Fluorescent pseudomonad mixtures mediate disease resistance in rice plants against sheath rot (<Emphasis Type="Italic">Sarocladium oryzae</Emphasis>) disease

Plant growth-promoting rhizobacterial (PGPR) strains were isolated from different agro-ecosystems of Tamil Nadu, India, and were tested for their efficacy against the sheath rot pathogen Sarocladium oryzae under in vitro, glasshouse and field conditions. Vigour and a relative performance index (RPI) were used to assay the growth promotion and antagonistic activity of Pseudomonas strains against S. oryzae under in vitro conditions. The results revealed the significant performance by strains Pf1, TDK1 and PY15 compared to other strains. Further, the combination of Pseudomonas strains Pf1, TDK1 and PY15 was more effective in reducing sheath rot disease in rice plants compared to individual strains under glasshouse and field conditions. Quantitative and native polyacrylamide gel electrophoresis (PAGE) analysis of peroxidase (PO), polyphenol oxidase (PPO) and chitinase activity in rice plants showed an increased accumulation of defence enzymes in the treatment with a combination of Pf1, TDK1 and PY15 compared to the treatment with individual strains and untreated controls. The present study revealed the probable influence of antagonism, plant growth promotion and induced systemic resistance (ISR) by the mixture of Pseudomonas bioformulations in enhancing the disease resistance in rice plants against sheath rot disease.

Biological control of rice root-knot nematode,Meloidogyne graminicola through mixture of Pseudomonas fluorescens strains

The plant growth promoting rhizobacterium, Pseudomonas fluorescens strains PF1, TDK1, and PY15 were evaluated individually and in combinations for their efficacy against root-knot nematode, Meloidogyne graminicola, in rice plants under in vitro, glass house and field conditions. Culture filtrates of these strains either individually or as mixture inhibited egg hatching and caused mortality of juveniles of M. graminicola in vitro. The efficacy was more pronounced when filtrates of the strain were used as mixtures than as individual strains. Mixtures of P. fluorescens strains signficantly reduced M. graminicola infestation when applied as bacterial suspensions through seed treatment. The higher activity of peroxidase and chitinase enzymes was observed in plants treated with P. fluorescens mixtures than the plants treated with individual strains, two strain mixtures and untreated control. In field trials on rice, talc formulations of the P. fluorescens strains individually as well as mixtures were evaluated as seed treatment, soil treatment and combination of both. A mixture of the three strains was the most effective when applied either as seed + soil treatment or as seed treatment alone. The introduced P. fluorescens strains survived endophytically on rice roots. The application of the P. fluorescens mixture PF1 + TDK1 + PY15 in seed + soil treatment resulted in higher grain yield which provided a 27.3% increase over the control followed by P. fluorescens mixture PF1 + TDK1 + PY15 in seed treatment alone, which increased the grain yield of rice by 24.7% compared to the control.     

Induction of peroxidase and polyphenol oxidase in Arachis hypogaea in response to treatment with Pseudomonas fluorescens and inoculation with Alternaria alternata

Abstract

The application of talc-formulation through seed, seed treatment plus foliar spray and foliar spray alone significantly reduced the leaf blight incidence both under greenhouse and field conditions. The groundnut plants treated with biocontrol agent and challenge inoculated with Alternaria alternata recorded significantly increased activity of Peroxidase isozyme (PO), Polyphenol oxidase isozymes (PPO) activity. Expression of PO2, PPO1 and PPO2 isoforms were found in all the plants treated with Pf1 while additional PO1, PPO3, PPO4 and PPO5 were observed in Pf1-treated plants followed by challenge inoculation with the pathogen.     

Plant growth promoting bacteria enhance water stress resistance in green gram plants

Plant growth promoting bacterial (PGPB) strains Pseudomonas fluorescens Pf1 and endophytic Bacillus subtilis EPB5, EPB22, EPB 31 were tested for their capacity to induce water stress related proteins and enzymes in green gram (Vigna radiata) plants. Among the different bacteria used, P. fluorescens Pf1 increased the vigour index, fresh weight and dry weight of green gram seedlings in vitro. Quantitative and qualitative analyses of stress-related enzymes indicated the greater activity of catalase and peroxidase in green gram plants bacterized with P. fluorescens Pf1 against water stress when compared to untreated plants. The greater accumulation of proline was recorded in Pf1 treated plants compared to untreated plants. The pot culture study revealed the greater resistance to water stress by green gram plants treated with P. fluorescens Pf1 compared to untreated plants. The greater activity of stress-related enzymes in green gram plants mediated by PGPB could pave the way for developing drought management strategies.     

Physiological Responses of Rice Plants Supplied with Silicon to Monographella albescens Infection

This study investigated the effect of silicon (Si) on the resistance of rice plants of the cv. ‘Primavera’ cultivar that were grown in a nutrient solution with 0 (?Si) or 2 mm (+Si) Si to leaf scald, which is caused by Monographella albescens. The leaf Si concentration increased in the +Si plants (4.8 decag/kg) compared to the ?Si plants (0.9 decag/kg), contributing to a reduced expansion of the leaf scald lesions. The extent of the cellular damage that was caused by the oxidative burst in response to the infection by M. albescens was reduced in the +Si plants, as evidenced by the reduced concentration of malondialdehyde. Higher concentrations of total soluble phenolics and lignin‐thioglycolic acid derivatives and greater activities of peroxidases (POX), polyphenoloxidases (PPO), phenylalanine ammonia‐lyases (PAL) and lipoxygenases (LOX) in the leaves of the +Si plants also contributed to the increased rice resistance to leaf scald. In contrast, the activities of chitinases and β‐1,3‐glucanases were higher in the leaves of the ?Si plants probably due to the unlimited M. albescens growth in the leaf tissues, as indicated by the larger lesions. The results of this study highlight the potential of Si in decreasing the expansion of the leaf scald lesions concomitantly with the potentiation of phenolic and lignin production, and the greater activities of POX, PPO, PAL and LOX rather than simply acting only as a physical barrier to avoid M. albescens penetration.     

Role of salicylic acid in systemic resistance induced by Pseudomonas fluorescens against Fusarium oxysporum f. sp. ciceri in chickpea

Selected isolates of Pseudomonas fluorescens (Pf1-94, Pf4-92, Pf12-94, Pf151-94 and Pf179-94) and chemical resistance inducers (salicylic acid, acetylsalicylic acid, DL-norvaline, indole-3-carbinol and lichenan) were examined for growth promotion and induced systemic resistance against Fusarium wilt of chickpea. A marked increase in shoot and root length was observed in P. fluorescens treated plants. The isolates of P. fluorescens systemically induced resistance against Fusarium wilt of chickpea caused by Fusarium. oxysporum f.sp. ciceri (FocRs1), and significantly (P = 0.05) reduced the wilt disease by 26-50% as compared to control. Varied degree of protection against Fusarium wilt was recorded with chemical inducers. The reduction in disease was more pronounced when chemical inducers were applied with P. fluorescens. Among chemical inducers, SA showed the highest protection of chickpea seedlings against wilting. Fifty two- to 64% reduction of wilting was observed in soil treated with isolate Pf4-92 along with chemical inducers. A significant (P = 0.05; r = -0.946) negative correlation was observed in concentration of salicylic acid and mycelial growth of FocRs1 and at a concentration of 2000 microg ml(-1) mycelial growth was completely arrested. Exogenously supplied SA also stimulated systemic resistance against wilt and reduced the disease severity by 23% and 43% in the plants treated with 40 and 80 microg ml(-1) of SA through root application. All the isolates of P. fluorescens produced SA in synthetic medium and in root tissues. HPLC analysis indicated that Pf4-92 produced comparatively more SA than the other isolates. 1700 to 2000 nanog SA g(-1) fresh root was detected from the application site of root after one day of bacterization whereas, the amount of SA at distant site ranged between 400-500 nanog. After three days of bacterization the SA level decreased and was found more or less equal at both the detection sites.     

PGPR mediates induction of pathogenesis-related (PR) proteins against the infection of blast pathogen in resistant and susceptible ragi [Eleusine coracana (L.) Gaertner] cultivars

The Pseudomonas fluorescens isolate 1 (Pf1) was found to protect the ragi [Eleusine coracana (L.) Gaertner] blast fungus, Pyricularia grisea. Induction of defense proteins viz. chitinase, β-1,3 glucanase, peroxidase (PO) and polyphenol oxidase (PPO) by the Pf1 isolate was studied against P. grisea. Chitinase in a resistant, susceptible and commonly used cultivar with and without challenge inoculation of P. grisea, revealed changes in the isoform pattern by UV illumination after staining the gel with fluorescent brightner 28. Native PAGE (polyacrylamide gel electrophoresis) of PO showed the single isoform in all the treatments including the control and a significant increase in the intensity of the band in the inoculated control and Pf1 treatment in all the varieties. Isoform analysis of PPO showed the induction of PPO in P. fluorescens treated plants challenged with P. grisea.     

Association of the Hydrolytic Enzyme Chitinase against Rhizoctonia solani in Rhizobacteria-treated Rice Plants

Twenty‐two strains of Pseudomonas fluorescens isolated from the rhizosphere soil of nine plant species were screened in vitro for their inhibitory effect on the mycelial growth of the rice sheath blight fungus, Rhizoctonia solani. Of the 22 strains, two promising strains (Pf1 and FP7) were assessed for their effect on seedling vigour and their ability to promote growth in vitro of four cultivars of rice. Both bacterial strains induced systemic resistance in rice cv. IR 50, which is susceptible to sheath blight. After inoculation of the sheaths with the pathogen, Pseudomonas‐treated plants showed an increase in chitinase activity significantly higher than that of untreated control plants. A twofold increase in chitinase activity occurred 2 days after inoculation of plants with the pathogen. Western blot analysis of chitinase indicated the expression of 28 and 38 kDa proteins in rice sheaths against R. solani. Increased induction of the pathogenesis‐related chitinase isoform in Pseudomonas‐treated rice in response to R. solani infection indicates that the induced chitinase has a definite role in suppressing disease development.     

Induced systemic resistance to tomato leaf curl virus and increased yield in tomato by plant growth promoting rhizobacteria under field conditions

The talc-based formulation of two Pseudomonas fluorescens strains (Pf1 and VPT10) and its mixture (with and without chitin) were tested against tomato leaf curl virus in tomato under greenhouse and field conditions. The mean percentage of tomato leaf curl virus infected plants were significantly lower (25%) with less symptom severity and delayed symptom expression up to nine additional days in Pseudomonas with chitin (VPT10 + chitin) treated tomato plants compared to non-bacterised control plants upon challenge inoculation with tomato leaf curl virus. Tomato leaf curl virus was partially purified and antiserum was developed. Using the antiserum the tomato leaf curl virus was detected in symptomatic leaves and in whitefly vector through direct antigen coating enzyme linked immunosorbent assay which revealed the low virus titre in Pseudomonas treated plants (VPT10 + chitin) and insect vector compared to untreated tomato plants. The results indicate the potentiality of plant growth promoting rhizobacteria strains and talc-powder formulations in the effective management of this tomato leaf curl virus in tomato under field conditions.     

Combinatorial effect of endophytic and plant growth promoting rhizobacteria against wilt disease of Capsicum annum L. caused by Fusarium solani

Combination of biocontrol agents that are compatible with each other is a strategic approach to control the plant disease and pest. The present study was designed to evaluate the protective effects of compatible endophytic bacterial strains (Bacillus subtilis; EPCO16 and EPC5) and rhizobacterial strain (Pseudomonas fluorescens; Pf1) against chilli wilt disease caused by Fusarium solani. Our results showed that B. subtilis (EPCO16 and EPC5) and P. fluorescens (Pf1) were compatible and effectively inhibited the growth of the F. solani. The application of endophytic and rhizobacterial strains, singly and in combination in green house and field conditions were found to be effective in controlling the chilli Fusarium wilt disease by inducing systemic resistance (ISR) as evidenced by enhanced activities of PO, PPO, PAL, β-1,3-glucanase, Chitinase and Phenolic involved in the synthesis of phytolaexins thereby promoting the growth of plants. However, combinations of EPCO16 + EPC5 + Pf1 bacterial strains were more effective than single agents. These findings suggest that synergistic interactions of biocontrol agents may be responsible for the management of chilli wilt disease caused by F. solani.     

Effect of plant growth-promoting Rhizobacteria and culture filtrate of Sclerotium rolfsii on phenolic and salicylic acid contents in chickpea (Cicer arietinum)

Two plant growth-promoting rhizobacteria (PGPR), viz., Pseudomonas fluorescens strain Pf4 and P. aeruginosa strain Pag, protected chickpea ( Cicer arietinum) plants from Sclerotium rolfsii infection when applied singly or in combination as seed treatment. Pag gave the best protection to the seedlings, applied either singly (mortality 16%) or in combination with Pf4 (mortality 17%) compared with 44% and 24% mortality in control and Pf4 treatment, respectively. The two PGPR strains induced the synthesis of specific phenolic acids, salicylic acid (SA), as well as total phenolics at different growth stages of chickpea seedlings with varied amount. The maximum amount of total phenolics was recorded in all the aerial parts of 4-week-old plants. Gallic, ferulic, chlorogenic, and cinnamic acids were the major phenolic acids detected in high-performance liquid chromatography (HPLC) analysis. Induction of such phenolic acids in the seedlings was observed up to 6 weeks in comparison with control. Salicylic acid (SA) was induced frequently during the first 3 weeks of growth only. Between the two strains, Pag was more effective in inducing phenolic acid synthesis applied either singly or in combination with strain Pf4 during the entire 6 weeks of growth of chickpea. In the presence of a culture filtrate of S. rolfsii, the two Pseudomonas strains induced more phenolic acids in treated than in non-treated and control plants. The occurrence of salicylic acid was frequent in the first 24 h, but infrequent at 48 and 96 h. Foliar spray of Pseudomonas strains also enhanced the phenolic acid content as well as total phenolics within 24 h of application. Gallic, chlorogenic, and cinnamic acids were consistently discerned in the treated leaves, whereas SA was absent even up to 96 h of application. Resistance in chickpea plants by Pseudomonas strains through induction of phenolic compounds as well as induced systemic resistance via SA-dependent pathway was evident.     

Combined application of botanical formulations and biocontrol agents for the management of Fusarium oxysporum f. sp. cubense (Foc) causing Fusarium wilt in banana

Plant products along with biocontrol agents were tested against Fusarium wilt of banana caused by Fusarium oxysporum f. sp. cubense (Foc). Of the 22 plant species tested, the leaf extract of Datura metel (10%) showed complete inhibition of the mycelial growth of Foc. Two botanical fungicides, Wanis 20 EC and Damet 50 EC along with selected PGPR strains with known biocontrol activity, Pseudomonas fluorescens 1, Pf1 and Bacillus subtilis, TRC 54 were tested individually and in combination for the management of Fusarium wilt under greenhouse and field conditions. Combined application of botanical formulation and biocontrol agents (Wanis 20 EC + Pf1 + TRC 54) reduced the wilt incidence significantly under greenhouse (64%) and field conditions (75%). Reduction in disease incidence was positively correlated with the induction of defense-related enzymes peroxidase (PO) and polyphenol oxidase (PPO). Three antifungal compounds (two glycosides and one ester) in D. metel were separated and identified using TLC, RP-HPLC (Reverse Phase-High Pressure Liquid Chromatography) and mass spectrometry. In this study it is clear that combined application of botanical formulations and biocontrol agents can be very effective in the management of Fusarium wilt of banana.     

Induction of systemic resistance by mixtures of antagonist bacteria for the management of crown rot complex on banana

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.     

Biological control of rice bacterial blight by plant-associated bacteria producing 2,4-diacetylphloroglucinol

Certain plant-associated strains of fluorescent Pseudomonas spp. are known to produce the antimicrobial antibiotic 2,4-diacetylphloroglucinol (DAPG). It has antibacterial, antifungal, antiviral, and antihelminthic properties and has played a significant role in the biological control of tobacco, wheat, and sugar beet diseases. It has never been reported from India and has not been implicated in the biological suppression of a major disease of the rice crop. Here, we report that a subpopulation of 27 strains of plant-associated Pseudomonas fluorescens screened in a batch of 278 strains of fluorescent pseudomonads produced DAPG. The DAPG production was detected by a PCR-based screening method that used primers Phl2a and Phl2b and amplified a 745-bp fragment characteristic of DAPG. HPLC, 1H NMR, and IR analyses provided further evidence for its production. We report also that this compound inhibited the growth of the devastating rice bacterial blight pathogen Xanthomonas oryzae pv. oryzae in laboratory assays and suppressed rice bacterial blight up to 59%-64% in net-house and field experiments. Tn5 mutants defective in DAPG production (Phl-) of P. fluorescens PTB 9 were much less effective in their suppression of rice bacterial blight.     

Genetic diversity in pseudomonads associated with cereal cultures infected with basal bacteriosis

The genetic properties of 45 pseudomonad strains isolated from cereal cultures exhibiting symptoms of basal bacteriosis have been investigated. Considerable genetic diversity has been demonstrated using DNA fingerprints obtained by amplification with REP, ERIC, and BOX primers. Restriction analysis of the 16S-23S internal transcribed spacer (ITS1) allowed the strains to be subdivided into two major groups. In a phylogenetic tree, the ITS1s of these groups fell into two clusters, which also included the ITS1 of Pseudomonas syringae ("Syringae" cluster) and the ITS1 of P. fluorescens, P. tolaasii, P. reactans, P. gingeri, and P. agarici ("Fluorescens" cluster) from the GenBank database. Comparison of the ITS1 divergence levels within the "Fluorescens" cluster suggests expediency of treating P. tolaasii, P. reactans, various P. fluorescens groups, and, possibly, P. gingeri and P. agarici as subspecies of one genospecies. The intragenomic heterogeneity of ITS1s was observed in some of the pseudomonad strains studied. The results of amplification with specific primers and subsequent sequencing of the amplificate suggest the possibility of the presence of a functionally active syrB gene involved in syringomycin biosynthesis in the strains studied.     

双价杀虫蛋白基因在荧光假单胞菌中的表达及增效

利用广宿主质粒载体pJMS6αlac将苏云金芽胞杆菌(Bacillus thuringiensis)杀虫晶体蛋白基因cry1Ac和cry2Aa基因分别及一起进行克隆,将重组质粒导入能在多种作物上定殖、对植物病菌有良好抑菌和防治作用的荧光假单胞菌(Pseudomonas fluorescens)P303菌株,分别得到工程菌株IPP101、IPP201和IPP202。PCRRFLP和Southern blot检测均证明目的基因已经导入了工程菌。SDSPAGE电泳显示工程菌中存在明显的Cry1Ac蛋白带;透射电镜观察发现含cry1Ac基因的两个菌株IPP101和IPP202中杀虫蛋白形成了典型的菱形晶体和蛋白包含体,而在野生P303菌株中均无这些结构。这些结果说明,工程菌中cry1Ac基因得到了很好表达。室内杀虫试验表明：工程菌对棉铃虫初孵幼虫的致死中浓度(LC50),只含cry1Ac的IPP101为000812mL/g饲料,只含cry2Aa的IPP201为002604mL/g饲料,含双基因的IPP202为000186mL/g饲料;HD73为000170mL/g饲料。cry1Ac和cry2Aa双基因表达产物具有显著增效作用,共毒系数达3328。     

Application of inorganic carrier-based formulations of fluorescent pseudomonads and Piriformospora indica on tomato plants and evaluation of their efficacy

Aims: Fluorescent pseudomonads are widely used as bioinoculants for improving plant growth and controlling phytopathogenic fungi. Piriformospora indica (Pi), a symbiotic root endophyte, also has beneficial effects on a number of plants. The present study focuses on the improvement of growth yields of tomato plants and control of Fusarium wilt using inorganic carrier‐based formulations of two fluorescent pseudomonad strains (R62 and R81) and Pi. Methods and Results: The inorganic carrier‐based formulations of pseudomonad strains and Pi were tested for plant growth promotion of tomato plants under glass house and field conditions. In controlled glass house experiments, 8·8‐fold increase in dry root weight and 8·6‐fold increase in dry shoot weight were observed with talcum powder‐based consortium formulation of R81 and Pi. Field trial experiments ascertained the glfass house results with a considerable amount of increase in plant growth responses, and amongst all the treatments, R81 + Pi treatment performed consistently well in field conditions with an increase of 2·6‐, 3·1‐ and 3·9‐fold increase in dry root weight, shoot weight and fruit yield, respectively. The fluorescent pseudomonad R81 and Pi also acted as biocontrol agents, as their treatments could control the incidence of wilt disease caused by Fusarium oxysporum f.sp. lycopersici in tomato plants under glass house conditions. Conclusions: The culture broths of pseudomonads R62, R81 and Pi were successfully used for development of talcum‐ and vermiculite‐based bioinoculant formulations. In controlled glasshouse experiments, the talcum‐based bioinoculant formulations performed significantly better over vermiculite‐based formulations. In field experiments the talcum‐based consortium formulation of pseudomonad R81 and Pi was most effective. Significance and Impact of the Study: This study suggests that the formulations of pseudomonad strains (R62 and R81) and Pi can be used as bioinoculants for improving the productivity of tomato plants. The application of such formulations is a step forward towards sustainable agriculture.     

Genomic and genetic analyses of diversity and plant interactions of Pseudomonas fluorescens

Background

Pseudomonas fluorescens are common soil bacteria that can improve plant health through nutrient cycling, pathogen antagonism and induction of plant defenses. The genome sequences of strains SBW25 and Pf0-1 were determined and compared to each other and with P. fluorescens Pf-5. A functional genomic in vivo expression technology (IVET) screen provided insight into genes used by P. fluorescens in its natural environment and an improved understanding of the ecological significance of diversity within this species.

Results

Comparisons of three P. fluorescens genomes (SBW25, Pf0-1, Pf-5) revealed considerable divergence: 61% of genes are shared, the majority located near the replication origin. Phylogenetic and average amino acid identity analyses showed a low overall relationship. A functional screen of SBW25 defined 125 plant-induced genes including a range of functions specific to the plant environment. Orthologues of 83 of these exist in Pf0-1 and Pf-5, with 73 shared by both strains. The P. fluorescens genomes carry numerous complex repetitive DNA sequences, some resembling Miniature Inverted-repeat Transposable Elements (MITEs). In SBW25, repeat density and distribution revealed ''repeat deserts'' lacking repeats, covering approximately 40% of the genome.

Conclusions

P. fluorescens genomes are highly diverse. Strain-specific regions around the replication terminus suggest genome compartmentalization. The genomic heterogeneity among the three strains is reminiscent of a species complex rather than a single species. That 42% of plant-inducible genes were not shared by all strains reinforces this conclusion and shows that ecological success requires specialized and core functions. The diversity also indicates the significant size of genetic information within the Pseudomonas pan genome.     

Enhancement of population densities of fluorescent pseudomonads in the rhizosphere of tomato plants by addition of acibenzolar-S-methyl

Fluorescent pseudomonad isolates G309 and CW2, in combination with the resistance inducer acibenzolar-S-methyl (ASM), improved control of fungal and bacterial diseases on tomato plants. The interactions of the bacteria in the presence of ASM showed that in vitro growth of Pseudomonas fluorescens G309 and Pseudomonas sp. strain CW2 was not affected in King's B broth supplemented with 10 and 20 microM ASM. Also, the bacterial cells were not able to utilize ASM as a nutrient source. In vitro production of the two antimicrobial secondary metabolites phenazine-1-carboxylic acid and 2-OH-phenazine by the isolate CW2 was not affected within 3 days from incubation. In contrary, addition of ASM at a concentration of 20 microM to King's B liquid medium significantly increased production of salicylic acid by isolate G309. When roots of tomato plants were treated with G309 or CW2 cell suspensions containing 20 microM ASM, the number of bacterial cells recovered from the rhizosphere was significantly higher in the combined treatments than in the single applications 5, 10, and 15 days after inoculation. However, ASM at a higher concentration (50 microM) did not appreciably enhance the population sizes of either bacterial isolate in the rhizosphere. Enhanced bacterial cell densities in the rhizosphere of tomato plants were also determined following simultaneous treatments of tomato roots with 10 and 20 microM ASM in combination with the transformed isolate G309-384 (mini-Tn5gfp), which encodes the green fluorescent protein.