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1.
Persistence of Bacillus thuringiensis parasporal protein-crystal in an arable soil (pH 5.2) incubated at ?0.01 MPa at 25°C, was monitored by radioisotope labelling (evolution of 14CO2 from soil with or without (±) organic supplement, inoculated with 14C-labelled crystal) and by bioassay (loss of insecticidal activity toward susceptible insect larvae, in soil ± CaCO3 ± organic supplement). The protein-crystal was decomposed by soil microorganisms. The extent of decomposition was significantly reduced (P < 0.05) by addition of a soluble organic supplement to soil. Insecticidal activity fell rapidly at exponential rates in all soil treatments. The loss rate did not diner significantly between soil and soil + CaCO3 (t0.5 = 2.7 days), but was significantly reduced in soil + organic supplement (t0.5 = 5.2 days), and in soil + CaCO3 + organic supplement (t0.5= 5.8 days).  相似文献   

2.
Reduction in the soil moisture from a potential of ?10 to?108 kPa reduced the growth of Gaeumannomyces graminis. The antagonist Bacillus cereus only spread through the soil or sand at potentials between ?10 and ?60 kPa. The spread of B. pumilus was not affected by the water potential within the range tested. Both B. cereus and B. pumilus were less antagonistic in the drier sand and soil. When the bacteria and the fungus were grown together the maximum growth of the fungus occurred at those potentials when bacterial antagonism was reduced but the fungus was still able to grow.  相似文献   

3.
The possible transfer of genes from Bacillus thuringiensis subsp. kurstaki (Btk) to indigenous Bacillus spp. was investigated in soil samples from stands of cork oak in Orotelli (Sardinia, Italy) collected 5 years after spraying of the stands with a commercial insecticidal preparation (FORAY 48B) of Btk. Two colonies with a morphology different from that of Btk were isolated and identified as Bacillus mycoides by morphological and physiological characteristics and by 16S rDNA analysis. Amplification by the polymerase chain reaction (PCR) of the DNA of the two isolated B. mycoides colonies with primers used for the identification of the Btk cry genes showed the presence of a fragment of 238 bp of the cry1Ab9 gene that had a similarity of 100% with the sequence of the cry1Ab9 gene present in GenBank, indicating that the isolates of B. mycoides acquired part of the sequence of this gene from Btk. No cells of Btk or B. mycoides carrying the 238-bp fragment of the cry1Ab9 gene were isolated from samples of unsprayed control soil. However, the isolates of B. mycoides were not able to express the partial Cry1Ab protein. Hybridization with probes for IS231 and the cry1Ab9 gene suggested that the inverted repeated sequence, IS231, was probably involved in the transfer of the 238-bp fragment from Btk to B. mycoides. These results indicate that transfer of genes between introduced Btk and indigenous Bacillus spp. can occur in soil under field conditions.  相似文献   

4.
The effects of two plant species—soybean (Glycine max), and sunflower (Helianthus annuus) on nutrient availability in two soils (an organically farmed soil, OF, and a native grassland soil, GS, both Alfisols)—were measured with anion exchange membranes (Plant Root Simulator, PRSTM probes) in a greenhouse study. Vegetation (especially sunflower) in the OF soil caused significant reductions in soil N and K availability (which was interpreted as due to uptake), and significant increases in P, S, Cu, Fe, Mn, and Zn availability. The increases in the latter case were consistent with the results of a previous study showing rhizosphere-enhanced mineralization of native soil organic matter in this soil. Vegetation had no significant effects on Ca, Mg, B, or Al availability in the OF soil and no significant effects on any measured nutrient in the GS soil. Collectively, these results show that the presence of plants can have either a negative or a positive effect on soil nutrient availability, and that plant uptake and soil nutrient availability are interdependent.  相似文献   

5.
The effects of temperature and water on the growth of the take-all fungus, Gaeumannomyces graminis var. tritici (Ggt), were examined in two factorial experiments. The first examined the effects of temperature and water potential on the growth of two isolates of Ggt on agar media, using osmotically-adjusted water potentials. The second experiment was concerned with the growth of the Ggt isolates in one sterile and two natural soils at two water regimes in the absence of a living host. Three temperatures (10, 18 and 26°C) were used in these experiments. A third experiment determined growth through soil.Growth was greatest at high temperatures and low water potential in axenic culture, but in unsterile soil growth at different temperatures and water potentials was strongly influenced by competition from the soil biota. The best temperature for growth in unsterile soil was 18°C. Growth at 26°C in unsterile soil was greatly reduced, this being attributed to more intense microbial competition. In sterile soil Ggt grew equally well at 18 and 26°C. At 10°C, both isolates of Ggt grew better in unsterile soil than in sterile soil.Under suitable conditions Ggt grew out readily from infected straw into unsterile soil (up to 5 cm in 10 days) in the absence of a host plant, forming melanized, hyaline and branched hyphae. These hyphae were infectious after dry storage for 5 months in the laboratory. Ggt thus appears to be a more successful soil inhabitant than is widely believed. Our experiments could explain many of the host-based concepts related to field expression of disease.The technique presented here could be of value for testing the suppressiveness or conduciveness of soils by measuring fungal growth in soil.  相似文献   

6.
A potassium-releasing bacterial strain Bacillus edaphicus NBT was examined for plant-growth-promoting effects and nutrient uptake on cotton and rape in K-deficient soil in pot experiments. Inoculation with bacterial strain B. edaphicus NBT was found to increase root and shoot growth of cotton and rape. Strain NBT was able to mobilize potassium efficiently in both plants when illite was added to the soil. In cotton and rape growing in soils treated with insoluble potassium and inoculated with strain NBT, the potassium content was increased by 30 and 26%, respectively. Bacterial inoculation also resulted in higher N and P contents of above ground plant components. The bacterial isolate was also able to colonize and develop in the rhizosphere soil of cotton and rape after root inoculation.  相似文献   

7.
Clay minerals have been shown to reduce the extent and rate of biodegradation of several compounds. Here, we investigated the ability of soil clays to protect proteins from biodegradation: the insecticidal protein from Bacillus thuringiensis subsp. tenebrionis (Btt toxin) and Bovine Serum Albumin (BSA). The two proteins adsorbed in large amounts (up to 0.24 g BSA g−1 clay and 0.74 g Btt toxin g−1 clay) and irreversibly to smectite clay particles from a vertisol. We measured the growth of a soil inoculum in the presence of each of proteins as the sole source of carbon. When clay was present in the medium, microbial growth was directly proportional to the amount of free protein (i.e. nonadsorbed). Hence, the two proteins were unavailable when adsorbed to clay. The clay had little influence on the ability of microorganisms to hydrolyse a soluble substrate. The inhibitory effect of clays on utilisation of BSA and Btt toxin was interpreted as being the result of the adsorption of the proteins to clay, which rendered the proteins unavailable for microbial utilisation.  相似文献   

8.
The efficacy of Bacillus cereus X5 as a potential biological control agent against root-knot nematodes was evaluated in vitro by examining second-stage juvenile mortality and egg hatching rate under addition of culture filtrate and in planta by application of bio-organic fertilizers enhanced with B. cereus X5, B. thuringiensis BTG, or Trichoderma harzianum SQR-T037 alone or together in greenhouse and field experiments. The biofumigation of the root-knot nematode-infested soil with organic materials (chicken manure, pig manure and rice straw) alone or in combination with B. cereus X5 was also conducted in greenhouse experiments. In laboratory, the filtrate of B. cereus X5 more effectively reduced egg hatching rates during the incubation period for 14 d and more effectively killed the second-stage juvenile during the incubation period of 24 h than that of B. thuringiensis BTG. The highest dry shoot weights for greenhouse tomatoes and field muskmelons were found in both the treatment consisting of the bio-organic fertilizer enhanced with the three biocontrol agents and the treatment consisting of the bio-organic fertilizer enhanced only with B. cereus X5. The two bio-organic fertilizers achieved better nematicidal effects than those enhanced only with B. thuringiensis BTG or T. harzianum SQR-T037. B. cereus X5 also enhanced effect of biofumigation, which resulted in increased plant biomass and reduced nematode counts in the roots and rhizosphere soil. Therefore, these results suggested that biological control of root-knot nematodes both in greenhouses and fields could be effectively achieved by using B. cereus X5 and agricultural wastes.  相似文献   

9.
The biocontrol agents Coniothyrium minitans and Bacillus subtilis MBI 600 were added separately to three soil types that had been either sterilised, pasteurised or left non-sterile. Applied as a conidial suspension of 1×106 cfu g−1 soil, C. minitans showed good survival in all sterilised, pasteurised and non-sterile soils, remaining at the numerical level at which it was applied for the duration of the 30 d experiment. Applied at a lower rate of 1×103 cfu g−1 soil, C. minitans proliferated in sterilised soil to numbers slightly over 1×106 cfu g−1 soil, whereas no increase was seen in pasteurised or non-sterile soils from this lower application rate. However, although C. minitans was not easily recovered on plates from non-sterile soil, it did survive at the lower numerical level in pasteurised soil, and was recoverable throughout the experiment at the rate at which it was applied. B. subtilis MBI 600 survived well following introduction as a cell suspension into sterilised soil at a rate of 1×106 cfu g−1 soil. Spores were formed rapidly and, after 14 d, the introduced microorganism survived in this form rather than as vegetative cells. However, in non-sterile soil, the introduced microorganism did not compete well and decreased in number, with spores being formed in low numbers. Survival of B. subtilis MBI 600 in pasteurised soil was variable, but resembled the survival seen in non-sterile soil more than that seen in sterilised soil. More B. subtilis MBI 600 spores were formed in pasteurised soil than in non-sterile soil, however, and may have been important for survival in pasteurised soil. In conclusion, this work has shown that the biocontrol agent C. minitans can survive well in soil irrespective of whether the soil has been pasteurised or not and shows good promise as a soil inoculant for control of Sclerotinia sclerotiorum. Although soil pasteurisation does improve establishment of B. subtilis MBI 600 compared to non-sterile soil, survival is relatively poor when applied as cells. The best survival of B. subtilis MBI 600 occurred as spores in sterilised soil, and spore applications to pasteurised soil in an integrated control strategy may allow sufficient establishment of the biocontrol agent to target pathogens causing damping-off.  相似文献   

10.
Characteristics of the predominant bacteria isolated in November and May from the forest soils of both dry and wet types under natural vegetation were studied.

Although Gram-negative rods were the most abundant bacteria in both soil types and in both seasons, their contents were less and other bacteria especially spore-forming ones increased in May.

Among Gram-negative rods in the soil of the dry type in November, the most predominant was those with nonchromogenic rods motile with polar flagella which grew in a simple synthetic media containing glucose or p-hydroxybenzoate and ammonium as the sole carbon and nitrogen sources. In May, the ratio of the bacteria which require amino nitrogen or those with more chomplex nutritional requirement increased. In the soil of moderately wet type, the difference in kinds of bacteria between the two seasons was not so clear as that in the dry type.

Most of Bacillus species obtained in these soils were those requiring amino acids or other growth factors among which B. cereus was most abundant.  相似文献   

11.
Our previous studies showed that, under P-limiting conditions, growth and P uptake were lower in the wheat genotype Janz than in three Brassica genotypes when grown in monoculture. The present study was conducted to answer the question if P mobilised by the Brassicas is available to wheat; leading to improved growth of wheat when intercropped with Brassicas compared to monocropped wheat. To assess if the interactions between the crops depend on soil type, the wheat genotype Janz and three Brassica genotypes (two canolas and one mustard) were grown for 6 weeks in monoculture or wheat intercropped with each Brassica genotype in an acidic and an alkaline soil with low P availability (with two plants per pot). Wheat grew equally well in the two soils, but the Brassicas grew better in the acidic than in the alkaline soil. In the acidic soil, monocropped Brassicas had a 3 to 4 fold greater plant dry weight (dw) and P uptake than wheat; plant dw and P uptake in wheat were decreased or not affected by intercropping and increased in the Brassicas. In the alkaline soil, dw and P uptake of the Brassicas was twice as high as in wheat, with intercropping having no effect on these parameters. The contribution of wheat to the total shoot dw and P uptake per pot was 4-21% and 32-40% in acidic and alkaline soil, respectively. Mycorrhizal colonisation was low in all genotypes in the acidic soil (1-6%). In the alkaline soil, mycorrhizal colonisation of monocropped wheat was 62%, but only 43-47% in intercropped wheat. Intercropping decreased P availability in the rhizosphere of wheat in the acidic soil but had no effect on rhizosphere P availability in the alkaline soil. Intercropping had a variable effect on rhizosphere microbial community composition (assessed by fatty acid methylester analysis (FAME) and ribosomal intergenic spacer amplification (RISA)), ranging from intercropping having no effect on the rhizosphere communities to intercropping resulting in a new and similar rhizosphere community composition in both genotypes. The results of this study show that intercropping with Brassicas does not improve growth and P uptake of wheat; thus there is no indication that P mobilised by the Brassicas is available to wheat.  相似文献   

12.
It is known that methylthio-s-triazines can be transformed to the corresponding 2-hydroxy derivatives through sulfoxides and sulfones in aerobic and flooded soil; however, production of sulfoxides and/or sulfones from methylthio-s-triazines by isolated s-triazine-degrading bacteria has not been reported yet. In the present study, a new bacterial strain, JUN7, was obtained from Japanese soil; the bacterium is capable of transforming simetryn to 2-methylsulfinyl 4,6-bis(ethylamino)-s-triazine (sulfoxide simetryn) and 2-hydroxy-4,6-bis(ethylamino)-s-triazine (2-hydroxy simetryn) in a Luria-Bertani (LB) medium. This is the first isolation of the specific microorganism that mediates sulfur oxidation of methylthio-s-triazines, as far as we know. Strain JUN7 could decrease other methylthio-s-triazines such as dimethametryn and prometryn, but not chlorinated s-triazines (atrazine, simazine, and terbuthylazine) and methoxy-s-triazine (atraton) in 1/10 LB medium. Strain JUN7 did not possessed gene atzA or trzN encoding triazine-degrading enzymes, suggesting that the strain may have another metabolic system. Characterizations of strain JUN7 based on comparative morphology, physiological classification, and comparison of the partial 16S rRNA sequence indicated that it is assigned as a Bacillus cereus species.  相似文献   

13.
Sudan is the fourth largest exporter of groundnuts in the world, yet little is known concerning the plant-rhizobial symbiosis. A study was made on the abundance of groundnut-nodulating rhizobia in the soils of Sudan as related to soil properties and the duration since groundnuts were last planted. Also, physiological, serological and nitrogen-fixing characteristics of Sudanese rhizobia are reported. All but one of 32 sites contained more than 300 rhizobia g?1 soil capable of forming nodules on siratro (Macroptilium atropurpureum). Several of these soils had never been planted to groundnut. A correlation matrix indicated no relationship was present between soil rhizobial populations and any of the measured soil properties, or between soil rhizobial populations and the time since groundnuts were last planted in the rotation. Individual isolates of Rhizobium from six legumes: groundnut (Arachis hypogaea), mung bean (Vigna radiata), lubia (Dolichos lablab), cowpea (Vigna unguiculata), pigeonpea (Cajanus cajan) and bambara groundnut (Voandzeia subterranea) were obtained from four locations in Sudan. All isolates were able to nodulate each of the six legumes when grown in sterile vermiculite. The isolates grew in 0.1% NaCl-amended media, but growth was variable in 2.0% amended media. Most isolates grew after exposure to moist heat for 15 min at 50°C. Optimum pH for growth was, in general, between pH 6 and 8. Agglutination reactions indicated isolates from groundnuts, as well as isolates from other legumes, belonged to several serological groupings. Some isolates formed a large number of nodules on a Sudanese groundnut cultivar, whereas other isolates formed only few nodules.  相似文献   

14.
Sprays of commercial insecticidal preparations of the bacterium, Bacillus thuringiensis subsp. kurstaki (Btk), usually a mixture of cells, spores and parasporal crystals, have been used for the last 10 yr in Sardinia (Italy) to protect cork oak forests against the gypsy moth (Lymantria dispar L.). Until now, the protective antilepidopteran efficacies of each of the various spray treatments rather than their effects on the environment have been evaluated. Consequently, the persistence of Btk and its toxin, released in sprays (FORAY 48B®), in soils of cork oak stands, located in Orotelli, Tempio Pausania and Calangianus (Sardinia), were investigated. In the Calangianus soil, the numbers of Btk remained essentially constant for 28 months (the longest time studied) after spraying, indicating that Btk was able to compete with the indigenous microbial community; the toxin was detected 28 months after spraying by immunological assay, but at a reduced concentration; and the larvicidal activity decreased essentially linearly to 14 months and then decreased markedly between 14 and 28 months. In the Tempio Pausania and Orotelli soils, cells of Btk were detected, whereas the toxin was not detected by immunological and larvicidal assays, 52 and 88 months (the longest times studied) after spraying, respectively. The numbers of Btk cells detected were probably too low to account for the presence of the toxin in all of the soils studied, as there was no correlation between numbers of Btk and toxin detected by immunological assays (correlation coefficient of −0.66) in the Calangianus soil. Our results indicated that Btk and its toxin introduced into soils in sprays can persist for long periods (at least 88 months for Btk and at least 28 months for its toxin).  相似文献   

15.
Genetically modified crops, which produce pesticidal proteins from Bacillus thuringiensis, release the toxins into soils through root exudates and upon decomposition of crop residues. Although the phenomena of gene transfer and emergence of resistance have been well documented, the fate of these toxins in soil has not yet been clearly elucidated. The aim of this study was to elucidate the adsorption and the desorbability of the Cry1Aa Bt insecticidal protein in contact with two sodium-saturated clays: montmorillonite and kaolinite. Because the toxin is released into soil in small quantities, it was assumed that it will be in a monomeric state in solution until it oligomerized on cell membranes. The originality of this study was to focus on the monomeric form of the protein. Specific sample conditions were required to avoid polymerisation. A pH above 6.5 and an ionic strength of at least 150 mM (NaCl) were necessary to keep the protein in solution and in a monomeric state. The adsorption isotherms obtained were of the L-type (low affinity) for both clays and fitted the Langmuir equation. The adsorption maximum of the toxin, calculated by the Langmuir nonlinear regression, decreased with increasing pH from 6.5, which was close to the isoelectric point, to 9. At pH 6.5, the calculated adsorption was 1.7 g g−1 on montmorillonite and 0.04 g g−1 on kaolinite. Desorbability measurements showed that a small fraction of toxin could be desorbed by water (up to 14%) and more by alkaline pH buffers (36 ± 7%), indicating that it was not tightly bound. Numerous surfactants were evaluated and the toxin was found to be easily desorbed from both clays when using zwitterionic and nonionic surfactants such as CHAPS, Triton-X-100, and Tween 20. This finding has important implications for the optimization of detection methods for Bt toxin in soil.  相似文献   

16.
Biological control agents like Bacillus subtilis offer an alternative and supplement to synthetic pesticides. Antibiotic production by biocontrol strains of B. subtilis can play a major role in plant disease suppression. Our current understanding of B. subtilis antibiosis comes from culture media measurements of antibiotic production and in vitro suppression of pathogens. Quantifying the antibiotic metabolite chemistry of B. subtilis biofilms growing on root surfaces provides a more accurate understanding of in vivo antibiotic production. An analytical method based on solid-phase extraction (SPE) with high-performance liquid chromatography (HPLC) and mass spectroscopy (MS) has been developed to quantify antibiotics produced by B. subtilis growing on plant roots. Cucumber (Cucumis sativus) was grown in composted soil and potting media inoculated with B. subtilis strain QST 713 (AgraQuest, USA). Two important B. subtilis antibiotics, surfactin and iturin A, were extracted from root and rhizosphere soil using acidified organic solvents followed by cleaning and concentration using SPE. HPLC and HPLC-MS were used to measure surfactin and iturin A. Rhizosphere concentrations of both antibiotics increased with plant age. For plants grown in peat-based potting media, surfactin concentrations increased from 9 μg g−1 root fresh weight (RFW) at 15 d to 30 μg g−1 RFW at 43 d. Iturin concentrations were 7 μg g−1 RFW at 15 d and 180 μg g−1 RFW at 43 d. In an initial field trial in a composted fine sandy loam, we demonstrated rhizosphere production of surfactin and iturin under competition and predation by the myriad macro- and microfauna existing in a fertile high-organic soil, with mature B. subtilis-inoculated cucumber roots yielding 33 μg g−1 RFW surfactin and 630 μg g−1 RFW iturin at 78 d.  相似文献   

17.
The effects of maize expressing the Bacillus thuringiensis Cry1Ab protein (Bt maize) on decomposition processes under three different European climatic conditions were assessed in the field. Farming practices using Bt maize were compared with conventional farming practices using near-isogenic non-Bt maize lines under realistic agricultural practices. The litter-bag method was used to study litter decomposition and nitrogen mineralization dynamics of wheat straw. After 4 months incubation in the field, decomposition and mineralization were mainly influenced by climatic conditions with no negative effect of the Bt toxin on decomposition processes.  相似文献   

18.
The ecology of bacteriophages attacking the thermophilic soil organism Bacillus stearothermophilus has been investigated. Phage was found in every soil tested including a sample from the geothermal area of Taupo. Since the numbers of plaque-forming units (pfu) in most soils were often low a technique for enriching soil with substrate for the host cells and factors promoting phage proliferation was developed; this enabled statistically significant numbers of pfu to be scored under a variety of test conditions. Phage development in soil was greatest at 45°C while proliferation at 55°C was usually much less pronounced. These results indicate that in soil these bacteria and their associated phages grow optimally at mesophilic rather than thermophilic temperatures. It is proposed that in the field the bulk of the metabolizing population of B. stearothermophilus is mesophilic in its biochemistry and that the ‘thermophilic’ strains of this organism held by many laboratories represent selection by the isolation procedure of a minority of atypical cells.  相似文献   

19.
Lineal extension of Gaeumannomyces graminis var. tritici hyphae along roots of intact wheat plants growing in soils was measured. Hyphal growth rates were lower in soils treated with NH4+-N than with NO3?-N. In a soil that is suppressive to the take-all disease, the controlling influence of NH4+-N was eliminated by soil fumigation (methyl bromide), and reintroduced to fumigated soil by additions of 1% nonsterile soil. Effects of fumigation on hyphal growth were absent in a nonsuppressive soil, and in NO3?-treatments of the suppressive soil. When inocula of selected groups of wheat rhizoplane microflora were reintroduced into a fumigated or a soil-reinoculated soil via a root-food base, the Pseudomonas spp. consistently appeared more suppressive in NH4+-N treatments than the general bacterial flora, Bacillus spp. spores, streptomycetes, and fungi.  相似文献   

20.
In acid soil, low pH, reduced availability of nutrients, and toxicity of Al and Mn limit plant growth and the survival and effectiveness of rhizobia. The symbiosis between legumes and rhizobia is particularly sensitive to acid soil stress. A pot experiment evaluated whether Bradyrhizobium japonicum strain growth on acidic agar media would predict ability to colonize the rhizosphere and form effective nodules in acidic soils. Three Indonesian strains of B. japonicum with similar effectiveness at neutral pH in sand culture but with different tolerance of acid soil stress factors in agar media, and an acid-tolerant commercial strain (CB1809) of comparable effectiveness, were tested in three acid soils using the Al tolerant soybean (Glycine max cv PI 416937). At 7 days after inoculation all strains had achieved large rhizosphere populations, but by day 14 the rhizosphere population of the acid-sensitive strain had decreased, while the more acid-tolerant strains increased. The acid-tolerant strains had significantly greater nodulation and symbiotic effectiveness than plants inoculated with the acid-sensitive strain. Laboratory prescreening of B. japonicum for acid, Al and Mn tolerance in acid media successfully identified strains which were symbiotically competent in low pH soils.  相似文献   

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