Biocontrol potential of five Burkholderia and Pseudomonas strains against Colletotrichum truncatum infecting chilli pepper

The effect of bacterial antagonism against the causal agent, Colletotrichum truncatum was assessed as a potential alternative in managing anthracnose in chilli pepper. Out of 104 contrasting bacterial colonies isolated from rhizosphere soil of a forest floor, five isolates caused the radial growth inhibition greater than 90% (Significant at p?<?0.05 level) of C. truncatum in dual cultures. Based on 16S rRNA analysis, these antagonistic bacterial isolates were identified as Pseudomonas aeruginosa, Burkholderia arboris, Burkholderia gladioli and Burkholderia rinojensis. The selected promising antagonists showed nearly 100% inhibition of the spore germination of C. truncatum in vitro. These antagonists produced antifungal compounds which are diffusible in nature. Microscopic studies of blackened fungal hyphae, which were subjected to antagonism showed many deformations such as thickening, swelling and malformation. In vivo study revealed that C. truncatum inoculated chilli pepper seeds treated with the five antagonists significantly inhibited the incidence of seed colonisation (p?<?0.05) by the pathogen. At the post emergence stage, the survival percentages and vigour indices of all the antagonists’ treatments, except B. rinojensis strain 1, were significantly higher compared to the untreated control. The efficacy of the selected antagonists in managing anthracnose fruit rot was 100% at the colour breaking stage of chilli pepper fruits. These bacterial antagonists had a negative effect on C. truncatum spore attachment and subsequent colonisation on chilli pepper leaves except in the treatment of B. arboris. The results of in-vitro and in-vivo studies, suggest that the screened antagonistic bacterial isolates are potential biocontrol agents and need to be further studied for the biochemical basis of their activity against C. truncatum.     

Plant Growth–Promoting Pseudomonas sp. Strains Reduce Natural Occurrence of Anthracnose in Soybean (Glycine max L.) in Central Himalayan Region

Biological control is an accepted important component of current plant disease management strategies. Introduction of bacterized seeds carrying bacterial isolates with proven growth-promotion capabilities and antagonistic characteristics offer a valid alternative to chemical protectants. Root colonization of disease-susceptible (PS 1024) and moderately resistant (PS1042) varieties of soyabean (Glycine Max L) by fluorescent pseudomonad (FLPs) strains GRP₃, PEn-4, PRS₁, and WRS-24 was studied in relation to natural occurrence of anthracnose caused by Colletotrichum dematium (Pers Ex Fr.) Grove. Rhizoplane population of FLPs was maintained at a critical level (5.3 cfu) up to 30 days of plant growth, followed by a steep decline. Indigenous FLPs population, however, remained nearly unchanged (3.0 to 2.4 log g⁻¹ root) between 30 days and 75 days of plant growth. The relative FLPs population in rhizosphere was lower than that in rhizoplane. Although intervarietal difference was observed, the root/shoot length remained unaffected. Compared to nonbacterized control, dry root weight was improved by FLPs treatment. Severity of foliar anthracnose was reduced significantly after FLPs treatment in the variety PS 1042. Because the point of FLPs treatment (seed bacterization) was away from the site of disease appearance (leaf), operation of induced systemic resistance in strains PEn-4 and GRP₃ appears imminent.     

Endophytic bacteria from tomato and chilli,their diversity and antagonistic potential against Ralstonia solanacearum

A total of 82 endophytic bacteria of tomato and chilli was isolated from different locations of tropical Islands of Andaman and Nicobar, India. Based on in vitro screening, 16 bacterial isolates that effectively inhibited Ralstonia solanacearum (a bacterial wilt pathogen) were characterised for their diversity and identified through Microbial Identification System (Biolog). Diversity analysed through BOX-PCR showed low similarity index among the antagonistic bacteria. Based on the in vitro antagonistic activities, the selected isolates were further characterised for siderophore, indole acetic acid production, phosphate solubilisation and other extracellular enzymes; it is found that most of the isolates were positive for these properties. The production of these metabolites may be responsible for the inhibition of the pathogen R. solanacearum. The isolates BECS3, BECS6 and BECS7 showed multiple attributes and demonstrated plant growth promotion properties through tomato- and chilli-based bioassay under greenhouse conditions. These bacterial inoculations were found to result in significant increase in root, shoot and biomass of both tomato and chilli. Hence, these isolates can be further formulated and used for field application.     

Trichoderma harzianum antagonistic activity and competition for seed colonization against seedborne pathogenic fungi of sunflower

This study investigated the antagonistic effects of Trichoderma harzianum isolate (TRIC8) on mycelial growth, hyphal alteration, conidial germination, germ tube length and seed colonization by the seedborne fungal pathogens Alternaria alternata, Bipolaris cynodontis, Fusarium culmorum and F. oxysporum, the causes of seedling rot in over 30% of sunflowers. The antagonistic effect of TRIC8 on mycelial growth of pathogens was evaluated on dual culture that included two inoculation assays: inoculation of antagonist at 48 h before pathogen (deferred inoculation) and inoculation at the same time with pathogen (simultaneous inoculation). TRIC8 inhibited mycelial growth of the fungal pathogens between 70·67 and 76·87% with the strongest inhibition seen with deferred inoculation. Alterations in hyphae were observed in all pathogens. Conidial germination of F. culmorum was inhibited by most of the fungal pathogens (38·28%) by TRIC8. Inhibition of germ tube length by the antagonist varied from 31·83 to 37·67%. In seed colonization experiments, TRIC8 was applied in combination with each pathogen to seeds of a sunflower genotype that is highly tolerant to downy mildew. Seed death was inhibited by TRIC8 and the antagonist did not allow growth of A. alternata, B. cynodontis and F. culmorum on seeds and inhibited the growth of F. oxysporum at the rate of 58·32%.     

Colonization of root tissues and protection against Fusarium wilt of rye (Secale cereale) by nonpathogenic rhizosphere strains of Fusarium culmorum

Colonization of rye (Secale cereale) tissues by nonpathogenic rhizosphere Fusarium culmorum isolates DEMFc2 and DEMFc5 and a pathogenic strain DEMFc37, and their effect on plant fresh weight were studied in pot experiments. Both rhizosphere isolates colonized the epidermis and the cortex but were not found in vessels, while the pathogen colonized all three layers of root cells. The numbers of pathogen CFU isolated from plant tissues were much higher than those of the rhizosphere isolates in spite of the same number of macroconidia used as inoculum (1 × 10⁵ g⁻¹ of soil). Inoculation of seedlings with DEMFc2 resulted in a 20% increase, with DEMFc5 in more than a 20% reduction, and with DEMFc37 in a 38% reduction of shoot fresh weight of 14-day-old plants. Pre-colonization of plants with (either of) the rhizosphere isolates and subsequent inoculation with the pathogen resulted in plant weights the same as those observed in plants inoculated with the rhizosphere strain alone. The disease severity index for shoots of plants pre-colonized with DEMFc2 was reduced from class 4 (86% diseased plants) observed for plants inoculated with the pathogen alone to class 2 (average of 8% diseased plants) when pre-treated with the rhizosphere strain. The CFU number of the pathogen isolated from the interior of roots of plants pre-colonized with the rhizosphere isolates was as low as 10% of the number isolated from plants inoculated with the pathogen alone. A study of in vitro interactions between the rhizosphere isolates and the pathogen suggests that changes in plant colonization by the pathogen and its effect on fresh weight of plants pre-colonized with the rhizosphere isolates were not connected with inhibition of its growth by a direct action of the rhizosphere isolates. The results suggest that strain DEMFc2 can be considered as a potential biocontrol agent.     

The plant growth‐promoting fungus Fusarium equiseti and the arbuscular mycorrhizal fungus Glomus mosseae stimulate plant growth and reduce severity of anthracnose and damping‐off diseases in cucumber (Cucumis sativus) seedlings

To alleviate the environmental contamination due to persistent chemical usage, approaches to integrated pest management were conceived. In this perspective, microbe–microbe interactions such as mycorrhizal relationships with other soil microbiota in the rhizosphere like the plant growth‐promoting fungi (PGPF) are particularly important. Better understanding of the interactions between beneficial microbial groups is imperative in the identification of possible synergistic or antagonistic effects to improve their practical usage as biocontrol agents or biofertilizers. In this study, the consequence of co‐inoculation of the arbuscular mycorrhizal fungus (AMF) Glomus mosseae (Gm) and the PGPF Fusarium equiseti (isolates GF18‐3 and GF19‐1) in terms of plant growth enhancement, root and rhizosphere colonisation, and development of anthracnose (Colletotrichum orbiculare) and damping‐off (Rhizoctonia solani AG‐4) diseases in cucumber plants was investigated under controlled conditions. The amendment of either GF18‐3 or GF19‐1 singly or in combination with Gm indicated a general tendency to significantly enhance the shoot dry weight (SDW) of cucumber plants at 4 weeks after planting (WAP). Similarly, Gm alone significantly enhanced SDW at 4 WAP. Gm showed a tendency to depress root colonisation by F. equiseti but such antagonistic effect was not observed in the rhizosphere soil. Both GF18‐3 and GF19‐1 significantly reduced percent root colonisation of Gm. However, these general tendencies may vary with the inoculum densities of AMF and PGPF. Both F. equiseti and Gm inoculated singly significantly increased percent of protection against anthracnose, but the combined inoculation was more effective in controlling the disease compared to single inoculation. The inoculation of the cucumber seedlings with GF18‐3, GF19‐1 or Gm, 6 or 12 days prior to damping‐off pathogen inoculation, increased percent of protection against damping‐off disease. This study shows that the co‐inoculation of F. equiseti and Gm resulted in additive effect on the suppression of anthracnose disease in cucumber.     

Development of microbial consortia as a biocontrol agent for effective management of fungal diseases in Glycine max L.

Thirty isolates of bacteria and six isolates of Trichoderma were isolated from fertile agricultural soil and evaluated for their antagonistic activity against phytopathogens like Macrophomina phaseolina and Sclerotinia sclerotiorum, under in vitro conditions. Different isolates showed varying degrees of antagonism. The three most antagonistic bacteria Pseudomonas aeruginosa (MBAA1), Bacillus cereus (MBAA2) and Bacillus amyloliquefaciens (MBAA3) and one fungi Trichoderma citrinoviride (MBAAT) were selected as the most effective isolates as biocontrol agents. The present study was undertaken to develop a plant growth promoting microbial consortium to reduce the disease incidence in Glycine max both under in vitro and in vivo conditions. Biocontrol attributes such as ammonia, siderophore, enzymes like β-1,3 glucanase, chitinase and cellulase were more potential in consortia in comparison to single isolates. Plants treated with consortia?+?pathogen showed lower disease incidence in comparison to single antagonist?+?pathogen and pathogen infested control (p?≤?0.05). Maximum disease control was observed in potted plants treated with S. sclerotiorum?+?MBAA1?+?MBAAT showing only 15.8% disease incidence in comparison to Sclerotinia infested control, in which disease incidence was 97%. Seed bacterised with MBAA1?+?MBAAT exhibited enhanced seed germination of G. max up to 68% along with subsequent increase in other plant growth parameters. Considerable increase in seedling vigour index (1863.2) and chlorophyll content (13.518?mg/g) was observed in seeds treated with MBAA1?+?MBAAT in plants infected with M. phaseolina.     

In Vitro and In Vivo Plant Growth Promoting Activities and DNA Fingerprinting of Antagonistic Endophytic Actinomycetes Associates with Medicinal Plants

Endophytic actinomycetes have shown unique plant growth promoting as well as antagonistic activity against fungal phytopathogens. In the present study forty-two endophytic actinomycetes recovered from medicinal plants were evaluated for their antagonistic potential and plant growth-promoting abilities. Twenty-two isolates which showed the inhibitory activity against at least one pathogen were subsequently tested for their plant-growth promoting activities and were compared genotypically using DNA based fingerprinting, including enterobacterial repetitive intergenic consensus (ERIC) and BOX repetitive elements. Genetic relatedness based on both ERIC and BOX-PCR generates specific patterns corresponding to particular genotypes. Exponentially grown antagonistic isolates were used to evaluate phosphate solubilization, siderophores, HCN, ammonia, chitinase, indole-3-acetic acid production, as well as antifungal activities. Out of 22 isolates, the amount of indole-3-acetic acid (IAA) ranging between 10–32 μg/ml was produced by 20 isolates and all isolates were positive for ammonia production ranging between 5.2 to 54 mg/ml. Among 22 isolates tested, the amount of hydroxamate-type siderophores were produced by 16 isolates ranging between 5.2 to 36.4 μg/ml, while catechols-type siderophores produced by 5 isolates ranging from 3.2 to 5.4 μg/ml. Fourteen isolates showed the solubilisation of inorganic phosphorous ranging from 3.2 to 32.6 mg/100ml. Chitinase and HCN production was shown by 19 and 15 different isolates, respectively. In addition, genes of indole acetic acid (iaaM) and chitinase (chiC) were successively amplified from 20 and 19 isolates respectively. The two potential strains Streptomyces sp. (BPSAC34) and Leifsonia xyli (BPSAC24) were tested in vivo and improved a range of growth parameters in chilli (Capsicum annuum L.) under greenhouse conditions. This study is the first published report that actinomycetes can be isolated as endophytes from within these plants and were shown to have antagonistic and plant growth promoting abilities. These results clearly suggest the possibility of using endophytic actinomycetes as bioinoculant for plant growth promotion, nutrient mobilization or as biocontrol agent against fungal phytopathogens for sustainable agriculture.     

Transmission of seed-borne infection of chilli by Burkholderia solanacearum and effect of biological seed treatment on disease incidence

Abstract

A survey of chilli fields in the state of Karnataka, India, showed the presence of bacterial wilt disease in important chilli growing regions. The disease incidence ranged from 26?–?32%. The pathogen was isolated from infected plant material and seeds. Infected plant material showed the release of milky white bacterial ooze. Burkholderia solanacearum was detected from chilli seeds by liquid assay and its identity was confirmed by biochemical tests, hypersensitive reaction and pathogenicity tests. Seed transmission of the pathogen up to 45% was observed in seeds artificially infested with the pathogen. Among different tissues of the seed, endosperm showed the presence of the pathogen. Biological seed treatment with antagonistic Pseudomonas fluorescens significantly (p?=?0.05) improved the seed quality parameters under laboratory conditions and drastically reduced the bacterial wilt incidence under field conditions. Seed-borne nature, transmission and effect of Pseudomonas fluorescens in both the forms of pure culture and formulation on seed quality parameters and bacterial wilt incidence are discussed in the present work.     

Biocontrol of Postharvest Grey Mould on Tomato by Yeasts

The fungal pathogen Botrytis cinerea causes severe rots on tomato fruit during storage and shelf life. Biological control of postharvest diseases of fruit may be an effective alternative to chemical control. Yeasts are particularly suitable for postharvest use, proving to be highly effective in reducing the incidence of fungal pathogens. Yeast fungi isolated from the surface of solanaceous plants were evaluated for their activity in reducing the postharvest decay of tomato caused by B. cinerea. Of 300 isolates, 14 strains of Rhodotorula rubra and Candida pelliculosa were found to be strongly antagonistic to the pathogen in vitro and were selected for further storage experiment. The antagonists were evaluated for their effect on the biological control of postharvest grey mould. Artificially wounded fruits were treated by means of a novel technique: small sterile discs of filter paper imbibed separately in suspensions of each yeast and the pathogen were superposed onto each wound. After 1‐week, 11 isolates were significantly effective in reducing the diameter of lesions by more than 60% compared to the control treated with B. cinerea alone. Total protection was obtained with the strain 231 of R. rubra on fruits challenged with pathogen spores. To our knowledge, R. rubra and C. pelliculosa have not been described as biocontrol agents against grey mould caused by B. cinerea. Our data demonstrate that the application of antagonistic yeasts represents a promising and environmentally friendly alternative to fungicide treatments to control postharvest grey mould of tomato.     

Effect of root exudates on the spore germiantion of rhizosphere and rhizoplane mycoflora of chilli (Capsicum annuum L.) cultivars

Summary From root exudates of three cultivars of chilli (Capsicum annuum L.) 12 amino acids and 7 sugars were detected. Methionine, d-1- phenylalanine, citrulline and d-xylose were detected only from the root exudates of resdistant cultivars. The root exudates of resistant variety inhibited spore germination of the pathogen (Fusarium oxysporum f. sp.capsici), but that of susceptible variety enhanced spore germiantion of the same. Spore germiantion of antagonistic fungi (Trichderma viride andAspergillus sydowi) was also influenced by the root exudates of resistant and susceptible varieties, but the influence was different.Spore germiantion of a number of rhizosphere fungi was studied and in general root exudate of susceptible cultivar enhanced spore germiantion of majority of fungi, but spore germination of antagonistic fungi against the pathogen was inhibited. However, root exudate of resistant cultivar stimulated spore germination of antagonistic fungi.     

Biocontrol Agent Talaromyces flavus Stimulates the Growth of Cotton and Potato

Beneficial plant-microbe interactions in the rhizosphere are primary determinants of plant health and soil fertility. Some antagonistic fungi have shown great effects toward the growth of plant crops. In this study, two major crops, cotton and potato, were selected to evaluate their growth promotion by the antagonistic fungus Talaromyces flavus. For each plant, five T. flavus isolates were selected from our fungal collection which had shown the highest antagonistic activities against the causal agent of wilt diseases on these plants. In the next step, for every crop, five isolates were used under greenhouse conditions. For evaluation of the plant growth promotion ability of T. flavus isolates, a split-plot trial was arranged in a randomized complete block design with four replications. The main factor was the method of application of T. flavus as a soil treatment, a seed treatment, and a combination of both methods. The subfactor was the use of different fungal isolates. Measured parameters were root length, crown length, plant height, plant fresh weight, and plant dry weight. Results showed that the maximum increase in the above parameters was mediated by the seed treatment method. The most effective isolate for cotton plants was TF-Co-M-23, which increased root length, plant height, plant fresh weight, and plant dry weight by 1.80-, 2.26-, 1.23-, and 1.19-fold, respectively. There were no significant differences among the various treatments affected by T. flavus in terms of crown length. The most effective isolate for potato plants was TF-Po-V-50, which increased root length, crown length, plant height, and plant dry weight by 1.71-, 1.09-, 1.45-, and 3.75-fold, respectively. The overall results of this study suggest that it may be possible to promote cotton and potato growth characteristics by using the antagonistic fungus T. flavus.     

Bacterial antagonists of Sunflower (Helianthus annuus L.) fungal pathogens

Bacteria isolated on nutrient agar and King's medium B from sunflower leaves, crown and roots inhibited in vitro growth of the leaf spot and wilt pathogens Alternaria helianthi, and Sclerotium rolfsii, respectively, and also the root rot pathogensRhizoctonia solani and Macrophomina phaseolina. Antagonistic bacteria from leaves were mainly actinomycetes and pigmented Gram-positive bacteria, while those from roots and crowns were identified asPseudomonas fluorescens-putida, P. maltophilia, P. cepacia, Flavobacterium odoratum andBacillus sp. In soil bioassays, when used as seed inoculum in the presence ofS. rolfsii, P. cepacia strain N24 increased significantly the percentage of seedling emergence. Bacterial strains which exhibited broad spectrum in vitro antagonistic activity were tested for colonisation of sunflower roots, when used as a seed inoculum. Good colonisers (10⁴ to 10⁶ bacteria/g root) were consistent in their ability to reduce disease and fungal wilt. A seedling having a primary root length < 5 cm with fewer lateral roots, necrosed cotyledons or crown and a wilted shoot indicated its diseased status. On an average, only 30% of seedlings were diseased when treated with the antagonistic strains, in the presence of the pathogen, while 60% of the seedlings were diseased in the presence of the pathogen alone. In microplots treated with strain N24, only 1 to 3% of the seedlings were wilted, while 14% of the seedlings were wilted in the presence of the pathogen alone. The results obtained show that bacterial antagonists of sclerotial fungi can be used as seed inocula to improve plant growth through disease suppression     

Impact of Plant Species and Site on Rhizosphere-Associated Fungi Antagonistic to Verticillium dahliae Kleb.

Fungi with antagonistic activity toward plant pathogens play an essential role in plant growth and health. To analyze the effects of the plant species and the site on the abundance and composition of fungi with antagonistic activity toward Verticillium dahliae, fungi were isolated from oilseed rape and strawberry rhizosphere and bulk soil from three different locations in Germany over two growing seasons. A total of 4,320 microfungi screened for in vitro antagonism toward Verticillium resulted in 911 active isolates. This high proportion of fungi antagonistic toward the pathogen V. dahliae was found for bulk and rhizosphere soil at all sites. A plant- and site-dependent specificity of the composition of antagonistic morphotypes and their genotypic diversity was found. The strawberry rhizosphere was characterized by preferential occurrence of Penicillium and Paecilomyces isolates and low numbers of morphotypes (n = 31) and species (n = 13), while Monographella isolates were most frequently obtained from the rhizosphere of oilseed rape, for which higher numbers of morphotypes (n = 41) and species (n = 17) were found. Trichoderma strains displayed high diversity in all soils, but a high degree of plant specificity was shown by BOX-PCR fingerprints. The diversity of rhizosphere-associated antagonists was lower than that of antagonists in bulk soil, suggesting that some fungi were specifically enriched in each rhizosphere. A broad spectrum of new Verticillium antagonists was identified, and the implications of the data for biocontrol applications are discussed.     

Development of rhizosphere and rhizoplane microflora of Aristida coerulescens in the Libyan desert

Summary Microflora of rhizosphere soil, rhizoplane and macerated root-portions of Aristida coerulescens, naturally occurring in the Libyan desert, were different in count and isolates, in the different root zones. A rhizosphere effect characteristic of each zone is shown. The root base contained the lowest numbers of microflora (bacteria and fungi) whilst the root tip included the highest counts. Distribution of most of the individual fungal species in the different root zones and root-surfaces is given in text.     

Microbial communities of Solanum tuberosum and magainin-producing transgenic lines

Antimicrobial peptide magainin II, isolated from the skin of the African clawed toad, has shown activity in vitro against a range of micro-organisms. Transgenic potato lines expressing a synthetic magainin gene show improved resistance to the bacterial plant pathogen, Erwinia carotovora. Culturable bacterial and fungal communities associated with magainin-producing potato plants were compared with those communities from the non-transgenic parental control and with another potato cultivar. Total numbers of aerobic bacteria recovered from the leaves of the magainin-producing line, its non-transgenic parent line and an unrelated cultivar did not differ significantly. There were no detectable differences in the numbers of Gram-positive and Gram-negative bacteria, pseudomonad populations or fungi recovered from foliage from the three plant lines. Bacterial populations recovered from the roots of a magainin-expressing plant line did not differ significantly from populations recovered from the unmodified parental line. Tubers from the magainin-expressing transgenic potatoes, however, had significantly lower total numbers of bacteria than tubers produced by unmodified plants. In vitro testing of rhizosphere isolates against magainin analogues found that bacterial isolates varied in their susceptibility to the peptides. There were no significant differences in the total numbers of fungi and yeasts recovered from the various plant lines, with one exception: higher numbers of fungi were recovered from roots of magainin-expressing plants than the unmodified control plants.     

Colletotrichum fructicola and C. siamense are involved in chilli anthracnose in India

Chilli anthracnose is a major problem in India and worldwide. In this study, we investigated the phylogenetic relationships of 52 fungal isolates associated with chilli anthracnose in southern India. All the 52 isolates were sequenced for partial ITS/5.8S rRNA and glyceraldehyde-3-phosphate dehydrogenase (gapdh) genes and showed affinities with Colletotrichum siamense and C. fructicola within Colletotrichum. gloeosporioides species complex. Further, a reduced subset of 17 selected isolates was made and in a maximum parsimony analysis of a multigene data-set including partial ITS/5.8S rRNA, actin (act), calmodulin (cal), chitin synthase (chs1), gapdh and β-tubulin (tub2) gene sequence data, these fungal isolates clustered with the type strain of C. fructicola, except for strain MTCC 3439 that showed phylogenetic affinities with C. siamense. The pathogenicity tests involving two representative isolates: UASB-Cg-14 and MTCC 3439, confirmed the involvement of C. fructicola and C. siamense in the development of disease symptoms on fresh chilli fruits. This is the first report of the association of C. fructicola and C. siamense in causing chilli anthracnose in India.     

Sugar Beet-Associated Bacterial and Fungal Communities Show a High Indigenous Antagonistic Potential Against Plant Pathogens

The aim of this study was to analyze microbial communities in/on sugar beet with special focus on antagonists toward plant pathogens. For this purpose, the composition of microorganisms isolated from the rhizosphere, phyllosphere, endorhiza, and endosphere of field-grown sugar beet plants was analyzed by a multiphasic approach at three different plant development stages at six locations in Europe. The analysis of microbial communities by Single Strand Conformation Polymorphism (SSCP) of 16S/18S rRNA clearly revealed the existence of discrete microenvironment- and site-specific patterns. A total of 1952 bacterial and 1344 fungal isolates screened by dual testing for antagonism toward the pathogens Aphanomyces cochlioides, Phoma betae, Pythium ultimum, and Rhizoctonia solani resulted in 885 bacterial (=45%) and 437 fungal (=33%) antagonists. In general, the indigenous antagonistic potential was very high and influenced by (a) the location, (b) the plant developmental stage, and (3) the microenvironment. Furthermore, we showed for the first time that the antagonistic potential was highly specific for each target pathogen. The majority of antagonistic microorganisms suppressed only one pathogen (bacteria: 664 = 75%; fungi: 256 = 59%), whereas the minority showed a broad host range (bacteria: 4 = 0.5%; fungi: 7 = 1.6%). The bacterial communities harbored the highest antagonistic potential against P. ultimum, whereas the fungal communities contained more antagonists against A. cochlioides and R. solani. In contrast to their high proportion, only a low diversity of antagonists at genotypic and species level was found. Novel antagonistic species, e.g., Subtercola pratensis or Microbacterium testaceum were found in the internal part of the sugar beet body.     

Cellular interactions between arbuscular mycorrhizal fungi and rhizosphere bacteria

Summary We have investigated whether direct physical interactions occur between arbuscular mycorrhizal (AM) fungi and plant growth promoting rhizobacteria (PGPRs), some of which are used as biocontrol agents. Attachment of rhizobia and pseudomonads to the spores and fungal mycelium ofGigaspora margarita has been assessed in vitro and visualized by a combination of electron and confocal microscopy. The results showed that both rhizobia and pseudomonads adhere to spores and hyphae of AM fungi germinated under sterile conditions, although the degree of attachment depended upon the strain.Pseudomonas fluorescens strain WCS 365 andRhizobium leguminosarum strains B556 and 3841 were the most effective colonizers. Extracellular material of bacterial origin containing cellulose produced around the attached bacteria may mediate fungal/bacterial interactions. These results suggest that antagonistic and synergistic interactions between AM fungi and rhizosphere bacteria may be mediated by soluble factors or physical contact. They also support the view that AM fungi are a vehicle for the colonization of plant roots by soil rhizobacteria.Abbreviations AM arbuscular mycorrhiza - PGPR plant growth promoting rhizobacteria - CBH cellobiohydrolase - DAPG 2,4-(diacetyl-phloroglucinol - TY triptone-yeast - LB Lauria-Bertani Dedicated to Prof. Dr. Dr. h.c. Eberhard Schnepf on the occasion of his retirement     

Antifungal and Root Surface Colonization Properties of GFP-Tagged Paenibacillus brasilensis PB177

Summary This study evaluates the potential of Paenibacillus brasilensis strain PB177 to inhibit phytopathogenic fungi commonly causing maize diseases and to colonize maize plants. In vitro assays demonstrated antagonistic activity against the fungal pathogens, Fusarium moniliforme and Diplodia macrospora. The PB177 strain was tagged with the gfp gene, encoding the green fluorescent protein (GFP) and GFP-tagged bacteria were detected attached to maize roots by stereo- and confocal microscopy. The GFP-tagged bacteria were also used to treat maize seeds before challenging the seeds with two phytopathogenic fungi. The results demonstrated that the bacterial cells are mobilized to the maize roots in the presence of the fungal pathogens. The ability of P. brasilensis PB177 to inhibit fungal growth in vitro and its capability of colonization of maize roots in vivo suggest a potential application of this strain as a biological control agent. This is the first report on the successful introduction of the GFP marker gene into a P. brasilensis strain, enabling the direct observation of these promising plant growth promoting bacteria on maize roots in situ.