Effects of plant age on disease development and virulence of Clavibacter michiganensis subsp. michiganensis on tomato

The effect of plant age at the time of inoculation on the severity of bacterial wilt and canker disease caused by Clavibacter michiganensis subsp. michiganensis (Cmm) was examined in six greenhouse experiments. The period during which inoculations led to wilt and death of tomato plants was defined. This period, designated ‘window of vulnerability’, ranged from transplanting to the 17‐ to 18‐leaf stage. Plants inoculated after this period expressed disease symptoms but did not wilt or die. No significant changes in disease incidence were observed when leaves of different ages were inoculated. Yield accumulation was significantly reduced in plants inoculated within the window of vulnerability compared with those inoculated after this period. Expression of virulence genes, viz. celA, encoding a secreted cellulase, and the serine protease‐encoding pat‐1, chpC and ppaA, was induced during the early stages after inoculation in plants inoculated within the window of vulnerability. Differences in Cmm population between plants inoculated within and outside of this period were insignificant after the first week post‐inoculation, indicating that differences in disease severity, yield loss and expression of virulence determinants are not correlated with Cmm population level. Results suggest that implementation of precautionary measures during the window of vulnerability to avoid secondary spread of Cmm will have a season‐long effect on plant mortality and may minimize, or even prevent, yield losses.     

Induction of systemic resistance byPseudomonas fluorescens Pf1 againstXanthomonas oryzae pv.Oryzae in rice leaves

When lower leaves of rice plants were inoculated with powder formulation of a saprophytic strain ofPseudomonas fluorescens, Pfl, upper leaves, in addition to the inoculated lower leaves, showed resistance to the rice bacterial blight pathogenXanthomonas oryzae pv.oryzae. When the leaves were challenge-inoculated withX. oryzae pv.oryzae 4 days afterP. fluorescens application on lower leaves, the disease intensity in upper leaves decreased from 6.7 to 1.1. When rice seeds were treated with the formulation ofP. fluorescens Pfl and sown, 30-day-old seedlings showed resistance toX. oryzae pv.oryzae and the disease intensity decreased from 6.8 to 1.2. The induced resistance was transient; leaves sprayed withP. fluorescens Pfl at 30 days after treatment and leaves of 60-day-old seedlings fromP. fluorescens-treated seeds did not show resistance to the pathogen. In field trials, seed treatment followed by foliar application of the powder formulation ofP. fluorescens Pfl effectively controlled rice bacterial blight and increased the yield. In the induced resistant leaves a sharp increase in lignification and activities of peroxidase, phenylalanine ammonia-lyase and 4-coumarate: CoA ligase was observed when the leaves were challenge-inoculated withX. oryzae pv.oryzae. An approximately threefold increase in lignin content, peroxidase activity and phenylalanine ammonia-lyase activity and a fivefold increase in 4-coumarate: CoA ligase activity were observed 5 days after challenge inoculation withX. oryzae pv.oryzae in rice leaves pretreated withP. fluorescens for 5 days. A similar increase in defense-related activities was not observed in susceptible interactions or inP. fluorescens-treated plants at later stages of interactions when no resistance to the pathogen was observed.     

Factors Influencing Transmission of Didymella rabiei (Ascochyta Blight) from Inoculated Seed of Chickpea Under Controlled Conditions

Ascochyta blight of chickpea (Cicer arietinum), caused by the fungus Didymella rabiei, has the potential to cause 100% crop loss in severe epiphytotics. Management of this disease often involves reducing sources of inoculum. The influence of sowing depth, host resistance, seed infection level and soil temperature on disease transmission was investigated in a series of glasshouse and growth room trials using seed artificially inoculated with D. rabiei. A positive correlation (R²=0.9992) was observed between rate of seed infection and the incidence of disease on seedlings. Disease transmission to seedlings was not significantly influenced by sowing depth (1, 3 and 6 cm) in separate trials on two cultivars. Susceptibility of the host showed no obvious influence on the frequency of disease transmission in two trials conducted using four cultivars ranging from highly susceptible to moderately susceptible/moderately resistant. Trials conducted in controlled conditions showed that there was no obvious relationship between soil temperature (5, 9, 14 and 19 °C) and the incidence of disease on seedlings.     

Effects of Host and Microbial Factors on Development of Clonostachys rosea and Control of Botrytis cinerea in Rose

Development of Clonostachys rosea in rose leaves and petals and control of Botrytis cinerea by the agent were investigated. C. rosea germinated, established endophytic growth, and sporulated abundantly whether the tissues were mature, senescent or dead when inoculated. Germination incidence was moderate on mature and senescent leaves (47% and 35%) and petals (31% and 43%), and high (>98%) on dead tissues. Sporulation of C. rosea in tissues inoculated when mature, senescent or dead averaged 41%, 61%, and 75% in leaves, and 48%, 87% and 53% in petals. When leaves were wounded with needles before inoculation, germination of C. rosea increased from 45–56% to 90–92%, but sporulation became high (>75%) regardless of wounds. When leaves were inoculated with C. rosea at 0–24h after wounding and subsequently with B. cinerea, germination of the pathogen was reduced by 25–41% and sporulation by 99%. A humid period prior to inoculation of senescent or dead leaves promoted communities of indigenous fungi, reduced sporulation of C. rosea and B. cinerea, and, in dead leaves, increased control of the pathogen associated with C. rosea. Applied at high density, isolates of indigenous Penicillium sp. and Alternaria alternata from rose interacted with C. rosea and reduced control of the pathogen by 16% and 21%, respectively. In conclusion, C. rosea markedly suppressed sporulation of B. cinerea in rose leaves and petals regardless of developmental stage, minor wounds, and natural densities of microflora. This versatility should allow C. rosea to effectively control inoculum production of B. cinerea in rose production systems.     

Reduction of Bacterial Speck (Pseudomonas syringae pv. tomato) of Tomato by Combined Treatments of Plant Growth-promoting Bacterium, Azospirillum brasilense, Streptomycin Sulfate, and Chemo-thermal Seed Treatment

Inoculation of tomato seeds with the plant growth-promoting bacterium Azospirillum brasilense, or spraying tomato foliage with A. brasilense, streptomycin sulfate, or commercial copper bactericides, separately, before or after inoculation with Pseudomonas syringae pv. tomato, the casual agent of bacterial speck of tomato, had no lasting effect on disease severity or on plant height and dry weight. Seed inoculation with A. brasilense combined with a single streptomycin foliar treatment and two foliar bactericide applications at 5-day intervals (a third or less of the recommended commercial dose) reduced disease severity in tomato seedlings by over 90% after 4 weeks, and significantly slowed disease development under mist conditions. A. brasilense did not induce significant systemic resistance against the pathogen although the level of salicylic acid increased in inoculated plants. Treatment of tomato seeds that were artificially inoculated with P. syringae pv. tomato, with a combination of mild chemo-thermal treatment, A. brasilense seed inoculation, and later, a single foliar application of a copper bactericide, nearly eliminated bacterial leaf speck even when the plants were grown under mist for 6 weeks. This study shows that a combination of otherwise ineffective disease management tactics, when applied in concert, can reduce bacterial speck intensity in tomatoes under mist conditions.     

非亲和性稻瘟病菌对水稻的诱导抗性

为研究稻瘟病菌Magnaporthe oryzae不同菌株间的相互作用,选择与单抗性基因系水稻IRBL5-M （携带抗性基因Pi5）表现为亲和性的菌株HN52与非亲和性的菌株HN119为研究对象,将其单独或混合接种到单抗性基因系水稻IRBL5-M中,并通过荧光显微镜观察接种后水稻叶鞘的发病情况及病斑面积,测定接种后水稻内相关抗性基因OsWRKY45、OsNPR1、OsPR10、OsMAPK2的表达量以及活性氧的变化。结果显示,相较于单独接种亲和性菌株,混合接种后单抗性基因系水稻IRBL5-M病斑发病面积减少;混合接种中亲和性菌株HN52菌丝侵染能力降低,侵染菌丝细胞间扩展率显著降低73.13%;同时单抗性基因系水稻IRBL5-M中OsWRKY45、OsNPR1、OsPR10和OsMAPK2抗性基因表达量显著增加,水稻叶片中活性氧含量增加,表明在菌株混合侵染过程中,非亲和性菌株可通过激发水稻的抗性反应来降低亲和性菌株对水稻的侵染程度。     

Antioxidant,ethylene and membrane leakage responses to powdery mildew infection of near-isogenic barley lines with various types of resistance

Leaves of powdery mildew-susceptible barley (Hordeum vulgare cv. Ingrid) and related near-isogenic lines bearing various resistance genes (Mla12, Mlg or mlo5) were inoculated with Blumeria graminis f. sp. hordei race A6. Fungal attack induced several-fold increases in ethylene emission and electrolyte leakage in leaves of susceptible Ingrid beginning 3 days after inoculation. Activities of peroxidase, superoxide dismutase, glutathione S-transferase, ascorbate peroxidase and glutathione reductase enzymes were induced markedly in susceptible leaves 5–7 days after inoculation. Similar, but less pronounced pathogen-induced changes were detected in inoculated leaves of Mla-type resistant plants that show hypersensitive cell death upon inoculation, and, to an even lesser extent, in the Mlg and mlo lines, where no visible symptoms accompanied the incompatible interaction. Glutathione content increased only in susceptible barley 7 days after inoculation. Catalase activity, total ascorbate content and redox state were not influenced by inoculation in any of the genotypes. The activity of dehydroascorbate reductase was significantly reduced 3–5 days after inoculation in the susceptible parental plants and after 5 days in Mla and Mlg lines, while it was stable in the mlo barley. Slightly elevated levels of H₂O₂ were observed in the inoculated resistant plants. In contrast, H₂O₂ content decreased in the susceptible line 7 days after pathogen attack. These data indicate that high levels of antioxidants are involved in the compatible interaction of susceptible barley and powdery mildew by protecting the pathogen from oxidative damage.     

Biocontrol agents in combination with Moringa oleifera extract for integrated control of Sclerotium-caused cowpea damping-off and stem rot

Damping-off and stem rot disease-causing Sclerotium rolfsii has been reported as a destructive soil-borne pathogen of numerous crops, especially in the tropics and subtropics. Trials were conducted to test the efficacy of biocontrol agents alone or combined with Moringa oleifera leaf extracts for the control of the disease. In the laboratory, PDA was amended with Moringa leaf extract, and mycelial growth of S. rolfsii was measured. In the greenhouse and field, Trichoderma Kd 63, Trichoderma IITA 508 and Bacillus subtilis were evaluated as seed treatments, soil drench or sprinkle, separately or combined with Moringa leaf extracts. Percentage disease incidence, severity and control were recorded. In the laboratory, the higher the extract concentration the less the mycelial growth and no mycelial growth occurred on extract at 15 or 20 g leaves 10 ml⁻¹ water. In the greenhouse, the highest disease control was observed at a Moringa extract concentration of 15 kg leaves 10 l⁻¹ water (w/v). Seed treatments using Trichoderma Kd 63, and soil sprinkle using Trichoderma IITA 508 had a significantly (P = 0.05) higher effect on a disease incidence than Bacillus. Disease severity followed the same pattern. Moringa seed treatment combined with Trichoderma soil sprinkle resulted in significantly more than 94% and 70% disease control in the greenhouse and field, respectively, with significant yield increase in the field. This is the first report of Moringa leaf extract combined with Trichoderma as an integrated control for Sclerotium damping-off and stem rot of cowpea in the field.     

Phoma black stem severity and phytoestrogen production in annual Medicago spp. is primarily determined by interaction of cultivar and pathogen isolate

Phoma black stem and leaf spot disease (caused by Phoma medicaginis) not only diminishes forage and seed yield but stimulates production of detrimental phytoestrogens in annual Medicago spp. This study aimed to evaluate relationships between disease development from five isolates of P. medicaginis on 16 cultivars with production of coumestrol and 4′-O-methylcoumestrol. In the presence of P. medicaginis, Sava had the highest coumestrol and 4′-O-methylcoumestrol (640 and 85 mg/kg, respectively) followed by Caliph (253 and 15 mg/kg, respectively). In the absence of P. medicaginis, Jemalong and Paragosa showed highest and lowest coumestrol (137 and 0 mg/kg, respectively). 4′-O-methylcoumestrol was not produced in disease-free plants, but coumestrol was. Disease incidence and severity on leaves and on petiole/stems, and consequent leaf yellowing severity ranged from 5%–98.7%, 0%–100%, 4.4%–98.7%, 1.7%–100%, and 0%–85%. Sava, Paraponto, Harbinger, and Serena were most susceptible, while Tornafield and Caliph were least susceptible. There was significant overall positive correlation of disease incidence/severity factors across cultivars (p < 0.01) with both coumestrol and 4′-O-methylcoumestrol. Jemalong was least responsive and Paragosa and Sava most responsive to coumestrol production following P. medicaginis inoculation. Coumestrol in inoculated Paragosa increased to 373 mg/kg in comparison with 0 mg/kg in controls. These findings are of critical importance towards developing less disease-susceptible annual Medicago spp. producing less detrimental phytoestrogens. Least susceptible cultivars like Tornafield and Caliph can be used to manage yield loss, whilst least responsive cultivars to phytoestrogen production like Caliph also can help to reduce phytoestrogen production.     

Experiments with some of the newer systemic fungicides on apple and marrow

In field trials against apple scab (caused by Venturia inaequalis) and powdery mildew (caused by Podosphaera leucotricha), improved control of one or other disease has been given by the newer systemic fungicides benomyl, triarimol and the thiophanates. However, in tests on apple seedlings in the greenhouse, biological evidence of translocation from individual deposits was generally limited to movement within the treated leaf. In greenhouse tests with cucumbers and marrows, using Oidium sp., there was again little evidence for movement of toxicant from a treated leaf, although effective disease control was readily obtained by root application of several compounds at low dosage. These results suggest that the systemic properties of the compounds evident when used as soil or seed treatments are of little account when they are applied as foliar sprays.     

Possibilities of biological and chemical control of Verticillium wilt in pepper

In pathogen populations in Serbia, the incidence, pathogenic and morphological characters ofVerticillium spp. were studied. Biological and chemical control ofVerticillium was investigated in pepper plants (Capsicum annuum L. cv. ‘Soroksari’) with the biofungicide Polyversum^® (Pythium oligandrum) and the conventional fungicides benomyl and propamocarb-hydrochloride. On the basis of macroscopic and microscopic characters of the isolates originating from eight localities in Serbia, it was established that they apparently belong to the speciesVerticillum dahliae. The isolates differed in their pathogenic characters. However, all of them caused marked wilting symptoms on pepper plants 40 days after inoculation, conducted when there were more than nine fully developed leaves on the primary stem. The fungicides were applied either before or after inoculation. Benomyl was the most efficient fungicide in wilt control (88.2% when applied after inoculation and 94.6% when applied before inoculation). Polyversum proved more efficient (66.6%) when applied before rather than after inoculation. Propamocarb-hydrochloride provided sufficient Verticillium wilt control; its efficacy and that of Polyversum were similar, and less efficient than benomyl, but still significantly different from the disease control.     

Assessment of recent outbreaks of <Emphasis Type="Italic">Dickeya</Emphasis> sp. (syn. <Emphasis Type="Italic">Erwinia chrysanthemi</Emphasis>) slow wilt in potato crops in Israel

Suspected Dickeya sp. strains were obtained from potato plants and tubers collected from commercial plots. The disease was observed on crops of various cultivars grown from seed tubers imported from the Netherlands during the spring seasons of 2004–2006, with disease incidence of 2–30% (10% in average). In addition to typical wilting symptoms on the foliage, in cases of severe infection, progeny tubers were rotten in the soil. Six strains were characterised by biochemical, serological and PCR-amplification. All tests verified the strains as Dickeya sp. The rep-PCR and the biochemical assays showed that the strains isolated from blackleg diseased plants in Israel were very similar, if not identical to strains isolated from Dutch seed potatoes, suggesting that the infection in Israel originated from the Dutch seed. The strains were distantly related to D. dianthicola strains, typically found in potatoes in Western Europe, and were similar to biovar 3 D. dadanti or D. zeae. This is the first time that the presence of biovar 3 strains in potato in the Netherlands is described. One of the strains was used for pathogenicity assays on potato cvs Nicola and Mondial. Symptoms appeared 2 to 3 days after stem inoculation, and 7 to 10 days after soil inoculation. The control plants treated with water, or plants inoculated with Pectobacterium carotovorum, did not develop any symptoms with either method of inoculation. The identity of Dickeya sp. and P. carotovorum re-isolated from inoculated plants was confirmed by PCR and ELISA.     

Effects of fungicides on Drechslera spp. and leaf surface filamentous saprophytic fungi on Lolium perenne

Small plots of Lolium perenne were treated between February and September 1979 with benomyl, triadimefon and chlorothalonil and the incidence and relative abundance of sporulating filamentous fungi measured on distinct lesions and senescent areas of leaves.
Benomyl reduced the levels of most saprophytes but increased the levels of the two Drechslera species present. Triadimefon reduced levels of Drechslera species but allowed an increase in levels of Phoma and Epicoccum and in the number of naturally senescing leaves devoid of obvious filamentous fungal growth. In chlorothalonil-treated plots, levels of all filamentous fungi except Epicoccum were reduced and, by controlling Drechslera species, the number of healthy leaves increased.     

Paraphoma root rot of alfalfa (Medicago sativa) in Inner Mongolia,China

Root rot symptoms were observed in fields of alfalfa in Chifeng city, Inner Mongolia, China in 2016. Disease incidences of seven alfalfa varieties planted in 2014 ranged from 56% to 95%, while incidence of Gongnong No. 1 planted in 2016 was 8%, 31% and 76% in 2016, 2017 and 2018, respectively. Paraphoma isolates were consistently recovered from black necrotic root tissues of diseased plants with a frequency of 77.1%. Based on morphological characters and phylogenetic analysis of rDNA internal transcribed spacer (ITS), elongation factor 1-α (EF1-α) and β-tubulin (TUB), this fungus was identified as Paraphoma radicina. Glasshouse pathogenicity experiments showed that P. radicina significantly reduced above- and below-ground biomass of alfalfa plants 2 months after inoculation. Paraphoma radicina infected 70% of the plants inoculated with a root dip in conidia, and these symptoms were consistent with the symptoms in the field. Paraphoma radicina was successfully reisolated from disease roots of the inoculated alfalfa plants. This is the first report of P. radicina as the causal agent of alfalfa root rot in China.     

Seed treatment of peas with fosetyl-Al against Aphanomyces euteiches

In 1983, seed dressing of peas with fosetyl-Al (Aliette) compared favourably with all other treatments on a field heavily contaminated with foot and root rot pathogens. Experiments carried out in 1983 and 1984 could not establish the reason for this phenomenon. Artificial inoculation with the most frequently isolated pathogens (Fusarium oxysporum, Phoma medicaginis var.pinodella, Pythium ultimum andFusarium solani f.sp.pisi) of sterilized soil before planting treated seeds did not reproduce the field observations, but glasshouse experiments using soil from the experimental field did. Glasshouse experiments in 1989, afterAphanomyces euteiches had been identified as a frequently occurring pea pathogen in the Netherlands, confirmed the favourable effect of fosetyl-Al as a seed treatment whenA. euteiches was in the pathogen flora.Samenvatting In 1983 bleek op een zwaar met voetrotpathogenen besmet veld zaaizaadbehandeling van erwten met fosetyl-Al (Aliette) gunstig af te steken bij alle andere behandelingen. De oorzaak kon toen niet worden gevonden. Kunstmatige besmetting van gesterilisserde grond vóór het planten van behandelde zaden met de meest geïsoleerde pathogenen (Fusarium oxysporum, Phoma medicaginis var.pinodella, Pythium ultimum enFusarium solani f.sp.pisi) gaf resultaten die niet overeenstemden met die van het proefveld, maar kasproeven met grond van het proefveld deden dat wel. Nadat was aangetoond dat het erwtepathogeenAphanomyces euteiches in Nederland veel voorkomt, bevestigden nieuwe kasproeven in 1989 het gunstige effect van fosetyl-Al als zaaizaadbehandeling wanneerA. euteiches deel uitmaakt van de pathogene bodemflora.     

Differential Control of Head Blight Pathogens of Wheat by Fungicides and Consequences for Mycotoxin Contamination of Grain

Fusarium head blight of wheat is caused by a disease complex comprised of toxigenic pathogens, predominantly Fusarium spp., and a non-toxigenic pathogen Microdochium nivale, which causes symptoms visually indistinguishable from Fusarium and is often included as a causal agent of Fusarium head blight. Four field trials are reported here, including both naturally and artificially inoculated trials in which the effect of fungicide treatments were noted on colonisation by Fusarium and Microdochium, and on the production of deoxynivalenol (DON) mycotoxin. The pathogen populations were analysed with quantitative PCR and samples were tested for the presence of the mycotoxin DON. Application of fungicides to reduce Fusarium head blight gave a differential control of these fungi. Tebuconazole selectively controlled F. culmorum and F. avenaceum and reduced levels of DON, but showed little control of M. nivale. Application of azoxystrobin, however, selectively controlled M. nivale and allowed greater colonisation by toxigenic Fusarium species. This treatment also lead to increased levels of DON detected. nobreak Azoxystrobin application two days post-inoculation increased the production of DON mycotoxin per unit of pathogen in an artificially inoculated field trial. This result indicates the potential risk of increased DON contamination of grain following treatment with azoxystrobin to control head blight in susceptible wheat cultivars. This is the first study to show differential fungicidal control of mixed natural pathogen populations and artificial inoculations in field trials.     

Genetic control of partial resistance to ‘collar’ and ‘root’ isolates of <Emphasis Type="Italic">Phoma macdonaldii</Emphasis> in sunflower

Phoma macdonaldii is one of the most important pathogens of sunflower (Heliantus annuus) in France. In order to determine the inheritance of resistance to the disease, five sunflower genotypes with wide genetic variability for resistance to two ‘collar’ and two ‘root’ Phoma isolates were crossed in a diallel programme. Four separate experiments were undertaken under controlled conditions. In each one, the response of parental genotypes and their F1 hybrids were evaluated with one of the four Phoma isolates. Analysis of variance was performed to determine the effects of genotype on disease severity score when inoculated with ‘collar’ or ‘root’ Phoma isolates and showed significant variability among parents and F1 hybrids for disease severity score. Diallel analysis showed that general combining ability (GCA) and specific combining ability (SCA) effects for resistance to ‘collar’ and ‘root’ Phoma isolates were highly significant for each of the four isolates indicating that both kinds of gene effects were important in controlling the resistance. The GCA/SCA ratios were more than one for three out of four isolates showing that additive genetic effects were more important than non-additive effects for resistance to three of the studied Phoma isolates. Hence, conventional breeding methods could be recommended to achieve genetic improvement to such ‘collar’ and ‘root’ Phoma isolates.     

Investigation into components of partial disease resistance,determined <Emphasis Type="Italic">in vitro</Emphasis>, and the concept of types of resistance to <Emphasis Type="Italic">Fusarium</Emphasis> head blight (FHB) in wheat

This work presents an analysis of the relationship between components of partial disease resistance (PDR) detected using in vitro detached leaf and seed germination assays, inoculated with Microdochium majus, and Fusarium head blight (FHB) resistance to Fusarium graminearum assessed using point inoculation, termed Type II resistance. Relationships between in vitro-determined PDR components and FHB resistance using techniques which inoculate the wheat spike uniformly, termed Type I resistance (incidence and severity), have been reported previously. In this study shorter incubation periods, longer latent periods and shorter lesion lengths in the detached leaf assay and higher germination rates in the seed germination assay were related to greater FHB resistance measured by single point inoculation (Type II), collectively explaining 54% of the variation. Overall the relationships observed for Type II FHB resistance were similar to previous findings for Type I resistances. However, the relative magnitude of effects of the individual PDR components determined in vitro varied between FHB disease resistance parameters. Resistance in seed germination and latent period in the detached leaf assay were more strongly related to resistance assessed by point inoculation (Type II) and severity-Type I as opposed to incubation period which was most strongly related to disease incidence-Type I. The results provide evidence that individual components of partial disease resistance differentially affect aspects of FHB disease progression in the wheat spike. This work supports the view that the current model of types of resistance is an oversimplification of the interacting mechanisms underlying expression of FHB resistance.     

Glomus intraradices, Pseudomonas alcaligenes, and Bacillus pumilus: effective agents for the control of root-rot disease complex of chickpea (Cicer arietinum L.)

The effects of Glomus intraradices, Pseudomonas alcaligenes and Bacillus pumilus on the root-rot disease complex caused by the root-knot nematode Meloidogyne incognita and the root-rot fungus Macrophomina phaseolina in chickpea was assessed by quantifying differences in the shoot dry mass, pod number, nodulation, and shoot content of chlorophyll, nitrogen, phosphorus and potassium. Inoculation of plants with G. intraradices, P. alcaligenes and B. pumilus alone and in combination significantly increased shoot dry mass, pod number, and content of chlorophyll, nitrogen, phosphorus and potassium in plants inoculated with pathogens over that in the uninoculated control plants. P. alcaligenes caused a greater increase in shoot dry mass, pod number, chlorophyll, nitrogen, phosphorus and potassium in plants with pathogens than did G. intraradices or B. pumilus. Combined application of G. intraradices, P. alcaligenes and B. pumilus to plants inoculated with pathogens caused a greater increase in shoot dry mass, pod number, nitrogen, phosphorus, and potassium than did an application of P. alcaligenes plus B. pumilus or of G. intraradices plus B. pumilus. Root colonization by G. intrardices was high when used alone, while inoculation with the pathogens reduced root colonization by G. intraradices. In the presence of P. alcaligenes and B. pumilus, root colonization by G. intraradices increased. In plants inoculated with just one antagonist, P. alcaligenes reduced galling and nematode multiplication the most, followed by G. intraradices, then B. pumilus. The greatest reduction in galling, nematode multiplication and root-rot was observed when both bacterial species and G. intraradices were applied together.