Mixtures of plant growth-promoting rhizobacteria for induction of systemic resistance against multiple plant diseases

Studies of induced systemic resistance using strains of plant growth-promoting rhizobacteria (PGPR) have concentrated on the use of individual PGPR as inducers against multiple diseases of a single crop. To date, few reports have examined the potential of PGPR strain mixtures to induce systemic resistance against diseases of several different plant hosts. The objective of this study was to select mixtures of compatible PGPR strains with the capacity to elicit induced systemic resistance in four hosts. The specific diseases and hosts tested in this study included: bacterial wilt of tomato (Lycopersicon esculentum) caused by Ralstonia solanacearum, anthracnose of long cayenne pepper (Capsicum annuum var. acuminatum) caused by Colletotrichum gloeosporioides, damping off of green kuang futsoi (Brassica chinensis var. parachinensis) caused by Rhizoctonia solani, and cucumber mosaic virus (CMV) on cucumber (Cucumis sativus). To examine compatibility, seven selected PGPR strains were individually tested for in vitro antibiosis against all other PGPR strains and against three of the tested pathogens (R. solanacearum, C. gloeosporioides, and R. solani). No in vitro antibiosis was observed among PGPR strains or against pathogens. Twenty-one combinations of PGPR and seven individual PGPR were tested in the greenhouse for induced resistance activity. Results indicated that four mixtures of PGPR and one individual strain treatment significantly reduced the severity of all four diseases compared to the nonbacterized control: 11 mixtures reduced CMV of cucumber, 16 mixtures reduced bacterial wilt of tomato, 18 mixtures reduced anthracnose of long cayenne pepper, and 7 mixtures reduced damping off of green kuang futsoi. Most mixtures of PGPR provided a greater disease suppression than individual PGPR strains. These results suggest that mixtures of PGPR can elicit induced systemic resistance to fungal, bacterial, and viral diseases in the four hosts tested.     

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

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

Evaluation of selected geocarposphere bacteria for biological control of Aspergillus flavus in peanut

Selected bacterial strains isolated from the region of peanut pod development (geocarposphere) and two additional bacterial strains were screened as potential biological control agents against Aspergillus flavus invasion and subsequent aflatoxin contamination of peanut in laboratory, greenhouse, and field trials. All 17 geocarposphere strains tested delayed invasion of young roots and reduced colonization by the fungus in a root-radicle assay used as a rapid laboratory prescreen. In a greenhouse study, seven bacterial strains significantly reduced pod colonization by A. flavus compared to the control. In a field trial, conducted similarly to the greenhouse assay, pods sampled at mid-peg from plants seed-treated with suspensions of either 91A-539 or 91A-550 were not colonized by A. flavus, and the incidence of pods invaded from plants treated with either 91A-539 or 91A-599 was consistently lower than nonbacterized plants at each of five sampling dates. At harvest, 8 geocarposphere bacterial strains significantly lowered the percentage of pods colonized (> 51%) compared to the control. Levels of seed colonization ranged from 1.3% to 45% and did not appear related to aflatoxin concentrations in the kernels.     

Screening rhizobacteria for biological control of Ralstonia solanacearum in Ethiopia

Bacterial wilt caused by Ralstonia solanacearum (Smith) has become a severe problem mainly on potato and tomato in Ethiopia and no effective control measure is available yet. To explore possibilities for the development of biological control for the disease, 118 rhizobacteria, most of them collected from Ethiopia, were screened against an Ethiopian R. solanacearum strain. On the basis of in vitro screening, six strains (RP87, B2G, APF1, APF2, APF3, and APF4) with good inhibitory effect were selected for in planta testing in a greenhouse. In the greenhouse, soil and tomato seedlings were treated with the antagonists and their effects studied. The study showed that APF1 and B2G strains significantly reduced disease incidence and increased weight of tomato plants. Area under disease progress curves (AUDPC) was reduced by 60% and 56% in plants inoculated with APF1 and B2G strains, respectively. Plant dry weight increase in plants inoculated with APF1 and B2G strains was 96% and 75%, respectively. APF1 was found to be the most beneficial strain in disease suppression and also growth promotion resulting in 63% dry weight increase compared to untreated control. The study revealed that APF1 and B2G strains are promising strains whose effectiveness under field conditions and their mode of action should be investigated.     

Plant growth-promoting rhizobacteria systemically protect Arabidopsis thaliana against Cucumber mosaic virus by a salicylic acid and NPR1-independent and jasmonic acid-dependent signaling pathway

Arabidopsis thaliana ecotype Columbia plants (Col-0) treated with plant growth-promoting rhizobacteria (PGPR) Serattia marcescens strain 90-166 and Bacillus pumilus strain SE34 had significantly reduced symptom severity by Cucumber mosaic virus (CMV). In some cases, CMV accumulation was also significantly reduced in systemically infected leaves. The signal transduction pathway(s) associated with induced resistance against CMV by strain 90-166 was determined using mutant strains and transgenic and mutant Arabidopsis lines. NahG plants treated with strains 90-166 and SE34 had reduced symptom severity indicating that the resistance did not require salicylic acid (SA). Strain 90-166 naturally produces SA under iron-limited conditions. Col-0 and NahG plants treated with the SA-deficient mutant, 90-166-1441, had significantly reduced CMV symptom severity with reduced virus accumulation in Col-0 plants. Another PGPR mutant, 90-166-2882, caused reduced disease severity in Col-0 and NahG plants. In a time course study, strain 90-166 reduced virus accumulation at 7 but not at 14 and 21 days post-inoculation (dpi) on the non-inoculated leaves of Col-0 plants. NahG and npr1-1 plants treated with strain 90-166 had reduced amounts of virus at 7 and 14 dpi but not at 21 dpi. In contrast, no decrease in CMV accumulation occurred in strain 90-166-treated fad3-2 fad7-2 fad8 plants. These data indicate that the protection of Arabidopsis against CMV by strain 90-166 follows a signaling pathway for virus protection that is independent of SA and NPR1, but dependent on jasmonic acid.     

Development of CMV-and TMV-resistant chili pepper: field perfermance and biosafety assessment

Both cucumber mosaic virus (CMV) and tobacco mosaic virus (TMV) coat protein (CP) genes have been transferred to chilli pepper (Capsicum annuum var. Longunt) cultivar 8212 by a modified procedure of Agrobacterium tumefaciens-mediated transformation using hypocotyl as the explant. PCR analysis revealed the presence of both CMV and TMV CP genes in at least 11 primary transformants out of 49 kanamycin-resistant chili pepper plants. Ten T1 lines from five independent transformation events were identified as putative homozygous transgenic lines based on the rooting assay of their T2 seedlings on the kanamycin-containing media. Integration and expression of CMV CP and TMV CP transgenes in one of the homozygous line, 16-13, were confirmed bySouthern blot, RT-PCR and western blot analyses. Line 16-13 was highly resistant to infection of homologous CMV and TMV strains in greenhouse conditions when successively challenged with CMV and TMV or challenged with TMV alone.Futhermore, field trials on T2, T3 and T4 progenies of Line 16-13 were performed on scales of 123, 300 and 10,000 plants, respectively, in consecutive years 1996, 1997 and 1998 with the permission of the Chinese government authority. The transgenic plants displayed delayed symptom development and significantly milder disease severity in field conditions when compared to untransformed chili pepper plants, resulting in 47 and 110% increase in pepper fruit yield in surveys conducted in 1997 and 1998 trials, respectively. Finally, quality analysis and biosafety assesment were performed on transgenic chili pepper fruit concurrently with the control fruit, and demonstrated that the transgenic chili pepper fruit is substantially equivalent to the non-transgenic pepper in terms of the quality and biosafety when consumed as a food additive.     

Real time expression of ACC oxidase and PR-protein genes mediated by Methylobacterium spp. in tomato plants challenged with Xanthomonas campestris pv. vesicatoria

Biotic stress like pathogenic infection increases ethylene biosynthesis in plants and ethylene inhibitors are known to alleviate the severity of plant disease incidence. This study aimed to reduce the bacterial spot disease incidence in tomato plants caused by Xanthomonas campestris pv. vesicatoria (XCV) by modulating stress ethylene with 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity of Methylobacterium strains. Under greenhouse condition, Methylobacterium strains inoculated and pathogen challenged tomato plants had low ethylene emission compared to pathogen infected ones. ACC accumulation and ACC oxidase (ACO) activity with ACO related gene expression increased in XCV infected tomato plants over Methylobacterium strains inoculated plants. Among the Methylobacterium spp., CBMB12 resulted lowest ACO related gene expression (1.46 Normalized Fold Expression), whereas CBMB20 had high gene expression (3.42 Normalized Fold Expression) in pathogen challenged tomato. But a significant increase in ACO gene expression (7.09 Normalized Fold Expression) was observed in the bacterial pathogen infected plants. In contrast, Methylobacterium strains enhanced β-1,3-glucanase and phenylalanine ammonia-lyase (PAL) enzyme activities in pathogen challenged tomato plants. The respective increase in β-1,3-glucanase related gene expressions due to CBMB12, CBMB15, and CBMB20 strains were 66.3, 25.5 and 10.4% higher over pathogen infected plants. Similarly, PAL gene expression was high with 0.67 and 0.30 Normalized Fold Expression, in pathogen challenged tomato plants inoculated with CBMB12 and CBMB15 strains. The results suggest that ethylene is a crucial factor in bacterial spot disease incidence and that methylobacteria with ACC deaminase activity can reduce the disease severity with ultimate pathogenesis-related protein increase in tomato.     

Use of Pseudomonas fluorescens-based formulations for management of tomato spotted wilt virus (TSWV) and enhanced yield in tomato

Strains of Pseudomonas fluorescens were investigated for biocontrol efficacy against tomato spotted wilt virus (TSWV) in tomato both alone and in mixtures. P. fluorescens strains applied to seed, soil and foliage or as a seedling dip significantly reduced TSWV, with a concomitant increase in growth promotion in both the glasshouse and field. Two native strains (CoP-1 and CoT-1) and one foreign strain (CHAO) reduced TSWV. In P. fluorescens-treated tomato plants, increased activity of polyphenol oxidase, β-1,3-glucanase and chitinase was observed, and induction of chitinase was confirmed by western blot analysis. Induction of new protein (18 kDa) detected by SDS-PAGE in P. fluorescens-treated tomato plants was not found in healthy and P. fluorescens-untreated virus inoculated control plants. Indirect ELISA clearly showed a reduction in viral antigen concentration in P. fluorescens-treated tomato plants corresponding to reduced disease ratings. All the P. fluorescens-treated tomato plants also showed enhanced growth and yield compared to control plants. Hence, plant growth promoting rhizobacteria (PGPR) could play a major role in reducing TSWV and increasing yield in tomato plants.     

Induction of defense responses in tomato against Pseudomonas syringae pv. tomato by regulating the stress ethylene level with Methylobacterium oryzae CBMB20 containing 1-aminocyclopropane-1-carboxylate deaminase

This study aimed to examine the induction of defense responses in tomato elicited by Methylobacterium oryzae CBMB20 as a consequence of reduced stress ethylene level possibly through its ACC deaminase activity. Significantly increased activities of pathogenesis-related (PR) proteins and defense enzymes such as β-1,3-glucanase, phenylalanine ammonia-lyase, peroxidase and polyphenol oxidase were noted in M. oryzae CBMB20 pretreated and challenged with Pseudomonas syringae pv. tomato (Pst) compared to either control or M. oryzae-treated tomato plants in both growth chamber and greenhouse conditions. Increased PR proteins and defense enzyme activities were correlated with the reduction of stress ethylene level. M. oryzae CBMB20 reduced the stress ethylene level about 27% and 55% when challenged with Pst, in growth chamber and greenhouse on day 7 respectively and the effect was comparable to that of the chemical ethylene biosynthesis inhibitor AVG, L-α-(2-aminoethoxyvinyl)-glycine hydrochloride. As a consequence of reduced stress ethylene level and its effect on defense response in crop plants, the disease severity was reduced 26% in M. oryzae CBMB20-treated plants challenged with pathogen. Therefore, inoculation of M. oryzae CBMB20 would induce the defense enzymes and contribute to the enhanced resistance of tomato plants against the pathogen Pst.     

Induced systemic resistance to tomato leaf curl virus and increased yield in tomato by plant growth promoting rhizobacteria under field conditions

The talc-based formulation of two Pseudomonas fluorescens strains (Pf1 and VPT10) and its mixture (with and without chitin) were tested against tomato leaf curl virus in tomato under greenhouse and field conditions. The mean percentage of tomato leaf curl virus infected plants were significantly lower (25%) with less symptom severity and delayed symptom expression up to nine additional days in Pseudomonas with chitin (VPT10 + chitin) treated tomato plants compared to non-bacterised control plants upon challenge inoculation with tomato leaf curl virus. Tomato leaf curl virus was partially purified and antiserum was developed. Using the antiserum the tomato leaf curl virus was detected in symptomatic leaves and in whitefly vector through direct antigen coating enzyme linked immunosorbent assay which revealed the low virus titre in Pseudomonas treated plants (VPT10 + chitin) and insect vector compared to untreated tomato plants. The results indicate the potentiality of plant growth promoting rhizobacteria strains and talc-powder formulations in the effective management of this tomato leaf curl virus in tomato under field conditions.     

Influence of arbuscular-mycorrhizal fungi, Rhizobium meliloti strains and PGPR inoculation on the growth of Medicago arborea used as model legume for re-vegetation and biological reactivation in a semi-arid mediterranean area

Medicago arborea can be used for re-vegetationpurposes under semiarid conditions. These woody legumes have the ability toforman association with arbuscular mycorrhizal (AM) fungi and rhizobial bacteria,which can be maximised by microorganisms producing certain stimulatingmetabolites acting as plant growth promoting rhizobacteria (PGPR). The effectsof single and combined inoculations using microorganisms with different andinteractive metabolic capacities, namely three Glomusspecies, two Rhizobium meliloti strains (a wild type, WTand its genetically modified derivative GM) and a plant growth promotingrhizobacterium, (PGPR), were evaluated. All three inoculated AM fungi affectedMedicago growth in different ways. Differences weremaintained when soil was co-inoculated with each of the rhizobial strains (WTorGM) and the PGPR. Mycorrhizal fungi were effective in all cases, but the PGPRonly affected plant growth specific microbial situations. PGPR increased growthof G. mosseae-colonised plants associated withRhizobium WT strain by 36% and those infected byG. deserticola when associated with the rhizobial GMstrainby 40%. The most efficient microbial treatments involved mycorrhizalinoculation, which was an indication of the AM dependency of this plantspecies.Moreover, PGPR inoculation was only effective when associated with specificmycorrhizal endophytes (G. mosseae plus WT andG.deserticola plus GM rhizobial strain). The reduced root/shoot (R/S)ratio resulting from PGPR inoculation, was an indication of more effective rootfunction in treated plants. AM colonisation and nodule formation wereunaffectedby the type of AM fungus or bacteria (rhizobial strain and/or PGPR). AM fromnatural soil were less infective and effective than those from the collection.The results supported the existence of selective microbial interactionsaffecting plant performance. The indigenous AM fungi appeared to be ineffectiveand M. arborea behaved as though it was highly dependentonAM colonisation, which implied that it must have a mycorrhizal association toreach maximum growth in the stressed conditions tested. Optimum growth ofmycorrhizal M. arborea plants was associated with specificmicrobial groups, accounting for a 355% increase in growth overnodulatedcontrol plants. The beneficial effect of PGPR in increasing the growth of awoody legume, such as M. arborea under stress, was onlyobserved with co-inoculation of specific AM endophytes. As a result of theinteraction, only shoot biomass was enhanced, but not as a consequence ofenhancing of the colonising abilities of the endophytes. The growthstimulation,occurring as a consequence of selected microbial groups, may be critical anddecisive for the successful establishment of plants under Mediterraneanclimaticand soil conditions.     

Induction of systemic resistance in tobacco against Tobacco mosaic virus by Bacillus spp.

Bacillus pumilus strain EN16 and Bacillus subtilis strain SW1 were tested for their systemic resistance and protection abilities against tobacco mosaic virus disease under greenhouse conditions. The results showed that strain EN16 and SW1 treatment significantly reduced mosaic symptoms and disease severity, resulting in 52 and 71% protection at 14 days of inoculation, respectively. A decreased amount of virus was detected in EN16- or SW1-treated tobacco plants by ELISA. Moreover, 5- and 7-day intervals between inducer treatment and pathogen inoculations were respectively required for strain EN16 and SW1 to induce optimal resistance. Further analysis on phenylalanine ammonia-lyase, peroxidase, polyphenol oxidase and pathogenesis-related (PR) proteins in tobacco showed that the amounts of defense enzymes and PR proteins significantly increased in Bacillus-treated plants challenged with pathogen when compared to control.     

Potential of biotic inducers on disease severity and variation of Cucumber mosaic cucumovirus in cucumber plants

Six bacterial strains (Bacillus subtilis, Bacillus polymyxa, Bacillus circulans, Pseudomonas putida, Pseudomonas fluorescens 2 and Pseudomonas fluorescens 8) and one fungal isolate (Trichoderma harzianum) were tested for their ability to protect Cucumis sativus L. cv. Beith Alpha against the disease development of Cucumber mosaic cucumovirus (CMV). Seed treatment with individual bacterial and fungal liquid cultures significantly and consistently reduced the disease severity (DS) of infected cucumber plants, after 14 days from CMV inoculation onto cotyledons. All seven biotic inducers reduced the CMV infection at the range of 16.6–39% and 0–46.5% under sterilised and non-sterilised soils, respectively. The effect of treatment with each biotic inducer has a significant difference in the percentage of CMV DS. In sterilised soil, B. circulans has a low percentage of DS (42%), while P. fluorescens 8 has a high DS percentage (62.9%). In non-sterilised soil, the low DS percentage was 30% for T. harzianum, while P. putida had the highest DS percentage (70%). On the other hand, CMV variability on Chenopodium amaranticolor revealed that all biotic treatments differed according to the local lesion number, similarity and morphology.     

Effects of Inoculation with a Mild Strain of Tomato Aspermy Virus on the Growth and Yield of Tomatoes and the Potential for Cross Protection

Mechanical inoculation of seedlings of 26 tomato cultivars with an attenuated strain of tomato aspermy virus (TAV-M, ATCC # PV361) resulted in development of only mild symptoms, which had largely disappeared by the time the infected plants had begun to set fruit. Yield trials revealed that plants inoculated with TAV-M when transplanted at the 6–8 leaf stage displayed no delay in fruit set and had yields only slightly less than uninoculated plants. Cross protection studies carried out in the greenhouse revealed that TAV-M has the potential to interfere with infection of tomato seedlings by a more severe TAV strain (ATCC # PV127). ‘Pik-Red’ seedlings inoculated with TAV-M showed a significant reduction in symptom expression when challenged with the Blencowe strain of TAV 21 days after the initial inoculation with TAV-M; interference was incomplete at 7 and 14 days.     

Effects of inoculation with plant growth promoting rhizobacteria (PGPRs) andSinorhizobium fredii on biological nitrogen fixation, nodulation and growth ofGlycine max cv. Osumi

We investigated the effects of three plant growth promoting rhizobacteria (PGPR), on Biological Nitrogen Fixation (BNF), nodulation and growth promotion by soybean (Glycine max) var. Osumi plants. The strains, Aur 6, Aur 9 and Cell 4, belong toPsedomonas fluorescens, Chryseobacterium balustinum andSerratia fonticola, respectively. Inoculation modes for the PGPRs andSinorhizobium fredii (carried out through irrigation), were examined. In the first mode, PGPRs andS. fredii were co-inoculated. In the second mode, we first inoculatedS. fredii and after the PGPRs, which were added 5 or 10 days later (each inoculation being an independent treatment). In the third mode, the PGPRs were inoculated first, and theS. fredii was inoculated 5 days later. We also included treatments inoculated with only the PGPRs (one PGPR per treatment) and only withS. fredii. Plants were maintained in a greenhouse under controlled environmental conditions, and were sampled 3 months after sowing. The results obtained showed the effects of the inoculation sequence. The most significant effects on growth parameters (stem plus leaf weight and fresh root weight) were found when inoculations with PGPR andS. fredii were at different times or when we inoculated only with PGPR and the plants were watered with nitrogen. Co-inoculation had no positive effects on any parameter, probably due to competition between the PGPR andS. fredii. Our results indicate that the inoculation modes with PGPR and rhizobia play a very important role in the effects produced. Thus, although plant growth promoting rhizobacteria may interact synergistically with root-nodulating rhizobia, plant growth promoting rhizobacteria selected for one crop should be assessed for potentially hazardous effects on other crops before being used as inoculants.     

Macromolecules Released by a Plant Growth-promoting Rhizobacterium as Elicitors of Systemic Resistance in Tomato to Bacterial and Fungal Pathogens

One of 500 rhizobacteria isolated from soil, rhizosphere and rhizoplane of healthy tomato plants was previously selected in laboratory, greenhouse and field tests as a good inducer of systemic resistance. This plant growth‐promoting rhizobacterium (PGPR) was identified as Bacillus cereus by fatty‐acid analysis. Bacillus cereus bacterial cells were removed from liquid culture by centrifugation and the supernatant repeatedly dialyzed (cut‐off = 12 000 daltons) against distilled water. Dialysates applied to roots protected tomato plants against leaf fungal and bacterial pathogens, evidence that macromolecules synthesized by the PGPR and released into the environment act as elicitors of systemic resistance.     

Interaction of Glomus mosseae and Paecilomyces lilacinus on Meloidogyne javanica of tomato

The effects of Glomus mosseae and Paecilomyces lilacinus on Meloidogyne javanica of tomato were tested in a greenhouse experiment. Chicken layer manure was used as a carrier substrate for the inoculum of P. lilacinus. The following parameters were used: gall index, average number of galls per root system, plant height, shoot and root weights. Inoculation of tomato plants with G. mosseae did not markedly increase the growth of infected plants with M. javanica. Inoculation of plants with G. mosseae and P. lilacinus together or separately resulted in similar shoots and plant heights. The highest root development was achieved when mycorrhizal plants were inoculated with P. lilacinus to control root-knot nematode. Inoculation of tomato plants with G. mosseae suppressed gall index and the average number of galls per root system by 52% and 66%, respectively, compared with seedlings inoculated with M. javanica alone. Biological control with both G. mosseae and P. lilacinus together or separately in the presence of layer manure completely inhibited root infection with M. javanica. Mycorrhizal colonization was not affected by the layer manure treatment or by root inoculation with P. lilacinus. Addition of layer manure had a beneficial effect on plant growth and reduced M. javanica infection.     

Impact of PGPR inoculation on growth and antioxidant status of wheat under saline conditions

Two plant growth‐promoting rhizobacterial (PGPR) strains, Bacillus subtilis SU47 and Arthrobacter sp. SU18, were found to tolerate 8% NaCl. Wheat co‐inoculated with these two PGPR strains, and grown under different salinity regimes (2–6 dS m^?1), showed an increase in dry biomass, total soluble sugars and proline content. Wheat sodium content was reduced under co‐inoculated conditions but not after single inoculation with either strain or in the control. The activity of antioxidant enzymes in wheat leaves decreased under salinity stress after PGPR co‐inoculation, suggesting these PGPR species could be used for amelioration of stress in wheat plants. Activity of three antioxidant enzymes in wheat grown with both PGPR strains was reduced, most notably that of catalase activity at a salinity of 6 dS m^?1, when compared with the control. The results indicate that co‐inoculation with B. subtilis and Arthrobacter sp. could alleviate the adverse effects of soil salinity on wheat growth.