Rhizobacteria of Maize and Their Antifungal Activities

During the growing season of 1984, the rhizobacteria (including organisms from the rhizosphere soil, the rhizoplane, and internal root zones) of 47 maize plants (two varieties) sampled from different locations in France and at different growth stages were inventoried. Isolates were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of their total cell proteins and were found to represent 352 different protein electrotypes. Maize seedlings were initially colonized by a small number of different strains. Densities reached up to 10⁸ CFU/g of root. Later in the season, the population density decreased but the heterogeneity of the rhizobacterial populations increased. Fluorescent pseudomonads represented up to 35% of the total rhizobacterial population and comprised 43 different electrotypes. Other bacteria regularly present were Xanthomonas maltophilia, Serratia liquefaciens, Pseudomonas paucimobilis, and Bacillus spp. There was a very low similarity between rhizobacterial populations of plants of the same cultivar (LG5) within one field at different growth stages and also between rhizobacterial populations of the cultivars LG5 and BRIO42 on the same field. Most electrotypes (76%) were found on a single occasion. None of the 352 electrotypes was present on all plants. In the 1985 analysis the rhizobacteria of maize seedlings (one variety) sampled from one field were characterized. They represented 236 different protein electrotypes. Thirty-three isolates showed antifungal activity against major maize pathogens; they comprised four Pseudomonas cepacia strains, producing pyrrolnitrin as well as another unknown antifungal compound.     

A Rapid Microplate Callus Bioassay for Assessment of Rhizobacteria for Biocontrol of Leafy Spurge ( Euphorbia esula L.)

Screening large collections of microorganisms for potential biological control activity on economically important weeds is often diY cult. Seeds required as indicator species are often not available in large supply and are highly variable in viability and germination, resulting in uneven seedling development. A bioassay system for rapidly assessing the phytotoxic eVects of rhizobacteria was developed based on leafy spurge ( Euphorbia esula L. ) callus tissue culture in multiple-well plates. Callus pieces (0.5 g) were placed in 24-well plates containing Gamborg's B5 medium, inoculated with rhizobacterial suspension and incubated at 27oC for 48 h. By rating inoculated callus for cellular damage, about 30% of rhizobacteria isolated from weedy Euphorbia spp. collected in Europe and North America were identified as being highly phytotoxic. Symptoms of phytotoxicity included growth reduction, discoloration and extensive cellular leakage. A high proportion of isolates found to be phytotoxic in tissue culture bioassays were similarly eVective in standard leafy spurge seedling bioassays. The method is rapid, host specific and more uniform compared with assays using seedlings, and should be adaptable to other weed species for screening microorganisms for potential biocontrol activity.     

Soil and root populations of fluorescent Pseudomonas spp. associated with seedlings and field-grown plants are affected by sorghum genotype

Sorghum [Sorghum bicolor (L.) Moench] is valued for bioenergy, feed and food. Potential of sorghum genotypes to support differing populations of root- and soil-associated fluorescent Pseudomonas spp. or Fusarium spp., in two soils, was assessed. Culturable pseudomonads were enumerated from roots and soil of sorghum (Redlan and RTx433) and wheat (Lewjain) seedlings repeatedly grown in cycled soils in the growth chamber. Pseudomonads and Fusarium spp. were assessed from roots and soil of field-grown sorghum along with biological control traits hydrogen cyanide (HCN) and 2,4-diacetylphlorogluconol (phl) production. After four 4-week cycles, soil associated with Redlan seedlings had greater numbers of fluorescent pseudomonads than Lewjain. In dryland field conditions, RTx433 roots had greater numbers of pseudomonads than Redlan before anthesis but similar numbers after. There were no differences in numbers of pseudomonads from dryland soil or roots or soil of irrigated plants. Percentages of HCN-producing root isolates and phl soil isolates declined on irrigated Redlan plants, but percentages of HCN-producers increased in dryland conditions. Redlan roots had greater percentages of Fusarium isolates in the Gibberella fujikuroi complex. Results indicated that sorghum genotype affected root-associated populations of fluorescent Pseudomonas spp. and Fusarium spp. across soil environments.     

Chemotaxis of deleterious rhizobacteria to velvetleaf (Abutilon theophrasti Medik.) seeds and seedlings

Abstract Intact seeds and seed and seedling root exudates of velvetleaf ( Abutilon theophrasti Medik.) were used as chemoattractants in experiments to determine the relative importance of chemotaxis in spermosphere and rhizosphere colonization by selected rhizobacteria. Results for soft-agar, capillary tube and soil chemotaxis assays indicated that selected deleterious rhizobacteria were specifically attracted to seed and seedling root exudates. Several amino acids and sugars detected in exudates were chemoattractants for these rhizobacteria. Using soil-chemotaxis assemblies, migration of rhizobacterial isolates through 2-cm distances of soil toward velvetleaf seeds or exudates was detected within 24 h. Isolates were not detected at the same site in soils without seeds or exudates until 72 h after inoculation. These results suggest that attraction of delecterious rhizobacteria toward seeds and seedling roots mediated by exudates (chemotaxis) might be the first step in establishment and subsequent colonization of biological control bacteria on weed seeds and seedling roots in soil.     

Soil conduciveness to take-all of wheat: Influence of the nitrogen fertilizers on the structure of populations of fluorescent pseudomonads

In a field cropped with wheat, a high and low level of soil conduciveness to take-all were induced by applying a nitrogen fertilizer with either calcium nitrate or ammonium sulphate. From these two soils, two representative populations of fluorescent pseudomonads were tested for their in situ behaviour. Take-all index and root dry weight were assessed on plants cropped in soils infested with Gaeumannomyces graminis var tritici (Ggt) and each bacterized with one of the isolates of fluorescent pseudomonads. The bacteria tested can be split into three groups: antagonists which reduce take-all, deleterious isolates which aggravate the disease and neutral without evident effect on the disease. The predominance of antagonistic fluorescent pseudomonads in the NH₄-treated soil and the predominance of deleterious ones in the NO₃-treated soil was confirmed after statistical analysis. The microbial impact on take-all must be more considered as the resulting effect of divergent activities of both rhizobacteria types than the only consequences of the presence of antagonistic pseudomonads. All the high cyanogenic pseudomonads were antagonists in situ and were more numerous in the NH₄-treated soil than in the NO₃-treated soil.     

Rhizobacteria of Cotton and Their Repression of Seedling Disease Pathogens

During the 1983 field season, the rhizobacteria (including organisms from rhizosphere soil and the root rhizoplane) of cotton plants at one location in Mississippi were inventoried at different plant growth stages. Isolates (1,000) were identified to the genus level and characterized for repression of Pythium ultimum and Rhizoctonia solani. Cotton seedlings were initially colonized by bacteria of many different genera, and populations quickly reached 10⁸ CFU/g of root tissue. As the season progressed, the bacterial populations declined as root mass increased and the roots became more woodlike in consistency. Fluorescent pseudomonads were the most numerous gram-negative rhizobacterial isolates of those that were randomly collected and identified, and they provided the largest number of isolates with fungal repressive activity. Several other gram-negative bacterial genera were recovered throughout the growing season, and some gram-positive bacteria were also isolated routinely, but at lower numbers. There was no correlation between the proportion of rhizobacterial isolates that possessed fungal repressive activity and the plant growth stage from which the isolates were obtained. Approximately twice as many bacterial isolates demonstrated fungal repression in the agar assay compared with the inplanta assay, and isolates were found more frequently with fungal repressive activity against P. ultimum than against R. solai.     

Screening of Rhizobacteria for Their Plant Growth Promotion Ability and Antagonism Against Damping off and Root Rot Diseases of Broad Bean (<Emphasis Type="Italic">Vicia faba</Emphasis> L.)

Development of microbial inoculants from rhizobacterial isolates with potential for plant growth promotion and root disease suppression require rigorous screening. Fifty-four (54) fluorescent pseudomonads, out of a large collection of rhizobacteria from broad bean fields of 20 different locations within Imphal valley of Manipur, were initially screened for antifungal activity against Macrophomina phaseolina and Rhizoctonia solani, of diseased roots of broad bean and also three other reference fungal pathogens of plant roots. Fifteen fluorescent pseudomonas isolates produced inhibition zone (8–29 mm) of the fungal growth in dual plate assay and IAA like substances (24.1–66.7 μg/ml) and soluble P (12.7–56.80 μg/ml) in broth culture. Among the isolates, RFP 36 caused a marked increase in seed germination, seedling biomass and control of the root borne pathogens of broad bean. PCR–RAPD analysis of these isolates along with five MTCC reference fluorescent pseudomonas strains indicated that the RFP-36 belonged to a distinct cluster and the PCR of its genomic DNA with antibiotic specific primers Phenazine-1-carboxylic acid and 2, 4-diacetyl phloroglucinol suggested possible occurrence of gene for the potent antibiotics. Overall, the result of the study indicated the potential of the isolate RFP 36 as a microbial inoculant with multiple functions for broad bean.     

Cyanide Production by Rhizobacteria and Potential for Suppression of Weed Seedling Growth

Rhizobacteria strains were characterized for ability to synthesize hydrogen cyanide and for effects on seedling root growth of various plants. Approximately 32% of bacteria from a collection of over 2000 isolates were cyanogenic, evolving HCN from trace concentrations to >30 nmoles/mg cellular protein. Cyanogenesis was predominantly associated with pseudomonads and was enhanced when glycine was provided in the culture medium. Concentrations of HCN produced by rhizobacteria were similar to exogenous concentrations inhibiting seedling growth in bioassays, suggesting that cyanogenesis by rhizobacteria in the rhizosphere can adversely affect plant growth. Growth inhibition of lettuce and barnyardgrass by volatile metabolites of the cyanogenic rhizobacteria confirmed that HCN was the major inhibitory compound produced. Our results suggest that HCN produced in the rhizospheres of seedlings by selected rhizobacteria is a potential and environmentally compatible mechanism for biological control of weeds. Received: 13 December 2000/Accepted: 6 February 2001     

In vitro Antagonism of Rhizobacteria Isolated from Coffea arabica L. against Emerging Fungal Coffee Pathogens

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

Evaluation of fluorescent pseudomonads for plant growth promotion,antifungal activity against Rhizoctonia solani on common bean,and biocontrol potential

Fluorescent pseudomonads are ubiquitous bacteria that are common inhabitants of the rhizosphere and are the most studied group within the genus Pseudomonas. Bacterial isolates (n = 103) from the rhizosphere of wheat and common bean were assessed as potential biocontrol agents in this study. Fungal inhibition tests were performed by a plate assay in which each isolate was tested directly for the production of hydrogen cyanide, protease, siderophore and cellulase. Production of DAPG was verified by using an analytical high performance liquid chromatography assay (HPLC). Plant growth promotion was assessed in phytochamber trials and biocontrol activity was evaluated in greenhouse trials. In all, 52 bacterial isolates with antifungal activity against Rhizoctonia solani were found. Of the 52 isolates, 41 were selected according to their high efficiency in in vitro antagonism, which was shown as inhibition zones in the dual-culture assay. Six of the 41 rhizobacteria, including isolates UTPF7, UTPF13, UTPF18, UTPF22, UTPF27 and strain CHA0 produced HCN. Production of protease enzyme was detected for all isolates excluding UTPF30 isolate. Although some stains appeared not to produce any compound with affinity for ferric iron, other isolates produced prolific amounts, creating a large zone of orange (up to 160 mm², i.e., UTPF16). Seventeen of 41 isolates of fluorescent pseudomonads including strain CHAO produced different amounts of DAPG ranging from 0.6 to 11.4 ng/10⁸ cfu. A total of 39 isolates induced statistically significant effects on plant growth compared with the non-treated control for at least one parameter. The predominant influence observed was increased root length. No bacteria could completely protect the plant against R. solani, although all isolates significantly increased fresh weight as compared to the infested control in greenhouse trials. Pseudomonas fluorescens isolates UTPF16 and UTPF26 significantly (P < 0.05) decreased the number of seedlings with damping-off symptoms in the means of the experiments.     

Effect of sorghum seedlings, and previous crop, on soil fluorescent Pseudomonas spp.

Hypotheses in which sorghum seedlings [Sorghum bicolor (L.) Moench] of different genotypes will differentially modify soil microorganisms and will affect subsequent planting of wheat (Triticum aestivum L.) seedlings, were tested. Wheat cultivar Lewjain, and sorghum genotypes Redlan and RTx433, were planted into soils previously planted with wheat or sorghum in growth chamber experiments. Total culturable fungi and oomycetes, and fluorescent Pseudomonas spp. numbers (cfu) were determined. Pseudomonads were screened for hydrogen cyanide (HCN) production, for the presence of the phlD gene for 2,4-diacetylphloroglucinol production (Phl) and for a region of the operon involved in phenazine-1-carboxylic acid (PCA) production. Pasteurized soils were inoculated with rifampicin-marked strains of Pseudomonas fluorescens then planted with Lewjain, Redlan and RTx433 to assess rhizosphere and soil colonization. Effects of plant species, sorghum genotype and previous crop on culturable fungi and oomycetes, and pseudomonad numbers (cfu g^?1 soil) were statistically significant. Soils planted with RTx433 or Lewjain had greater numbers of fungal cfu than soils planted with Redlan. When Lewjain seedlings were grown in soil previously planted with RTx433, there were greater numbers of fungal cfu than when Lewjain was planted into Redlan soil. Wheat planted into wheat soil resulted in statistically significantly fewer numbers of pseudomonads than when planted into sorghum soil. Overall, percentages of HCN-producing pseudomonads increased, especially when wheat seedlings were planted in wheat soil. For most treatments, percent of isolates with Phl declined, except when Redlan was planted into Redlan soil, which resulted in increased Phl isolates. When rifampicin-marked P. fluorescens isolates were applied to pasteurized soil, sorghum seedlings sustained rhizosphere and soil populations similar to those on wheat. Sorghum genotypes may differ in associations with soil microorganisms, suggesting that they may differentially affect numbers of fluorescent pseudomonads in cropping systems.     

Predicting the host range of Nystalea ebalea: Secondary plant chemistry and host selection by a surrogate biological control agent of Schinus terebinthifolia

The safety of weed biological control depends upon the selection and utilization of the target weed by the agent while causing minimal harm to non-target species. Selection of weed species by biological control agents is determined by the presence of behavioral cues, generally host secondary plant compounds that elicit oviposition and feeding responses. Non-target species that possess the same behavioral cues as found in the target weed may be at risk of damage by classical biological control agents. Here we conducted host range tests and examined secondary plant compounds of several test plant species. We studied the specialist herbivore Nystalea ebalea (Lepidoptera: Notodontidae) a Neotropical species, present in Florida as a surrogate biological control agent of the weed, Brazilian peppertree Schinus terebinthifolia, invasive in Florida and Hawaii. We found that the larvae had the greatest survival when fed the target weed, the Neotropical species Spondias purpurea, the Florida native species Rhus copallinum, and the ornamental Pistacia chinensis. Reduced survival and general larval performance were found on the native species Metopium toxiferum and Toxicodendron radicans. Both the volatiles and the allergen urushiols were chemically characterized for all species but urushiol diversity and concentration best predicted host range of this herbivore species. These results provide insight into host selection and utilization by one oligophagous Schinus herbivore. Other potential biological control agents may also be sensitive to plants that contain urushiols and if so, they may pose minimal risk to these native species.     

Phylogeny of HCN synthase-encoding hcnBC genes in biocontrol fluorescent pseudomonads and its relationship with host plant species and HCN synthesis ability

Hydrogen cyanide (HCN) is a broad-spectrum antimicrobial compound involved in biological control of root diseases by many plant-associated fluorescent pseudomonads. The HCN synthase is encoded by three biosynthetic genes (hcnA, hcnB, and hcnC), but little is known about the diversity of these genes in fluorescent Pseudomonas spp. and in other bacteria. Here, the partial hcnBC sequence was determined for a worldwide collection of biocontrol fluorescent Pseudomonas spp. Phylogenies based on hcnBC and deduced protein sequences revealed four main bacterial groups, but topological incongruences were found between hcnBC and rrs-based phylogenies, suggesting past lateral transfer of hcnBC among saprophytic root-colonizing pseudomonads. Three of the four groups included isolates from different countries and host plants. Yet, these groups corresponded to distinct, ecologically-adapted populations of HCN-producing biocontrol fluorescent pseudomonads, as indicated by high hcnBC distinctness ratio values and the differences in production levels of HCN in vitro found between groups. This is in accordance with previous results on catabolic properties and biocontrol abilities of these strains. HCN synthase gene diversity may thus reflect the adaptive radiation of HCN+ biocontrol fluorescent pseudomonads. Positive correlations were found between HCN production in vitro and plant protection in the cucumber/Pythium ultimum and tomato/Fusarium oxysporum f. sp. radicis-lycopersici pathosystems.     

Enhanced suppression of plant growth through production of L-tryptophan-derived compounds by deleterious rhizobacteria

Plant-growth-suppressive activity of deleterious rhizobacteria (DRB) may be due to production of metabolites absorbed through roots. Auxins produced in high concentrations in the rhizosphere by DRB contribute to reduced root growth. Selected DRB able to produce excessive amounts of auxin compounds for suppression of weed seedling growth may be effective for biological control of weeds. The objectives to this study were to assess the ability of DRB originating from weed seedlings to synthesize auxins from L-tryptophan (L-TRP), determine effects of DRB with or without L-TRP on seedling root growth, and characterize auxins produced from L-TRP using high performance liquid chromatography (HPLC). Auxins expressed as indole-3-acetic acid (IAA)-equivalents were produced by 22.8% of the DRB tested based on a colorimetric method. Under laboratory conditions, a DRB isolate classified as Enterobacter taylorae with high auxin-producing potential (72 mg L^–1 IAA-equivalents) inhibited root growth of field bindweed (Convolvulus arvensis L.) by 90.5% when combined with 10^–5 M L-TRP compared with non-treated control. Auxin derivatives produced by E. taylorae from L-TRP in broth culture after 24 h incubation identified by HPLC included IAA (102 g L^–1), indole-3-aldehyde (IALD; 0.4 g L^–1), and indole-3-lactic acid (ILA; 7.6 g L^–1). Results suggest that providing L-TRP with selected auxin-producing DRB to increase phytotoxic activity against emerging weed seedlings may be a practical biological control strategy.     

Interaction of rhizobacteria with leafy spurge (Euphorbia esula L.) callus tissue cells

The interaction of two rhizobacterial isolates,Pseudomonas fluorescens isolate LS 102 andFlavobacterium balustinum isolate LS 105, with leafy spurge cells at the cellular level was studied using scanning and electron microscopy. Leafy spurge callus tissue inoculated with either isolate showed considerable changes compared to non-inoculated tissue. The attachment of rhizobacteria to cell surfaces was associated with the elaboration of fibrillar material which may anchor bacteria to surfaces and contribute to mediation of the phytotoxic effect caused by rhizobacteria. At the ultracellular level, inoculated callus tissue showed numerous cell alterations including esiculation and convolution of the plasmalemma, cell wall degradation and disorganization of the cytoplasm, similar to those detected in the whole plant. It is concluded that callus tissue may provide an excellent working model to investigate the mode and/or mechanism of action of potential biocontrol agents on their host plants.     

Structural and functional diversity of rhizobacteria associated with Rauwolfia spp. across the Western Ghat regions of Karnataka, India

The present study carried out with denaturing gradient gel electrophoresis of DNA extracted from rhizosphere soils of Rauwolfia spp. collected from Western Ghat (WG) regions of Karnataka indicated that Pseudomonas sp. was prevalently found followed by Methylobacterium sp., Bacillus sp. and uncultured bacteria. A total of 200 rhizobacteria were isolated from 58 rhizosphere soil samples comprising of 15 different bacterial genera. The Shannon Weaver diversity index (H′) and Simpson’s diversity index (D) were found to be 2.57 and 0.91 for cultivable bacteria, respectively. The total species richness of cultivable rhizobacteria was high in Coorg district comprising 15 bacterial genera while in Mysore district, four bacterial genera were recorded. Rarefaction curve analysis also indicated the presence of higher species richness in samples of Shimoga and Coorg. All the rhizobacteria were screened for their multiple plant growth promotion and disease suppression traits. The results revealed that 70 % of the isolates colonized tomato roots, 42 % produced indole acetic acid, 55 % solubilized phosphorus, while 43, 22, 27, 19, 40, 15 and 44 % produced siderophore, salicylic acid, hydrogen cyanide, chitinase, phytase, cellulase and protease, respectively. Rhizobacterial isolates showing antagonistic activity against Fusarium oxysporum and Aspergillus flavus were 53 and 33 %, respectively. Plant growth promotion studies revealed that most of the isolates increased percent germination with significantly higher vigour index as compared to untreated control. Most predominant rhizobacteria found in the rhizospheres of Rauwolfia spp. of WG regions are potential PGPR which can serve as biofertilizers and biopesticides.     

Screening for fluorescent pseudomonades,isolated from the rhizosphere of tomato,for antagonistic activity toward <Emphasis Type="Italic">Clavibacter michiganensis</Emphasis> subsp<Emphasis Type="Italic">. michiganensis</Emphasis>

Bacterial canker of tomato, caused by Clavibacter michiganensis subsp. michiganensis, continues to be a problem for tomato growers in the Souss-Massa Draa valley, South of Morocco. Assuming that biological control is an alternative for the management of this disease, a total of 303 fluorescent pseudomonads strains isolated from roots and rhizospheric soil of tomato plants were in vitro tested against C. michiganensis subsp. michiganensis. Fluorescent pseudomonads strains which showed the highest antagonistic properties were thereafter investigated for their ability to colonize tomato roots. Our results showed that fluorescent pseudomonads are more represented in rhizospheric soils. However, the most efficient fluorescent pseudomonads isolates were found in the rhizoplane soil and the endorhizosphere. Among 42 spontaneous antibiotic resistant mutants obtained by treatment of the wild-type isolates with five antibiotics (rifampicine, nalidixic acid, ampicilline and chloramphenicol), 28 completely colonized the roots of all tomatoes seedlings used in this investigation. The 42 wild type isolates were then used for in vivo screening with the cotyledon test. Using this test, eight isolates from 42 tested induced a significant decrease of disease incidence and disease symptoms. The eight efficient isolates were then tested for their effectiveness in the protection of tomato plants in pots under greenhouse conditions. Results obtained showed that all tested isolates applied as seed and root treatments reduced significantly (P ≤ 0.001) the incidence of bacterial canker.     

The Ability of Plant Compost Leachates to Control Black Mold (Aspergillus niger) and to Induce the Accumulation of Antifungal Compounds in Onion Following Seed Treatment

Onion seeds treated with leachates of composts prepared from alfalfa and sunflower stalks, at the dosages of 10% and 20% respectively, were inoculated with Aspergillus niger van Tieghem, causal agent of onion black mold disease. The ability of the leachates to induce the production of antifungal compounds and to control black mold were tested at seedling and set stages. Leachates from both composts were able to reduce disease incidence in sets, but not disease severity in onion seedlings. Extracts from treated seedlings and sets were fractionated by thin layer chromatography for their content of antifungal compounds. There were no significant differences between the fractions of alfalfa and sunflower compost leachates in the inhibition of the mycelium growth of A. niger, with the exception of one fraction. The presence of fluorescent pseudomonads and Pantoae agglomerans [synonym: Erwinia herbicola (Löhnis)] bacteria was determined in both leachates. The population of P. agglomerans was higher in the sunflower compost leachate compared to the alfalfa leachate. The tested strains of both bacteria were able to inhibit mycelium growth of the fungal pathogen in agar tests. This study suggests the possible role of beneficial bacteria in the induction of antifungal compounds in onion against A. niger during seedling and set stages.     

Manipulation of Rhizosphere Bacterial Communities to Induce Suppressive Soils

Naturally occurring disease-suppressive soils have been documented in a variety of cropping systems, and in many instances the biological attributes contributing to suppressiveness have been identified. While these studies have often yielded an understanding of operative mechanisms leading to the suppressive state, significant difficulty has been realized in the transfer of this knowledge into achieving effective field-level disease control. Early efforts focused on the inundative application of individual or mixtures of microbial strains recovered from these systems and known to function in specific soil suppressiveness. However, the introduction of biological agents into non-native soil ecosystems typically yielded inconsistent levels of disease control. Of late, greater emphasis has been placed on manipulation of the cropping system to manage resident beneficial rhizosphere microorganisms as a means to suppress soilborne plant pathogens. One such strategy is the cropping of specific plant species or genotypes or the application of soil amendments with the goal of selectively enhancing disease-suppressive rhizobacteria communities. This approach has been utilized in a system attempting to employ biological elements resident to orchard ecosystems as a means to control the biologically complex phenomenon termed apple replant disease. Cropping of wheat in apple orchard soils prior to re-planting the site to apple provided control of the fungal pathogen Rhizoctonia solani AG-5. Disease control was elicited in a wheat cultivar-specific manner and functioned through transformation of the fluorescent pseudomonad population colonizing the rhizosphere of apple. Wheat cultivars that induced disease suppression enhanced populations of specific fluorescent pseudomonad genotypes with antagonistic activity toward R. solani AG-5, but cultivars that did not elicit a disease-suppressive soil did not modify the antagonistic capacity of this bacterial community. Alternatively, brassicaceae seed meal amendments were utilized to develop soil suppressiveness toward R. solani. Suppression of Rhizoctonia root rot in response to seed meal amendment required the activity of the resident soil microbiota and was associated with elevated populations of Streptomyces spp. recovered from the apple rhizosphere. Application of individual Streptomyces spp. to soil systems provided control of R. solani to a level and in a manner equivalent to that obtained with the seed meal amendment. These and other examples suggest that management of resident plant-beneficial rhizobacteria may be a viable method for control of specific soilborne plant pathogens.     

Community Profiling of Culturable Fluorescent Pseudomonads in the Rhizosphere of Green Gram (Vigna radiata L.)

Study on microbial diversity in the unexplored rhizosphere is important to understand their community structure, biology and ecological interaction with the host plant. This research assessed the genetic and functional diversity of fluorescent pseudomonads [FP] in the green gram rhizophere. One hundred and twenty types of morphologically distinct fluorescent pseudomonads were isolated during vegetative as well as reproductive growth phase of green gram. Rep PCR, ARDRA and RISA revealed two distinct clusters in each case at 75, 61 and 70% similarity coefficient index respectively. 16S rRNA partial sequencing analysis of 85 distantly related fluorescent pseudomonads depicted Pseudomonas aeruginosa as the dominant group. Out of 120 isolates, 23 (19%) showed antagonistic activity towards phytopathogenic fungi. These bacterial isolates showed varied production of salicylic acid, HCN and chitinase, 2, 4-diacetylphloroglucinol (DAPG), phenazine-1-carboxylic acid (PCA) and pyoluteorin (PLT). Production efficiency of inherent level of plant growth promoting (PGP) traits among the 120 isolates demonstrated that 10 (8%) solubilised inorganic phosphates, 25 (20%) produced indoles and 5 (4%) retained ACC deaminase activity. Pseudomonas aeruginosa GGRJ21 showed the highest production of all antagonistic and plant growth promoting (PGP) traits. In a greenhouse experiment, GGRJ21 suppressed root rot disease of green gram by 28–93% (p = 0.05). Consistent up regulation of three important stress responsive genes, i.e., acdS, KatA and gbsA and elevated production efficiency of different PGP traits could promote GGRJ21 as a potent plant growth regulator.