Producing Plant Growth Promoting Rhizobacteria Research Articles

Decolorization, degradation and detoxification of mutagenic dye Methyl orange by novel biofilm producing plant growth-promoting rhizobacteria

Discharge of untreated dyeing wastewater nearby water-bodies is one of major causes of water pollution. Generally, bacterial strains isolated from industrial effluents and/or contaminated soils are used for the bioremediation of Methyl orange (MO), a mutagenic recalcitrant mono-azo dye, used in textiles and biomedical. However, MO degradation by biofilm producing plant growth-promoting rhizobacteria (BPPGPR) was not studied yet. In this study, 19 out of 21 BPPGPR strains decolorized 96.3–99.9% and 89.5–96.3% MO under microaerophilic and aerobic conditions, respectively from Luria-Bertani broth (LBB) followed by yeast-extract peptone and salt-optimized broth plus glycerol media within 120 h of incubation at 28 °C. Only selected BPPGPR including Pseudomonas fluorescens ESR7, P. veronii ESR13, Stenotrophomonas maltophilia ESR20, Staphylococcus saprophyticus ESD8, and P. parafulva ESB18 were examined for process optimization of MO decolorization using a single factor optimization method. This study showed that under optimal conditions (e.g., LBB, 100 mg L−1 MO, pH 7, incubation of 96 h, 28 °C), these strains could remove 99.1–99.8% and 97.6–99.5% MO under microaerophilic and aerobic conditions, respectively. Total azoreductase and laccase activities responsible for biodegradation were also remarkably activated in the biodegraded samples under optimal conditions, while these activities were repressed under unfavorable conditions (e.g., 40 °C and 7.5% NaCl). This study confirmed that MO was degraded and detoxified by these bacterial strains through breakage of azo bond. So far, this is the first report on bioremediation of MO by the BPPGPR strains. These BPPGPR strains are highly promising to be utilized for the bioremediation of dyeing wastewater in future.

Efficacy of rhizobacterial exopolysaccharides in improving plant growth, physiology, and soil properties.

The current study investigated the influence of exopolysaccharides (EPSs) producing plant growth-promoting rhizobacteria (PGPR) on the growth, physiology, and soil properties. The pre-isolated and compatible EPS producing PGPR strains were first screened based on improvement in soil aggregates in an incubation study. The screened strains (Rhizobium phaseoli strain Mn-6, Pseudomonas bathysetes strain LB5, and unidentified strain R2) were then employed in pot study for assessing improvements in maize growth, physiology, and soil properties. Eight treatments including T1 = control, T2 = Mn-6, T3 = R2, T4 = LB5, T5 = Mn-6 + R2, T6 = Mn-6 + LB5, T7 = R2 + LB5, and T8 = Mn-6 + R2 + LB5 were applied in completely randomized design (CRD) hexa replicated (half for root and half for soil, and yield attributes). The results depicted that among various treatments, the application of PGPR strain Mn-6 increased plant height, root length, root fresh and dry weight, root length density, SPAD value, leaf areas index, photosynthesis rate, transpiration, and stomatal conductance by 24, 79, 72, 90, 49, 35, 23, 21, 75, and 77%, respectively, compared with non-inoculated treatment. Similarly, significant improvement in maize yield and soil physical properties was also observed in response to the application of EPS-producing PGPR. Therefore, it is concluded that the application of EPS producing PGPR is an effective strategy to improve plant growth, physiology, yield, and soil physical properties. Moreover, EPS-producing PGPR should be exploited in field studies for their potential in improving plant growth and soil properties.

Efficacy of Indole Acetic Acid and Exopolysaccharides-Producing Bacillus safensis Strain FN13 for Inducing Cd-Stress Tolerance and Plant Growth Promotion in Brassica juncea (L.)

Untreated wastewater used for irrigating crops is the major source of toxic heavy metals and other pollutants in soils. These heavy metals affect plant growth and deteriorate the quality of edible parts of growing plants. Phytohormone (IAA) and exopolysaccharides (EPS) producing plant growth-promoting rhizobacteria can reduce the toxicity of metals by stabilizing them in soil. The present experiment was conducted to evaluate the IAA and EPS-producing rhizobacterial strains for improving growth, physiology, and antioxidant activity of Brassica juncea (L.) under Cd-stress. Results showed that Cd-stress significantly decreased the growth and physiological parameters of mustard plants. Inoculation with Cd-tolerant, IAA and EPS-producing rhizobacterial strains, however, significantly retrieved the inhibitory effects of Cd-stress on mustard growth, and physiology by up regulating antioxidant enzyme activities. Higher Cd accumulation and proline content was observed in the roots and shoot tissues upon Cd-stress in mustard plants while reduced proline and Cd accumulation was recorded upon rhizobacterial strains inoculation. Maximum decrease in proline contents (12.4%) and Cd concentration in root (26.9%) and shoot (29%) in comparison to control plants was observed due to inoculation with Bacillus safensis strain FN13. The activity of antioxidant enzymes was increased due to Cd-stress; however, the inoculation with Cd-tolerant, IAA-producing rhizobacterial strains showed a non-significant impact in the case of the activity of superoxide dismutase (SOD), peroxidase (POX) and catalase (CAT) in Brassica juncea (L.) plants under Cd-stress. Overall, Bacillus safensis strain FN13 was the most effective strain in improving the Brassica juncea (L.) growth and physiology under Cd-stress. It can be concluded, as the strain FN13 is a potential phytostabilizing biofertilizer for heavy metal contaminated soils, that it can be recommended to induce Cd-stress tolerance in crop plants.

Diversity analysis of ACC deaminase producing bacteria associated with rhizosphere of coconut tree (Cocos nucifera L.) grown in Lakshadweep islands of India and their ability to promote plant growth under saline conditions

ACC deaminase producing Plant growth promoting rhizobacteria (PGPR) offers a great promise for ameliorating the negative impacts of salinity stress manifested on plants. In this context, 28 rhizospheric bacteria associated with ACC deaminase potential (198−1069 nmol α-ketobutyrate mg protein−1 h−1) were isolated from 5 different islands of Lakshadweep, union territory, India- Agatti, Kavaratti, Bangaram, Kadmat, and Thinnakara islands using DF-minimal medium. The diversity of cultivable ACC deaminase producing bacteria was analysed by PCR-RFLP (Restriction Fragment Length Polymorphism) method using three endonucleases AluI, MspI and HaeIII which led to the grouping of these isolates into six clusters at 80 % similarity index. Subsequently, isolates were functionally characterized for various PGP traits such that indole-3-acetic acid (IAA) production (∼10−80 μg mL−1); 16 isolates had phosphate solubilizing potential ranging from ∼19 to 88 P mg L−1 ; siderophore and ammonia production abilities were observed in 5 and 24 isolates, respectively while two strains tolerated up to 8% NaCl. Phylogenetic analysis of 16S rRNA gene sequences of representative strain from each cluster revealed that twenty-eight ACC deaminase producing PGPR belong to eight distinct genera: Pseudomonas, Bacillus, Azospirillum, Azotobacter, Escherichia, Paenibacillus, Burkholderia, and Klebsiella. Two isolates, CO1 (Pseudomonas putida) and CO8 (Bacillus paramycoides) were evaluated for plant growth promoting effects on French bean (Phaseolus vulgaris) under salinity (100 mM NaCl) stress. Both the selected isolates in consortium form significantly increased the root length, shoot length, root fresh and dry weight, shoot fresh and dry weight of French bean seedlings exposed to salinity stress, compared to non-inoculated control plants. The co-inoculation with selected strains CO1 and CO8 has significantly improved chlorophyll concentration, relative water content, membrane stability index, gas exchange parameters including net photosynthesis rate (PN), stomatal conductance (gs), transpiration rate (E) and water use efficiency of French bean plants by ∼100 %, ∼85 %, ∼40 %, ∼198 %, ∼80 %, ∼70 % and ∼75 %, respectively under saline conditions in comparison with non-inoculated plants. Moreover, the consortium treated French bean plants showed lower levels of stress-induced ethylene by 38 %, electrolyte leakage and Malondialdehyde (MDA) content by ∼15 % under salt stress compared to non-inoculated ones. This study unveiled the potential of halotolerant strains, Pseudomonas putida and Bacillus paramycoides as French bean biofertilizers in mitigating the adverse effects of salinity in plant growth in sustainable agriculture.

Growth-Stimulatory Effect of Quorum Sensing Signal Molecule N-Acyl-Homoserine Lactone-Producing Multi-Trait Aeromonas spp. on Wheat Genotypes Under Salt Stress.

Salinity is one of the major threats to agricultural productivity worldwide. Soil and plant management practices, along with inoculation with plant-beneficial bacteria, play a key role in the plant’s tolerance toward salinity stress. The present study demonstrates the potential of acyl homoserine lactone (AHL)-producing plant growth promoting rhizobacteria (PGPR) strains of Aeromonas sp., namely, SAL-17 (accession no. HG763857) and SAL-21 (accession no. HG763858), for growth promotion of two wheat genotypes inherently different for salt tolerance potential. AHLs are the bacterial signal molecules that regulate the expression of various genes in bacteria and plants. Both Aeromonas spp., along with innate plant-growth-promoting (PGP) and salt tolerance traits, showed AHL production which was identified on tandem mass spectrometry as C6-HSL, 3-OH-C5-HSL, 3-OH-C6-HSL, 3-oxo-C7-HSL C10-HSL, 3-oxo-C10-HSL, 3-OH-C10-HSL, 3-oxo-C12-HSL and C6-HSL, and 3-oxo-C10-HSL. The exogenous application of purified AHLs (mix) significantly improved various root parameters at 200 mM NaCl in both salt-sensitive (SSG) and salt-tolerant (STG) genotypes, where the highest increase (≈80%) was observed where a mixture of both strains of AHLs was used. Confocal microscopic observations and root overlay assay revealed a strong root colonization potential of the two strains under salt stress. The inoculation response of both STG and SSG genotypes was evaluated with two AHL-producing strains (SAL-17 and SAL-21) and compared to non-AHL-producing Aeromonas sp. SAL-12 (accession no. HG763856) in saline (EC = 7.63 ms/cm2) and non-saline soil. The data reveal that plants inoculated with the bacterial consortium (SAL-21 + SAL-17) showed a maximum increase in leaf proline content, nitrate reductase activity, chlorophyll a/b, stomatal conductance, transpiration rate, root length, shoot length, and grain weight over non-inoculated plants grown in saline soil. Both STG and SSG showed relative effectiveness toward inoculation (percent increase for STG: 165–16%; SSG: 283–14%) and showed a positive correlation of grain yield with proline and nitrate reductase activity. Furthermore, principal component analysis (PCA) and categorical PCA analysis clearly showed an inoculation response in both genotypes, revealing the effectiveness of AHL-producing Aeromonas spp. than the non-AHL-producing strain. The present study documents that the consortium of salt-tolerant AHL-producing Aeromonas spp. is equally effective for sustaining the growth of STG as well as SSG wheat genotypes in saline soil, but biosafety should be fully ensured before field release.

Alleviation of Cadmium Adverse Effects by Improving Nutrients Uptake in Bitter Gourd through Cadmium Tolerant Rhizobacteria

Cadmium is acute toxicity inducing heavy metal that significantly decreases the yield of crops. Due to high water solubility, it reaches the plant tissue and disturbs the uptake of macronutrients. Low uptake of nutrients in the presence of cadmium is a well-documented fact due to its antagonistic relationship with those nutrients, i.e., potassium. Furthermore, cadmium stressed plant produced a higher amount of endogenous stress ethylene, which induced negative effects on yield. However, inoculation of 1-amino cyclopropane-1-carboxylate deaminase (ACCD), producing plant growth promoting rhizobacteria (PGPR), can catabolize this stress ethylene and immobilized heavy metals to mitigate cadmium adverse effects. We conducted a study to examine the influence of ACCD PGPR on nutrients uptake and yield of bitter gourd under cadmium toxicity. Cadmium tolerant PGPRs, i.e., Stenotrophomonas maltophilia and Agrobacterium fabrum were inoculated solely and in combination with recommended nitrogen, phosphorus, and potassium fertilizers (RNPKF) applied under different concentration of soil cadmium (2 and 5 mg kg−1 soil). Results showed that A. fabrum with RNPKF showed significant positive response towards an increase in the number of bitter gourds per plant (34% and 68%), fruit length (19% and 29%), bitter gourd yield (26.5% and 21.1%), N (48% and 56%), and K (72% and 55%) concentration from the control at different concentrations of soil cadmium (2 and 5 mg kg−1 soil), respectively. In conclusion, we suggest that A. fabrum with RNPKF can more efficaciously enhance N, K, and yield of bitter gourd under cadmium toxicity.

Deciphering metal toxicity responses of flax (Linum usitatissimum L.) with exopolysaccharide and ACC-deaminase producing bacteria in industrially contaminated soils

Rapid industrialization is the main reason of heavy metals contamination of soil colloids and water reservoirs. Heavy metals are persistent inorganic pollutants; deleterious to plants, animals and human beings because of accumulation in food chain. The aim of the current work was to evaluate the role of indole acetic acid (IAA), exopolysaccharide (EPS) and ACC-deaminase producing plant growth promoting rhizobacteria (PGPR) i.e .B. gibsonii PM11 and B. xiamenensis PM14 in metal phytoremediation of metals, their survival and plant growth promotion potential in metal polluted environment as well as alterations in physio-biochemical responses of inoculated L. usitatissimum plants towards heavy metal toxicity. Two bacterial strains Bacillus gibsonii (PM11) and Bacillus xiamenensis (PM14), previously isolated from sugarcane's rhizosphere, were screened for metal tolerance (50 mg/l to 1000 mg/l) and plant growth promoting traits like IAA, ACC-deaminase, EPS production and nitrogen fixing ability under metal stress. The response of flax plant (Linum usitatissimum L.) was analyzed in a pot experiment containing both industrially contaminated and non-contaminated soils. Experiment was comprised of six different treatments, each with three replicates. At the end of the experiment, role of metal tolerant plant growth promoting bacterial inoculation was elucidated by analyzing the plant growth parameters, chlorophyll contents, antioxidative enzymes, and metal uptake both under standard and metal contaminated rhizospheres. Results revealed that root and shoot length, plant's fresh and dry weight, proline content, chlorophyll content, antioxidant enzymatic activity was increased in plants inoculated with plant growth promoting bacteria as compared to non-inoculated ones both in non-contaminated and industrial contaminated soils. In current study, inoculation of IAA, EPS and ACC-deaminase producing bacteria enhances plant growth and nutrient availability by minimizing metal-induced stressed conditions. Moreover, elevated phytoextraction of multi-metals from industrial contaminated soils by PGPR inoculated L. usitatissimum plants reveal that these strains could be used as sweepers in heavy metals polluted environment.

ACC-deaminase producing plant growth promoting rhizobacteria and biochar mitigate adverse effects of drought stress on maize growth.

Availability of good quality irrigation water is a big challenge in arid and semi arid regions of the world. Drought stress results in poor plant growth and low yield; however, the rhizobacteria, capable of producing 1-aminocyclopropane-1-carboxylate (ACC)-deaminase are likely to improve crop growth and productivity under drought stress. Similarly, biochar could also ameliorate the negative impacts of drought stress. Therefore, this pot experiment was conducted to evaluate the role of ACC-deaminase producing plant growth promoting rhizobacteria (PGPR) alone and in combinations with timber-waste biochar in improving maize growth under drought stress. The ACC-deaminase producing rhizobacteria, Pseudomonas aeruginosa, Enterobacter cloacae, Achromobacter xylosoxidans and Leclercia adecarboxylata were studied along with two rates (0.75 and 1.50% of the soil weight) of biochar under three moisture levels i.e., normal moisture, mild drought stress and severe drought stress. The E. cloacae in conjunction with higher rate of biochar produced a significant improvement i.e., up to 60, 73, 43, 69, 76 and 42% respectively, in grain yield plant-1, photosynthetic rate, stomatal conductance, chlorophyll a, total chlorophyll and carotenoids contents of maize as compared to the control under mild drought stress. Similarly, A. xylosoxidans with higher rate of biochar also enhanced grain yield plant-1, photosynthetic rate, stomatal conductance, chlorophyll a, total chlorophyll and carotenoids contents of maize up to 200, 213, 113, 152, 148 and 284%, respectively over control under severe drought stress. In conclusion, combination of ACC-deaminase containing PGPR, A. xylosoxidans and biochar (0.75%) proved an effective technique to improve maize growth and productivity under drought stress.

ACC Deaminase Producing PGPR Bacillus amyloliquefaciens and Agrobacterium fabrum along with Biochar Improve Wheat Productivity under Drought Stress

Drought stress retards wheat plant’s vegetative growth and physiological processes and results in low productivity. A stressed plant synthesizes ethylene which inhibits root elongation; however, the enzyme 1-Aminocyclopropane-1-Carboxylate (ACC) deaminase catabolizes ethylene produced under water stress. Therefore, the ACC deaminase producing plant growth promoting rhizobacteria (PGPR) can be used to enhance crop productivity under drought stress. Biochar (BC) is an organically active and potentially nutrient-rich amendment that, when applied to the soil, can increase pore volume, cation exchange capacity and nutrient retention and bioavailability. We conducted a field experiment to study the effect of drought tolerant, ACC deaminase producing PGPR (with and without timber waste BC) on plant growth and yield parameters under drought stress. Two PGPR strains, Agrobacterium fabrum or Bacillus amyloliquefaciens were applied individually and in combination with 30 Mg ha−1 BC under three levels of irrigation, i.e., recommended four irrigations (4I), three irrigations (3I) and two irrigations (2I). Combined application of B. amyloliquefaciens and 30 Mg ha−1 BC under 3I, significantly increased growth and yield traits of wheat: grain yield (36%), straw yield (50%), biological yield (40%). The same soil application under 2I resulted in greater increases in several of the growth and yield traits: grain yield (77%), straw yield (75%), above- and below-ground biomasses (77%), as compared to control; however, no significant increases in chlorophyll a, b or total, and photosynthetic rate and stomatal conductance in response to individual inoculation of a PGPR strain (without BC) were observed. Therefore, we suggest that the combined soil application of B. amyloliquefaciens and BC more effectively mitigates drought stress and improves wheat productivity as compared to any of the individual soil applications tested in this study.