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1.
Sclerotia are the primary over wintering inoculum of Sclerotinia sclerotiorum (Lib.) de Bary. The effects of tillage on the primary inoculum are not well understood. The purpose of this research was to study sclerotial viability over time and between burial depths in soil, to identify bacteria colonizing and degrading the sclerotia, and determine whether these bacteria may be utilized as biological control agents. Correlation analysis indicated that a significant negative relationship existed between sclerotial viability and elapsed temporal factors (R2=−0.68, P<0.0001), and depth of burial (R2=−0.58, P<0.0001). After twelve months, sclerotia on the soil surface had the highest viability (57.5%), followed by those at the 5 cm depth (12.5%), and only 2.5% of those placed at the 10 cm depth remained viable. A significant negative relationship between sclerotial viability and bacterial populations also existed (R2=−0.60, P<0.0001). Two hundred and sixty-eight bacteria were isolated from sclerotia, 29 of which showed strong in vitro antagonism to the mycelial growth of S. sclerotiorum. Biodiversity of the inhibitory bacterial isolates was minimal on sclerotia from the soil surface and within all depths sampled at three months (i.e. in January). All burial depths within the April and July sampling dates produced bacterial diversities that were distinct from each other.  相似文献   

2.
Dried sclerotia of Sclerotium delphinii rotted in moist soil whereas those of Sclerotium cepivorum. Botrytis cinerea and Botrytis tulipae did not. A number of fungi invaded dried sclerotia of S. delphinii in soil, the principal coloniser found in the first sampling being Trichodermu hamatum. Leakage of 14C compounds from dried labelled sclerotia placed in water was rapid and was little affected by variation in leakage temperature from 1 to 25°C or by prolonging the drying period beyond a day. Leakage from dried sclerotia which were allowed to imbibe water through a small part of their surface was much reduced. Sclerotia which were redried after leakage leaked again when returned to water but with all four fungi the first of three leakage cycles gave the highest 14C levels. Loss in dry weight in the first leakage cycle was greater with S. delphinii than with the other three fungi and this may be related to the poor survival of dried sclerotia of S. delphinii in moist soil. Substances lost during leakage appear to originate from within sclerotial hyphae rather than from the hyphal free space.  相似文献   

3.
Ecological and morphological characteristics of the sclerotia of R. solani produced in soil were compared among Japanese isolates belonging to five anastomosis groups (AG). Twenty-three out of 24 isolates produced sclerotia in soil and the number and size of sclerotia varied considerably among isolates within the same AG. Sclerotia from each group ranged in size from 0.25 mm to more than 2.5mm dia, with most between 0.5–1.0mm dia. Sclerotia formed in soil were smaller than those formed in pure culture. Sclerotia of each AG were tested for viability after 30–270 days incubation in artificially-inoculated soil. Sclerotia produced in soil were brown to dark-brown and these were divided into two distinct groups on external morphology: firm and irregularly globose to subglobose with either a pitted or smooth surface in one group, and rather soft with an indefinite shape in the other. On the basis of internal morphology, the sclerotia fell into three groups. Sclerotia of AG-1 were composed of three well-defined layers; the central cells had dense contents and were surrounded by an outer layer of empty cells which were bordered by a darkly-pigmented mucilaginous surface-layer. Sclerotia of AG-2, Type-1 had only two well-defined layers; The darkly-pigmented mucilaginous layer was absent. Sclerotia of the other AG were constructed of very loosely-arranged brown cells without any well-defined zones.  相似文献   

4.
The development and survival of the mycoparasite Coniothyrium minitans associated with sclerotia of the plant pathogen Sclerotinia sclerotiorum was studied in pasteurised and non-sterile (untreated) soil. Using scanning electron microscopy, developing pycnidia were first seen within the sclerotial medulla at 7 days post-inoculation with the mycoparasite in pasteurised soil. However, by 14 days post-inoculation, pycnidia had developed fully in both pasteurised and non-pasteurised treatments, and conidial droplets were exuded onto the outer surface of the infected sclerotia. Thirty days post-inoculation, irrespective of soil treatment, the majority of the sclerotial medulla had been converted to pycnidia, with the sclerotial rind remaining largely intact. The pycnidia and dried intact droplets were still observed 6 months post-inoculation with C. minitans, although the conidia on the outer surface of the dried droplets had largely collapsed by this stage. Germinability studies at 10 months post-inoculation showed that approximately 13% of the conidia in dried droplets were still viable. This work shows the potential for infected sclerotia of S. sclerotiorum to provide a unique reservoir for the survival of C. minitans.  相似文献   

5.
Factors affecting longevity of sclerotia in sandy clay loam (s.c.l.) and sandy loam (s.l.) were examined, using sclerotia from a laboratory culture of S. sclerotiorum and from natural infestations on beans and lettuce.Survival of sclerotia from culture and lettuce was compared in s.l. Recovery and viability were less, and incidence of Fusarium, Mucor and Trichoderma spp. greater, in sclerotia from lettuce than from culture. Rinds of sclerotia from lettuce were more perforated than those from culture.Burial of sclerotia at 4 cm for 35 weeks reduced recovery of sclerotia to zero in s.c.l. and by 50% in s.l. At the soil surface recovery was reduced by 55% in s.c.l. and by 10% in s.l. Less than 50% of sclerotia recovered were viable. Neither a chloropicrin-methyl bromide fumigant nor a tomato compost treatment affected recovery or viability. Fumigation increased incidence of Trichoderma spp. and decreased incidence of Fusurium and Mucor spp. isolated from sclerotia.Apothecia were produced over 6 weeks in s.c.l. and over 20 weeks in s.l. Production was increased by the low rate of fumigant in s.c.l. and by tomato compost in s.l.  相似文献   

6.
At least 75% of the sclerotia of Macrophomina phaseolina survived for 1 yr in most natural soils kept at 26°C and at 50–55% of the soil moisture holding capacity (m.h.c.). Although survivability was reduced in a very acid soil (pH 4.5) collected under a pine stand, 33% of the sclerotia survived for 1 yr. Soil pH had very little or no effect on sclerotial survivability. Of three organic amendments tested (alfalfa hay, chitin, pine needles) only ground alfalfa hay at 0.8% (w/w) reduced survivability of sclerotia in soil by about 75% in a year. Alfalfa hay at 0.4% reduced survivability by 36%. Various N sources added at 200 μg Ng?1 soil had no effect on survival. Of 13 fungicides tested, only benomyl and captan at 20 μg a.i. g?1 soil appreciably reduced populations of sclerotia in soil.Soil temperature and moisture content were the two most important factors affecting survivability of sclerotia. At ?5 or 5°C the biggest drop in sclerotial survivability occurred when the soil was incubated moist (at 50% m.h.c. or more). At 26°C the biggest drop occurred in air-dried soil (2–3% m.h.c.) and survivability was decreased to some extent at 15 and 30% m.h.c. Survivability also dropped rapidly in moist soil (50–55% m.h.c.) exposed to four cycles each having 3-week freezing (?5°C) and 1 week thawing (26°C). Sclerotia in air-dried soil (2–3% m.h.c.) continuously kept at ?5°C maintained nearly complete survivability after 16 weeks. Sclerotia survived almost 80–90% in moist soil (50–55% m.h.c.) kept for 16 weeks at 26°C or in moist soil exposed to four cycles each having 3-week thawing (26°C) and 1-week freezing (?5°C).  相似文献   

7.
Exposure of sclerotia of Macrophomina phaseolina to 0 and 33% relative humidity (r.h.) for 12 weeks and of Sclerotium cepivorum to 0, 33 and 55% r.h. for 20 weeks did not reduce their germinability on agar. Exposure to 78% r.h. caused high loss of germinability in M. phaseolina and complete loss in S. cepivorum. After 7-day exposures respective moisture contents of sclerotia of M. phaseolina and S. cepivorum were 1 and 2% at 0% r.h.; and 10 and 14% at 78% r.h. M. phaseolina sclerotia held at 0% and 33% r.h. in desiccators for several times up to 12 days did not decrease in subsequent survivability in moist soil, unlike sclerotia held at 78% r.h. for 4 days.More sclerotia of M. phaseolina were colonized by fungi and Streptomyces spp. on alkaline soil than on acid soil. On alkaline soil twice as many sclerotia were colonized after exposure to 0% r.h. as after exposure to 33, 55 and 78% r.h. Colonization of S. cepivorum sclerotia was as high on acid as on alkaline soil and 3 times as high on sclerotia treated at 0% r.h. as on those treated at higher r.h. Attempts to ascertain the effects of colonization on sclerotial viability were unsuccessful. Incubation of sclerotia of M. phaseolina in moist Rumsford sandy loam (50% m.h.c.) for 20 weeks reduced survivability by 43%. At room temperature, alternate drying and wetting of soil containing sclerotia did not appreciably affect survivability of either pathogen. Survivability of S. cepivorum sclerotia was highest when the sclerotia were incubated in air-dried soil (2–3% m.h.c.) for 20 weeks.Incidence of white rot on onion seedlings transplanted to S. cepivorum-infested soil was higher in soil that had been air-dried for 20 weeks than in soil that had been alternately wetted and dried. Sclerotia that were exposed to 0% r.h. for 7 days before soil incubation produced little white rot.  相似文献   

8.
The colonization of Macrophomina phaseolina sclerotia by microbial parasites was evaluated in unsterilized field soil at different levels of soil moisture (0,-5, and-10 kPa) and temperature (20, 30, and 40°C). The maximum colonization of sclerotia was recorded in soil held at-5 or-10 kPa at 30–40°C. Trichoderma harzianum isolate 25–92 and Pseudomonas fluorescens isolate 4–92 were recorded as potential sclerotial parasites, and they significantly (P=0.05) reduced the germination of sclerotia by 60–63%. Cells of P. fluorescens and buffer-washed conidia of T. harzianum were completely agglutinated at 28°C with crude agglutinin of M. phaseolina. The ability of different antagonists to parasitize the sclerotia were correlated with the agglutination ability of the antagonists.  相似文献   

9.
Germinability and virulence of sclerotia of Sclerotium rolfsii were assessed after 50 days of exposure of 14C-labeled sclerotia to soil at 0, −5 and −15 kPa and pH 6.9, or to soil at 15, 25 or 30 °C, pH 5 or 8 and −1 kPa. Evolution of 14CO2 accounted for the greatest share of endogenous carbon loss from sclerotia under all soil conditions, except in water-saturated soil (0 kPa), in which sclerotial exudates contributed the major share of carbon loss. Total evolution of 14CO2 from sclerotia in soil at −15 kPa (42.4% of total 14C) and at −5 kPa (38%) was significantly higher than at 0 kPa (23.8%). Evolution of 14CO2 in soil at 25 or 30 °C was more rapid than at 15 °C with regardless of pH. Loss of endogenous carbon by sclerotia was the greater after 50 days of exposure to soil at 0 kPa, or at 25 or 30 °C and pH 8, than at other soil conditions. Sclerotia exposed to water-saturated soil (0 kPa) showed a more rapid decline in nutrient independent germinability, viability and virulence, than to those exposed to −5 or −15 kPa. Sclerotia became dependent on nutrient for germination and lost viability and virulence within 30–40 days in soil at 25 or 30 °C, pH 8. However, more than 60% of sclerotia retained viability in soil at 15 °C regardless of pH, even after 50 days. Radish shoot growth was increased significantly by the sclerotia that had been exposed to soil at 0 kPa, or to soil at 25 or 30 °C and pH 8 for 50 days. In conclusion, carbon loss by sclerotia during incubation on soil at different pH levels, temperatures and water potentials was inversely correlated with sclerotial ability to infect radish seedlings. The relationship between carbon loss by sclerotia and radish shoot length was positive.  相似文献   

10.
A single injection of 0.2 ml diallyl disulphide (DADS) at 0.156% (v/v) into soil containing naturally-produced sclerotia of Sclerotium cepivorum and maintained in the laboratory at 15°C stimulated sclerotial germination and reduced sclerotial numbers by 67%; ungerminated sclerotia remained viable. Higher concentrations of DADS had no additional effect except that at 20% (v/v), germination was slightly inhibited. A similar reduction in sclerotial numbers was obtained when the mixture of soil and sclerotia was exposed to DADS vapour. Four, monthly applications of DADS at 0.2 ml 0.15% (v/v) per application did not give a further reduction.The effect of DADS was temperature dependant, with a reduction in sclerotial numbers of 65 and 9% at 15 and 5°C respectively.  相似文献   

11.
Crop rotation has been used for the management of soilborne diseases for centuries, but has not often been planned based on scientific knowledge. Our objective was to generate information on Sclerotium rolfsii dynamics under different crop or intercrop activities, and design and test a research approach where simple experiments and the use of models are combined to explore crop sequences that minimize Southern blight incidence.The effect of seventeen green manure (GM) amendments on sclerotia dynamics was analyzed in greenhouse and field plot experiments during two years. The relative densities of viable sclerotia 90 days after winter GM (WGM) incorporation were generally lower than after summer GM (SGM) incorporation, with average recovery values of 60% and 61% for WGM in the field, 66% and 43% for WGM in the greenhouse, and 162% to 91% for SGM in the greenhouse, in 2009 and 2010, respectively. Sclerotia survival on day d after GM amendment was described by the model Sf = Si × exp(−b × d), relating initial (Si) and final (Sf) sclerotia densities. Relative decay rates of the sclerotia (b) in SGM amended soil were largest for alfalfa (0.0077 ± 0.0031 day−1) and sudangrass (0.0072 ± 0.0030 day−1). In WGM amended soil, the largest b values were for oat (0.0096 ± 0.0024 day−1), wheat (0.0090 ± 0.0024 day−1) and alfalfa (0.0087 ± 0.0023 day−1).The effect of three cropping sequences (sweet pepper–fallow, sweet pepper–black oat and sweet pepper–onion) on sclerotia dynamics was analyzed in microplot experiments, and the data were used to calibrate the model Pf = Pi/(α + βPi), relating initial (Pi) and final (Pf) sclerotia densities. Median values for the relative rate of population increase at low Pi (1/α, dimension less) and the asymptote (1/β, number of viable sclerotia in 100 g of dry soil) were 8.22 and 4.17 for black oat (BO), 1.13 and 8.64 for onion (O), and 6.26 and 17.93 for sweet pepper (SwP).By concatenating the two models, sclerotia population dynamics under several crop sequences were simulated. At steady state, the sequence SwP–O–Fallow–BO resulted in the lowest long-term sclerotia density (7.09 sclerotia/100 g soil), and SwP–Fallow in the highest (17.89 sclerotia/100 g soil). The developed methodology facilitates the selection of a limited number of rotation options to be tested in farmers’ fields.  相似文献   

12.
Mulching of Macrophomina phaseolina-inksted soil (moist or dry) with transparent polyethylene sheets during the hot days of May increased temperature of wet soil at 5 cm from 37°C (unmulched) to 52°C (mulched) and of dry soil from 52°C (unmulched) to 65°C (mulched). At 20 cm mulching increased temperature from 30°C to 41°C (wet) and from 38°C to 42°C (dry). In artificially-infested soil. the sclerotia of M. phaseolina were eradicated at 5 cm by a mulch treatment for 1 week and at 20 cm depth 50% sclerotia lost viability in wet soil but were not affected in dry soil. In a naturally infested soil (5–7 sclerotia g?1), which gave 20% infection on Vigna, the sclerotia were reduced to such an extent that after 1 week mulching no disease was observed on Vigna.  相似文献   

13.
The effects of three Coniothyrium minitans isolates (Conio, IVT1 and Contans®), applied to soil as conidial suspensions or as maizemeal-perlite (MP) inocula (Conio), on apothecial production and infection of Sclerotinia sclerotiorum sclerotia were assessed in two soil pot bioassays and two novel box bioassays in the glasshouse at different times of the year. C. minitans isolate Conio applied as either MP or ground MP at full rate (106-107 cfu cm−3 soil) consistently decreased the carpogenic germination, recovery and viability of sclerotia and increased C. minitans infection of the sclerotia of S. sclerotiorum by in comparison with either MP or conidial suspension treatments applied at lower rates (103-104 cfu cm−3 soil). Additionally, when applied at the same rate, MP inoculum of C. minitans was consistently more effective at reducing carpogenic germination than a conidial suspension. The effect of MP and ground MP at full rate on carpogenic germination was expressed relatively early as those sclerotia recovered before apothecia appeared on the soil surface already had reduced numbers of apothecial initials. In general, there were few differences between the isolates of C. minitans applied as conidial suspensions. Box bioassays carried out at different times of the year indicated that temperature and soil moisture influenced both apothecial production and mycoparasitism. Inoculum concentration of C. minitans and time of application appear to be important factors in reducting apothecial production by S. sclerotiorum.  相似文献   

14.
Rhizoctonia solani causes worldwide losses in numerous crops. Sclerotia of R. solani remain viable for several years in soil and are an important source of primary infection. In this study the effect of soil incorporation of Kraft pine lignin, a side product of the paper industry, on viability of R. solani AG1-1B sclerotia was investigated. The efficacy of lignin was assessed in a sandy loam (Oppuurs) and a silt loam soil (Leest) collected from commercial fields in Belgium. Evaluating sclerotial viability after 4 weeks incubation in the two soils amended with 1% (w/w) Kraft pine lignin demonstrated a soil-dependent effect. In Leest soil the addition of lignin resulted in a significantly reduced sclerotial viability, together with an increased mycoparasitism by Trichoderma spp.; in Oppuurs soil, on the other hand, only a slight and insignificant reduction in sclerotial viability was observed. Based on phospholipid fatty acid analysis, different changes in microbial community structure upon lignin amendment were detected in the two soils. Both amended soils showed a significant increase in Gram negative bacteria. In Leest soil this increase was accompanied with a significantly higher increase in fungi and actinomycetes compared with Oppuurs soil. In addition, Kraft pine lignin resulted in both soils in a small but significant increase in manganese peroxidase activity and this increase tended to be higher in Leest soil. Manganese peroxidase produced by lignin-degrading basidiomycetes has previously been shown to degrade melanin, which protects the sclerotia against biotic and abiotic stress. We hypothesize that lignin-degrading fungi increased the susceptibility of the sclerotia to sclerotial antagonists such as Trichoderma, Gram negative bacteria and actinomycetes. Clearly, the effect observed here did not rely on the stimulation of one microbial group, but is the result of an interaction of different groups.  相似文献   

15.
The morphology and chemical composition of fungal sclerotium-like grains collected from the buried humic horizon of volcanic ash soil in Mt. Myoko were examined using a scanning electron microscope (SEM), energy dispersion xray micro-analyzer (EDX), and electron probe micro-analyzer (EPMA). A high C content, low level of Si and high concentration of Al characterized the grains considered to correspond to the sclerotia of Cenoccocum graniforme. The 27Al MAS NMR spectrum of the sclerotia showed the existence of 6- and 4-coordinated Al. X-ray diffraction analyses supported the presence of amorphous Al (ex. Al-humus complex) in the sclerotia.  相似文献   

16.
Aspects of the biology of C. minitans and its potential for control of S. sclerotiorum were investigated.Temperatures below 7°C resulted in comparatively slow rates of germination and infection of sclerotia by C. minitans. The optimum temperature for germination, growth, infection of sclerotia, and destructive parasitism by C. minitans was 20°C. The optimum relative humidity for germination, growth and infection by C. minitans was above 95%.Autumn inoculations with suspensions of conidia, pycnidia and mycelium of C. minitans in the field resulted in negligible numbers of sclerotia remaining viable after 1 month. With culture-grown sclerotia 2 months were required for a similar reduction of sclerotial viability. In the absence of C. minitans mulching had no significant effect on sclerotial viability. In the presence of C. minitans mulching did, however, influence the viability and infection by C. minitans of culture-grown sclerotia. Populations of field sclerotia also differed from culture-grown sclerotia in that they harboured an internal population of microorganisms, which included C. minitans, and had a lower level of viability at the commencement of the treatments.A winter application of C. minitans did not result in significant infection of sclerotia nor in a reduction in viability of sclerotia. This failure is believed to have resulted from low temperatures and dry conditions.  相似文献   

17.
Summary The mass loss and N dynamics of barley stems and leaves, placed on the soil surface or buried, were examined over two summers. There was little difference in mass loss or N dynamics in straw placed 7.5 or 15 cm deep. However, the surface straw lost mass much more slowly and immobilized more N for a longer time than the buried straw. Filter paper had a slow rate of mass loss initially, but once started, lost mass much more rapidly than either the barley stems or leaves. Loss of mass was closely correlated with the cellulose loss in straw, whether buried or placed on the soil surface. The sustained rate of mass loss was 6.3 and 7.0% month-1, respectively, for surface and incorporated leaves compared with 3.5 and 4.3% month-1, for surface and incorporated stems. The greater loss sustained by the leaves was attributed to a lower lignin content rather than a higher N content, because the addition of N to the straw after 30 days in the field failed to increase CO2 evolution. Maximum net N immobilization occurred within 30 days for all the barley straw, except for the stems placed on the ground surface, which did not reach maximum N immobilization until the second summer. Immobilization and mineralization of N were estimated for a 3000 kg ha-1 grain crop. Surface straw immobilized 3.8 kg N ha-1 in the 1st year and 9 kg N ha-1 in the 2nd year, whereas incorporated straw immobilixed 3.5 kg N hs-1 in the 1st year and mineralized 4.5 kg N ha-1 in the 2nd year. Thus, in Alaska, residue management does not affect N fertilizer requirements in the 1st year, but an additional 13.5 kg N ha-1 is required for surface residues in the 2nd year.  相似文献   

18.
Mulching of Sclerotium oryzae infested soil (moist or dry) with polyethylene sheets during hot summer days of May and June increased the soil temperature at 5 cm from 36°C (unmulched) to 48°C (wet) and from 44 to 52°C (dry) and at 20cm from 32 to 38°C (wet) and from 35 to 39°C (dry). In artificially-infested soil, the sclerotia were not eradicated but 95–100% loss in viability was observed at 5 cm by a mulch treatment for 1 week and at 20 cm by mulching for 8 weeks. Mulching effects were not influenced by moisture content of soil or by amendments with lucerne or wheat straw. Mulching of naturally-infested soil at a second site did not eradicate S. oryzae but reduced sclerotial viability by 93%.  相似文献   

19.
The ability of Trichoderma harzianum isolate 203 to attack the soil-borne plant pathogen Sclerotium rolfsii is apparently connected with the production by the isolates of chitinase and β-(1,3)-glucanase inside the attacked sclerotia during parasitism.SEM and TEM micrographs show that the mycoparasite degraded walls of sclerotial cells and the attacked cells lost their cytoplasmic contents. It is assumed that T. harzianum utilizes sclerotial cell contents thus enabling it to sporulate intensively on the sclerotial surface and inside the digested cells.  相似文献   

20.
ABSTRACT

The neutral monosaccharide composition of forest soils differs from that of non-forest soils suggesting there is an accumulation of microbial saccharides. Ectomycorrhizal (ECM) fungi can be responsible as the fungi are typical in forest soils. We investigated neutral saccharides of ECM fungal sclerotia to determine what part it might play in the origin of forest soil polysaccarides. Sclerotial grain (SG) was collected from the O, A1 and A2 horizons of a soil of subalpine forest of Mt. Ontake, central Japan. Neutral saccharides in soil and SG were analyzed by two step hydrolysis with sulfuric acid and gas-chromatography of alditol acetate derivatives. Saccharides accounted for 6.0?16% of the SG by carbon content. The SG contained predominantly easily hydrolysable (EH)-glucose, which accounted for 75–85% of the composition depending on grain size and the soil horizon, followed by mannose (7.7?15%), galactose (2.2?4.8%) and non-easily hydrolysable (NEH)-glucose (1.7?6.1%). The SG contained all of these sugars irrespective of its size. The SG collected from the A1 and A2 horizons contained all sugar components found in that from the O horizon, except for fucose in that from A2 horizon. The monosaccharide composition of SG indicates that accumulation of ECM fungal sclerotial polysaccharides might have been responsible for enlarging the molar ratios of (galactose + mannose) /(arabinose + xylose) and EH-glucose/NEH-glucose of forest soils. The proportions of SG saccharides relative to soil saccharides were 3.6, 1.2, and 0.83% for the O, A1 and A2 horizons, respectively. These levels of the proportion are considerable as ECM fugal sclerotia are the products of a limited species among hundreds and thousands of microbial species inhabiting forest soils. The sclerotia forming ECM fungal species such as Cenococcum geophilum may be key sources of forest soil polysaccharides.  相似文献   

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