Survival of sclerotia of Macrophomina phaseolina and Sclerotium cepivorum after drying and wetting treatments

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.     

Diallyl-disulphide to reduce the numbers of sclerotia of Sclerotium cepivorum in soil

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.     

Parasitism of sclerotia of Sclerotium rolfsii by trichoderma harzianum

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.     

Polyethylene mulching of soil to reduce viability of sclerotia of Sclerotium oryzae

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%.     

Survival of sclerotia of sclerotinia sclerotiorum in soil

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.     

Survival of Coniothyrium minitans associated with sclerotia of Sclerotinia sclerotiorum in soil

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.     

Some factors affecting survival of sclerotia of Macrophomina phaseolina in soil

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).     

Spatial distribution of the nematode biocontrol agent Pasteuria penetrans as influenced by its soil habitat

Root-knot nematodes belonging to the Meloidogyne genus are ubiquitous plant-parasitic pests, especially on vegetables. The Pasteuria penetrans bacterium is an obligate parasite of nematodes, parasitizing most of the Meloidogyne species. Spatial distributions of Meloidogyne javanica populations infested or not by P. penetrans and of bacterial populations were studied in a vegetable plot naturally infested by these organisms. It was observed that distributions of M. javanica populations, of populations infested by P. penetrans, and of free bacteria populations were not overlapped. Soil factors involved were investigated. Soil texture and water flow in porosity are concerned, as they directly influence the level of the pool of bacteria and then the chances of both organisms to meet. The soil solution has a direct effect on the attachment of the bacterium on the nematode cuticle.     

Reduction in viability of sclerotia of Macrophomina phaseolina with polyethylene mulching of soil

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.     

Solubility of soil phosphorus as influenced by urea

The objective of this study was to investigate possible ways of mobilizing residual fertilizer P as a result of local pH elevation caused by urea hydrolysis. The response of water-soluble P (P_w) and dissolved organic C (DOC) to urea hydrolysis was monitored in three cultivated soils and at two P levels for up to 127–135 d and compared with corresponding changes in soils limed with Ca(OH)₂. Hydrolysis of urea was complete in 8–15d during which soil pH increased by 1–1.5 units at the maximum. Subsequently, the pH decreased to or below the original level owing to nitrification. Mobilization of soil P was enhanced substantially in parallel with the increase in pH, the peak P_w occurring simultaneously with the highest pH value. In all urea-treated soils, P_w remained at an elevated level for at least 60d. As compared to urea, elevation of soil pH with Ca(OH)₂ had only a minor and inconsistent influence on P_w. In mobilization of soil P, the urea-induced increase in pH and a simultaneous production of NH₄⁺ ions proved to be superior to liming with Ca(OH)₂. It was hypothesized that when an acid soil is amended with urea, phosphate is first displaced by OH^? ions, resulting in elevated solution P concentrations. A simultaneous dissolution of organic matter contributes to the persistence of high P concentration by competition for sorption sites on Fe and Al oxides, and thus retards the resorption of P.     

Properties of soil aggregates as influenced by tillage practices

Abstract. Changes in aggregate stability, density, and porosity as well as the water retention and nutrient contents of different aggregate size fractions due to intensive tillage were investigated. Three soils from Vicarello, Fagna and Gambassi in North Central Italy which had been under permanent vegetation, minimum or conventional tillage for more than seven years were studied. The aggregates on conventionally tilled plots were slightly denser and less porous than those on the untilled or minimum-tilled plots. The aggregates were less stable under conventional tillage on all soils. Conventional tillage reduced the proportion or macro-aggregates by 22% at Vicarello and 35% at Gambassi. There were no differences in macro-aggregate proportions between minimum- and conventionally tilled plots at Fagna. The potential of the dry aggregates to distintegrate upon contact with water was greatest in the conventionally tilled and least in the untilled treatments. The proportions of dry macro-aggregates (> 0.25 mm) in the untilled and tilled plots were 90 and 71%, respectively. The soil of the tilled plots contained less carbon and nitrogen than that of the untilled plots in all aggregate size fractions. The silt-plus-clay contents of the aggregates accounted for between 65 and 93% of variability in the water they retained at small potentials while organic carbon contents accounted for between 71 and 90% of variability in the stability of the aggregates irrespective of the tillage treatments.     

Nitrate leaching as influenced by soil tillage and catch crop

Because of public and political concern for the quality of surface and ground water, leaching of nitrate is of special concern in many countries. To evaluate the effects of tillage and growth of a catch crop on nitrate leaching, two field trials were conducted in spring barley (Hordeum vulgare L.) under temperate coastal climate conditions. On a coarse sand (1987–1992), ploughing in autumn or in spring in combination with perennial ryegrass (Lolium perenne L.) as a catch crop was evaluated. Furthermore, rotovating and direct drilling were included. The experiment was conducted on a 19-year-old field trial with continuous production of spring barley. On a sandy loam (1988–1992), ploughing in autumn or in spring in combination with stubble cultivation and perennial ryegrass, in addition to minimum tillage, was evaluated in a newly established field trial. For calculation of nitrate leaching, soil water isolates from depths of 0.8 or 1.0 m were taken using ceramic cups. No significant effect of tillage was found on the coarse sand; however, a significant effect of tillage was found on the sandy loam, where leaching from autumn ploughed plots without stubble cultivation was 16 kg N ha⁻¹ year⁻¹ higher than leaching from spring ploughed plots. Leaching was significantly less when stubble cultivation in autumn was omitted. Leaching on both soil types was significantly reduced by the growth of a catch crop which was ploughed under in autumn or in spring. It was concluded that soil cultivation increased leaching on the sandy loam but not on the coarse sand, and that the growth of perennial ryegrass as a catch crop reduced leaching on both soil types, particularly when ryegrass was ploughed under in spring.     

Aluminium chemistry of the soil solution in an acid forest soil as influenced by percolation rate and soil structure

The predicted activity of Al in the soil solutions of acid forest soils often differs from that observed in the field. We have investigated the influence of soil structure and flow rate of the soil solution on the aluminum release to explain this divergence. Disturbed and undisturbed samples of soil were collected from the A and B horizons of a dystric cambisol at Waldstein (Fichtelgebirge, Germany). The samples were irrigated with solutions mixed according to field data on throughfall or soil solution composition with pH 3.5 with flow rates of 4 mm d^?1, 12 mm d^?1 and 36 mm d^?1. The percolates were analysed for major ions. Resulting relations between pH and pAl were compared with batch experiments. In neither the A horizon nor in the B horizon did soil structure influence the relation between pH and pAl. The apparent equilibrium between pH and pAl was described as the pK_app value with pK_app= pAl—a pH (where a is an empirical constant). It was found that the pK_app values for the column percolates were in the range of variation of those found in batch experiments. Flow rate had no influence on pK_app at 4 and 12 mm d^?1. At 36 mm d^?1 a significant increase of pK_app was observed. This relative undersaturation of Al was more pronounced in the A horizon than in the B horizon. When flow is fast Al release into the percolating soil solution might be limited by diffusion.     

Survival of Escherichia coli in soil with modified microbial community composition

Understanding the survival and persistence of Escherichia coli in soil with different microbial composition is essential for the accuracy of water quality assessment and microbial source tracking. This microcosm experiment was conducted to investigate the survival pattern of three E. coli strains (originated from soil, dog feces and human feces, separately) in soil with modified microbial community composition. Bile salt No. 3 (BS3) of progressively increased density (0.05%, 0.15%, 0.30% and 0.50%) was added into sandy loam soils and incubated for 90 days. Laboratory cultured E. coli were then inoculated into soil and incubated for another 150 days to monitor their survival pattern. Change of bacterial community diversity by BS3 was detected by both cultivation based and cultivation independent (PCR-Denaturing Gradient Gel Electrophoresis) methods. In general, progressively increased BS3 concentration resulted in decreased CFU counts both at 10 days and 90 days incubation. DGGE analysis indicated only a slight change in bacterial community composition at 10 days but a significant change at 90 days. Cluster analysis suggested that BS3 treatment grouped separately from controls. Survival of E. coli in soil was significantly influenced by the complexity of the microbial community, as die-off rate of E. coli progressively declined with the reduction of microbial community diversity. Differential survival of E. coli under different soil microbial stress highlights the importance of incorporating biotic factors in predictive models for water quality management and microbial source tracking study.     

Relationships between soil aggregation and mycorrhizae as influenced by soil biota and nitrogen nutrition

 The effect of the form of N nutrition on soil stability is an important consideration for the management of sustainable agricultural systems. We grew soybean [Glycine max (L.) Merr.] plants in pot cultures in unsterilized soil, and treated them by (1) inoculating them with Bradyrhizobium japonicum, fertilizing with (2) nitrate or (3) ammonia, or (4) by providing only minimum N amendment for the controls. The soils were sampled at 3-week intervals to determine changes in water-stable soil aggregates (WSA), soil pH, the development of roots, arbuscular mycorrhizal (AM) soil and root colonization, and selected functional groups of soil bacteria. The soil fauna was assayed at the end of the experiment (9 weeks). WSA was correlated positively with root and AM soil mycelium development, but negatively with total bacterial counts. Soil arthropod (Collembola) numbers were negatively correlated with AM hyphal length. Soils of nodulated and ammonia-fertilized plants had the highest levels of WSA and the lowest pH at week 9. Sparse root development in the soils of the N-deficient, control plants indicated that WSA formation was primarily influenced by AM hyphae. The ratio of bacterial counts in the water-stable versus water-unstable soil fractions increased for the first 6 weeks and then declined, while counts of anaerobic bacteria increased with increasing WSA. The numbers of soil invertebrates (nematodes) and protozoans did not correlate with bacterial counts or AM soil-hyphal lengths. Soil pH did not affect mycorrhiza development, but actinomycete counts declined with decreasing soil pH. AM fungi and roots interacted as the factors that affect soil aggregation, regardless of N nutrition. Received: 20 December 1997     

Changes in soil aggregation and glomalin-related soil protein content as affected by the arbuscular mycorrhizal fungal species Glomus mosseae and Glomus intraradices

Arbuscular mycorrhizal (AM) fungi are key organisms of the soil/plant system, influencing soil fertility and plant nutrition, and contributing to soil aggregation and soil structure stability by the combined action of extraradical hyphae and of an insoluble, hydrophobic proteinaceous substance named glomalin-related soil protein (GRSP). Since the GRSP extraction procedures have recently revealed problems related to co-extracting substances, the relationship between GRSP and AM fungi still remains to be verified. In this work the hypothesis that GRSP concentration is positively correlated with the occurrence of AM fungi was tested by using Medicago sativa plants inoculated with different isolates of Glomus mosseae and Glomus intraradices in a microcosm experiment. Our results show that (i) mycorrhizal establishment produced an increase in GRSP concentration - compared to initial values - in contrast with non-mycorrhizal plants, which did not produce any change; (ii) aggregate stability, evaluated as mean weight diameter (MWD) of macroaggregates of 1-2 mm diameter, was significantly higher in mycorrhizal soils compared to non-mycorrhizal soil; (iii) GRSP concentration and soil aggregate stability were positively correlated with mycorrhizal root volume and weakly correlated with total root volume; (iv) MWD values of soil aggregates were positively correlated with values of total hyphal length and hyphal density of the AM fungi utilized.The different ability of AM fungal isolates to affect GRSP concentration and to form extensive and dense mycelial networks, which may directly affect soil aggregates stability by hyphal enmeshment of soil particles, suggests the possibility of selecting the most efficient isolates to be utilized for soil quality improvement and land restoration programs.     

Metabolism of fluoroacetate by a soil Pseudomonas SP. and Fusarium solani

Enzymes (haloacetate halidohydrolases) capable of cleaving the C-F bond of fluoroacetate and some other organofluorine compounds have been isolated and partially purified from a soil pseudomonad and from the common soil fungus Fusarium solani. Both enzymes readily released F^? from monofluoroacetate and fluoroacetamide but were without effect on a wide range of other organic F-compounds. The enzymes also cleaved the C-Cl and C-Br bonds in mono-chloroacetate and monobromoacetate. Inorganic F^? acted as a competitive inhibitor of the enzymes. The molecular weights of both enzymes were about 62,000. Thus the properties of the halidohydrolases from both organisms were similar in many respects but the bacterial enzyme was more stable at 55°C and exhibited an unusual difference in temperature coefficient (Q₁₀ value) over its higher (30–55°C) and lower (15–30°C) temperature ranges.     

Enzymatic activities and microbial biomass in soil as influenced by metalaxyl residues

A laboratory experiment was conducted to study the impact of metalaxyl application at different concentration levels on microbial biomass and the biochemical activities in soil. A dissipation study of metalaxyl highlighted 52.5-56.8% loss of metalaxyl due to the presence of microbial activity. However, a small but significant decline in microbial biomass was observed on 60 d of incubation period. Metalaxyl showed a highly significant effect in decreasing total N and organic C content in soil from 0 to 30 d of incubation. Dehydrogenase, phosphatase, urease, arylsulphatase and β-glucosidase activities were monitored in metalaxyl treated soils. Except urease, all the enzymatic activities initially increased and then decreased. Urease activity showed a continuous gradual decrease throughout the experimental period. Thus, metalaxyl might influence the growth and development of crop-plants, since it has direct impact on nutrient recycling and energy flow in soil.     

Earthworm burrowing in laboratory microcosms as influenced by soil temperature and moisture

Earthworm burrows contribute to soil macroporosity and support diverse microbial communities. It is not well known how fluctuations in soil temperature and moisture affect the burrowing activities of earthworms. The objective of this experiment was to evaluate the maximum depth and length of burrows created by the endogeic earthworm Aporrectodea caliginosa (Savigny) and the anecic earthworm Lumbricus terrestris L. for a range of temperatures (5–20 °C) and soil water potentials (−5 and −11 kPa). The laboratory microcosm was a plexiglass chamber (45 cm high, 45 cm wide) containing 0.14 m² of pre-moistened soil and litter, designed to house a single earthworm for 7 days. Earthworm mass, surface casting and burrowing activities were affected significantly by soil temperature, moisture and the temperature×moisture interaction. Burrow length and maximum burrow depth increased with increasing temperature, but there was less burrowing in wetter soil (−5 kPa) than drier soil (−11 kPa). Weight gain and surface casting, however, were greater in soil at −5 kPa than −11 kPa. Our results suggest more intensive feeding and limited burrowing in wetter soil than drier soil. Earthworms inhabiting the non-compacted, drier soil may have pushed aside particles without ingesting them to create burrows. The result was that earthworms explored a larger volume of soil, deeper in the chamber, when the soil was drier. How these burrowing activities may affect the community structure and activity of soil microorganisms and microfauna in the drilosphere remains to be determined.