety of tests. However, no evidence was obtained to suggest that intact Allium seedlings exude antibiotic compounds in sufficient quantity to cause inhibition of bacteria in the soil or in assay tests in the laboratory. Germination of sclerotia of Sclerotium cepivorum was induced by Allium extracts which were too dilute to cause antibiotic responses in laboratory tests using a number of bacteria. Synthetic methyl-methanethiolsulphinate had little effect on germination of sclerotia. The specific reversal, by Allium species and their extracts, of the mycostatic effect of unsterile soil on sclerotia of S. cepivorum does not appear to be due to antibiotic effects.     

Long-term effects of sheep grazing on giant hogweed (Heracleum mantegazzianum)

A meadow dominated by a mature, flowering stand of Giant Hogweed (Heracleum mantegazzianum) was grazed by sheep in the years 1987–1993. After two years Giant Hogweed cover was much reduced and a typical meadow vegetation was established. By 1993 Giant Hogweed was completely eliminated though total species diversity was much reduced. Soil sampled from the grazed area developed no Giant Hogweed seedlings in a germination test and contained no viable seeds of the species. In contrast soil from adjacent stands produced numerous seedlings with a peak emergence from samples taken after the winter. Seeds collected in October showed a viability in tetrazolium test of 88%. Germination averaged 22% after storage at room temperature and 25% following three weeks treatment at –18°C. It was concluded that the persistence of Giant Hogweed seeds in Danish meadow soils is less than seven years.     

Effect of soil solarization on corn stalk rot

Seven weeks solarization of irrigated soil raised its temperature by 11.5°C over non-solarized soil at 10 cm depth and effectively controlled weeds (98.5%), stalk borer (8.9%) and stalk rot disease (69.1%) in corn. Solarization also reduced symptoms of Fusarium moniliforme and Macrophomina phaseolina significantly by 64.2% and 78.4%, respectively, and completely controlled M. phaseolina in corn cultivars, viz. Pool-10, Shaheen and Gauher. Whereas symptoms of F. moniliforme were observed in these cultivars, Fusarium graminearum was not observed except in two cultivars, Shaheen and Akbar. Growth of crop planted in solarized plots was better and it yielded almost one to three times more grains in cultivars under test. Soil analysis immediately following solarization revealed that essential elements were readily available in simpler forms, which may have increased pest resistance and reduced stalk breakage.     

Population dynamics of Macrophomina phaseolina in relation to disease management: A review

Macrophomina phaseolina (Tassi) Goid. causes seedling blight, charcoal rot, leaf blight, stem and pod rot on over 500 plant species in different parts of the world. The pathogen survives as sclerotia formed in host tissues which are released into the soil as tissue decay. Low soil moisture is considered the more important predisposing factor for M. phaseolina-induced diseases than high temperature. The intensity of the disease on a crop is related to the population of viable sclerotia in the soil and abiotic factors. The influence of various management strategies in reducing the number of viable propagules of the pathogen in the soil has been studied in order to minimize the impact of the disease. Any management approach that reduces inoculum density in the soil may reduce disease incidence on the host. However, to reduce inoculum density, quantitative determination of viable propagules from soil is necessary in order to understand the effect of management strategies on the population dynamics of this pathogen. Considerable work has been done on organic amendments, changing crop sequences with tolerant crops, fumigants, herbicides and tillage in managing M. phaseolina populations in the soil and the resulting disease. Solarization has been used in controlling M. phaseolina in different countries where this pathogen is causing disease on economically valuable crops. However, this method of soil disinfestation was effective in eliminating viable populations at the top soil layer although by combining other approaches its effectiveness was improved at lower soil depth. Use of biological control agents with or without organic amendments or after solarization has emerged to be a practical management approach in the control of M. phaseolina. In this paper, an attempt has been made to review those research findings where the influence of various management approaches on survival of M. phaseolina mainly sclerotia have been investigated.     

Effect of Moisture and Temperature on the Survival of Sclerotia of Sclerotium rolfsii in Soil

The sclerotia of Sclerotium rolfsii Sacc. survived in natural soil for 225 days under controlled moisture at 50% water holding capacity (WHC) after which there was a progressive reduction in the population of viable sclerotia. At 390 days only 48% were recovered. Sclerotia survived well at moisture contents upto 75% WHC but at 100% the population declined rapidly and none were recovered after 60 days. The contents of the sclerotia were found to lyse without germination leaving hollow rinds. Such lysis was found to be favoured between 25 and 40°C. At and below 20°C no such lysis was recovered and more than 80% sclerotia were recovered even after 60 days.     

Carpogenic Germination of Sclerotia of Sclerotinia sclerotiorum after Periods of Conditioning in Soil

Periods of conditioning in soil reduced the length of the resting period needed before sclerotia of Sclerotinia sclerotiorum could germinate to form apothecia. Curves for germination of sclerotia were fitted by a form of the log-logistic equation and from this equation the time taken for 50% germination (x₅₀) was calculated. These x₅₀ values were used as the basis for comparing germinability of sclerotia collected from infected sunflower plants and others conditioned in soil, or moist vermiculite for various times. Sclerotia from sunflower roots germinated sooner than those from the stem cavities. Germinability increased with the length of the conditioning period. Conditioning in soil was more effective than in moist vermiculite.     

Fungistatic activity of the sternal gland secretion of the dampwood termite <Emphasis Type="Italic">Zootermopsis angusticollis</Emphasis>

Summary In vivo and in vitro studies indicate that cuticular chemicals from the ventral region of the abdomen where the sternal gland of the dampwood termite Zootermopsis angusticollis is located have fungistatic properties. Germination rates of conidia of the entomopathogenic fungus Metarhizium anisopliae were significantly reduced from 91% (controls) to 38.5% after nymphs walked over conidia-seeded agar medium, but did not differ from controls when the sternal gland and surrounding cuticle were sealed with nail polish. In vitro studies show that germination of fungal conidia was also significantly reduced following incubation with cuticular extracts of either sternal or tergal segments suggesting that cuticular exudates in general may have antifungal properties. Extracts of sternites had greater fungistatic activity than extracts of tergites, but the difference was not statistically significant. Extracts of the sternal gland significantly reduced germination rates by up to 9%. Germination rates were significantly reduced when conidia were incubated with n-hexanoic acid, or its vapor. n-Hexanoic acid has been recovered from whole body extracts of Zootermopsis nevadensis and may indeed be a component of the sternal gland of Z. angusticollis. Here we suggest that sternal gland secretions in termites may have had the original function of controlling microbes within the nest and their prominent role in communication may have evolved secondarily.Received 18 April 2003; revised 20 November and 17 December 2003; accepted 19 January 2004.     

STUDIES OF THE BIOLOGY OF SCLEROTIUM CEPIVORUM BERK.

Sclerotia from 6-week-old pure cultures of Sclerotium cepivorum germinated immediately in soil only after abrasion of their rinds, but after burial in soil for a month or more, unabraded sclerotia became capable of germination.
Marked stimulation of germination occurred in the presence of host plants (onion, leek and shallot). Little or no germination occurred in soil alone or in the presence of non-host plants (barley, cabbage, carrot and white clover). Sclerotial germination was observed in a number of soils of widely differing pH and over a wide range of soil water content. Germination of sclerotia on uninjured onion roots was greatest at the tip region. On artificially injured roots sclerotial germination was enhanced but the effect of position was lost.
Sclerotial germination was independent of contact between roots and sclerotia. It was induced by root extracts of all Allium spp. tested, but of no other plants. Boiling or autoclaving root extracts did not destroy the active principle and it is concluded that under field conditions sclerotia are induced to germinate by a thermostable chemical substance from Allium roots.
The process of germination of sclerotia is described.     

The Role of White Mold-Infected White Bean (Phaseolus vulgaris L.) Seeds in the Dissemination of Sclerotinia sclerotiorum (Lib.) de Bary

Sclerotinia sclerotiorum survived in infected seeds of white beans as dormant mycelium in testa and cotyledons. The rate of survival averaged 85 to 89% and did not change appreciably over a 3-year period. When the infected bean seeds were sown in soil or sand, 88 to 100% failed to germinate. The seeds that failed to germinate, depending on the severity of seed infection, were rotted by S. sclerotiorum. In place of each seed, 3 to 6 sclerotia were formed. A low percentage of these sclerotia germinated carpogenically with or without preconditioning, (2.5 and 11.5% respectively). Myceliogenic germination of sclerotia with and without preconditioning was 35.5% and 70.5% on water agar and 81.0% and 93.0% on glucose agar, respectively. Both, preconditioning and nonpreconditioned sclerotia which were scattered on soil surface could germinate myceliogenically and infect bean leaves by contact. It is therefore, concluded that dormant mycelia in the infected seeds play an important role notonly in dissemination of the fungus but also in epidemiology of the disease.     

The effect of species of Allium on germination in soil of sclerotia of Sclerotium cepivorum Berk

Sixty-three different plants, from a wide range of families, were tested for their effects on germination of sclerotia of Sclerotium cepivorum. Germination was stimulated only by members of the genus Allium. Water extracts of Allium species also stimulated germination of sclerotia, but extracts of other plants, and a number of nutrient solutions, had little effect. There was no evidence of inhibition of sclerotial germination by plants which failed to elicit a germination response.     

Germination ecology and seed population dynamics of Digitalis purpurea

Summary The germination ecology and the dynamics of the generative reproduction in populations of Digitalis purpurea L. were investigated in the field as well as in experiments. Germination of fresh seeds in the dark on moist filter paper appeared to differ between populations. These differences were eliminated when a moist natural soil functioned as germination substrate. An interaction between the spectral composition of light and the germination substrate was present. Germination in gradients of light, temperature and soil moisture revealed some clear-cut results. Germination proved to be strongly dependent on the percentage of vegetation cover. During two years of burial in litter bags, the number of buried viable seeds did not decrease. From one generation of seeds produced in a natural population, 18% was introduced into the buried seed bank, 10% germinated in autumn and 24% was present as a enforced dormant surface seed bank in late autumn.The results are discussed in relation to secondary succession. can be derived from Milton (1936), Salisbury (1942) and Thompson and Grime (1979). Soil disturbance and germination seem to be correlated in D. purpurea (Grime 1979). The purpose of this study is to analyse the dormancy and germination behaviour of D. purpurea in relation to the relevant environmental factors in order to explain the mechanisms of entry into, and the escape of D. purpurea seeds from a seed bank. Furthermore, an attempt will be made to quantify seed rain as well as the fate of different germinating and non-germinating seed rain fractions in space and time per unit area, in different stages of succession.     

Biocontrol of Macrophomina root-rot disease of jute by an antagonistic organism,Aspergillus versicolor

Summary Aspergillus versicolor was grown in soil-compost medium at pH 4.0 for 10 days under diffused light to getA. versicolor-grown soil which was found to control experimental infection in jute byMacrophomina phaseolina to the extent of 56% in pot-culture experiment.     

Factors Affecting the Parasitic Activity of Gliocladium virens on Sclerotia of Sclerotinia sclerotiorum and a Note on its Host Range

Aspects of the biology of Gliocladium virens and parasitism of sclerotia of Sclerotinia sclerotiorum in soil were studied. G. virens parasitized and decayed sclerotia of S. sclerotiorum, S. minor, Botrytis cinerea, Sclerotium, rolfsii and Macrophomina phaseolina on laboratory media and caused a reduction in survival of sclerotia of S. sclerotiorum in soil. It was active over a broad range of soil moisture levels and over the entire agricultural soil pH range. The main factor limiting its use as a biological control agent was its temperature requirements.     

Survival of Sclerotia of Rhizoctonia solani AG3PT and Effect of Soil‐Borne Inoculum Density on Disease Development on Potato

Sclerotia produced by a single isolate of Rhizoctonia solani AG3PT were buried in small plot experiments to investigate the effects of sclerotial production method, soil type and burial depth on sclerotial viability in field soil. The factor with the greatest effect on sclerotial viability, defined as the percentage of sclerotia germinating on agar following retrieval, in all experiments was the duration of burial. After 18 months, on average across all experiments, 20% of retrieved sclerotia were viable. A comparison between sclerotia produced in vitro on malt yeast extract agar and in vivo using micropropagated tubers in field soil found no significant differences between the two production methods on sclerotial viability. Burial in field soil at 20‐cm depth was found to significantly reduce sclerotial viability to 50% compared to 60% at 5 cm. In two pot experiments, amending the growing medium and soil with increasing inoculum densities of R. solani was found to increase stem number, stem canker and black scurf severity regardless of whether this soil‐borne inoculum was derived from mycelium or sclerotia. Black scurf incidence and severity were assessed 30–32 days posthaulm destruction and found to be similar for a range of sclerotial soil‐borne inoculum densities (1.0 × 10^?1 g/kg d.w. soil to 6 × 10^?3 g/kg d.w. soil). The significance of these findings in relation to pathogen survival, detection in soil and disease development is discussed.     

La dormance embryonnaire chez Taxus baccata: Influence de la composition du milieu liquide sur I'induction de la germination Par

Embryo dormancy of Taxus baccata var. fastigiata is eliminated when cultured continuously in nutritive liquid medium. An equivalent percentage of germination is obtained when the embryos are transferred to agar medium after 8 days of liquid culture. There is no morphological development of the embryo during the period in the liquid medium. But we have ascertained that water-soluble germination inhibitors present in the embryo are leached out into the medium, permitting germination. Germination is totally absent when the embryos are cultured continuously in distilled water, alone or with minerals; incidental in sucrose solution; and maximal when the medium contains sucrose and Ca²⁺ or K⁺ ions. The extent of germination on agar medium depends upon the composition of the liquid medium in which the embryos are cultured for the initial 8 days. But this preliminary culture in the liquid medium does not always remove the endogenous inhibitors, irrespective of its composition. This can be achieved only in the presence of sucrose; and this process can be made more effective by the addition of Ca²⁺ ions.     

Increasing in vitro germination of Musa balbisiana seed

Seeds of Musa balbisiana were soaked in water for five days prior to excision of embryos. Embryos with their longitudinal axis laid flat and half-way embedded on agar-solidified medium produced the highest germination and the most desirable plantlet characteristics. Germination in vitro was 94% within 7 days compared to 50% after 54 days for greenhouse-sown seeds.     

Germination requirements and responses to leaf litter of four species of eucalypt

We studied how the responses of four species of eucalypt to leaf litter related to their germination responses to light and water availability. Two of the species (Eucalyptus obliqua and E. baxteri) have a mesic distribution, while the other two (E. oleosa, and E. incrassata) are more xerophytic. We studied the effect of litter on emergence of the four species in a glasshouse experiment. Litter did not affect the emergence of E. incrassata and E. oleosa, but enhanced the emergence of E. obliqua and E. baxteri. Litter increased the seedling mortality of all four species. Germination responses to light and water availability were studied in growth cabinets under controlled conditions. The germination of E. obliqua and E. baxteri was substantially lower under fluorescent light than in darkness, but that of E. oleosa and E. incrassata was not affected by the light environment. The germination of E. obliqua and E. baxteri was significantly reduced by reduced water potential (a). Substantial germination of E. oleosa and E. incrassata occurred even at very low a (less than –1.05 MPa). We conclude that both the shade and the humid micro-environment provided by litter may have contributed to the emergence responses of the four species to litter, and these responses may correspond to ecological adaptations to the different environments in which they live.