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1.
The efficacy of eight fungal and ten endophytic bacterial isolates were tested for their ability to inhibit the growth of Pythium aphanidermatum, the causal agent of chilli damping-off. In vitro studies revealed that Trichoderma viride (TVA) and endophytic Pseudomonas fluorescens (EBL 20-PF) showed the highest inhibition of mycelial growth (71.5%; 76.7%) of P. aphanidermatum. Both the antagonists were compatible with each other and they were tested alone and together in vivo for the control of P. aphanidermatum. Besides, the induction of defense-related enzymes such as peroxidase (PO), polyphenoloxidase (PPO), phenylalanine ammonia-lyase (PAL), PR-protein like β-1, 3-glucanase and the accumulation of phenolics in chilli seedlings due to the application of bioagents were also studied. Combined application of talc-based formulation of bio-agents and challenge inoculated with P. aphanidermatum recorded maximum induction of defense-related enzymes, PR-proteins and accumulation of phenolics compared with individual application. This study suggest that the increased induction of defense-related enzymes (four to fivefold) and phenolic content (sixfold) due to the combination treatment of bioagents might have involved in the reduction of damping-off incidence and in turn increased the plant growth and yield of chilli.  相似文献   

2.
Among nine native bacterial strains isolated from banana fruit surface and rhizosphere and six bacterial strains introduced from the culture collection, three native strains viz., non-fluorescent Pseudomonas (NFP6), Pseudomonas fluorescens (Pf3a), and Bacillus subtilis (BS1); and two bacterial strains from culture collection viz., Azospirillum (AS1) and Azotobacter (AZ1) have recorded maximum inhibition of mycelial growth of crown rot pathogens (Lasiodiplodia theobromae and Colletotrichum musae) under in vitro condition. When these effective bacterial strains were treated on banana fruits under in vivo, significant reduction of crown rot disease and increased shelf life of banana was observed. However, bacterial strains applied as three way combinations (NFP6 + Pf3a + BS1) had greater effect compared with individual and two way combination of bacterial antagonist treatments. The effect of crown rot disease reduction was also comparable to that of fungicide Benomyl (0.1%) both under cold and room temperature storage conditions. Besides, the induction of defense-related enzymes such as phenylalanine ammonia-lyase (PAL), peroxidase (PO), polyphenoloxidase (PPO), and the accumulation of phenolics in banana fruit due to the application of bacterial antagonists were also studied at five different time intervals viz. 0th, 1st, 3rd, 5th and 7th days after treatment. When banana fruits treated with bacterial antagonists (individually and also in different combinations) and challenge-inoculated with crown rot pathogens, up to fourfold increase in defense-related enzymes and 3.6 fold increase in phenolic content was observed compared with control. The activity of these defense-related enzymes and phenolic content had gradually increased from 1st day after treatment to 3rd after treatment and reached their peak on 5th day after treatment. Among the bacterial antagonists which have been applied individually and in different combinations, the banana fruits treated with three-way antagonist mixture, i.e., NFP6 + Pf3a + BS1 recorded maximum induction of defense-related enzymes and accumulation of phenolics compared with individual and two-way combination of antagonist mixtures. This study suggest that the increased induction of defense-related enzymes and phenolic content due to the treatment of banana fruits with bacterial antagonists might have involved in the reduction of crown rot severity and in turn increased the shelf life of banana fruits.  相似文献   

3.
Stem rot is a serious disease in Jerusalem artichoke (JA). To reduce the impact of this disease on yield and quality farmers often use fungicides, but this control method can be expensive and leave chemical residues. The objective of this study was to evaluate the efficacy of two biological control agents, Trichoderma harzianum T9 and Bacillus firmus BSR032 for control of Sclerotium rolfsii under field conditions. Four accessions of JA (HEL246, HEL65, JA47, and JA12) were treated or notreated with T. harzianum T9 and B. firmus BSR032 in a 4 × 2 × 2 factorial experiment in two fields (environments), one unfertilized and one fertilized. Plants were inoculated with S. rolfsii and disease was evaluated at 3-day intervals for 46 days. T. harzianum T9 and B. firmus BSR032 reduced disease incidence by 48% and 49%, respectively, whereas T. harzianum T9 + B. firmus BSR032 reduced disease incidence by 37%. The efficacy of T. harzianum T9 and B. firmus BSR032 for control of S. rolfsii was dependent on environments and genotypes. The expression of host plant resistance also depended on the environment. However, HEL246 showed consistently low disease incidence and severity index in both environments (fertilized and unfertilized). Individually, T. harzianum T9, B. firmus BSR032, or host plant resistance control stem rot caused by S. rolfsii in JA. However, no combination of these treatments provided more effective control than each alone.  相似文献   

4.
Induction of defense-related enzymes, such as phenylalanine ammonia lyase (PAL), peroxidase (PO), polyphenol oxidase (PPO), superoxide dismutase (SOD) and catalase (CAT) due to bacterial antagonists viz., Pseudomonas fluorescens (Pf1) and Bacillus subtilis (EPCO16) and plant-derived lipoxygenase volatile compound hexanal, were studied in mango fruits against Lasiodiplodia theobromae causing stem-end rot disease. The results showed increased induction of all the defense-related enzymes in mango fruits 3–5 days after dipping treatment with combination of bacterial antagonists and hexanal when compared to untreated control treatment and treatment with fungicide carbendazim in storage condition. The increased activity was observed up to 3 days after treatment and thereafter declined. Further, increased expression of specific isoforms of PO, PPO, SOD and CAT were also observed in the treatment effect of P. fluorescens (0.5%)?+?hexanal (0.02%) treated fruits against L. theobromae. From the results obtained, it is inferred that due to the enhancement of defense-related enzymes via the phenylpropanoid pathway and due to secretion of secondary metabolites that would play significant role in hindering the pathogen quiescence and further invasion in mango fruits and thereby prevent the fruit rot.  相似文献   

5.
Rhizobacteria isolated from the rhizosphere soil were evaluated for their ability to control rhizome rot in turmeric (Curcuma longa L). These isolates were characterised as Pseudomonas fluorescens and Bacillus subtilis. Under in vitro condition, two isolates, namely P. chlororaphis (PcPA23) and B. subtilis (BsCBE4), showed maximum inhibition of mycelial growth of Pythium aphanidermatum, were found effective in reducing rhizome rot of turmeric both under greenhouse and field conditions and increased the plant growth and rhizome yield. Both the isolates were further tested for its ability to induce production of defense-related enzymes and chemicals in plants. Increased activities of phenylalanine ammonia lyase, peroxidase, polyphenol oxidase, chitinase and β-1,3-glucanase were observed in PcPA23 and BsCBE4 pre-treated turmeric plants challenged with P. aphanidermatum. Moreover, higher accumulation of phenolics was noticed in plants pre-treated with PcPA23 and BsCBE4 challenged with P. aphanidermatum. Thus, the present study shows that in addition to direct antagonism and plant growth promotion, induction of defense-related enzymes involved in the phenyl propanoid pathway collectively contributed to enhance resistance against invasion of Pythium in turmeric.  相似文献   

6.
This study is focused on isolation and characterisation of 2,4-diacetylphloroglucinol (2,4-DAPG)-producing Pseudomonas fluorescens isolates from different soils of groundnut-based cropping systems in Andhra Pradesh. In our studies, 21 isolates of P. fluorescens were isolated and confirmed through various biochemical tests, of which five were tested positive for 2,4-DAPGproduction with specific primers. Biocontrol potential of these isolates on groundnut stem rot pathogen (Sclerotium rolfsii) was determined through in vitro dual culture assays. The eight isolates were found effective against S. rolfsii (up to 75% inhibition) in dual culture method. All the five 2,4-DAPG-producing Plant Growth-Promoting Rhizobacteria isolates were highly antagonistic to S. rolfsii. Genetic diversity of these P. fluorescens isolates was determined by random amplification of polymorphic DNA analysis. Overall, our results suggest that the prevalence of 2,4-DAPG-producing fluorescent Pseudomonads in different crop rhizospheres of groundnut-based cropping systems.  相似文献   

7.
Abstract

Identification of individual phenolic acids of Sorghum vulgare Pers. cv. M.P. after interaction with Sclerotium rolfsii Sacc. using high performance liquid chromatograph (HPLC) showed the presence of phenolics namely tannic, gallic, ferulic, chlorogenic and cinnamic acids in varying amounts. After 72 h inoculation with S. rolfsii, a maximum amount of ferulic acid (166.6 µg g?1 fresh wt) was present in the collar of inoculated plants, followed by leaves and roots and its level decreased gradually with time. Similarly, the presence of chlorogenic acid was traced after 48 h, while that of cinnamic acid was traced after 72 h of inoculation. Reddish-brown pigmentation at the collar region of inoculated plants was also observed along with the high content of tannic acid. Among other phenolics, the presence of gallic acid was recorded consistently and maximum accumulation (139.3 µg g?1 fresh wt) was noticed at the zone of interaction (collar region) after 72 h of inoculation. In contrast, maximum lignin deposition was observed at collar region after 96 h of inoculation. Induction of phenolic acids in S. vulgare along with the lignin deposition and red pigmentation at collar region is considered a key biomarker in the non-host-pathogen interaction in the S. valgare–S. rolfsii pathosystem.  相似文献   

8.
Abstract

An attempt was made to study the biocontrol efficacy of antagonistic microorganisms in phylloplane of rose cv. Edward to manage the black spot (Diplocarpon rosae) disease. Eight antagonistic microorganisms were tested in vivo against the black spot pathogen. Among these, Trichoderma viride and Pseudomonas fluorescens pf1 reduces the mycelial growth significantly. These two biocontrol agents were evaluated for their ability to induce defense-related enzymes and chemicals in plants. Increased activity of phenylalanine ammonia lyase (PAL), peroxidase (PO), polyphenoloxidase (PPO) and total phenolics were recorded in all the biocontrol agents treated leaves. P. fluorescens Pf1 recorded early and increased synthesis of the entire defense-related enzymes and total phenol within 6 days. The application of biocontrol agents induced the defense-related enzymes involved in phenyl propanoid pathway in addition to direct antagonism, which collectively contribute for enhanced resistance against invasion of Diplocarpon rosae in rose.  相似文献   

9.
Biochemical analysis was carried out for 10 isolates of Pseudomonas fluorescens. All isolates were found positive for siderophore and indole acetic acid production (except Pf IX) and phosphate solubilisation (except Pf VII). Biochemically efficient strains (Pf I, Pf IV, Pf VII and Pf IX) were selected for management of root rot, collar rot and damping off caused by Phytophthora parasitica, Pythium sp. and Fusarium solani. The activity of defence-related enzymes like esterase, peroxidase and polyphenol oxidase was also detected by polyacrylamide gel electrophoresis (PAGE). Consistent appearance of esterase isozyme bands was visualised, in the range of 0.193–0.349. The highest Rm value 0.349 was observed on Pf IV. Whereas, for peroxidase, Rm value ranged between 0.302 and 0.373 and for polyphenol oxidase, it was in the range of 0.211–0.800. P. fluorescens Pf IV was found significantly effective to arrest the per cent mycelial growth of P. parasitica (55.20%), Pythium sp. (65.33%) and F. solani (64.67%). Among bioagents, seed treatment and soil drenching with Pf IV at 15 and 30 days after sowing were found effective to reduce per cent disease incidence (30.55%) at 120 days after emergence. Seed treatment with copper oxychloride at 3g/kg seed and metalaxyl at 2 g/kg seed were also found effective.  相似文献   

10.
Biological control of charcoal root rot disease caused by Macrophomina phaseolina in chickpea was studied by using Streptomyces sp. S160. This biocontrol agent (BCA) inhibited the mycelial growth of M. phaseolina by 50 % in vitro and significantly reduced charcoal rot incidence in the greenhouse by 33.3 %. The greenhouse experiment revealed that seed treatment along with soil application supported the highest germination (88.6 %), vigor index (7326.91) and reduced root rot incidence (12.5 %) in comparison to seed treatment and soil application alone. BCA enhanced the growth and helped in inducing resistance against charcoal rot disease of chickpea caused by M. phaseolina by increasing activity of defense-related enzymes in chickpea plants, leading to the synthesis of defense chemicals in plants. BCA (Streptomyces sp. S160) was also characterized and identified by using polyphasic approaches including 16S rDNA sequencing.  相似文献   

11.
Abstract

Sclerotinia sclerotiorum is a cosmopolitan fungal pathogen causing stem and collar watery soft rot of cauliflower. Previous management of this disease with chemical pesticides caused hazardous results which lead to use of more eco-friendly microbial approaches. In the present study, consortia of Trichoderma harzianum TNHU27 and Pseudomonas aeruginosa PJHU15 were assessed for their ability in controlling Sclerotinia rot in cauliflower. The seedlings of cauliflower were challenged singly and in consortium with these two compatible microbes. The plants were evaluated upon challenge inoculation with S. sclerotiorum for changes in total phenolic content, the activity of defense and antioxidative enzymes. The microbial consortium comprising of T. harzianum and P. aeruginosa significantly enhanced the defense responses of the plant in comparison to pathogen challenged and unchallenged control. The study elucidates that plant beneficial microbes in the consortium may provide superior protection by induction of faster and enhanced defense responses in comparison to unchallenged and single microbe challenged plants under pathogen challenged conditions.  相似文献   

12.
The study was conducted to examine efficacy of a rhizospheric microbial consortium comprising of a fluorescent Pseudomonas (PHU094), Trichoderma (THU0816) and Rhizobium (RL091) strain on activation of physiological defense responses in chickpea against biotic stress caused by the collar rot pathogen Sclerotium rolfsii. Results of individual microbes were compared with dual and triple strain mixture treatments with reduced microbial load (1/2 and 1/3rd, respectively, of individual microbial load compared to single microbe application) in the mixtures. Periodical studies revealed maximum activities of phenylalanine ammonia lyase [E.C. 4.1.3.5] and polyphenol oxidase [E.C. 1.14.18.1] and accumulation of total phenol content in chickpea in the triple microbe consortium treated plants challenged with the pathogen compared to the single microbe and dual microbial consortia. Similarly, the expression of the antioxidant enzymes superoxide dismutase [E.C.1.15.1.1] and peroxidase [E.C.1.11.1.7] was also highest in the triple microbial consortium which was correlated with lesser lipid peroxidation in chickpea under the biotic stress. Histochemical staining clearly showed maximum and uniform lignification in vascular bundles of chickpea stem sections treated with the triple microbes. The physiological responses were directly correlated with the mortality rate as least plant mortality was recorded in the triple microbe consortium treated plants. The results thus suggest an augmented elicitation of stress response in chickpea under S. rolfsii stress by the triple microbial consortium in a synergistic manner under reduced microbial load.  相似文献   

13.
Exudates from sclerotia of two Sclerotium rolfsii isolates (one causing collar rot in Cicer arietinum, isolate VC971, and the other leaf spots in Rauvolfia serpentina, isolate VL016) were assayed for their antifungal activity against 26 fungi consisting of plant parasites as well as saprophytes. Spore germination of all the test fungi was affected by the exudates reaching 100% in some cases. Foliar spray with exudates of isolate VL016 significantly reduced disease incidence of balsam (Impatiens balsamina) powdery mildew caused by Erysiphe cichoracearum and pea (Pisum sativum) powdery mildew caused by Erysiphe pisi, under field conditions. Characterization of exudates from 25 isolates of S. rolfsii revealed pH ranging from 3.8 to 5.3 and colour from light yellow to deep yellow. Among the phenolic acids found in the exudates were tannic, gallic, caffeic, vanillic, ferulic, chlorogenic and cinnamic acids. Oxalic acid was also found in varied amounts. Among the phenolic acids, ferulic acid was found to be present at high concentration in exudates of most isolates (3.9–153.4 μg/ml). The antioxidant properties of phenolics, which generally inhibit fungal morphogenesis including spore germination along with the antifungal nature of some phenolics are chiefly attributed to the inhibitory effect of sclerotial exudates of S. rolfsii. Additionally, both the isolates VC971 and VL016 showed almost similar antifungal activities despite they are of different origin and thereby demonstrate the antifungal nature of sclerotial exudates.  相似文献   

14.
Abstract

Sclerotium rolfsii Sacc. the causal agent of collar and root rot of chili (Capsicum annuum L.), is one of the most important soil-borne pathogens in Sri Lanka. Bacillus subtilis CA32r, a stable spontaneous kanamycin resistant isolate, showing antagonism in a Petri plate assay, was selected for greenhouse pot experiments to control S. rolfsii. An ethyl acetate extract of the culture filtrate of B. subtilis CA32r inhibited radial colony growth as well as germination of sclerotia of the pathogen in vitro, indicating the presence of antifungal compound(s) in the culture extract. B. subtilis CA32r was investigated for its effectiveness as a biological control agent against S. rolfsii infecting transplanted chilli seedlings in greenhouse pot experiments. CA32r significantly decreased the disease incidence in terms of lesion development on stem base and roots depending on the mode of the bacterial application. Seed bacterization and soil application alone did not protect chilli plants, but root bacterization prior to the transplanting of seedlings significantly decreased the disease incidence caused by S. rolfsii. However, even in the combined treatment, seed bacterization and soil application, did not protect chilli plants. The best protection was achieved by combination of root bacterization prior to transplant and soil application of CA32r. Root bacterization resulted in maintaining higher numbers of bacteria at the collar region of chilli plants and may have shielded the most vulnerable area from the pathogen, resulting enhanced protection. Since the application of CA32r resulted in a significant reduction of the number of viable S. rolfsii propagules in the soil indicates that B. subtilis CA32r possesses not only protective but also eradicative potential.  相似文献   

15.
Two non-conventional chemicals, ZnSO4 (10−4 mM) and oxalic acid (4 mM) were tested (alone as well as in combination with seeds bacterized with Pseudomonas syringae strain PUR46 and vermicompost substitution in the potting soil), for their ability to suppress collar rot of chickpea (Cicer arietinum) caused by Sclerotium rolfsii under greenhouse conditions. ZnSO4 and oxalic acid were applied as pre-inoculation foliar spray on chickpea and subsequently challenged with S. rolfsii. Both the chemicals provided significant protection to chickpea compared to control (100% plant mortality) when used alone as well as in combination with PUR46 and vermicompost. However, ZnSO4 was more effective than oxalic acid against S. rolfsii. Amongst the treatments tried, plant mortality was least when ZnSO4 was used in combination with seed bacterization with PUR46 and 25% vermicompost substitution. The findings indicate the utility of integration of the above factors in managing collar rot efficiently.  相似文献   

16.
Trichoderma harzianum and Trichoderma viride, which were grown on modified media of rice straw waste, were used as suitable natural media for determining the population of the bioagents and stimulating the production of antimicrobial substances, i.e. toxins, enzymes at different periods (1, 2, 3, 4, 5 and 6 weeks). Also, the evaluation of synergistic effect between biocontrol fungi and bacteria may play a key role in the natural process of biocontrol. Synergism can occur when different agents are applied together and cell wall degrading enzymes produced by fungi can increase the efficacy of bacteria. The evaluation of the bioagents formulated on rice straw waste has influenced the percentage of infection of root rot on bean plants under green house and field conditions. The formulation of the Trichoderma spp. to reduce the incidence of the diseases caused by soil-borne fungi in the field has great importance in the biocontrol of the diseases. This work was aimed to determine and improve the efficacy of application and formulation of Trichoderma spp. on the rice straw waste against root-rot disease of bean under green house and field conditions.  相似文献   

17.
The effects of native isolates of Pseudomonas fluorescens, Azospirillum brasilense, and Trichoderma harzianum on rooting and acclimatization of in vitro-grown shoots and plantlets of tea were evaluated. In vitro bacterization of P. fluorescens failed to establish, while both T. harzianum and A. brasilense retarded shoot growth, eventually overtaking shoot cultures in in vitro rooting. Acclimatization of rooted plantlets in soil amended with bioinoculants, either individually or in various combinations, promoted plantlet survival. Moreover, efficiency of nutrient uptake of plantlets was higher in the presence of microorganisms. Root rot or wilting of tissue culture-derived plants was not observed in bioinoculant-treated plants, as they possessed relatively higher activities of defense enzymes, including peroxidase and phenylalanine ammonia lyase.  相似文献   

18.
The objective of this study was to screen Streptomyces spp. for biological control of root and stem rot (Sclerotium rolfsii) and bacterial wilt (Ralstonia solanacearum), the very destructive diseases of chili pepper in Thailand. About 265 isolates of Streptomyces spp. were tested for their inhibitory effects on S. rolfsii mycelial growth on dual culture plates. Then, 14 promising isolates were further tested for their effects on R. solanacearum growth. Three effective isolates further identified as S. mycarofaciens SS-2-243, S.philanthi RL-1-178 and S. philanthi RM-1-138 were selected and proved to produce both antifungal and antibacterial substances in the culture medium. S. philanthi RM-1-138 strongly inhibited seed germination and seedling growth of chili pepper in laboratory tests. Therefore, it was not used in the following studies. When tested in greenhouse conditions, the efficacy of S. philanthi RL-1-178 in suppressing Sclerotium root and stem rot of chili pepper was approximately equal to that of Trichoderma harzianum NR-1-52 or that of carboxin treatment. S. mycarofaciens SS-2-243 and S. philanthi RL-1-178 suppressed Ralstonia wilt of chili pepper in a way that was similar to streptomycin sulfate treatment and it was observed that T. harzianum NR-1-52 had no effect on the bacterial wilt. Under field conditions where the soil was inoculated with two pathogens, the results showed that S. philanthi RL-1-178 could protect the chili pepper plants from S. rolfsii and R. solanacearum infection better than S. mycarofaciens SS-2-243 or T. harzianum NR-1-52. S. philanthi RL-1-178 treatment resulted in 58.75% survival of chili pepper plants and its efficacy was not significantly different from the carboxin-and-streptomycin sulfate treatment.  相似文献   

19.
The objective of this research was to evaluate Trichoderma harzianum strain T22 as a biocontrol agent of collar and root rot caused by different Calonectria pauciramosa isolates. Thus, the microsclerotia-forming ability and virulence of twenty C. pauciramosa isolates were assessed. Microsclerotia production varied partially among the isolates and dual culture with T22 on carnation leaf agar revealed isolates with both high and low microsclerotia-forming ability. Inoculation tests on red clover (Triflolium pratense) demonstrated its susceptibility to the pathogen. On red clover, the degree of virulence and T22 effects in controlling infections were highly variable among the isolates tested. A nursery trial performed on Feijoa sellowiana seedlings confirmed previous results, clearly indicating virulence variability among C. pauciramosa isolates. For three isolates tested in nursery trial, T22 effectiveness in controlling infection was inversely related to their degree of virulence. Overall, T. harzianum strain T22 showed good antagonist activity in reducing microsclerotia production on carnation leaf and the incidence and severity of collar and root rot on both selected hosts. This data could be crucial in developing integrated pest management strategies in ornamental plant nurseries.  相似文献   

20.
Abstract

Fluorescent pseudomonads based bioformulation was evaluated for their ability to control Macrophomina root rot disease in mungbean (Vigna mungo). P. fluorescens isolate Pf1 showed the maximum inhibition in mycelial growth of Macrophomina phaseolina under in vitro conditions. Bioformulation of Pf1 with chitin was effective in reducing the root rot incidence in green gram both under glasshouse and field conditions. The rhizosphere colonization of P. fluorescens was observed appreciable with the green gram plants. However, Pf1 amended with chitin colonized effectively. Furthermore, the induction of defence-related enzymes and chemicals in plants by Pf1 amended with or without chitin and neem were tested. Increased accumulation of defence enzymes viz., phenylalanine ammonia lyase (PAL), peroxidase (PO), polyphenol oxidase (PPO), chitinase, β-1,3-glucanse and phenolics were observed in Pf1 bioformulation amended with chitin, pre-treated plants challenge inoculated with M. phaseolina under glasshouse conditions. The present study reveals that in addition to direct antagonism and plant-growth promotion, PGPR strains amended with chitin bioformulation induced defence-related enzymes and pathogenesis related (PR) proteins which collectively enhance the resistance in green gram against the infection of M. phaseolina.  相似文献   

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