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1.
Bacillus spp. has emerged as the most effective alternative to synthetic chemical fungicides. To get a better insight in the antagonistic potential of Bacillus strains, rhizospheric soil samples of healthy tomato plants from Indo-gangetic plain regions of India were analysed. A total of 108 Bacillus strains were obtained from preliminary screening. Potent strains identified on the basis of in vitro antagonistic and biochemical assays were subjected to diversity analysis using 16S-rDNA, BOX and ERIC-PCR. Furthermore, the four best performing antagonistic Bacillus strains under in vitro plant growth promotion and antagonistic assay were selected for pot experiment. In field study, Bacillus amyloliquefaciens MB101 and Bacillus subtilis MB14 showed drastic reduction in disease index by 55.7 and 41.74% with significant elevation in fruit yield up to 220 and 184 qha–1, respectively. The present study was successful in selecting effective Bacillus strains by performing phenotypic and genotypic characterisation of Bacillus strains that can be used as an integral component of integrated disease management of tomato root rot and damping-off.  相似文献   

2.
Abstract

The antibacterial efficacy of hydrogen peroxide encapsulated in micelles (mH2O2) against biofilms was compared with that of hydrogen peroxide alone and of three commercially available aqueous biocides. The activity of mH2O2 on 24-h biofilms of reference strains of Staphylococcus spp. and Pseudomonas aeruginosa was tested in a static microtiter plate model. The biofilms were incubated with mH2O2 (17% v/v H2O2, 2% lactic acid, 0.3% phytoextract, H2O) and its individual ingredients and compared with three aqueous biocides at different concentrations and times of exposure. After 5-min exposure, 10% mH2O2 (corresponding to 1.7% v/v H2O2) achieved > 8 log10 reductions against all the test strains, while 1.7% H2O2 achieved a maximum of 1.5 log10 reduction. After 5-min exposure, none of the commercially available biocides tested showed themselves to be capable of completely eliminating the test strains embedded in biofilms. Hydrogen peroxide encapsulated in micelles demonstrated enhanced activity against planktonic cells and biofilms of Staphylococcus spp. and P. aeruginosa.  相似文献   

3.
In vitro antagonistic effects of rhizobacteria associated with Coffea arabica L. against some fungal coffee pathogens were studied. The aims were to screen indigenous coffee‐associated isolates for their inherent antagonistic potential against major coffee wilt diseases induced by Fusarium spp. Antagonistic effects, siderophore, HCN and lytic enzyme production were determined on standard solid media. Chemical methods were employed to categorize the major types of siderophores. From a total of 212 rhizobacterial isolates tested, over 10 % (all Pseudomonas and Bacillus spp.) exhibited remarkable inhibition against Fusarium spp. One isolate AUPB24 (P. chlororaphis) showed maximum inhibition of mycelial growth against all fungal pathogens tested, whereas other isolates were mostly inhibitory to F. stilboides and F. oxysporum. The isolate AUBB20 (B. subtilis) was most antagonistic to F. xylarioides. Of the rhizobacterial isolates tested, 67 % produced siderophores and 35 % produced HCN. Many strains (all Pseudomonas spp.) produced siderophores of the hydroxamate type and only a small proportion produced those of the catecholate type. Few antagonists showed chitinase activity. The production of siderophores and HCN by Pseudomonas spp., lipase and protease by all antagonists and β‐1,3‐glucanase by several Bacillus spp. could be considered the major mechanisms involved in the inhibition of fungal growth. The in vitro results provide the first evidence of an antagonistic effect of coffee‐associated rhizobacteria against the emerging fungal coffee pathogens F. stilboides and F. xylarioides and indicate the potential of both bacterial groups for biological control of coffee wilt diseases.  相似文献   

4.
Bacillus subtilis KS1 was isolated from grape berry skin as a biological control agent against grapevine fungal diseases. KS1 was identified as a new strain of B. subtilis according to morphological, biochemical, and genetic analyses. In vitro bioassay demonstrated that KS1 suppressed the growth of Botrytis cinerea (the casual agent of grape grey mold) and Colletotrichum gloeosporioides (the casual agent of grape ripe rot). The biocontrol activity of KS1 against grapevine fungal diseases in vineyards was evaluated over a 3-year span (from 2007 to 2009). Downy mildew, caused by Plasmopara viticola, was reduced on berry skins and leaves by treatment with KS1. The KS1 genome possesses ituD and lpa-14 genes, both of which play a role in iturin A production followed by iturin A production in the culture. In contrast, mutants lacking both genes lost the antagonistic activity against B. cinerea and C. gloeosporioides and the activity in iturin A production, suggesting that the antagonistic activity of KS1 against grapevine fungal pathogens may depend on iturin A production. As KS1 showed tolerance to various chemical pesticides, chemical pesticides could be applied before and/or after KS1 treatment in vineyards. Due to its potential as a biological control agent against grape downy mildew, KS1 is expected to contribute to the further improvement of integrated pest management systems and to potentially reduce the amount of chemical fungicides applied in vineyards.  相似文献   

5.
Antifouling (AF) paints are used to prevent the attachment of living organisms to the submerged surfaces of ships, boats and aquatic structures, usually by the release of biocides. Apart from copper, organic booster biocides are the main active components in AF paints, but their use can have a negative impact on the marine environment. The direct effects of biocides on marine bacteria are poorly known. This work investigates the impact of two biocides, viz. diuron and tolylfluanid, on the growth and the viability of marine microorganisms and on their ability to form biofilms. The biocides in solution were found to inhibit growth of two strains of marine bacteria, viz. Pseudoalteromonas and Vibrio vulnificus, at a high concentration (1000 μg ml?1), but only a small effect on viability was observed. Confocal laser scanning microscopy (CLSM) showed that the booster biocides decreased biofilm formation by both bacteria. At a concentration of 10 μg ml?1, the biocides inhibited cell attachment and reduced biofilm thickness on glass surfaces. The percentage of live cells in the biofilms was also reduced. The effect of the biocides on two diatoms, Fragilaria pinnata and Cylindrotheca closterium, was also evaluated in terms of growth rate, biomass, chlorophyll a content and attachment to glass. The results demonstrate that diuron and tolylfluanid are more active against diatoms than bacteria.  相似文献   

6.
The properties of new B. subtilis strains GM2 and GM5, isolated from potato rhizosphere and possessing high antimicrobial activity, were studied. The potential of the strains for their use as probiotics was characterized. The strains were resistant to bile and to a wide range of the ambient pH. B. subtilis strains GM2 and GM5 possessed proteolytic and phytate-hydrolyzing activity and proved to be safe for model animals. The strains were characterized by antagonistic properties against phytopathogenic micromycetes, as well as against pathogenic and opportunistic enterobacteria. B. subtilis GM2 and GM5 were concluded to be promising strains for use as probiotics.  相似文献   

7.
Aims: To identify bacilli, lactic acid bacteria and bifidobacteria that inhibit the growth of Brachyspira hyodysenteriae. Methods and Results: A total of 80 isolates were obtained from various porcine intestinal compartments using selective conditions and grouped into 15 similarity clusters based on whole‐cell protein profiles. Random amplified polymorphic DNA PCR patterns identified 24 genotypes. 16S rDNA sequencing assigned all genotypes, except eight aerobes, to established species (Bacillus subtilis, Enterococcus faecium, Lactobacillus salivarius, Lactobacillus mucosae, Lactobacillus reuteri, Lactobacillus amylovorus, Bifidobacterium thermophilum). According to their minimum inhibitory concentrations, four strains (Ent. faecium, Lact. reuteri, Lact. amylovorus, Bif. thermophilum) were susceptible to all clinically relevant antibiotics. Two lactobacilli showing multiresistance harboured the erm(B) determinant. A cross‐section of eight representative strains was examined for growth suppression of two strains of Brach. hyodysenteriae, the aetiological agent of swine dysentery, and compared with intestinal strains derived from other animal sources. The Brachyspira strains were inhibited by strains of Lact. salivarius, Bif. thermophilum, Ent. faecium and B. subtilis. Conclusions: Three porcine strains of Ent. faecium, Bif. thermophilum and B. subtilis were found to be suitable as probiotic candidates because of their well‐established identity, antibiotic susceptibility and antagonistic activity. Significance and Impact of the Study: For the first time, antagonistic activity of well‐characterized porcine strains against Brach. hyodysenteriae is presented. These findings suggest that certain intestinal strains might have a potential as probiotic feed additives for prevention of swine dysentery.  相似文献   

8.
The antagonistic activity of Bacillus subtilis strain G1 was tested against various isolates of Aspergillus flavus in vitro. A talc-based powder formulation of B. subtilis strain G1 was prepared and evaluated to control A. flavus infection and aflatoxin B1 contamination in groundnut under greenhouse and field conditions. The results showed that B. subtilis strain G1 could inhibit the growth of all isolates of A. flavus tested in dual culture assay and the growth inhibition ranged from 93 to 100%. Results of greenhouse and field experiments indicated that B. subtilis strain G1 when applied to groundnut as seed treatment and soil application significantly suppressed A. flavus population in the soil, A. flavus infection and aflatoxin B1 content in kernels and increased the pod yield. These studies show that B. subtilis strain G1 has potential as a biocontrol agent for control of aflatoxin contamination in groundnut.  相似文献   

9.
A sensitive and specific assay was developed to detect and biological control of bacterial leaf spot of pumpkin, Xanthomonas cucurbitae was identified on the basis of the morphological, biochemical and molecular assay. The antibiotic sensitivity of the isolate showed that, Carbenicillin revealed highest antibacterial activity with 29 ± 0.00 mm zone of inhibition against isolated bacterial strain. Isolated bacterial strains from soil were also identified by biochemical and molecular characterisation. By analysing morphological and biochemical characteristics and 16S rDNA of three bacterial strains isolated from soil was matched 96% with Bacillus subtilis, 98% with Bacillus brevis and 97% with Pseudomonas fluorescens strain. They were subjected to the antagonistic activity against Xanthomonas cucurbitae by disc diffusion method. Among them, B. subtilis showed significant positive antagonistic activity with 17.0 ± 0.28 mm zone of inhibition against Xanthomonas cucurbitae. The presence of DNA sequence related to the hrpD gene successfully amplified in some isolates of Xanthomonas cucurbitae.  相似文献   

10.
Thirty seven bacterial cultures isolated from soil samples obtained from different locations were tested for their antagonistic activity against some fungal pathogens, viz., Sclerotium rolfsii, Fusarium oxysporum and Rhizoctonia solani, causal agents of collar rot of sunflower, wilts and root rots, respectively. Among them, 5 bacterial strains, viz., A1 6 (Bacillus sphaericus), K1 24 (Pseudomonas fluorescens), M1 42 (Bacillus circulans), M1 66 (Bacillus brevis) and T1 22 (Bacillus brevis) showed positive antagonistic activity. M1 66 was the most effective in inhibiting mycelial growth of S. rolfsii in vitro followed by M1 42, T1 22, K1 24 and A1 6. Only one bacterial strain i.e. M1 42 exhibited antagonistic activity against F. oxysporum, and none of the bacterial strains gave positive activity against R. solani. Furthermore, antimicrobial activities of all the 5 strains were checked against different test organisms. These strains showed their extensive inhibition effect particularly against gram-positive test bacteria (Staphylococcus aureus and Bacillus subtilis) and the test fungal strain (Candida albicans). On the other hand, B. brevis M1 66 and B. brevis T1 22 strains had an inhibitory effect against gram positive and gram-negative test bacteria (Escherichia coli and Proteus vulgaris) as well as the test fungal strain.  相似文献   

11.
This study was devoted to increasing the production of fungal cell wall degrading enzymes by Bacillus subtilis JF419701 to enhance its efficiency in the biological control process. In dual culture, B. subtilis JF419701 showed the highest antagonistic effect of the 256 bacterial strains tested against six soil-borne pathogens, Alternaria alternata, Exserohilum rostratum, Fusarium oxysporum, Macrophomina phaseolina, Pythium ultimum and Rhizoctonia solani. The production potentiality of the enzymes α-1,3-glucanase, β-1,3-glucanase, chitinase and protease by B. subtilis JF419701 was studied in vitro. Results proved that the maximum production of enzymes by this bacterium was achieved after a two-day incubation period at a slightly alkaline pH (8). The addition of colloidal chitin or S-glucan to the growth media enhanced the production of all the enzymes except protease, which was stimulated by casein. This study therefore recommends that to obtain an efficient and strong bioagent culture of B. subtilis JF419701, it is necessary to grow this micro-organism on a specific medium containing either chitin or its derivatives at pH 8 for two days.  相似文献   

12.
Features of phosphate-mobilizing bacteria Bacillus subtilis IMB B-7023 and its streptomycin-resistant strain were investigated. While cultivated in medium with glucose and glycerophosphate, the growth rate of the antibiotic-marked strain was approximately similar to this parameter for Bacillus subtilis IMB B-7023 but cell sizes were 1.3-fold less. Both strains significantly stimulated the germinating of plant seeds, attached to their roots, and insignificantly differed in antagonistic activity toward phytopathogens and quantitative content of cell fatty acids and phosphatase activity. Streptomycin-resistant strain may be used for monitoring of Bacillus subtilis introduced to agroecosystem.  相似文献   

13.
Sclerotinia sclerotiorum is an important plant pathogen with worldwide distribution that causes severe economic losses of various agricultural crops such as soybean. This fungus is normally controlled with synthetic chemical fungicides that pose risks to the environment, and can be harmful to human health, and they can also induce resistance in pests. The aim of this study was to investigate the potential of Trichoderma asperelloides as a biocontrol agent towards white mold disease on soybeans crops. The antagonism of two strains of T. asperelloides (T25 and T42) isolated from soil samples was determined in-vitro by dual-culture confrontation testing on nine S. sclerotiorum strains obtained from sclerotia collected on diseased soybean plants. The mycelial growth and inhibition of carpogenic and ascospore germination by T. asperelloides extracts, as well as the efficacy of these on white mold control in soybeans were evaluated. Both strains of T. asperelloides exhibited high potential of antagonism. Methanolic and ethyl acetate extracts of the two T. asperelloides strains showed excellent growth inhibition (60–100%) on all of the pathogens tested. The ethyl acetate extracts of both T. asperelloides strains exhibited the highest efficacy against carpogenic germination, decreasing by 20–30% the number of ascospores per apothecium. Strains of T. asperelloides tested were more efficient in controlling white mold than two commercial products made from Trichoderma harzianum. The new strains of T. asperelloides have potential for successful biological control of white mold disease of soybean crops in the field.  相似文献   

14.
The biocontrol potential of Bacillus subtilis isolated from cow dung microflora was investigated in vitro and in vivo against two postharvest yam pathogenic fungi, Fusarium oxysporum and Botryodiplodia theobromae. B. subtilis strains inhibited the growth of F. oxysporum and B. theobromae in vitro in liquid medium in the range of 49.3–56.6% and in solid medium in the range of 31.0–36.0%, in comparison to the corresponding growth of fungi without bacterial inoculation. The interaction between B. subtilis CM1 and F. oxysporum was also studied by scanning electron microscopy. Chitinase production was demonstrated in vitro when B. subtilis was grown in the presence of colloidal chitin as the sole carbon source in a liquid medium. In vivo study showed that B. subtilis strains inhibited the growth of fungi (F. oxysporum and B. thobromae) up to 83% in wound cavities of yam tubers.  相似文献   

15.
Novel strains of rhizobacteria, Pseudomonas fluorescens Pf 9A‐14, Pseudomonas sp. Psp. 8D‐45 and Bacillus subtilis Bs 8B‐1, showed broad‐spectrum antagonistic activity and provided suppression of Pythium damping‐off and root rot of cucumber. Their biocontrol potential was further investigated for suppression of additional seedling diseases of cucumber (Phytophthora capsici) and radish (Rhizoctonia solani). Bacterial strains were also characterised for production of antibiotics, metabolites, volatiles, phytohormones and lytic enzymes. Seed and pre‐plant applications of all three antagonistic bacteria as cell suspension and talc or irradiated peat formulations to the infested potting mix provided overall high level of suppression of Phytophthora damping‐off and root rot of cucumber (66–85% healthy seedlings) and relatively low level of suppression of Rhizoctonia damping‐off of radish (18–38% healthy seedlings). Bacterial treatments also resulted in higher plant fresh masses. Seed coating with irradiated peat formulation of a mixture of three bacteria resulted in superior control of Phytophthora damping‐off and root rot of cucumber and much higher plant fresh masses. The presence of genes for biosynthesis of phenazine‐1‐carboxylic acid, 2,4‐diacetylphloroglucinol, pyrrolnitrin and pyoluteorin was confirmed in Pseudomonas strains, and that of fengycin, bacillomycin, bacilysin, surfactin and iturin A in B. subtilis Bs 8B‐1. All three strains produced HCN, salicylic acid, indole‐3‐acetic acid, protease and β‐1,3‐glucanase. Both Pseudomonas strains produced siderophores and only P. fluorescens Pf 9A‐14 showed phosphate solubilisation and chitinase activity. All three strains inhibited pathogen growth by producing volatiles, and gas chromatography–mass spectrometry analysis revealed eight compounds in Pf 9A‐14, 10 in Bs 8B‐1 and 4 in Psp 8D‐45, some with known antifungal activity. The antagonistic and plant‐growth promotion activities of these strains might be due to production of antibiotics, metabolites, lytic enzymes or phytohormones.  相似文献   

16.
This study determines the inhibitory effect of Stevia rebaudiana leaf extracts and its purified bioactive compound ‘stevioside’ against food‐related pathogens. The S. rebaudiana solvent extracts (1000 μg/mL) displayed antibacterial activity to Serratia marcescens, Klebsiella pneumoniae, Bacillus cereus, Pseudomonas aeruginosa, B. subtilis, Alcaligenes denitrificans and Salmonella typhimurium. Of the six solvents, ethanol and acetone extracts displayed the highest zone of inhibition. The bioactive compound from S. rebaudiana was purified by solvent extraction, thin‐layer chromatography followed by structural characterization by spectroscopy evidence. Purified stevioside prevented the growth of tested bacterial species, i.e. B. subtilis, K. pneumoniae and S. typhimurium. Significant zone of inhibition (12 mm) was observed against B. cereus which proposes potential application of stevioside in foods to increase their shelf life.  相似文献   

17.
Indigenous strains isolated from rhizosphere may contain highly competent genotypes to enhance the plant growth and often perform better than the introduced isolates. The present study deals with the characterisation of plant growth-promoting (PGP) attributes and antagonistic activity of Azotobacter chroococcum AZO2 against Macrophomina phaseolina causing charcoal rot disease and their effect on the growth of sesame (Sesamum indicum L.). Eight strains of Azotobacter were isolated from sesame rhizosphere on nitrogen-free medium, which exhibited significant PGP parameters such as phosphate solubilisation, indole acetic acid and siderophore production. The strain A. chroococcum AZO2 (EU274299) was characterised by 16S rDNA gene sequencing. Amplification of 781 bp nif H gene confirms nitrogenase activity of all the strains. A. chroococcum AZO2 exhibited strong antagonistic activities against M. phaseolina causing 81% colony growth inhibition and resulted in hyphal perforations, empty cell (halo) formation, hyphal twisting, shrinking and lysis of fungal mycelia along with degeneration of sclerotia. A. chroococcum AZO2 produced chitinase that caused degradation and digestion of the cell wall component of M. phaseolina. Different vegetative and reproductive parameters of sesame were found to be enhanced significantly upon application of A. chroococcum AZO2 + half doses of chemical fertilisers. A. chroococcum AZO2 was also found to be an effective root coloniser, plant growth promoter and potential antagonistic bacterium. It can be concluded that A. chroococcum AZO2 strain bears the characteristics of technological applications for inoculant preparation and growth enhancement of sesame besides being utilised as a better PGP bacterium as well as an effective agent for biocontrol of M. phaseolina.  相似文献   

18.
Aims: To analyse the effects of plipastatin operon disruption and constitutive expression of surfactin operon in Bacillus subtilis 168 on surfactin productivity, in vitro invasive growth and antagonism against fungi. Methods and Results: The srfA native promoter was replaced by the constitutive promoter PrepU in B. subtilis 168 after integration of a functional sfp gene. Moreover, the plipastatin synthesis was further disrupted in the B. subtilis 168 derivatives. In liquid media, an earlier and higher expression of PrepU, than that found with PsrfA, led to a specific surfactin production fivefold higher after 6 h of culture. On solid media, not only the invasive growth and the haemolytic activity but also the antifungal activity of the constitutive strains were improved when compared to the parental strain BBG111. As expected, the disruption of the plipastatin operon strongly reduced in vitro antifungal properties but, interestingly, enhanced specific surfactin production (1·47 g g?1 of biomass), spreading behaviour and haemolytic activity of the strains. Conclusions: This work demonstrates for the first time the interdependency of surfactin and plipastatin regarding their biosynthesis as well as their influence on the biological activities of the producing strain. Significance and Impact of the Study: The constitutive overproduction of surfactin enhances the invasive growth and the in vitro antagonistic activity of the mutant strain. Both properties are known to play an important role in the biocontrol of plant diseases. Plipastatin operon disruption increases the surfactin productivity of mutant strains. These mutants are interesting for use in continuous bioprocesses for surfactin production or in bioremediation.  相似文献   

19.
Potential sources of mortality of marine invertebrate larvae are numerous and include predation and diseases caused by marine microorganisms. Extracts from the eggs of 11 coral species were evaluated for their ability to deter surface attachment and inhibit the growth of two marine tolerant laboratory bacteria and 92 bacterial strains isolated from seawater and the surface of coral colonies on the Great Barrier Reef (GBR). Extracts of the eggs of Montipora digitata inhibited the growth of the two laboratory bacteria, Vibrio harveyii and Bacillus subtilis, and one bacterial isolate from the mucus of the coral Favia pallida in disc diffusion and liquid culture assays. No other microbial strains (n=91) from the surface of corals and the reef environment were inhibited by M. digitata extracts. No antibacterial activity was found in the egg extracts of the remaining ten coral species and none of the extracts inhibited surface attachment of various bacteria. Extrapolation of estimated surface concentrations of the biologically active extract of M. digitata suggests that the level of the growth inhibitory compounds may be sufficient to deter microbial growth in situ.  相似文献   

20.
The potential of biosurfactant PS to permeabilize bacterial cells of Pseudomonas aeruginosa, Escherichia coli, and Bacillus subtilis on growing (in vivo) and resting (in vitro) cells was studied. Biosurfactant was shown to have a neutral or detrimental effect on the growth of Gram-positive strains, and this was dependent on the surfactant concentration. The growth of Gram-negative strains was not influenced by the presence of biosurfactant in the media. Cell permeabilization with biosurfactant PS was shown to be more effective with B. subtilis resting cells than with Pseudomonas aeruginosa. Scanning-electron microscopy observations showed that the biosurfactant PS did not exert a disruptive action on resting cells such that it was detrimental to the effect on growing cells of B. subtilis. Low critical micelle concentrations, tender action on nongrowing cells, and neutral effects on the growth of microbial strains at low surfactant concentrations make biosurfactant PS a potential candidate for application in different industrial fields, in environmental bioremediation, and in biomedicine.  相似文献   

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