Herbicide-tolerant endophytic bacteria of rice plants as the biopriming agents for fertility recovery and disease suppression of unhealthy rice seeds.

BACKGROUND: Dirty panicle disease (DPD) caused by several fungal phytopathogens results in damage and depreciation of rice seeds. Unhealthy rice seeds with DPD are potent reservoirs of pathogens and unable to be used as seed stock as they can spread the disease in the paddy fields leading to the severe loss of rice yield and quality. In this study, we aim to search for beneficial endophytes of commercially cultivated rice plants and utilize them as biostimulants in seed biopriming for fertility recovery and disease suppression of unhealthy rice seeds. RESULTS: Forty-three bacterial endophytes were isolated from rice plants grown in the herbicide-treated paddy fields. Five isolates of these endophytes belonging to the genus Bacillus show excellent antifungal activity against fungal pathogens of DPD. Based on germination tests, biopriming unhealthy rice seeds by soaking in bacterial suspensions for 9 or 12 h was optimal as evidenced by the lowest disease incidence and longer shoot and root lengths of seedlings germinated, compared with controls made of non-treated or hydroprimed healthy and unhealthy seeds. Pot experiments were carried out to evaluate the impact of seed biopriming, in which the percentage of healthy rice yield produced by rice plants emerging from bioprimed seeds was not significantly different, compared to the controls originating respectively from non-treated healthy seeds and chemical fungicide-treated unhealthy seeds. CONCLUSION: Biopriming of unhealthy rice seeds with herbicide-tolerant endophytic bacteria could recover seed fertility and protect the full life cycle of emerging rice plants from fungal pests. With our findings, seed biopriming is a straightforward approach that farmers can apply to recover unhealthy rice seed stock, which enables them to reduce the cost and use of agrochemicals in the commercial production of rice and to promote green technology in sustainable agriculture.

Bio-Priming with Bacillus Isolates Suppresses Seed Infection and Improves the Germination of Garden Peas in the Presence of Fusarium Strains.

Seed infection caused by Fusarium spp. is one of the major threats to the seed quality and yield of agricultural crops, including garden peas. The use of Bacillus spp. with multiple antagonistic and plant growth-promoting (PGP) abilities represents a potential disease control strategy. This study was performed to evaluate the biocontrol potential of new Bacillus spp. rhizosphere isolates against two Fusarium strains affecting garden peas. Six Bacillus isolates identified by 16S rDNA sequencing as B. velezensis (B42), B. subtilis (B43), B. mojavensis (B44, B46), B. amyloliquefaciens (B50), and B. halotolerans (B66) showed the highest in vitro inhibition of F. proliferatum PS1 and F. equiseti PS18 growth (over 40%). The selected Bacillus isolates possessed biosynthetic genes for endoglucanase (B42, B43, B50), surfactin (B43, B44, B46), fengycin (B44, B46), bacillomycin D (B42, B50), and iturin (B42), and were able to produce indole-3-acetic acid (IAA), siderophores, and cellulase. Two isolates, B. subtilis B43 and B. amyloliquefaciens B50, had the highest effect on final germination, shoot length, root length, shoot dry weight, root dry weight, and seedling vigor index of garden peas as compared to the control. Their individual or combined application reduced seed infection and increased seed germination in the presence of F. proliferatum PS1 and F. equiseti PS18, both after seed inoculation and seed bio-priming. The most promising results were obtained in the cases of the bacterial consortium, seed bio-priming, and the more pathogenic strain PS18. The novel Bacillus isolates may be potential biocontrol agents intended for the management of Fusarium seed-borne diseases.

Seed Bio-priming of wheat with a novel bacterial strain to modulate drought stress in Daegu, South Korea.

Wheat is one of the major cereal crop grown food worldwide and, therefore, plays has a key role in alleviating the global hunger crisis. The effects of drought stress can reduces crop yields by up to 50% globally. The use of drought-tolerant bacteria for biopriming can improve crop yields by countering the negative effects of drought stress on crop plants. Seed biopriming can reinforce the cellular defense responses to stresses via the stress memory mechanism, that its activates the antioxidant system and induces phytohormone production. In the present study, bacterial strains were isolated from rhizospheric soil taken from around the Artemisia plant at Pohang Beach, located near Daegu, in the South Korea Republic of Korea. Seventy-three isolates were screened for their growth-promoting attributes and biochemical characteristics. Among them, the bacterial strain SH-8 was selected preferred based on its plant growth-promoting bacterial traits, which are as follows: abscisic acid (ABA) concentration = 1.08 ± 0.05 ng/mL, phosphate-solubilizing index = 4.14 ± 0.30, and sucrose production = 0.61 ± 0.13 mg/mL. The novel strain SH-8 demonstrated high tolerance oxidative stress. The antioxidant analysis also showed that SH-8 contained significantly higher levels of catalase (CAT), superoxide dismutase (SOD), and ascorbic peroxidase (APX). The present study also quantified and determined the effects of biopriming wheat (Triticum aestivum) seeds with the novel strain SH-8. SH-8 was highly effective in enhancing the drought tolerance of bioprimed seeds; their drought tolerance and germination potential (GP) were increased by up to 20% and 60%, respectively, compared with those in the control group. The lowest level of impact caused by drought stress and the highest germination potential, seed vigor index (SVI), and germination energy (GE) (90%, 2160, and 80%, respectively), were recorded for seeds bioprimed with with SH-8. These results show that SH-8 enhances drought stress tolerance by up to 20%. Our study suggests that the novel rhizospheric bacterium SH-8 (gene accession number OM535901) is a valuable biostimulant that improves drought stress tolerance in wheat plants and has the potential to be used as a biofertilizer under drought conditions.

Seed bio-priming enhanced salt stress tolerance of maize (Zea mays L.) seedlings by regulating the antioxidant system and miRNA expression.

Maize (Zea mays) is moderately sensitive to salt stress. Therefore, increasing salinity in soil causes the arrestment of physiological processes and retention of growth and development, consequently leading to yield loss. Although many strategies have been launched to improve salt stress tolerance, plant growth-promoting rhizobacteria (PGPR) are considered the most promising approach due to being more environmentally friendly and agronomically sustainable than chemicals. Therefore, this study aims to investigate the potential of Bacillus spp. and the role of microRNA-mediated genetic regulation in maize subjected to seed bio-priming application to mitigate salt stress effects. To this end, maize seeds were bio-primed with the vegetative form of B. pumilus, B. licheniformis, and B. coagulans both individually or combined, subsequently treated to NaCl, and the seedlings were screened morphologically, physiologically, and transcriptionally. The study revealed that seed bio-priming with B. licheniformis reduced the stress effects of maize seedlings by increasing catalase (CAT) and ascorbate peroxidase (APX) activities by 2.5- and 3-fold, respectively, tolerating the decrease in chlorophyll content (CC), upregulating miR160d expression which led to a 36% increase in root fresh weight (RFW) and a 39% increase in shoot fresh weight (SFW). In conclusion, Bacillus spp. successfully alleviated salt stress effects on maize by modulating antioxidant enzymes and miRNA expression.

Efficacy of Seed-Biopriming with Trichoderma spp. and Foliar Spraying of ZnO-Nanoparticles Induce Cherry Tomato Growth and Resistance to Fusarium Wilt Disease.

Several microbes that cause plant diseases drastically lower the production of agriculture and jeopardize the safety of the world's food supply. As a result, sustainable agriculture requires disease management tactics based on modern, eco-friendly techniques as alternatives to various agrochemicals. The current study aimed to assess the antifungal activity of ZnO-nanoparticles against Fusarium solani in-vitro, and the ability of two antagonistic Trichoderma isolates, Trichoderma viride and Trichoderma harzianum, to produce antifungal secondary metabolites and identify them using gas chromatography-mass spectrometry, and to evaluate the combined effects of foliar spray of ZnO-nanoparticles and bioprimed seeds of cherry tomato (Solanum lycopersicum L.) with two antagonistic Trichoderma isolates against Fusarium wilt disease caused by Fusarium solani in greenhouse conditions. The results revealed that, in-vitro, the highest concentration of ZnO nanoparticles (3000 ppm) resulted in the greatest decrease in Fusarium solani mycelial growth (90.91% inhibition). The scanning electron microscopy demonstrated the evident distortion in Fusarium solani growing mycelia treated with ZnO-nanoparticles, which might be the source of growth suppression. Additionally, twenty-eight bioactive chemical compounds were isolated and identified from Trichoderma spp. ethyl acetate crude extracts using gas chromatography-mass spectrometry. In a greenhouse experiment, the combination of bioprimed cherry tomato plants with Trichoderma harzianum and foliar spraying of ZnO-nanoparticles at 3000 ppm was the most effective interaction treatment for reducing disease severity index (23.4%) and improving the vegetative growth parameters, micronutrient contents (Mn, Zn, and Fe in leaves), and chlorophyll content (SPAD unit), as well as stimulating phenylalanine ammonia-lyase activity of cherry tomato leaves at 75 days after sowing. In conclusion, the antifungal potential of seed-biopriming with antagonistic Trichoderma isolates and the foliar spraying of ZnO-nanoparticles can boost cherry tomato growth and confer resistance to Fusarium wilt caused by Fusarium solani.

The Effects of Seed Pretreatment with Endophytic Bacteria Bacillus subtilis on the Water Balance of Spring and Winter Wheat Seedlings under Short-Time Water Deficit.

We investigated the effect of pre-sowing seed treatment with endophytic Bacillus subtilis 10-4 (B. subtilis) on spring and winter wheat (Triticum aestivum L.; cultivars Ekada-70 (Ek) and Scepter (Sc), respectively) growth and tolerance under 1-24 h of drought stress, modulated by 12% polyethylene glycol 6000 (PEG). The results showed that drought decreased transpiration intensity (TI), root relative water content (RWC), osmotic potential (Ψπ) of cell sap, and induced proline accumulation and electrolyte leakage (EL) in both wheat cultivars. It was revealed that Sc was more responsive to PEG and B. subtilis treatments than Ek. Under drought, Ek did not significantly change root length, shoot height, or dry biomass. The pretreatment of wheat plants with B. subtilis performed significantly better under drought conditions through the enhanced TI, RWC, and Ψπ of the cell sap in comparison with the plants treated with 12% PEG alone. B. subtilis also reduced stress-caused EL, especially in the Sc cultivar. Under water deficit wheat seedlings, pretreated with B. subtilis, have a higher proline accumulation in comparison to untreated stressed plants. Taken together, our results demonstrate the crucial role of endophytic B. subtilis in ameliorating the adverse effects of water stress on the water balance of both winter and spring wheat cultivars.

Compatible Consortium of Endophytic Bacillus halotolerans Strains Cal.l.30 and Cal.f.4 Promotes Plant Growth and Induces Systemic Resistance against Botrytis cinerea.

Evaluating microbial-based alternatives to conventional fungicides and biofertilizers enables us to gain a deeper understanding of the biocontrol and plant growth-promoting activities. Two genetically distinct Bacillus halotolerans strains (Cal.l.30, Cal.f.4) were evaluated for the levels of their compatibility. They were applied individually or in combination under in vitro and greenhouse conditions, using seed bio-priming and soil drenching as inoculum delivery systems, for their plant growth-promoting effect. Our data indicate that application of Cal.l.30 and Cal.f.4 as single strains and as a mixture significantly enhanced growth parameters of Arabidopsis and tomato plants. We investigated whether seed and an additional soil treatment with these strains could induce the expression of defense-related genes in leaves of young tomato seedling plants. These treatments mediated a long lasting, bacterial-mediated, systemic-induced resistance as evidenced by the high levels of expression of RP3, ACO1 and ERF1 genes in the leaves of young tomato seedlings. Furthermore, we presented data showing that seed and soil treatment with B. halotolerans strains resulted in an effective inhibition of Botrytis cinerea attack and development on tomato leaves. Our findings highlighted the potential of B. halotolerans strains as they combine both direct antifungal activity against plant pathogens and the ability to prime plant innate immunity and enhance plant growth.

Biopriming of Maize Seeds with a Novel Bacterial Strain SH-6 to Enhance Drought Tolerance in South Korea.

Maize is the third most common cereal crop worldwide, after rice and wheat, and plays a vital role in preventing global hunger crises. Approximately 50% of global crop yields are reduced by drought stress. Bacteria as biostimulants for biopriming can improve yield and enhance sustainable food production. Further, seed biopriming stimulates plant defense mechanisms. In this study, we isolated bacteria from the rhizosphere of Artemisia plants from Pohang beach, Daegu, South Korea. Twenty-three isolates were isolated and screened for growth promoting potential. Among them, bacterial isolate SH-6 was selected based on maximum induced tolerance to polyethylene glycol-simulated drought. SH-6 showed ABA concentration = 1.06 ± 0.04 ng/mL, phosphate solubilizing index = 3.7, and sucrose concentration = 0.51 ± 0.13 mg/mL. The novel isolate SH-6 markedly enhanced maize seedling tolerance to oxidative stress owing to the presence of superoxide dismutase, catalase, and ascorbate peroxidase activities in the culture media. Additionally, we quantified and standardized the biopriming effect of SH-6 on maize seeds. SH-6 significantly increased maize seedling drought tolerance by up to 20%, resulting in 80% germination potential. We concluded that the novel bacterium isolate SH-6 (gene accession number (OM757882) is a biostimulant that can improve germination performance under drought stress.

Modulation of plant growth and antioxidative defense system through endophyte biopriming in cotton (Gossypium spp.) and non-host crops.

Seed biopriming is very promising in improving seed health by mitigating various biotic and abiotic stresses. In this study, the effect of biopriming with cotton endophytes on seed germination and other growth parameters in host and non-host crops like wheat, sorghum, cowpea and chick pea was examined. The endophytes were antagonistic to cotton pathogens Corynespora cassiicola and Fusarium solani under in vitro. Among the eight endophytes, CFR-1 and CEL-48 were highly efficient with inhibition rates of 66.16% and 64.24% respectively against C. cassiicola, whereas CFL-34 was efficient against F. solani with more than 50% inhibition. Seed biopriming enhanced seed germination in cotton and non-host crops whereas seed vigor index was highest in bio-primed cotton. Moreover, growth promotion parameters were also enhanced upon endophyte biopriming. Total sugar content ranged from 5.46 to 7.54 mg/g F.W in cotton and highest was found in CFL-34 treated wheat (8.64 mg/g FW). There was an increase of 10-30% soluble protein in bioprimed cotton over control. Interestingly, the antioxidant potential in all the bio-primed crops was improved with increased catalase and peroxidase activity. Specific activity of catalase ranged from 0.42 to 1.90 µmol/min/mg protein in cotton, while highest activity was reported in CEL-48 primed wheat. The findings of this investigation emphasizes seed biopriming with endophytes for sustainable plant health management.

Characterization and selection of endophytic actinobacteria for growth and disease management of Tea (Camellia sinensis L.).

Endophytic microbes are vital for nutrient solubilization and uptake, growth, and survival of plants. Here, 88 endophytic actinobacteria (EnA) associated with five tea clones were isolated, assessed for their diversity, plant growth promoting (PGP), and biocontrol traits, and then used as an inoculant for PGP and disease control in host and non-host plants. Polyphasic methods, including phenotypic and genotypic characteristics led to their identification as Streptomyces, Microbacterium, Curtobacterium, Janibacter, Rhodococcus, Nocardia, Gordonia, Nocardiopsis, and Kribbella. Out of 88 isolates, 35 (39.77%) showed antagonistic activity in vitro against major fungal pathogens, viz. Fusarium oxysporum, Rhizoctonia solani, Exobasidium vexans, Poria hypobrunnea, Phellinus lamaensis, and Nigrospora sphaerica. Regarding PGP activities, the percentage of isolates that produced indole acetic acid, siderophore, and ammonia, as well as P-solubilisation and nitrogen fixation, were 67.05, 75, 80.68, 27.27, 57.95, respectively. A total of 51 and 42 isolates showed chitinase and 1-aminocyclopropane-1-carboxylic acid deaminase activity, respectively. Further, two potent Streptomyces strains KA12 and MA34, selected based on the bonitur scale, were screened for biofilm formation ability and tested in vivo under nursery conditions. Confocal laser scanning microscopy and the crystal violet staining technique revealed that these Streptomyces strains can form biofilms, indicating the potential for plant colonization. In the nursery experiment, they significantly enhanced the shoot and root biomass, shoot and root length, and leaf number in host tea plants. Additionally, treatment of tomato seeds by KA12 suppressed the growth of fungal pathogen Fusarium oxysporum, increased seed germination, and improved root architecture, demonstrating its ability to be used as a seed biopriming agent. Our results confirm the potential of tea endophytic actinobacterial strains with multifarious beneficial traits to enhance plant growth and suppress fungal pathogens, which may be used as bioinoculant for sustainable agriculture.

Biopriming of Durum Wheat Seeds with Endophytic Diazotrophic Bacteria Enhances Tolerance to Fusarium Head Blight and Salinity.

There is growing interest in the use of bio inoculants based on plant growth-promoting bacteria (PGPB) to promote plant growth under biotic and abiotic stresses. Here, we provided a detailed account of the effectiveness of a number of endophytic PGPB strains, isolated from the roots of the halophyte Salicornia brachiata in promoting durum wheat growth and enhancing its tolerance to salinity and fusarium head blight (FHB) disease. Bacillus spp. strains MA9, MA14, MA17, and MA19 were found to have PGPB characteristics as they produced indole-3-acetic acid, siderophores, and lytic enzymes, fixed free atmospheric nitrogen, and solubilized inorganic phosphate in vitro. Additionally, the in vivo study that involved in planta inoculation assays under control and stress conditions indicated that all PGPB strains significantly (p < 0.05) increased the total plant length, dry weight, root area, seed weight, and nitrogen, protein, and mineral contents. Particularly, the MA17 strain showed a superior performance since it was the most efficient in reducing disease incidence in wheat explants by 64.5%, in addition to having the strongest plant growth promotion activity under salt stress. Both in vitro and in vivo assays showed that MA9, MA14, MA17, and MA19 strains were able to play significant PGPB roles. However, biopriming with Bacillus subtilis MA17 offered the highest plant growth promotion and salinity tolerance, and bioprotection against FHB. Hence, it would be worth testing the MA17 strain under field conditions as a step towards its commercial production. Moreover, the strain could be further assessed for its plausible role in bioprotection and growth promotion in other crop plants. Thus, it was believed that the strain has the potential to significantly contribute to wheat production in arid and semi-arid regions, especially the salt-affected Middle Eastern Region, in addition to its potential role in improving wheat production under biotic and abiotic stresses in other parts of the world.

Endophytic Bacillus subtilis antagonize soil-borne fungal pathogens and suppress wilt complex disease in chickpea plants (Cicer arietinum L.).

The present study aimed to identify potential endophytic bacteria antagonistic against three soil-borne fungal pathogens, Rhizoctonia solani, Sclerotium rolfsii, and Fusarium oxysporum f.sp. ciceri causing root rot, collar rot, and fungal wilt diseases in chickpea plants, respectively. A total of 255 bacterial endophytes were isolated from the leaves, stems, and roots of seven different crop plants (chickpea, tomato, wheat, berseem, mustard, potato, and green pea). The dual culture-based screening for antifungal properties indicated that three endophytic isolates had strong inhibition (>50%) against all three pathogens tested. Based on morphological, biochemical, and molecular characterization, the selected isolates (TRO4, CLO5, and PLO3) were identified as different strains of Bacillus subtilis. The bacterial endophytes (TRO4 and CLO5) were positive for plant growth promoting (PGP) traits viz., ammonia, siderophore, and indole-3-acetic acid (IAA) production. The bio-efficacy of the endophytes (TRO4, CLO5, and PLO3) was tested by an in planta trial in chickpea pre-challenged with R. solani, S. rolfsii, and F. oxysporum f.sp. ciceri. The B. subtilis strains TRO4 and CLO5 were found to be effective in reducing percent disease incidence (p ≤ 0.05) and enhancing plant growth parameters. The different root parameters viz. root length (mm), surface area (cm2), root diameter (mm), and root volume (cm3) were significantly (p ≤ 0.05) increased in TRO4 and CLO5 inoculated chickpea plants. Confocal Scanning Laser Microscopy showed heavy colonization of bacteria in the roots of endophyte-inoculated chickpea plants. The inoculation of endophytic Bacillus subtilis strains TRO4 and CLO5 in chickpea plants through seed biopriming reduced the accumulation of superoxide, enhanced the plant defense enzymes, and induced the expression of Pathogenesis-Related (PR) genes. Semi-quantitative analysis of defense-related genes showed differential activation of PR genes (60srp and IFR) by endophyte inoculation. The results of the present study reveal the antagonistic potential of B. subtilis strains TRO4 and CLO5 against three major soil-borne fungal pathogens and their ability to suppress wilt complex disease in chickpea plants. This is the first report on the simultaneous suppression of three major soil-borne fungal pathogens causing wilt complex in chickpea plants by endophytic B. subtilis strains.

Plant Growth-Promoting Bacterial Consortia as a Strategy to Alleviate Drought Stress in Spinacia oleracea.

Drought stress is one of the most severe abiotic stresses affecting soil fertility and plant health, and due to climate change, it is destined to increase even further, becoming a serious threat to crop production. An efficient, eco-friendly alternative is the use of plant growth-promoting bacteria (PGPB), which can promote plant fitness through direct and indirect approaches, protecting plants from biotic and abiotic stresses. The present study aims to identify bacterial consortia to promote Spinacia oleracea L. cv Matador's seed germination and protect its seedlings from drought stress. Eight PGPB strains belonging to the Bacillus, Azotobacter, and Pseudomonas genera, previously characterized in physiological conditions, were analyzed under water-shortage conditions, and a germination bioassay was carried out by biopriming S. oleracea seeds with either individual strains or consortia. The consortia of B. amyloliquefaciens RHF6, B. amyloliquefaciens LMG9814, and B. sp. AGS84 displayed the capacity to positively affect seed germination and seedlings' radical development in both standard and drought conditions, ameliorating the plants' growth rate compared to the untreated ones. These results sustain using PGPB consortia as a valid ameliorating water stress strategy in the agro-industrial field.

Seed Biopriming with Microbial Inoculant Triggers Local and Systemic Defense Responses against Rhizoctonia solani Causing Banded Leaf and Sheath Blight in Maize (Zea mays L.).

Plant growth promoting rhizobacteria Pseudomonas aeruginosa strain MF-30 isolated from maize rhizosphere was characterized for several plant growth stimulating attributes. The strain MF-30 was also evaluated for antifungal properties against Rhizoctonia solani causing banded leaf and sheath blight in maize (Zea mays L.) under in vitro conditions and was found to have higher mycelial growth suppression in the culture suspension (67.41%) followed by volatile organic compounds (62.66%) and crude extract (51.20%) in a dual plate assay. The endophytic and epiphytic colonization ability was tested using Green Fluorescent Protein (GFP)-tagging. Visualization through confocal scanning laser microscope clearly indicated that strain MF-30 colonizes the root and foliar parts of the plants. Further, the effects of seed bio-priming with P. aeruginosa MF-30 was evaluated in the induction and bioaccumulation of defense-related biomolecules, enzymes, natural antioxidants, and other changes in maize under pot trial. This not only provided protection from R. solani but also ensured growth promotion under pathogenic stress conditions in maize. The maximum concentration of hydrogen peroxide (H2O2) was reported in the root and shoot of the plants treated with R. solani alone (8.47 and 17.50 mmol mg-1 protein, respectively) compared to bioagent, P. aeruginosa MF-30 bio-primed plants (3.49 and 7.50 mmol mg-1 protein, respectively). Effects on total soluble sugar content, total protein, and total proline were also found to enhanced significantly due to inoculation of P. aeruginosa MF-30. The activities of anti-oxidative defense enzymes phenylalanine ammonia lyase (PAL), ascorbate peroxidase, peroxidase, superoxide dismutase, and catalase increased significantly in the plants bio-primed with P. aeruginosa MF-30 and subsequent foliar spray of culture suspension of MF-30 compared to pathogen alone inoculated plants. qRT-PCR analysis revealed that seed bio-priming and foliar application of P. aeruginosa MF-30 significantly increased the expression of PR-1 and PR-10 genes with the simultaneous decrease in the disease severity and lesion length in the maize plants under pathogenic stress conditions. A significant enhancement of shoot and root biomass was recorded in MF-30 bio-primed plants as compared to untreated control (p < 0.05). Significant increase in plant growth and antioxidant content, as well as decreased disease severity in the P. aeruginosa MF-30 bio-primed plants, suggested the possibility of an eco-friendly and economical means of achieving antioxidants-rich, healthier maize plants.

Restructuring the Cellular Responses: Connecting Microbial Intervention With Ecological Fitness and Adaptiveness to the Maize (Zea mays L.) Grown in Saline-Sodic Soil.

Salt stress hampers plant growth and development. It is now becoming one of the most important threats to agricultural productivity. Rhizosphere microorganisms play key roles in modulating cellular responses and enable plant tolerant to salt stress, but the detailed mechanisms of how this occurs need in-depth investigation. The present study elucidated that the microbe-mediated restructuring of the cellular responses leads to ecological fitness and adaptiveness to the maize (Zea mays L.) grown in saline-sodic soil. In the present study, effects of seed biopriming with B. safensis MF-01, B. altitudinis MF-15, and B. velezensis MF-08 singly and in consortium on different growth parameters were recorded. Soil biochemical and enzymatic analyses were performed. The activity and gene expression of High-Affinity K+ Transporter (ZmHKT-1), Sodium/Hydrogen exchanger 1 (zmNHX1), and antioxidant enzymes (ZmAPX1.2, ZmBADH-1, ZmCAT, ZmMPK5, ZmMPK7, and ZmCPK11) were studied. The expression of genes related to lateral root development (ZmHO-1, ZmGSL-1, and ZmGSL-3) and root architecture were also carried out. Seeds bioprimed with consortium of all three strains have been shown to confer increased seed germination (23.34-26.31%) and vigor indices (vigor index I: 38.71-53.68% and vigor index II: 74.11-82.43%) as compared to untreated control plant grown in saline-sodic soil at 30 days of sowing. Results indicated that plants treated with consortium of three strains induced early production of adventitious roots (tips: 4889.29, forks: 7951.57, and crossings: 2296.45) in maize compared to plants primed with single strains and untreated control (tips: 2019.25, forks: 3021.45, and crossings: 388.36), which was further confirmed by assessing the transcript level of ZmHO-1 (7.20 folds), ZmGSL-1 (4.50 folds), and ZmGSL-3 (12.00 folds) genes using the qPCR approach. The uptake and translocation of Na+, K+, and Ca2+ significantly varied in the plants treated with bioagents alone or in consortium. qRT-PCR analysis also revealed that the ZmHKT-1 and zmNHX1 expression levels varied significantly in the maize root upon inoculation and showed a 6- to 11-fold increase in the plants bioprimed with all the three strains in combination. Further, the activity and gene expression levels of antioxidant enzymes were significantly higher in the leaves of maize subjected seed biopriming with bioagents individually or in combination (3.50- to 12.00-fold). Our research indicated that ZmHKT-1 and zmNHX1 expression could effectively enhance salt tolerance by maintaining an optimal Na+/K+ balance and increasing the antioxidant activity that keeps reactive oxygen species at a low accumulation level. Interestingly, up-regulation of ZmHKT-1, NHX1, ZmHO-1, ZmGSL-1, and ZmGSL-3 and genes encoding antioxidants regulates the cellular responses that could effectively enhance the adaptiveness and ultimately leads to better plant growth and grain production in the maize crop grown in saline-sodic soil.

Seed Biopriming with Salt-Tolerant Endophytic Pseudomonas geniculata-Modulated Biochemical Responses Provide Ecological Fitness in Maize (Zea mays L.) Grown in Saline Sodic Soil.

Under changing climate, soil salinity and sodicity is a limiting factor to crop production and are considered a threat to sustainability in agriculture. A number of attempts are being made to develop microbe-based technologies for alleviation of toxic effects of salts. However, the mechanisms of salt tolerance in agriculturally important crops are not fully understood and still require in-depth study in the backdrop of emerging concepts in biological systems. The present investigation was aimed to decipher the microbe-mediated mechanisms of salt tolerance in maize. Endophytic Pseudomonas geniculate MF-84 was isolated from maize rhizosphere and tagged with green fluorescent protein for localization in the plant system. Confocal microphotographs clearly indicate that MF-84 was localized in the epidermal cells, cortical tissues, endodermis and vascular bundles including proto-xylem, meta-xylem, phloem and bundle sheath. The role of P. geniculate MF-84 in induction and bioaccumulation of soluble sugar, proline and natural antioxidants enzymes in maize plant was investigated which lead not only to growth promotion but also provide protection from salt stress in maize. Results suggested that application of P. geniculate MF-84 reduces the uptake of Na+ and increases uptake of K+ and Ca2+ in maize roots indicative of the role of MF-84 in maintaining ionic balance/homeostasis in the plant roots under higher salt conditions. It not only helps in alleviation of toxic effects of salt but also increases plant growth along with reduction in crop losses due to salinity and sodicity.