Appraising the potential of EPS-producing rhizobacteria with ACC-deaminase activity to improve growth and physiology of maize under drought stress.

Rhizobacteria containing 1-aminocyclopropane-1-carboxylic acid-deaminase (ACC-deaminase) and exopolysaccharides (EPS) activity are important to induce stress tolerance in plants. The present study was conducted to screen and characterize plant growth-promoting rhizobacteria (PGPR) with ACC-deaminase and EPS-producing activity for improving maize growth under drought stress. Eighty-five rhizobacterial strains were isolated from the rain-fed areas, among those 69 isolates were able to utilize ACC and 31 strains were found positive for EPS production. These strains containing ACC-deaminase and/or EPS-producing activity were subjected to drought tolerance assay by inducing water stress in media using polyethylene glycol 6000. Based on results of the drought tolerance bioassay, 12 most prominent strains were selected to evaluate their growth-promoting abilities in maize under water-stressed conditions by conducting jar trial. The impact of strains on maize growth parameters was variable. Strains with co-existence of ACC-deaminase and EPS-producing activity showed comparatively better results than those with either ACC-deaminase or EPS-producing activity only. These strains were also significantly better in improving the plant physiological parameters including photosynthesis rate, stomatal conductance, vapor pressure, water-use efficiency and transpiration rate. The strain D3 with co-existence of ACC-deaminase and EPS-producing activity was significantly better in colonizing maize roots, improving plant growth and physiological parameters. The strain was named as Bacillus velezensis strain D3 (accession number MT367633) as confirmed through results of 16S rRNA partial gene sequencing. It is concluded that the strains with co-existence of ACC-deaminase and EPS-producing activity could be better suited for improving crop growth and physiology under drought stress.

Preliminary study on phosphate solubilizing Bacillus subtilis strain Q3 and Paenibacillus sp. strain Q6 for improving cotton growth under alkaline conditions.

BACKGROUND: Low phosphorus availability limits crop production in alkaline calcareous soils in semi-arid regions including Pakistan. Phosphate solubilizing bacteria may improve crop growth on alkaline calcareous soils due to their ability to enhance P availability. METHODS: Twenty rhizobacterial isolates (Q1-Q20) were isolated from rhizosphere of cotton and characterized for their growth promoting attributes in vitro. The selected phosphate solubilizing isolates were further screened for their ability to improve cotton growth under axenic conditions (jar trial). The phosphorus solubilization capacities of selected strains were quantified and these strains were identified through 16S rDNA sequencing. RESULTS: Isolates Q2, Q3, Q6, Q7, Q8, Q13 and Q14 were able to solubilize phosphate from insoluble sources. Most of these isolates also possessed other traits including catalase activity and ammonia production. The growth promotion assay showed that Q3 was significantly better than most of the other isolates followed by Q6. Maximum root colonization (4.34 × 106 cfu g-1) was observed in case of isolate Q6 followed by Q3. The phosphorus solubilization capacities of these strains were quantified, showing a maximum phosphorus solubilization by Q3 (optical density 2.605 ± 0.06) followed by the Q6 strain. The strain Q3 was identified as Bacillus subtilis (accession # KX788864) and Q6 as Paenibacillus sp. (accession # KX788865) through 16S rDNA sequencing. DISCUSSION: The bacterial isolates varied in their abilities for different growth promoting traits. The selected PGPR Bacillus subtilis strain Q3 and Paenibacillus sp. strain Q6 have multifarious growth promoting traits including ability to grow at higher EC and pH levels, and phosphorus solubilizing ability. These strains can efficiently colonize cotton roots under salt affected soils and help plants in phosphorus nutrition. It is concluded that both strains are potential candidates for promoting cotton growth under alkaline conditions, however further investigation is required to determine their potential for field application.

Synergistic use of biochar, compost and plant growth-promoting rhizobacteria for enhancing cucumber growth under water deficit conditions.

BACKGROUND: Limited information is available about the effectiveness of biochar with plant growth-promoting rhizobacteria (PGPR) and compost. A greenhouse study was conducted to evaluate the effect of biochar in combination with compost and PGPR (Pseudomonas fluorescens) for alleviating water deficit stress. Both inoculated and un-inoculated cucumber seeds were sown in soil treated with biochar, compost and biochar + compost. Three water levels - field capacity (D0), 75% field capacity (D1) and 50% field capacity (D2) - were maintained. RESULTS: The results showed that water deficit stress significantly suppressed the growth of cucumber; however, synergistic use of biochar, compost and PGPR mitigated the negative impact of stress. At D2, the synergistic use of biochar, compost and PGPR caused significant increases in shoot length, shoot biomass, root length and root biomass, which were respectively 88, 77, 89 and 74% more than in the un-inoculated control. Significant improvements in chlorophyll and relative water contents as well as reduction in leaf electrolyte leakage demonstrated the effectiveness of this approach. Moreover, the highest population of P. fluorescens was observed where biochar and compost were applied together. CONCLUSION: These results suggest that application of biochar with PGPR and/or compost could be an effective strategy for enhancing plant growth under stress. © 2017 Society of Chemical Industry.

Effectiveness of halo-tolerant, auxin producing Pseudomonas and Rhizobium strains to improve osmotic stress tolerance in mung bean (Vigna radiata L.)

Halo-tolerant, auxin producing bacteria could be used to induce salt tolerance in plants. A number of Rhizobium and auxin producing rhizobacterial strains were assessed for their ability to tolerate salt stress by conducting osmoadaptation assay. The selected strains were further screened for their ability to induce osmotic stress tolerance in mung bean seedlings under salt-stressed axenic conditions in growth pouch/jar trials. Three most effective strains of Rhizobium and Pseudomonas containing ACC-deaminase were evaluated in combination, for their ability to induce osmotic stress tolerance in mung bean at original, 4, and 6 dS m-1 under axenic conditions. Results showed that sole inoculation of Rhizobium and Pseudomonas strains improved the total dry matter up to 1.4, and 1.9 fold, respectively, while the increase in salt tolerance index was improved up to 1.3 and 2.0 fold by the Rhizobium and Pseudomonas strains, respectively. However, up to 2.2 fold increase in total dry matter and salt tolerance index was observed due to combined inoculation of Rhizobium and Pseudomonas strains. So, combined application of Rhizobium and Pseudomonas strains could be explored as an effective strategy to induce osmotic stress tolerance in mung bean.

Efficacy of Rhizobium and Pseudomonas strains to improve physiology, ionic balance and quality of mung bean under salt-affected conditions on farmer's fields.

High ethylene concentration under different environmental stresses such as salinity is one of the contributing factors for premature senescence of different plant parts. Plants under salinity stress produce increased levels of ethylene which inhibit the plant growth and physiology thus deteriorating the quality of the produce. Some plant growth promoting rhizobacteria (PGPR) have the ability to improve quality of crops through reduction in detrimental effects of salinity on plant growth and physiology by lowering endogenous level of ethylene along with other mechanisms. Two field trials were conducted to evaluate the Rhizobium and Pseudomonas containing ACC-deaminase for their efficacy to reduce the effect of salinity on physiology, ionic and nutrient balance of mung bean. Results showed that salinity stress adversely affected the physiological parameters of mung bean. It decreased the CO(2) assimilation, stomatal conductance of water, relative water content, photosynthetic rate, transpiration rate and chlorophyll contents in mung bean but inoculation of either Rhizobium or Pseudomonas alone reduced adverse effect of salinity significantly. However, co-inoculation with Rhizobium and Pseudomonas was the most effective treatment and it diluted the adverse effects of salinity on relative water contents and CO(2) assimilation rate thus improving the photosynthetic rate, water use efficiency and chlorophyll content over the un-inoculated control. Co-inoculation improved the ionic balance and also increased the phosphorus and protein concentration in grain of mung bean. The results suggested that these strains could be effectively used to improve the growth, physiology and quality of mung bean under salt-affected conditions.

Effectiveness of halo-tolerant, auxin producing Pseudomonas and Rhizobium strains to improve osmotic stress tolerance in mung bean (Vigna radiata L.).

Halo-tolerant, auxin producing bacteria could be used to induce salt tolerance in plants. A number of Rhizobium and auxin producing rhizobacterial strains were assessed for their ability to tolerate salt stress by conducting osmoadaptation assay. The selected strains were further screened for their ability to induce osmotic stress tolerance in mung bean seedlings under salt-stressed axenic conditions in growth pouch/jar trials. Three most effective strains of Rhizobium and Pseudomonas containing ACC-deaminase were evaluated in combination, for their ability to induce osmotic stress tolerance in mung bean at original, 4, and 6 dS m(-1) under axenic conditions. Results showed that sole inoculation of Rhizobium and Pseudomonas strains improved the total dry matter up to 1.4, and 1.9 fold, respectively, while the increase in salt tolerance index was improved up to 1.3 and 2.0 fold by the Rhizobium and Pseudomonas strains, respectively. However, up to 2.2 fold increase in total dry matter and salt tolerance index was observed due to combined inoculation of Rhizobium and Pseudomonas strains. So, combined application of Rhizobium and Pseudomonas strains could be explored as an effective strategy to induce osmotic stress tolerance in mung bean.

Inducing salt tolerance in mung bean through coinoculation with rhizobia and plant-growth-promoting rhizobacteria containing 1-aminocyclopropane-1-carboxylate deaminase.

Twenty-five strains of plant-growth-promoting rhizobacteria (PGPR) containing 1-aminocyclopropane-1-carboxylate (ACC) deaminase and 10 strains of rhizobia were isolated from rhizosphere soil samples and nodules of mung bean. They were screened in separate trials under salt-stressed axenic conditions. The three most effective strains of PGPR (Mk1, Pseudomonas syringae ; Mk20, Pseudomonas fluorescens ; and Mk25, Pseudomonas fluorescens biotype G) and Rhizobium phaseoli strains M1, M6, and M9 were evaluated in coinoculation for their growth-promoting activity at three salinity levels (original, 4 dS·m(-1), and 6 dS·m(-1)) under axenic conditions. The results showed that salinity stress significantly reduced plant growth but inoculation with PGPR containing ACC deaminase and rhizobia enhanced plant growth, thus reducing the inhibitory effect of salinity. However, their combined application was more effective under saline conditions, and the combination Mk20 × M6 was the most efficient for improving seedling growth and nodulation. The effect of high ethylene concentrations on plant growth and the performance of these strains for reducing the negative impact of saline stress was also evaluated by conducting a classical triple-response bioassay. The intensity of the classical triple response decreased owing to inoculation with these strains, with the root and shoot lengths of inoculated mung bean seedlings increasing and stem diameter decreasing, which is a typical response to the dilution in a classical triple response bioassay. Thus, coinoculation with PGPR containing ACC deaminase and Rhizobium spp. could be a useful approach for inducing salt tolerance and thus improving growth and nodulation in mung bean under salt-affected conditions.

Fertilizer-dependent efficiency of Pseudomonads for improving growth, yield, and nutrient use efficiency of wheat (Triticum aestivum L.).

Acquisition of nutrients by plants is primarily dependent on root growth and bioavailability of nutrients in the rooting medium. Most of the beneficial bacteria enhance root growth, but their effectiveness could be influenced by the nutrient status around the roots. In this study, two 1-aminocyclopropane-1-carboxylate (ACC)-deaminase containing plant-growth-promoting rhizobacteria (PGPR), Pseudomonas fluorescens and P. fluorescens biotype F were tested for their effect on growth, yield, and nutrient use efficiency of wheat under simultaneously varying levels of all the three major nutrients N, P, and K (at 0%, 25%, 50%, 75%, and 100% of recommended doses). Results of pot and field trials revealed that the efficacy of these strains for improving growth and yield of wheat reduced with the increasing rates of NPK added to the soil. In most of the cases, significant negative linear correlations were recorded between percentage increases in growth and yield parameters of wheat caused by inoculation and increasing levels of applied NPK fertilizers. It is highly likely that under low fertilizer application, the ACC-deaminase activity of PGPR might have caused reduction in the synthesis of stress (nutrient)-induced inhibitory levels of ethylene in the roots through ACC hydrolysis into NH(3) and alpha-ketobutyrate. The results of this study imply that these Pseudomonads could be employed in combination with appropriate doses of fertilizers for better plant growth and savings of fertilizers.

Application of redox mediator to accelerate selenate reduction to elemental selenium by Enterobacter taylorae.

Acceleration of bacterial reduction of selenate [Se(VI)] to insoluble elemental Se [Se(0)] plays an important role in Se bioremediation. Anthraquinone-2,6-disulfonate (AQDS), a redox mediator, was assessed for its ability to enhance the reduction of Se(VI) (2000 microg/L) to Se(0) by Enterobacter taylorae in various media. The results showed that addition of AQDS did not increase Se(VI) reduction in the media containing 50 and 250 mg/L yeast extract, suggesting that E. taylorae cannot directly use anthrahydroquinone-2,6-disulfonate (AHQDS, a reduced form of AQDS) to respire Se(VI). An increase of yeast extract concentration from 50 to 250 mg/L in the medium dramatically enhanced the AQDS function for rapid reduction of selenite [Se(IV)] to Se(0). During an 8-day experiment, 85-91% of the added Se was reduced to Se(0) in the AQDS-amended medium in comparison to formation of 46% of Se(0) in the medium without AQDS. These results show that redox mediators have great application potential in bioremediation of Se in Se-contaminated water.

Fate of colloidal-particulate elemental selenium in aquatic systems.

Bacterial reduction of selenate [Se(VI)] to elemental Se [Se(0)] is considered an effective bioremediation technique to remove selenium (Se) from agricultural drainage water. However, the fate of the newly formed Se(0) in aquatic systems is not known when it flows out of the treatment system. A set of laboratory experiments was conducted to determine the fate of the colloidal-particulate Se(0) in a water column and in a water-sediment system. Results showed that the newly formed colloidal-particulate Se(0) followed two removal pathways in aquatic systems: (i) flocculation-sedimentation to the bottom of the water and (ii) oxidation to selenite [Se(IV)] and Se(VI). During 58 d of the experiments, 51% of the added Se(0) was precipitated to the bottom of the water and 47% was oxidized to Se(IV) in the water column. In the water-sediment system, Se(IV) in the water accounted for 21 to 25% of the added Se(0). Adsorption of Se(IV) to the bottom sediment resulted in a relatively low amount of Se(IV) in the water. This study indicates that the newly formed Se(0) may be an available form of Se for uptake by organisms if it flows to aquatic systems from a treatment site. Therefore, an effective bioremediation system for removing Se from drainage water must reduce Se(VI) to Se(0) and remove Se(0) directly from the drainage water.

Factors affecting reduction of selenate to elemental selenium in agricultural drainage water by Enterobacter taylorae.

Microbial reduction of selenate [Se(VI)] to elemental selenium [Se(0)] is a useful technique for removing Se from agricultural drainage water. A series of batch experiments were conducted in the laboratory to determine the effects of yeast extract (50-1000 mg/L), salinity (EC, 5-75 dS/m), and NO(3)(-) (5-100 mg/L) on the removal of Se(VI) (2000 microg/L) from drainage water by Enterobacter taylorae. Results showed that relatively high amounts of yeast extract (500 mg/L) were needed for E. taylorae to effectively reduce Se(VI) to Se(0). During a 7-day experiment, approximately 95% of added Se(VI) was reduced to Se(0) in the low-salinity drainage water (5 dS/m) with NO(3)(-) values of 5-50 mg/L. In the high-salinity drainage water (50-75 dS/m), reduction of Se(VI) to Se(0) was limited. E. taylorae was also capable of reducing Se(VI) to Se(0) in the San Joaquin Valley drainage water, with a reduction of the added Se(VI) to Se(0) (73.8%) and Se(-II) (20%). This study suggests that E. taylorae may be used to treat Se(VI)-contaminated drainage water in the field.

Fate of selenate metabolized by Enterobacter taylorae isolated from rice straw.

Rice straw has been successfully tested as an effective organic source and a carrier of selenate [Se(VI)]-reducing bacteria to remove Se(VI) from agricultural drainage water. In this study, an Se(VI)-reducing bacterium identified as Enterobacter taylorae was isolated from rice straw and used to remove Se(VI) from a 0.5% tryptic soy broth (TSB) and high-salt (15.5 dS m(-)(1)) synthetic agricultural drainage water containing Se(VI) in a range of 500-5000 microg/L. Results showed that E. taylorae reduced 81-94% of the added Se(VI) to elemental Se [Se(0)] in the 0.5% TSB solution during a 5-day experiment. In the high-salt drainage water, Se(VI) reduction was rapid during a 9-day experiment. On the final day of the experiment, Se(0) [75%] and Se(-II) [19%] were the major forms of Se in the drainage water with small amounts of Se(VI), Se(IV), and volatile Se released. The pathway of Se(VI) reduction in the drainage water followed the order Se(VI) --> selenite [Se(IV)] --> Se(0) --> selenide [Se(-II)]. This study suggests that E. taylorae may be used to remediate high-salt Se(VI)-contaminated agricultural drainage water.