Influence of inoculation with plant growth promoting rhizobacteria (PGPR) on tomato plant growth and nematode reproduction under greenhouse conditions

Numerous species of soil bacteria which flourish in the rhizosphere of plants or around plant tissues stimulate plant growth and reduce nematode population by antagonistic behavior. These bacteria are collectively known as PGPR (plant growth promoting rhizobacteria). The effects of six isolates of PGPR Pseudomonas putida, Pseudomonas fluorescens, Serratia marcescens, Bacillus amyloliquefaciens, Bacillus subtilis and Bacillus cereus, were studied on tomato plant growth and root knot nematode reproduction after 45 days from nematode infection. The highest number of shoot dry weight/g (43.00 g) was detected in the plant treated with S. marcescens; then P. putida (34.33 g), B. amyloliquefaciens (31.66 g), P. fluorescens (30.0 g), B. subtilis (29.0 g), B. cereus (27.0 g) and nematode alone (untreated) 20 g/plant. While the highest number of plant height was observed when plant was treated with S. marcescens, P. fluorescens, P. putida, B. amyloliquefaciens and P. putida 52.66, 50.66, 48 and 48 cm respectively. No significant differences were seen between previous treatments but only had significant differences compared with untreated plant. The highest number of fruit/plant was observed when plants were treated with S. marcescens (10.66), then B. amyloliquefaciens (8.66), P. putida (8), P. fluorescens (8) and B. cereus (7.66). No significant differences between the last 4 treatments, but all had significant differences compared with untreated plants. The highest weight of plant yield (g) was observed with S. marcescens (319.6 g/plant) and the lowest weight of plant yield was observed in plants treated with nematode alone (untreated). On the other hand, the lowest numbers of J₂/10 g of soil (78), galls/root, (24.33) galls/root, egg masses/root (12.66) and egg/egg masses were observed in the plants treated with S. marcescens.     

Biological control of sunflower necrosis virus disease with powder and liquid formulations of plant growth promoting microbial consortia under field conditions

Sunflower necrosis virus disease (SNVD) was reported to cause a significant damage to sunflower production in India. Based on the biocontrol efficacy against SNVD observed in the previous greenhouse experiments, two plant growth promoting microbial consortia (PGPMCs) viz., PGPMC-1 consisting of Bacillus licheniformis strain MML2501 + Bacillus sp. strain MML2551 + Pseudomonas aeruginosa strain MML2212 + Streptomyces fradiae strain MML1042 and PGPMC-2 consisting of B. licheniformis MML2501 + Bacillus sp. MML2551 + P. aeruginosa MML2212 were used in the present study. Powder and liquid formulations of the above plant growth promoting microorganisms (PGPMs) were evaluated along with farmers’ practice (imidacloprid + mancozeb) and control in farmers’ fields. Significant disease reduction, increase of seed germination, plant height and yield parameters with an additional seed yield of 840 kg/ha, an additional income of Rs. 10,920/ha and benefit cost ratio of 6.1 were recorded following treatment with a powder formulation of PGPMC-1 compared to control. Further, liquid formulation of PGPMC-1 significantly reduced SNVD up to 51.4% compared to control. Besides, this treatment improved the seed germination (24.4%), plant height (61.3%), yield parameters with an additional seed yield of 936 kg/ha and an additional income of Rs. 12,168/ha. Benefit cost ratio was calculated as 6.8 in this treatment compared to 4.6 and 3.7, respectively for PGPMC-2 liquid formulation and farmers’ practice compared to control. This is the first report of PGPMCs mediated biological control of SNVD under field conditions.     

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.     

Experimental and mathematical simulation of plant growth promoting rhizobacteria and plant interaction under cadmium stress

Bacterial inoculants of the commercially available plant growth promoting rhizobacteria (PGPR) Arthrobacter mysorens 7, Flavobacterium sp. L30, and Klebsiella mobilis CIAM 880 were selected to obtain ecologically safe barley crop production on cadmium (Cd) polluted soils. All the PGPR immobilized 24–68% soluble cadmium from soil suspension. A. mysorens 7 and K. mobilis CIAM 880 were highly resistant to Cd and grew in up to 1 and 3 mmol CdCl₂ on DAS medium respectively. All PGPR were able to fix nitrogen (276–1014 nmol mg^–1 bacterial DW) and to produce indole acetic acid (IAA) (126–330 nmol mg^–1 bacterial DW) or ethylene (4.6–13.5 nmol bacterial DW). All the PGPR actively colonized barley root system and rhizosphere and significantly stimulated root elongation of barley seedlings (up to 25%), growing on soil containing 5 or 15 mg Cd kg^–1 of soil. Created in the simulation mathematical model confirms our hypothesis that PGPR beneficial effect on barley growing under Cd-stress is a complex process. One of mechanisms underlying this effect might be increase of bacterial migration from rhizoplane to rhizosphere, where PGPR bind soluble free Cd ions in biologically unavailable complex forms. Among the studied PGPR K. mobilis CIAM 880 was the most effective inoculant. Inoculation with K. mobilis CIAM 880 of barley plants growing on Cd contaminated soil (5 mg Cd kg^–1 of soil) under field conditions increased by 120% grain yield and 2-fold decreased Cd content in barley grain. The results suggest that the using K. mobilis CIAM 880 is an effective way to increase the plant yield on poor and polluted areas.     

Effect of plant growth promoting rhizobacteria on bacterial canker of tomato

Use of plant growth promoting rhizobacteria in managing bacterial canker disease of tomato was studied in the present work. Tomato seeds were treated with PGPR strains viz., Bacillus pumilus INR7, Bacillus pumilus SE34, Bacillus pumilus T4, Bacillus subtilis GBO3, Bacillus amyloliquefaciens IN937a and Brevibacillus brevis IPC11 were subjected for seed germination and seedling vigor. Among the PGPR strains tested, only three strains (IN937a, GBO3 and IPC11) which showed enhancement in the seed quality parameters like seed germination and seedling vigor, were further subjected for estimation of one of the defence-related enzymes, Phenylalanine Ammonia Lyase (PAL) with total phenol contents. The same three strains were recorded for maximum disease protection under greenhouse conditions. The level of PAL and total phenol contents increased significantly upon the PGPR treatment. The rate of reduction in the bacterial canker disease incidence was directly proportional to the amount of increased level of PAL and total phenol content. The possible uses of these PGPR strains in effective management of bacterial canker of tomato were discussed in the present work.     

Sunflower inoculation with Azospirillum and other plant growth promoting rhizobacteria

The bacterial microflora from sunflower rhizosphere (Helianthus annus L.) and one strain ofAzospirillum lipoferum of a different origin were screened for their ability to promote sunflower growth in a 6-day germination test and in pot experiments. TwoAzospirillum lipoferum strains and oneXanthomonas maltophilia strain produced the best responses. These strains were chosen for field testing.     

植物根际促生菌对3种土传真菌病害病原的抑制作用

【目的】获取促生同时可防治3种土传真菌病害(Fusarium oxysporum、Sclerotinia sclerotiorum和Rhizoctonia solani)的生防菌,并明确其抑菌效果。【方法】利用前期研究获得的17株促生菌,采用平板对峙法测定其对病原真菌的拮抗作用及对菌丝生长的抑制作用。【结果】可有效拮抗立枯丝核菌的生防菌有6株,其中促生菌株FX2和LM4-3的抑制率达73.82%;拮抗尖孢镰刀菌的生防菌有7株,其中FX2的抑制率达到66.81%;拮抗油菜菌核病菌的生防菌有4株,其中菌株LHS11的抑制率高达85.71%。菌株LHS11和JM170通过次生代谢物抑制病原真菌。所有的生防菌对病原菌的菌丝生长均有一定的抑制作用。【结论】筛选得到对3种真菌病害病原具有较好生防作用的菌株LHS11和FX2。     

Biocontrol of Alternaria triticina by plant growth promoting rhizobacteria on wheat

Abstract

Eighteen isolates of fluorescent pseudomonads and Bacillus spp. were isolated from Alternaria triticina suppressive soils of wheat fields. These isolates were evaluated in the laboratory and greenhouse for the biocontrol of A. triticina. Six isolates were considered to have potential for the biocontrol of A. triticina on the basis of antibiotic sensitivity, fluorescence produced by Pseudomonas, inhibitory effect on A. triticina and root colonization of wheat roots by these isolates. These six isolates (Pa22, Pf27, Pa28, B25, B28, and B30) were further tested for their biocontrol potential against A. triticina on wheat in a pot test. Out of six isolates, isolate B28 was best in improving wheat growth of A. triticina inoculated plants. Isolate B28 also caused higher reduction in percentage infected leaf area caused by A. triticina while isolate Pa22 was found best in improving growth of plants without A. triticina.     

Cytokinin production by plant growth promoting rhizobacteria and selected mutants

One of the proposed mechanisms by which rhizobacteria enhance plant growth is through the production of plant growth regulators. Five plant growth promoting rhizobacterial (PGPR) strains produced the cytokinin dihydrozeatin riboside (DHZR) in pure culture. Cytokinin production by Pseudomonas fluorescens G20-18, a rifampicin-resistant mutant (RIF), and two TnphoA-derived mutants (CNT1, CNT2), with reduced capacity to synthesize cytokinins, was further characterized in pure culture using immunoassay and thin layer chromatography. G20-18 produced higher amounts of three cytokinins, isopentenyl adenosine (IPA), trans-zeatin ribose (ZR), and DHZR than the three mutants during stationary phase. IPA was the major metabolite produced, but the proportion of ZR and DHZR accumulated by CNT1 and CNT2 increased with time. No differences were observed between strain G20-18 and the mutants in the amounts of indole acetic acid synthesized, nor were gibberellins detected in supernatants of any of the strains. Addition of 10(-5) M adenine increased cytokinin production in 96- and 168-h cultures of strain G20-18 by approximately 67%. G20-18 and the mutants CNT1 and CNT2 may be useful for determination of the role of cytokinin production in plant growth promotion by PGPR.     

Integrated biological control of bacterial speck and spot of tomato under field conditions using foliar biological control agents and plant growth-promoting rhizobacteria

Integration of foliar bacterial biological control agents and plant growth promoting rhizobacteria (PGPR) was investigated to determine whether biological control of bacterial speck of tomato, caused by Pseudomonas syringae pv. tomato, and bacterial spot of tomato, caused by Xanthomonas campestris pv. vesicatoria and Xanthomonas vesicatoria, could be improved. Three foliar biological control agents and two selected PGPR strains were employed in pairwise combinations. The foliar biological control agents had previously demonstrated moderate control of bacterial speck or bacterial spot when applied as foliar sprays. The PGPR strains were selected in this study based on their capacity to induce resistance against bacterial speck when applied as seed and soil treatments in the greenhouse. Field trials were conducted in Alabama, Florida, and California for evaluation of the efficacy in control of bacterial speck and in Alabama and Florida for control of bacterial spot. The foliar biological control agent P. syringae strain Cit7 was the most effective of the three foliar biological control agents, providing significant suppression of bacterial speck in all field trials and bacterial spot in two out of three field trials. When applied as a seed treatment and soil drench, PGPR strain Pseudomonas fluorescens 89B-61 significantly reduced foliar severity of bacterial speck in the field trial in California and in three of six disease ratings in the field trials in Alabama. PGPR strains 89B-61 and Bacillus pumilus SE34 both provided significant suppression of bacterial spot in the two field trials conducted in Alabama. Combined use of foliar biological control agent Cit7 and PGPR strain 89B-61 provided significant control of bacterial speck and spot of tomato in each trial. In one field trial, control was enhanced significantly with combined biological control agents compared to single agent inoculations. These results suggest that some PGPR strains may induce plant resistance under field conditions, providing effective suppression of bacterial speck and spot of tomato, and that there may be some benefit to the integration of rhizosphere-applied PGPR and foliar-applied biological control agents.     

Adsorption kinetics of Pb and Cd by two plant growth promoting rhizobacteria

A bench study was carried out to characterize the kinetics of two plant growth promoting rhizobacteria (PGPR) Azotobacter chroococcum and Bacillus megaterium to adsorb heavy metals from solution. Adsorption of Pb²⁺ and Cd²⁺ by bacterial cells was processed quickly with an equilibration achieved within 5 min. The adsorptions were fitted well with Freundlich and Langmuir isotherm models. The comparison of isotherm parameters indicated that A. chroococcum had a stronger capacity to bind metal ions than B. megaterium, with an average increase of 59.8% for Pb²⁺ and 75.6% for Cd²⁺, respectively. Both bacteria had a stronger affinity to Pb²⁺ than Cd²⁺ since Pb²⁺ was more easily bound with the phosphoryl groups on the cell surface than Cd²⁺. This demonstrated that the presence of bacteria in the rhizosphere may result in the reduction of mobile ions in soil solution.     

Fusarial wilt control and growth promotion of pigeon pea through bioactive metabolites produced by two plant growth promoting rhizobacteria

The bioactive metabolites produced by two plant growth promoting rhizobacteria strains, a Pseudomonas aeruginosa strain RRLJ 04 and a Bacillus cereus strain BS 03, which showed growth promotion and disease control in pigeon pea against Fusarium udum, were isolated and screened for their efficacy to control fusarial wilt of pigeon pea under gnotobiotic and nursery condition. Bioactive metabolites viz., BM 1 and BM 2 from RRLJ 04 and BM 3 from BS 03 also showed in vitro antibiosis against F. udum. Seeds treated with 50 μl seed^?1 of BM 1, 30 μl seed^?1 of BM 2 and 70 μl seed^?1 of BM 3 and grown in pathogen infested soil showed suppression of wilt disease besides growth enhancement. Per cent disease control was 90 % with BM 2 application as compared to 87 and 83 %, respectively in BM 1 and BM 3 after 90 days of growth. BM 2 treated plants were more resistant to the pathogen as compared to the other fractions tested. Mycelial dry weight was found to be reduced on treatment with the bioactive metabolites. Formation of chlamydospore-like structures was observed in the pathogen mycelium treated with BM 3. The analytical studies confirmed that two of these metabolites are phenazine derivatives.     

A natural plant growth promoter calliterpenone from a plant Callicarpa macrophylla Vahl improves the plant growth promoting effects of plant growth promoting rhizobacteria (PGPRs)

Experiments were conducted to evaluate the efficacy of calliterpenone, a natural plant growth promoter from a shrub Callicarpa macrophylla Vahl., in enhancing the growth and yield promoting effects of plant growth promoting rhizobacteria (PGPRs), in menthol mint (Mentha arvensis L).This study is based on our previous results indicating the microbial growth promotion by calliterpenone and assumption that application of calliterpenone along with PGPRs will improve the population of PGPRs resulting in higher impacts on plant growth and yield. Of the 15 PGPRs (identified as potent ones in our laboratory), 25 μl of 0.01 mM calliterpenone (8.0 μg/100 ml) was found to be useful in improving the population of nine PGPRs in culture media. The five selected strains of PGPRs exhibiting synergy with calliterpenone in enhancing growth of maize compared to PGPR or calliterpenone alone were selected and tested on two cultivars (cvs. Kosi and Kushal) of M. arvensis. Of the five strains, Bacillus subtilis P-20 (16S rDNA sequence homologous to Accession No NR027552) and B. subtilis Daz-26 (16SrDNA sequence homologuos to Accession No GU998816) were found to be highly effective in improving the herb and essential oil yield in the cultivars Kushal and Kosi respectively when co-treated with calliterpenone. The results open up the possibilities of using a natural growth promoter along with PGPRs as a bio-agri input for sustainable and organic agriculture.     

Role of allelochemicals in plant growth promoting rhizobacteria for biocontrol of phytopathogens

Soil borne fungal diseases pose serious constraints on agro-productivity. Biological control is non-hazardous strategy to control plant pathogens and improve crop productivity. PGPR (plant growth promoting rhizobacteria) have long been used as plant disease control agents. PGPR produced a wide range of secondary compounds that may act as signals—that is, allelochemicals that include metabolites, siderophores, antibiotics, volatile metabolites, enzymes and others. Their mode of action and molecular mechanisms provide a great awareness for their application for crop disease management. The present review highlights the role of PGPR strains, specifically referring to allelochemicals produced and molecular mechanisms. Further research to fine tune combinations of allelochemicals, plant-microbe–pathogen interaction will ultimately lead to better disease control.     

Biological control of Aphis gossypii on pepper plant using Coccinella algerica

To calibrate biological control in pepper plant against Aphis gossypii using Coccinella algerica different experimentation were made. The consumption of C. algerica during the larval stage was enumerated in laboratory conditions. The densities (1, 4, 8 larvae/plant) to L2 stage of C. algerica necessarily to decrease aphid's populations were determined in plant pepper and the daily consumption in the pepper plant at the different densities was calculated. The density with 8 larvae at semi natural conditions is higher significantly. In protected crop of pepper, different releases to C. algerica were made with eggs, second stage larva and with ladybird before infestation with A. gossypii. A strategy to introduce L2 stage is preferred than the release with punters of C. algerica at different densities. At the same, the Introduce L2 stage is compared to release the C. algerica ladybird before the infestation of A. gossypii in the protected crops. The activity of ladybird has been maintained in the protected crop and it was shown when the infestation appeared.     

Salinity stress responses in the plant growth promoting rhizobacteria,Azospirillum spp

In order to adapt to the fluctuations in soil salinity/osmolarity the bacteria of the genusAzospirillum accumulate compatible solutes such as glutamate, proline, glycine betaine, trehalose, etc. Proline seems to play a major role in osmoadaptation. With increase in osmotic stress the dominant osmolyte inA. brasilense shifts from glutamate to proline. Accumulation of proline inA. brasilense occurs by both uptake and synthesis. At higher osmolarityA. brasilense Sp7 accumulates high intracellular concentration of glycine betaine which is taken up via a high affinity glycine betaine transport system. A salinity stress induced, periplasmically located, glycine betaine binding protein (GBBP) of ca. 32 kDa size is involved in glycine betaine uptake inA. brasilense Sp7. Although a similar protein is also present inA. brasilense Cd it does not help in osmoprotection. It is not known ifA. brasilense Cd can also accumulate glycine betaine under salinity stress and if the GBBP-like protein plays any role in glycine betaine uptake. This strain, under salt stress, seems to have inadequate levels of ATP to support growth and glycine betaine uptake simultaneously. ExceptA. halopraeferens, all other species ofAzospirillum lack the ability to convert choline into glycine betaine. Mobilization of thebet ABT genes ofE. coli intoA. brasilense enables it to use choline for osmoprotection. Recently, aproU-like locus fromA. lipoferum showing physical homology to theproU gene region ofE. coli has been cloned. Replacement of this locus, after inactivation by the insertion of kanamycin resistance gene cassette, inA. lipoferum genome results in the recovery of mutants which fail to use glycine betaine as osmoprotectant.     

丛枝菌根真菌与根围促生细菌相互作用的效应与机制

丛枝菌根(arbuscular mycorrhiza,AM)真菌是植物活体营养专性共生菌,广泛存在于陆地各生态系统中.研究表明,AM真菌与根围促生细菌(plant growth promoting rhizobacteria,PGPR)之间的相互作用,尤其是它们之间的协同作用不仅影响植物养分吸收利用、病原物发生发展、土壤理化特性与生物修复等,而且对于可持续农、林、牧业生产、稳定生态系统都具有十分重要的意义.因此,近年来给予众多关注和研究.综述了AM真菌与PGPR之间的相互影响及其可能的作用机制,以及AM真菌与PGPR协同改善植物营养和生长、协同抑制病原菌、协同修复土壤方面的作用,旨在总结AM真菌与PGPR相互作用的效应与机制方面的最新研究进展,为今后研究发展提供依据.     

Effect of plant growth promoting rhizobacteria containing ACC-deaminase on maize (Zea mays L.) growth under axenic conditions and on nodulation in mung bean (Vigna radiata L.)

AIMS: This study was conducted to test the hypothesis that the bacterial strains possessing 1-aminocyclopropane-1-carboxylic acid (ACC)-deaminase activity may also promote growth of inoculated plants and could increase nodulation in legumes upon co-inoculation with rhizobia. METHODS AND RESULTS: Several rhizobacteria were isolated from maize rhizosphere through enrichment on ACC as a sole N source. Purified isolates were screened for growth promotion in maize under axenic conditions and for in vitro ACC-deaminase activity. A significant positive correlation was observed between in vitro ACC-deaminase activity of bacterial cells and root elongation. None of the isolates produced auxins. Bradyrhizobium japonicum produced less amount of auxins but did not carry ACC-deaminase activity. Results of pot experiment revealed that co-inoculation with Bradyrhizobium and plant growth promoting rhizobacteria (PGPR) isolates enhanced the nodulation in mung bean compared with inoculation with Bradyrhizobium alone. CONCLUSIONS: It is highly expected that inoculation with rhizobacteria containing ACC-deaminase hydrolysed endogenous ACC into ammonia and alpha-ketobutyrate instead of ethylene. Consequently, root and shoot growth as well as nodulation were promoted. SIGNIFICANCE AND IMPACT OF THE STUDY: The ACC-deaminase trait could be employed as an efficient tool to screen effective PGPR, which could be successfully used as biofertilizers to increase the growth of inoculated plants as well as nodulation in legumes.