Efficacy of Pseudomonas fluorescens and Paecilomyces lilacinus against Meloidogyne graminicola infecting rice under system of rice intensification

The talc-based formulations of the plant growth promoting rhizobacterium, Pseudomonas fluorescens and egg parasitic fungi, Paecilomyces lilacinus, were evaluated as seed treatment, soil application and combination of both for the management of M. graminicola in fields of rice grown under system of rice intensification. Both the bioformulations significantly reduced the root invasion and soil populations of M. graminicola but P. fluorescens was most effective when applied as seed cum soil application and seed treatment alone. Effect of these treatments was comparable with the standard chemical carbofuran application. The introduced P. fluorescens survived significantly in rice roots when applied as seed cum soil application and seed application alone than as soil application. There was significant increase in phenol, peroxidase and chitinase accumulation in plants treated with P. fluorescens. Application of bioagents had positive influence on growth parameters such as plant height, root length, shoot weight, root weight and number of tillers per hill. Application of P. fluorescens as seed cum soil treatment resulted in higher grain yield, which was 20.6%–26.9% increase over control followed by P. fluorescens as seed treatment alone that increased grain yield of rice by 10.7%–11.2% than control. However, economic returns per investment was higher when P. fluorescens was applied as seed treatment alone (1:8.8–1:12.0 incremental cost benefit ratio) followed by the P. fluorescens as seed cum soil treatment (1:6.2–1:9.7 incremental cost benefit ratio).     

Pseudomonas fluorescens enhances resistance and natural enemy population in rice plants against leaffolder pest

Plant growth promoting fluorescent pseudomonad strains Pf1, TDK1 and PY15 were evaluated for their efficacy against leaffolder ( Cnaphalocrocis medinalis ) pest in rice plants under field conditions individually and in combinations. Application of mixtures of Pseudomonas fluorescens strains Pf1, TDK1 and PY15 significantly reduced the leaffolder damage in rice plants compared with untreated control. Interestingly, natural enemy population in plots treated with P. fluorescens was greater than the chemical and untreated controls. Further, support for these results was gathered by assaying activities of polyphenol oxidase (PPO) and lipoxygenase (LOX) under glasshouse conditions. The results showed the higher activity of PPO and LOX in plants treated with P. fluorescens mixtures (Pf1 + TDK1 + PY15) than the plants treated with individual strains, chemical and untreated controls. Further, fluorescent pseudomonad mixtures increased the rice yield compared with individual strain and non-bacterized treatments. The present study reveals that in addition to plant growth promotion, plant growth-promoting rhizobacterial (PGPR) strains-mediated induction of PPO and LOX in rice plants could be involved in enhanced natural enemy populations and resistance mechanisms against leaffolder attack.     

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.

Effect of PGPR on population growth parameters of cotton aphid

Plant-growth-promoting rhizobacteria (PGPR) help plants by increasing plant growth and defending them against pathogens and pests. There is considerable research supporting the use of these bacteria in plant pathogens management, but the number of research papers that have focused on their effect on insect pests control is quite few. In this research, we examined the ability of four native strains of Pseudomonas fluorescens to manage the cotton aphid, Aphis gossypii, on greenhouse-grown cucumbers in Iran. The strains UTPF (University of Tehran Pseudomonas fluorescens) 68, UTPF1, UTPF6 and PF169 of P. fluorescens were used as seed treatment to evaluate their impact on both cucumber growth and cotton aphid population growth parameters. The strains UTPF68 and PF169 of this bacterium influenced phenology of cucumber by decreasing the Growing Degree Days of flowering period. Also, the plants treated with these bacterial strains had a significant increase in yield weight of approximately 58%. In the other part of the study, various effects on developmental parameters of aphid were recorded among bacterial strains, but only PF169 caused significant decreasing effect on r_m value of aphid, which is the most important factor in aphid’s life table. We conclude that the PF169 treatment had significant negative effects on population growth rate of A. gossypii. The PF169 could be reliable in a control programme for A. gossypii on cucumber plants in greenhouses.     

Induction of phenylpropanoid metabolism by Pseudomonas fluorescens against tomato spotted wilt virus in tomato

Pseudomonas fluorescens (two native strains, one collection strain and their strain mixtures in all possible combinations) when applied through seed, seedling dip, soil and on leaf significantly reduced the tomato spotted wilt virus (TSWV) disease. InP. fluorescens-treated plants, the peroxidase and phenylalamine ammonia-lyase activity increased. Accumulation of phenolic compounds and lignin were shown to be increased in theP. fluorescens-treated plants. Isoperoxidase native PAGE indicated that the peroxidase isoforms in tomato plants induced by fluorescent pseudomonads were different from the control plants; this suggests that the general phenylpropanoid pathway is probably stimulated in tomato plants treated which in turn led to significant reduction in TSWV.     

Biological control of onion leaf blight disease by bulb and foliar application of powder formulation of antagonist mixture

Abstract

Three antagonists: Pseudomonas fluorescens (Pf1), Bacillus subtilis and Trichoderma viride, were tested alone and in combination for suppression of onion leaf blight (Alternaria palandui) disease under glasshouse and field conditions. The average mean of disease reduction was 24.81% for single strains and 42.44% for mixtures. In addition to disease suppression, treatment with a mixture of antagonists promoted plant growth in terms of increased plant height and ultimately bulb yield. Though seed treatment of either single strain or strain mixtures alone could reduce the disease, subsequent application to root, leaves or soil further reduced the disease and enhanced the plant growth. The mixture consisting of Pseudomonas fluorescens Pf1 plus Bacillus subtilis plus Trichoderma viride was the most effective in reducing the disease and in promoting plant growth and bulb yield in greenhouse and field tests.     

Evaluation of Some Fungi and Bacteria for Potential Control of Safflower Rust

Some fungal and bacterial isolates applied as soil and seed treatments in greenhouse trials gave a good protection against Puccinia carthami Cda. on safflower, significantly reducing the infection from rust-infested seeds. Trichoderma viride and T. harzianum added alone and in combination as air-dried inoculum to the soil were the most effective fungal isolates. However T. viride at two higher rates reduced the rate of emergence of safflower seedlings. Among bacteria, Bacillus subtilis, B. cereus, isolates of Pseudomonas fluorescens and B. thuringiensis reduced rust infection when added both as soil drench and as seed treatment. Good results in the biocontrol of P. carthami were also obtained with some combination antagonist treatments such as T. viride + B. cereus, T. viride + P. fluorescens (16), T. viride + T. harzianum + B. cereus and T. viride +, T. harzianum + P. fluorescens (16). Fungal isolates added as soil treatment increased seedling length, whereas no similar effects were observed when these isolates were applied as seed treatment.     

Biological control of the wheat root rot caused by Fusarium graminearum using some PGPR strains in Saudi Arabia

The aim of this study was to evaluate the efficacy of selected bacterial strains against the wheat soil‐borne pathogen Fusarium graminearum under greenhouse conditions. The most potent isolates were 3 isolates out of 18 isolates, which have numbers 3, 9 and 10 with in vitro inhibition index 42.5%, 41.3% and 46.3% respectively. Isolates 3 and 10 were selected for the following experiments. Isolates 3 and 10 were identified as Bacillus subtilis MAA03 and Pseudomonas fluorescens MAA10, respectively according to International Identification Keys and, confirmed by using Biolog system and 16S rDNA where the strains exhibited more than 99.5% sequence identity. Their close taxonomic relationship was further documented by phenotypic similarities. The using of B. subtilis and P. fluorescens separately or in mixture as biocontrol agent against F. graminearum on wheat significantly increased the final germination percent, the mean daily germination and germination index of wheat cultivar, while the mean germination time was significantly decreased relative to infested control. The final infection percent, the mean daily infection and infection index were decreased significantly, while the mean infection time was significantly increased relative to infested control. The use of P. fluorescens as biocontrol agent was the most efficient than B. subtilis or in mixture and the best treatment was seed coating. The application of B. subtilis and P. fluorescens separately or in combination significantly affected the growth parameters of wheat cultivar Tabuki, the root length was significantly increased in seed coating and seed soaking treatments, while non‐significantly decreased in case of soil drench treatment relative to infested control. Shoot length was significantly decreased in case of seed coating treatment relative to infested control. The shoot fresh and dry weights were significantly increased in seed coating and seed soaking treatments relative to infested control. The root fresh and dry weights were significantly increased in seed coating and seed soaking treatments relative to infested control. The number of leaves was significantly increased in all treatments relative to infested control.     

A new microbial consortia containing entomopathogenic fungus,Beauveria bassiana and plant growth promoting rhizobacteria,Pseudomonas fluorescens for simultaneous management of leafminers and collar rot disease in groundnut

The virulence of 20 isolates of Beauveria bassiana (Balsamo) Vuillemin to larvae of the leafminer, Aproaerema modicella, was tested in the laboratory. Leafminer larvae were sprayed with a standard concentration of 1×10⁸ condia/mL of each B. bassiana isolate. All the B. bassiana isolates tested were pathogenic to A. modicella and the mortality varied between 16.7 and 68.9%. Beauveria bassiana isolate B2 was found to be the most virulent followed by isolate B4 which resulted in 59% mortality. Beauveria isolate B2 was selected for dose–response mortality studies with four different doses (1×10², 1×10⁴, 1×10⁶ and 1×10⁸ conidia/mL). Among the various doses tested, 1×10⁸ conidia/mL produced the highest mortality (70.7%). In addition, morphogenesis of the insect pest in all stages, larval, pupal and adult was greatly affected due to fungal infection. Further, B. bassiana isolate B2 and two Pseudomonas fluorescens strains, TDK1 and Pf1 were tested alone and in combination for suppression of groundnut leafminer and collar rot disease and promotion of plant growth and yield both under glasshouse and field conditions. The mixture of B. bassiana and P. fluorescens strains significantly reduced the leafminer and collar rot disease incidences when applied as talc-based formulation through seed, soil and foliar application under glasshouse and field conditions.     

Three native Pseudomonas fluorescens strains tested under growth chamber and field conditions as biocontrol agents against damping-off in alfalfa

Alfalfa (Medicago sativa) is one of the most important crops used in Uruguay for livestock feeding. Seedling diseases, particularly damping-off, are a critical factor which limits its establishment. Three native Pseudomonas fluorescens strains, UP61.2, UP143.8 and UP148.2, previously isolated from Lotus corniculatus, were evaluated to determine their efficacy as biological control agents for alfalfa seedling diseases in the field. Their compatibility with the alfalfa-Sinorhizobium meliloti symbiosis was also assessed. In growth chamber conditions seed inoculation with Pseudomonas strains did not affect different parameters of alfalfa-rhizobium symbiosis as shown by nodulation rate and shoot dry weight of plants. The presence of the commercial inoculant strains of S. meliloti did not impair colonization by the P. fluorescens and vice versa. In field trials the dynamics of rhizobial rhizospheric populations were not affected by the presence of P. fluorescens. Each P. fluorescens strain successfully colonized alfalfa roots at adequate densities for biocontrol activity. Results showed that P. fluorescens strains provided a 10–13% increase in the number of established plants relative to the control, an intermediate result compared to the fungicide treatment (24%). The alfalfa above-ground biomass was increased by 13% and 15–18% in the presence of the fungicide and P. fluorescens strains, respectively. Therefore, results from this study demonstrated that the three P. fluorescens strains provided effective control against soil-borne pathogens and suggest a potential use in the development of a commercial inoculant to be applied for the control of legume seedling diseases.     

Evaluation of Pseudomonas fluorescens for Suppression of Sheath Rot Disease and for Enhancement of Grain Yields in Rice (Oryza sativa L.)

Pseudomonas fluorescens strains antagonistic to Sarocladium oryzae, the sheath rot (Sh-R) pathogen of rice (Oryza sativa L.), were evaluated in greenhouse and field tests for suppression of Sh-R severity and enhancement of grain yields of rice. Imprints of rice seedlings and a direct-observation technique of staining roots with fluorochromes confirmed the association of P. fluorescens with roots and the ability of the strain to move along shoot tips. In greenhouse tests, P. fluorescens-treated rice plants (cv. IR 20) showed a 54% reduction in the length of Sh-R lesions. In three field tests, treatment with P. fluorescens reduced the severity of Sh-R by 20 to 42% in five rice cultivars. Bacterization of rice cultivars with P. fluorescens enhanced plant height, number of tillers, and grain yields from 3 to 160%.     

Understanding the molecular basis of plant growth promotional effect of Pseudomonas fluorescens on rice through protein profiling

Background  

Plant Growth Promoting Rhizobacteria (PGPR), Pseudomonas fluorescens strain KH-1 was found to exhibit plant growth promotional activity in rice under both in-vitro and in-vivo conditions. But the mechanism underlying such promotional activity of P. fluorescens is not yet understood clearly. In this study, efforts were made to elucidate the molecular responses of rice plants to P. fluorescens treatment through protein profiling. Two-dimensional polyacrylamide gel electrophoresis strategy was adopted to identify the PGPR responsive proteins and the differentially expressed proteins were analyzed by mass spectrometry.     

Pectolytic enzymes activity and pathogenicity ofGaeumannomyces graminis var.tritici and related Phialophora-like fungi

Gaeumannmyces graminis var.tritici (Ggt), Phialophora sp. (lobed hyphopodia) andPhialophora graminicola vere grown in a liquid medium with pectin and on autoclaved wheat roots (root media) and the activity of pectolytic enzymes in culture filtrates was measured. Most strains of the fungi exhibited polygalacturonate trans-eliminase activity but no pectin methylesterase activity was detected.Ggt polygalacturonase was found in culture filtrates from all the media used whilePhialophora sp. did not exhibit activity of this enzyme in the unbuffered root media. No polygalacturonase activity was demonstrated forP. graminicola. A correlation was found (r=0.548) betweenin vitro polygalacturonase activity and the pathogenicity ofGgt to wheat seedlings.     

Effects of Pseudomonas fluorescens DF57 on growth and P uptake of two arbuscular mycorrhizal fungi in symbiosis with cucumber

 The effect of Pseudomonas fluorescens DF57 on growth and P uptake of two arbuscular mycorrhizal (AM) fungi in symbiosis with cucumber plants was studied in compartmentalised growth systems. Hyphae of Glomus intraradices Schenck & Smith (BEG87) or G. caledonium (Nicol. & Gerd.) Trappe & Gerdeman (BEG15) grew into lateral root-free compartments. Non-mycorrhizal plants served as control. The soil in half of the growth units of each mycorrhizal treatment was inoculated with P. fluorescens DF57. P. fluorescens DF57 enhanced hyphal length density of one of the AM fungi, G. caledonium, but this was not reflected in a higher hyphal transport of P from the root-free soil to the plant. The total P content was higher in plants grown in symbiosis with G. intraradices than in plants in the other treatments. G. caledonium and P. fluorescens DF57 had a synergistic effect in that total P content in plants inoculated with G. caledonium was higher in the presence than in the absence of P. fluorescens DF57. Accepted: 7 January 1999     

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.     

Mycolytic effect of fluorescent Pseudomonas in biocontrolling of fungal phytopathogenic Curvularia lunata,Fusarium oxysporum,Alternaria padwickii and Rhizoctonia solani

About 50 bacterial strains, each of Pseudomonas fluorescens, from different rhizospheric soil of different plants were screened for antagonistic activity against Curvularia lunata, Fusarium oxysporum, Alternaria padwickii, Rhizoctonia solani causing black kernel, kernel spotting, root rots, stackburn and sheath blight diseases of rice (Oryza sativa L.). Out of the 50 isolates, 15 isolates were found to be effective in lysing the cell wall of the above-mentioned putative pathogens tested in vitro. These Pseudomonas isolates produced mycolytic enzymes, viz. β-1,3-glucanases, β-1,4-glucanases and lipases. P. fluorescens PAK1 and PAK12 among the strains were more effective for the production of these enzymes while PAK12 produce good level of β-1,3-glucanases, β-1,4-glucanases and lipases against tested fungal pathogens. These findings demonstrate a mechanism of antagonism by P. fluorescens against different fungal plant pathogens.     

Variability in Chemical Composition and Antimicrobial Activity of Mentha pulegium L. Essential Oil,Cultivated under Different Plant Growth Promoting Rhizobacteria

The aim of present work was to study the essential oil chemical composition and antimicrobial activity of cultivated Mentha pulegium L. under different plant growth promoting rhizobacteria (Pseudomonas fluorescens, Bradyrhizobium sp. and Sinorhizobium meliloti) individually and in consortium. Yield, in plants inoculated with Bradyrhizobium sp. and S. meliloti in consortium, increase significantly relative to control plants. GC and GC/MS analyses pointed to a qualitative and quantitative variability of components. The investigated essential oils were clustered into three chemotypes: piperitenone/1,8-cineol (40.9/29.4 %) chemotype in plants inoculated with Bradyrhizobium sp. individually, S. meliloti individually, and Bradyrhizobium sp. and S. meliloti in consortium, piperitone/menthone (41.8/33.8 %) chemotype in plants inoculated with P. fluorescens individually, P. fluorescens and Bradyrhizobium sp. in consortium, and P. fluorescens and S. meliloti in consortium and pulegone/menthol (47.9/31.5 %) chemotype in control plants. The antimicrobial activity, carried out by the disc diffusion method and the determination of the Minimum Inhibitory Concentration (MIC) against ten microorganisms, varied significantly according to the tested microorganism and the rhizobacterial species used individually or in consortium (inhibition zone: 8.5–33.5 mm; MIC: 0.25–2.5 μL/mL). Our findings provided useful indications to select interesting chemotype within M. pulegium, especially in perspective of its cultivation.     

A new bio-formulation containing plant growth promoting rhizobacterial mixture for the management of sheath blight and enhanced grain yield in rice

Three plant growth promoting rhizobacterial (PGPR) strains, PF1,FP7 and PB2, were tested alone and in combinations for suppression ofrice sheath blight disease and promotion of plant growth underglasshouse and field conditions. The mixture of PGPR strainssignificantly reduced the sheath blight incidence when applied as eitherbacterial suspension through seed, root, foliar and soil application inglasshouse conditions, or as talc-based formulation under fieldconditions, compared to the respective individual strains. The averagemean of disease reduction was 29.2% for single strains and45.1% for mixtures. In addition to disease suppression, treatmentwith mixture of PGPR strains promoted plant growth in terms of increasedplant height and number of tillers, and ultimately grain yield. Theaverage increases in yield for single strains were 17.7%, and25.9% in case of mixture. Mixture of three PGPR strains reduceddisease and promoted growth to a level equivalent to two strainmixtures. Though seed treatment of either single strain or strainmixtures alone could reduce the disease, subsequent application to root,leaves or soil further reduced the disease and enhanced the plantgrowth. The mixture consisting of PF1 plus FP7 was the most effective inreducing the disease and in promoting plant growth and grainyield.     

Effect of Pseudomonas fluorescens RB4 and Bacillus subtilis 189 on the phytoremediation potential of Catharanthus roseus (L.) in Cu and Pb-contaminated soils

The remediation of heavy metal-contaminated soils has become a critical issue due to toxic effects of these metals on living organisms. The current research was conducted to study the effect of Pseudomonas fluorescens RB4 and Bacillus subtilis 189 on the growth and phytoremediation potential of Catharanthus roseus in Cu- and Pb-contaminated soils. The bacterial strains exhibited significantly higher level of water-extractable Pb and Cu in Pb, Cu, and Cu+Pb-contaminated. The P. fluorescens RB4 inoculated plants, produced 102%, 48%, and 45% higher fresh weight (FW) in soils contaminated with Cu, Pb, and both elements, respectively, as compared to un-inoculated control plants. Similarly, B. subtilis 189 inoculated plants produced 108%, 43%, and 114% more FW in the presence of Cu, Pb, and both elements. The plants co-cultivated with both bacteria exhibited 121%, 102%, and 177% higher FW, in Cu, Pb, and both elements contaminated soils, as compared to respective un-inoculated control. Co-cultivation of P. fluorescens RB4, B. subtilis 189, and P. fluorescens RB4 + B. subtilis 189 resulted in higher accumulation of Cu and Pb in shoots of the C. roseus grown in contaminated soils as compared to un-inoculated control. Bacterial treatments also improved the translocation and metal bioconcentration factors. The growth and phytoextraction capability of C. roseus was improved by inoculation of P. fluorescens RB4 and B. subtilis 189.     

Endophytic bacteria mediate plant resistance against cotton bollworm

Abstract

The efficacy of endophytic bacterial strains was evaluated in cotton against American bollworm infestation under greenhouse conditions. Among the 103 endophytic bacterial strains, the Bacillus strains (EPCO 102 and EPCO 16) and Pseudomonas fluorescens strain Pf1 significantly reduced the bollworm incidence. Talc-based bioformulation of EPCO 102, EPCO 16 and P. fluorescens Pf1 with and without chitin in inducing systemic resistance was tested against bollworm. The application of the bioformulation through seed, soil and foliar spray significantly reduced the bollworm incidence. The amendment of chitin in the formulation further reduced the pest incidence. Maximum bollworm reduction by endophytic bacterial strains EPCO 102, EPCO 16 and Pf1 strain with chitin was recorded. In addition, endophytic bacterial bioformulation with chitin induced more and timely activities of chitinase, β-1, 3-glucanase, peroxidase, polyphenol oxidase, phenylalanine ammonia-lyase and phenol in cotton plants infested with Helicoverpa armigera.