Direct and Specific Assessment of Colonisation of Wheat Rhizoplane by Pseudomonas fluorescens Pf29A

The efficacy of fluorescent pseudomonads as suppressors of soil-borne diseases is linked to their ability to colonise plant roots. Monitoring the dynamics of biocontrol agents in the rhizosphere should improve the irreliability. We designed a pair of Sequenced Characterised Amplified Region (SCAR) primers specific to Pseudomonas fluorescens Pf29A, based on a specific 700 bp RAPD product selected in a previous work. Primer specificity was tested with DNA samples extracted from rhizospheric soil and rhizoplane of wheat plants grown in two different non-sterile soils. We assessed the total population of Pf29A by PCR and the culturable population by counting a tetracycline-resistant Pf29A transformant producing Green Fluorescent Protein (GFP), on selective medium 5 days after inoculation of non-sterile soil. SCAR primers were specific for Pf29A in both soils. We evaluated the limit of detection to 14.2 fg of target DNA, equivalent to 242 Pf29A cells per cm of wheat root. Culturable populations of Pf29A transformant accounted for 13% and 4% of the total populations 5 days after treatment with 10³ and 10⁷ CFU of transformed Pf29A per gram of soil. The SCAR derived sequence is a good candidate to develop a strain specific and sensitive PCR-quantification of Pf29A available for population dynamic studies in fields. We confirm that only a small proportion of the total Pf29A rhizosphere population is culturable.     

Effect of Organic Management of Soils on Suppressiveness to <Emphasis Type="Italic">Gaeumannomyces graminis</Emphasis> var. <Emphasis Type="Italic">tritici</Emphasis> and its Antagonist, <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis>

Organic management of soils is generally considered to reduce the incidence and severity of plant diseases caused by soil-borne pathogens. In this study, take-all severity on roots of barley and wheat, caused by Gaeumannomyces graminis var. tritici, was significantly lower in organically-managed than in conventionally-managed soils. This effect was more pronounced on roots of barley and wheat plants grown in a sandy soil compared to a loamy organically-managed soil. Fluorescent Pseudomonas spp. and in particular phlD⁺ pseudomonads, key factors in the take-all decline phenomenon, were represented at lower population densities in organically-managed soils compared to conventionally-managed soils. Furthermore, organic management adversely affected the initial establishment of introduced phlD⁺ P. fluorescens strain Pf32-gfp, but not its survival. In spite of its equal survival rate in organically- and conventionally-managed soils, the efficacy of biocontrol of take-all disease by introduced strain Pf32-gfp was significantly stronger in conventionally-managed soils than in organically-managed soils. Collectively, these results suggest that phlD⁺ Pseudomonas spp. do not play a critical role in the take-all suppressiveness of the soils included in this study. Consequently, the role of more general mechanisms involved in take-all suppressiveness in the organically-managed soils was investigated. The higher microbial activity found in the organically-managed sandy soil combined with the significantly lower take-all severity suggest that microbial activity plays, at least in part, a role in the take-all suppressiveness in the organically-managed sandy soil. The significantly different bacterial composition, determined by DGGE analysis, in organically-managed sandy soils compared to the conventionally-managed sandy soils, point to a possible additional role of specific bacterial genera that limit the growth or activity of the take-all pathogen.     

Concentration-dependent effect of salicylic acid application on wheat seedling resistance to take-all fungus, <Emphasis Type="Italic">Gaeumannomyces graminis</Emphasis> var. <Emphasis Type="Italic">tritici</Emphasis>

Application of 0.1 and 0.2 mM salicylic acid (SA) significantly reduced take-all disease caused by Gaeumannomyces graminis var. tritici (Ggt) and increased the root and shoot lengths and biomass, whereas 0.5 and 1 mM SA had no significant effect. The effective SA concentrations also increased the activities of soluble peroxidase (SPOX) and cell-wall-bound peroxidase (CWPOX) and the concentration of total phenolic compounds. SPOX activity was highest at days 4 and 3 in healthy roots and those inoculated with Ggt, respectively, and that of CWPOX at day 6 in both healthy and inoculated roots. The concentration of phenolic compound was also highest at day 3 in both healthy roots and those inoculated with Ggt. The results indicate that the protective effect of SA depends on certain concentrations which increase peroxidase activity and phenolic compounds accumulation in the wheat roots; higher SA concentrations did not differ from the controls.     

Interactions Between Strains of 2,4-Diacetylphloroglucinol-Producing Pseudomonas fluorescens in the Rhizosphere of Wheat

ABSTRACT Strains of fluorescent Pseudomonas spp. that produce the antibiotic 2,4-diacetylphoroglucinol (2,4-DAPG) are among the most effective rhizobacteria controlling diseases caused by soilborne pathogens. The genotypic diversity that exists among 2,4-DAPG producers can be exploited to improve rhizosphere competence and biocontrol activity. Knowing that D-genotype 2,4-DAPG-producing strains are enriched in some take-all decline soils and that P. fluorescens Q8r1-96, a representative D-genotype strain, as defined by whole-cell repetitive sequence-based polymerase chain reaction (rep-PCR) with the BOXA1R primer, is a superior colonizer of wheat roots, we analyzed whether the exceptional rhizosphere competence of strain Q8r1-96 on wheat is characteristic of other D-genotype isolates. The rhizosphere population densities of four D-genotype strains and a K-genotype strain introduced individually into the soil were significantly greater than the densities of four strains belonging to other genotypes (A, B, and L) and remained above log 6.8 CFU/g of root over a 30-week cycling experiment in which wheat was grown for 10 successive cycles of 3 weeks each. We also explored the competitive interactions between strains of different genotypes inhabiting the same soil or rhizosphere when coinoculated into the soil. Strain Q8r1-96 became dominant in the rhizosphere and in nonrhizosphere soil during a 15-week cycling experiment when mixed in a 1:1 ratio with either strain Pf-5 (A genotype), Q2-87 (B genotype), or 1M1-96 (L genotype). Furthermore, the use of the de Wit replacement series demonstrated a competitive disadvantage for strain Q2-87 or strong antagonism by strain Q8r1-96 against Q2-87 in the wheat rhizosphere. Amplified rDNA restriction analysis and sequence analysis of 16S rDNA showed that species of Arthrobacter, Chryseobacterium, Flavobacterium, Massilia, Microbacterium, and Ralstonia also were enriched in culturable populations from the rhizosphere of wheat at the end of a 30-week cycling experiment in the presence of 2,4-DAPG producers. Identifying the interactions among 2,4-DAPG producers and with other indigenous bacteria in the wheat rhizosphere will help to elucidate the variability in biocontrol efficacy of introduced 2,4-DAPG producers and fluctuations in the robustness of take-all suppressive soils.     

Interaction ofPseudomonas fluorescens withRhizobium for their effect on the management of peanut root rot

A biocontrol agent (Pseudomonas fluorescens) and a phytostimulator (Rhizobium) have been shown to have beneficial effects on plant growth and health. The study of plants inoculated withPseudomonas andRhizobium requires special attention because of the possibility that these agents may influence each other. Our study was conducted to test the effect of these inoculants on co-inoculation in peanut to control root rot, a severe soilborne disease caused byMacrophomina phaseolina. One fluorescent pseudomonad strain, Pf 1, which effectively inhibited the mycelial growth ofM. phaseolina underin vitro conditions, was studied for its compatibility with the biofertilizer bacterial strainRhizobium TNAU 14. Dual culture and colorimetric studies indicated the existence of a positive interaction between the microbial inoculants. However, glasshouse and field studies showed seed treatment and soil application ofPseudomonas fluorescens Pf 1 to be the most effective treatment in reducing root rot incidence and improving the crop vigor index, in comparison with treatments in which both inoculants were applied. http://www.phytoparasitica.org posting Feb. 11, 2002.     

Frequency, Diversity, and Activity of 2,4-Diacetylphloroglucinol-Producing Fluorescent Pseudomonas spp. in Dutch Take-all Decline Soils

ABSTRACT Natural suppressiveness of soils to take-all disease of wheat, referred to as take-all decline (TAD), occurs worldwide. It has been postulated that different microbial genera and mechanisms are responsible for TAD in soils from different geographical regions. In growth chamber experiments, we demonstrated that fluorescent Pseudomonas spp. that produce the antibiotic 2,4-diacetylphloroglucinol (2,4-DAPG) play a key role in the natural suppressiveness of two Dutch TAD soils. First, 2,4-DAPG-producing fluorescent Pseudomonas spp. were present on roots of wheat grown in both of the TAD soils at densities at or above the threshold density required to control take-all of wheat; in a complementary take-all conducive soil, population densities of 2,4-DAPG-producing Pseudomonas spp. were below this threshold level. Second, introduction of 2,4-DAPG-producing strain SSB17, a representative of the dominant geno-typic group found in the Dutch TAD soils, into the take-all conducive soil at population densities similar to the densities of indigenous 2,4-DAPG producers found in TAD soils provided control of take-all similar to that observed in the TAD soil. Third, a mutant of strain SSB17 deficient in 2,4-DAPG production was not able to control take-all of wheat, indicating that 2,4-DAPG is a key determinant in take-all suppression. These results show that in addition to the physicochemically different TAD soils from Washington State, 2,4-DAPG-producing fluorescent Pseudomonas spp. are also a key component of the natural suppressiveness found in Dutch TAD soils. Furthermore, it is the first time since the initial studies of Gerlagh (1968) that at least part of the mechanisms and microorganisms that operate in Dutch TAD soils are identified. Although quantitatively similar, the genotypic composition of 2,4-DAPG-producing Pseudomonas spp. varied between the Dutch TAD soils and the TAD soils from Washington State.     

Bacillus sp. L324-92 for Biological Control of Three Root Diseases of Wheat Grown with Reduced Tillage

ABSTRACT Strain L324-92 is a novel Bacillus sp. with biological activity against three root diseases of wheat, namely take-all caused by Gaeumannomyces graminis var. tritici, Rhizoctonia root rot caused by Rhizoctonia solani AG8, and Pythium root rot caused mainly by Pythium irregulare and P. ultimum, that exhibits broad-spectrum inhibitory activity and grows at temperatures from 4 to 40 degrees C. These three root diseases are major yieldlimiting factors for wheat in the U.S. Inland Pacific Northwest, especially wheat direct-drilled into the residue of a previous cereal crop. Strain L324-92 was selected from among approximately 2,000 rhizosphere/rhizoplane isolates of Bacillus species isolated from roots of wheat collected from two eastern Washington wheat fields that had long histories of wheat. Roots were washed, heat-treated (80 degrees C for 30 min), macerated, and dilution-plated on (1)/(10)-strength tryptic soy agar. Strain L324-92 inhibited all isolates of G. graminis var. tritici, Rhizoctonia species and anastomosis groups, and Pythium species tested on agar at 15 degrees C; provided significant suppression of all three root diseases at 15 degrees C in growth chamber assays; controlled either Rhizoctonia root rot, takeall, or both; and increased yields in field tests in which one or more of the three root diseases of wheats were yield-limiting factors. The ability of L324-92 to grow at 4 degrees C probably contributes to its biocontrol activity on direct-drilled winter and spring wheat because, under Inland Northwest conditions, leaving harvest residues of the previous crop on the soil surface keeps soils cooler compared with tilled soils. These results suggest that Bacillus species with desired traits for biological control of wheat root diseases are present within the community of wheat rhizosphere microorganisms and can be recovered by protocols developed earlier for isolation of fluorescent Pseudomonas species effective against take-all.     

Effect of Foliar Application of Urea on Rhizosphere and Rhizoplane Microflora of Wheat1)

The effect of urea leaf treatment on plant growth and rhizosphere microorganisms was studied under controlled environmental conditions with spring wheat at 2 growth stages, and at 2 inorganic nutrient levels of the soil. The increase in dry weight of shoots and roots was inhibited by the urea treatment. Treatment of seedlings also resulted in a delay in the formation of tillers and nodal roots, especially in low fertilized soil. A temporary increase of rhizoplane bacteria was demonstrated after 7–9 days due to a stimulation of fast growing strains. A quantitative effect on rhizoplane fungi was only apparent, on day 9 after treatment of seedlings, as a decrease in total numbers and coincided with the increase in numbers of bacteria and actinomycetes. At this time the numbers of fungi known to be early root colonizers like Penicillium spp. and Mortierella spp. decreased after initially being stimulated, while numbers of late colonizers like Fusarium spp. and Trichoderma spp. increased after 9 days after initially being reduced in comparison with the control. Investigation of the rhizosphere effect (R/S) of wheat by the soil dilution plate method and fluorescence microscopy showed a 200% increase in total numbers of micro-organisms in the rhizosphere soil when the former method was used, but a 10–30% reduction when fluorescence microscopy was employed.     

Field testing putative biological controls of take-all: rationale and results1

In seeking biological control of the wheat take-all fungus (Gaeumannomyces graminis var. tritici) by introduced organisms, the demonstration of satisfactory field performance is proving a formidable hurdle. A novel experimental design incorporating small plots, 37 x 31 cm, has been used in UK at Rothamsted and Woburn since 1983 to test different kinds of control for take-all and to explore some of the problems of providing adequate field tests of putative biocontrol agents. Three years of bacterial treatments, using different plots each year, provided no evidence of effective control of the disease. Of the few significant treatment effects, most occurred in spring and were temporary: at Woburn they were mostly decreases in take-all and at Rothamsted mostly decreases in growth of the wheat plant without concomitant changes in take-all.     

Biocontrol of soil-borne fungal plant diseases by 2,4-diacetylphloroglucinol-producing fluorescent pseudomonads with different restriction profiles of amplified 16S rDNA

Fluorescent pseudomonads producing the antimicrobial compound 2,4-diacetylphloroglucinol (Phl) are being studied extensively for use as biocontrol agents of soil-borne fungal diseases. Some of them can produce pyoluteorin (Plt) in addition to Phl, whereas others synthesise only Phl. Here, a collection of seven Phl⁺ Plt^- pseudomonads, seven Phl⁺ Plt⁺ pseudomonads and seven Phl^- biocontrol pseudomonads were compared for protection of plant roots against fungal pathogens. The seven Phl⁺ Plt⁺ pseudomonads were identical by restriction analysis of amplified spacer ribosomal DNA (spacer ARDRA), whereas the Phl⁺ Plt^- pseudomonads and especially the Phl^- biocontrol pseudomonads were quite diverse by spacer ARDRA. Collectively, the Phl⁺ Plt^- pseudomonads proved superior to the Phl⁺ Plt⁺ pseudomonads and the Phl^- biocontrol pseudomonads for protection of tomato against Fusarium crown and root rot (in rockwool microcosms) or cucumber against Pythium damping-off (in non-sterile soil microcosms). There was no correlation between protection in vivo and inhibition of the corresponding fungal pathogen on plates. However, there was a significant correlation between the amount of Phl produced on plates and protection of tomato against Fusarium crown and root rot, but not with protection of cucumber against Pythium damping-off. Interestingly, the minority of strains unable to produce HCN, an extracellular protease, or both, were among those unable to protect plants in both pathosystems. A seedling assay was developed to compare pseudomonads for suppression of Fusarium crown and root rot in vitro, and a significant correlation was found between disease severity in vitro and in vivo. Overall, results suggest that promising biocontrol pseudomonads may be identified based on the ability to produce Phl and/or specific ARDRA-based fingerprints.     

Assessment of in vivo screening systems for potential biocontrol agents of Gaeumannomyces graminis

A screening programme was used to search for biocontrol agents against Gaeumannomyces graminis causing take-all disease of wheat. Of the 1800 rhizosphere microorganisms tested, 10% controlled the disease in a secondary screen. The 30 most effective isolates were further investigated for mode of action. Although 72% of the sites sampled for antagonistic microbes were planted to continuous cereals, they yielded only 23% of the most effective isolates. Of all the isolates selected, 63% belonged to the genera Bacillus, Pseudomonas and Penicillium; Beauveria and Rhodococcus were also antagonistic. Fluorescent pseudomonads, all producing siderophores in low-iron medium, accounted for 23% of the isolates. Over 50% of strains produced β-glucanases and chitinases. Less than 50% of the strains selected by the in vivo screen inhibited G. graminis in agar plate tests. In the gnotobiotic system used, the Pseudomonas strains were faster in colonizing the wheat roots than the majority of the Bacillus and fungal strains.     

Interactions between the biocontrol agent Pseudomonas fluorescens CHA0 and Thielaviopsis basicola in tobacco roots observed by immunofluorescence microscopy

Pseudomonas fluorescens CHA0 protects plants from damage caused by several soilborne fungi. In this work, immunofluorescence microscopy was used to investigate the colonization of tobacco roots by CHA0 and its physical relationship with the black root rot fungus Thielaviopsis basicola . The pseudomonad colonized the rhizoplane shortly after planting of tobacco seedlings in sterile soil microcosms, in which it had been introduced as soil inoculant. CHA0 was found between and inside cells in the epidermis and the cortex, as well as in the xylem vessels, within 4–7 days after planting of seedlings. The presence of CHA0 delayed the colonization of the interior of tobacco roots by T. basicola compared with the treatment in which only the fungus had been inoculated. Likewise, the pseudomonad reduced the extent of black root rot from 82% to 28%. However, CHA0 was seldom found in contact with the mycelium of T. basicola or in its vicinity, indicating that direct colonization of the mycelium of T. basicola by CHA0 was not required for protection of tobacco against black root rot. Overall, the results suggest that the interior of the root is a key site for implementation of the strain's biocontrol activity against soilborne plant-pathogenic fungi.     

Diversity and Ecology of Biocontrol Pseudomonas spp. in Agricultural Systems

ABSTRACT Diverse Pseudomonas spp. may act as biological controls of plant pathogens, but the ecology of those natural populations is not well understood. And, while biocontrol potential has been identified in multiple pseudomonad strains, the linkages between genotype and phenotype have yet to be fully delineated. However, intensive studies of one class of biocontrol strains, i.e., those that can produce 2,4-diacetylphloroglucionl (DAPG), have provided new insights into the diversity, distribution, and interactions of biocontrol pseudomonads. Those studies also laid the foundation for future research and development of pseudomonad-based biocontrol strategies. Over the past several years, numerous studies have also revealed that biocontrol pseudomonads are widely distributed in agricultural soils, and that multiple crop and soil factors can affect their abundance and activities. Recent work has shown that a variety of farm management practices that reduce soilborne disease pressure can also alter the rhizosphere abundance of DAPG producers in complex ways. Such studies provide support for the hypothesis of an ecological feedback mechanism whereby a native biocontrol population increase and subsequently reduce root disease severity following infection. It is well established that complex biological interactions can take place among bio-control pseudomonads, plant pathogens, their hosts, and other members of the microbial community. The net result of such interactions likely dilutes biocontrol efficacy at the field scale. Nonetheless, inoculation can be effective, and several successful applications of biocontrol pseudomonads have been developed. Future applications of microbial ecology research will hopefully improve the consistency and efficacy of bio-control mediated by Pseudomonas spp. Current applications and future opportunities for improving pseudomonad-based biological control are discussed.     

Soil receptivity toFusarium solani f. sp.pisi and biological control of root rot of pea

Potential antagonists ofFusarium solani f. sp.pisi (Fsp) were selected from soil samples with varying degrees of receptivity to this pathogen. They were tested against Fsp isolate 48 (Fs48), in increasingly complex systems. Most species testedin vitro were able to antagonize Fs48. No relation could be establishedin vitro between the receptivity of the soil from which an isolate originated and its antagonism to Fs48. In soils naturally infested with pea root rot pathogens, which were stored humid at 4°C for a period longer than a year, various isolates ofFusarium, Gliocladium andPenicillium spp. were able to reduce root rot. After sterilization of these soils, onlyGliocladium roseum isolates, added at 10⁵ conidia g^–1 dry soil, significantly reduced disease severity and prevented root weight losses caused by Fs48 at 10⁴ conidia g^–1 dry soil. In soils in which the biota were activated by growing peas before the assays, doses of 10⁶ and 10⁷ ofG. roseum were required to reduce root rot. In these soils, the antagonistic effects of fluorescent pseudomonad strains from soil of low receptivity to Fsp were variable. Some strains of fluorescent pseudomonads, from soil moderately receptive to Fsp and from highly infested soils, were also able to reduce root rot. Disease suppression by pseudomonad strains was more evident in the absence than in the presence ofAphanomyces euteiches in the root rot pathogen complex. The role of receptiveness of the soil with regard to potential antagonists is discussed.     

Wheat Genotype-Specific Induction of Soil Microbial Communities Suppressive to Disease Incited by Rhizoctonia solani Anastomosis Group (AG)-5 and AG-8

ABSTRACT The induction of disease-suppressive soils in response to specific cropping sequences has been demonstrated for numerous plant-pathogen systems. The role of host genotype in elicitation of the essential transformations in soil microbial community structure that lead to disease suppression has not been fully recognized. Apple orchard soils were planted with three successive 28-day cycles of specific wheat cultivars in the greenhouse prior to infestation with Rhizoctonia solani anastomosis group (AG)-5 or AG-8. Suppressiveness to Rhizoctonia root rot of apple caused by the introduced isolate of R. solani AG-5 was induced in a wheat cultivar-specific manner. Pasteurization of soils after wheat cultivation and prior to pathogen introduction eliminated the disease suppressive potential of the soil. Wheat cultivars that induced disease suppression enhanced populations of specific fluorescent pseudomonad genotypes with antagonistic activity toward R. solani AG-5 and AG-8, but cultivars that did not elicit a disease suppressive soil did not modify the antagonistic capacity of this bacterial community. When soils were infested prior to the initial wheat planting, all cultivars were uniformly susceptible to R. solani AG-8. However, when pathogen inoculum was added after three growth-cycles, wheat root infection during the fourth growth-cycle varied in a cultivar specific manner. The same wheat cultivar-specific response in terms of transformation of the fluorescent pseudomonad community and subsequent suppression of Rhizoctonia root rot of apple was observed in three different orchard soils. These results demonstrate the importance of host genotype in modification of indigenous saprophytic microbial communities and suggest an important role for host genotype in the success of biological control.     

荧光假单胞菌Tn5诱变菌株防治小麦全蚀病的初步研究

 利用生物技术(转座子Tn5诱变)对荧光假单胞菌进行遗传改良获得对小麦全蚀病有更好防治效果的基因工程菌株。将大肠杆菌S17-1中psup2021值粒上的转座子Tn5转移到荧光假单胞菌CN12菌系的基因组内获得5100个Tn5突变体,转化频率为1.2×10^-7。与自然菌系CN12比较,7个突变体对全蚀病菌的离体拮抗作用提高到160-250%,其中3个突变体对温室苗期全蚀病防治效果也显著提高(p=0.01)。初步研究证明:(1)荧光假单胞菌中可能存在两类功能不同的拮抗作用基因(暂称为抗生基因和调控基因);(2)在目前抗病基因缺乏或难于应用的情况下,利用微生物或其有用基因达到防病增产的目的可能是一条有希望的途径。     

枯草芽胞杆菌YB-05对小麦抗病性相关防御酶系的诱导作用

本文研究了生防菌枯草芽胞杆菌YB-05和病原菌小麦全蚀病菌GGT007对小麦体内防御酶活性的影响,探讨其诱导小麦抗病性机理。以苯丙氨酸解氨酶(PAL)、过氧化氢酶(CAT)、过氧化物酶(POD)、超氧化物歧化酶(SOD)和多酚氧化酶(PPO)5种防御酶作为小麦抗病性反应指标,于不同时段测定各防御酶活性;以PD培养基为对照,测定生防菌YB-05及小麦全蚀病菌GGT007对小麦叶片和根部抗性相关酶的影响。结果表明,小麦经生防菌与病原菌混合处理、病原菌处理、生防菌处理后,叶片和根部与植物防御抗病相关的PPO、POD、SOD、PAL、CAT防御酶活性均比对照组高,其中生防菌与病原菌混合处理后抗性相关酶活最高,叶片中PAL、POD、SOD、PPO、CAT酶活峰值达到46.705、16 829.274、104.687、97.44和1 259.565U/g,为对照组的1.74、2.44、2.27、2.40和2.42倍。根部PAL、POD、SOD、PPO、CAT酶活峰值达到131.536、56 424.79、1 977.04、22.564和206.241U/g,为对照组的1.65、1.52、2.57、2.07、1.74倍。表明枯草芽胞杆菌YB-05和小麦全蚀病菌GGT007均能诱导小麦叶片和根部的防御酶活性增强,两者共同处理后小麦叶片和根部5种防御酶活性高于单独处理,说明枯草芽胞杆菌YB-05和小麦全蚀病菌GGT007共同诱导具有协同增效作用。     

Mechanisms of inhibition of Gaeumannomyces graminis var. tritici by fluorescent pseudomonads

The take-all fungus Gaeumannomyces graminis var. tritici reduced the weight of wheat plants grown in tubes containing sterilized sand and plant nutrient solution. Fluorescent pseudomonads, when added to the tubes, inhibited the take-all fungus and increased plant weight. Iron (as FeNaEDTA) had no effect on the inhibition of the fungus by the pseudomonads. Some of the pseudomonads produced a compound in the wheat rhizosphere with a UV absorption peak at 365 nm, but the inhibition of G. graminis by pseudomonads was not proportional to the UV absorbance of rhizosphere extracts. A yellow crystalline compound, absorbing at 365 nm, was extracted from broth cultures and shown to be toxic to G. graminis under acid conditions. This compound is considered to be phenazine-I-carboxylic acid.     

Application of Pseudomonas fluorescens isolates to wheat as potential biological control agents against take-all

Two isolates of Pseudomonas fluorescens (2–79 and 13–79) from the USA were evaluated in the UK as biological control agents against Gaeumannomyces graminis var. tritici , the cause of take-all in wheat. Biological control agents were applied as seed coatings in carboxymethyl cellulose (CMC) to seven wheat trials sown in 1987 and 1988 on fen peat and clay soils, and as peat-based and microgranule formulations in one of these trials. In a trial of spring wheat on fen peat, all treatments with biological control agents reduced the percentage take-all infection of crown roots and seminal roots, but the effects of only one isolate were statistically significant ( P <0·05). Effects of biological control agents on infection rates in five other trials were not significant. In the trial in which application methods were compared, peat-based inoculum initially appeared most effective but none of the treatments reduced take-all significantly throughout the season. Application of biological control agents was associated with yield increases in several trials; these were not consistently associated with effects on take-all. These results suggest that the isolates of P. fluorescens have potential to reduce take-all and increase yields of wheat in the UK, but the beneficial effects are inconsistent. There is a need to develop isolates which reliably control severe take-all in a variety of soil types.