The wheat growth-promoting traits of Ochrobactrum and Pantoea species, responsible for solubilization of different P sources, are ensured by genes encoding enzymes of multiple P-releasing pathways.

Production and release of organic acids and phosphatase enzymes by microbes are important for inorganic and organic phosphorus cycling in soil. The presence of microorganisms with corresponding traits in the plant rhizosphere lead to improved plant P uptake and ultimately growth promotion. We studied the potential of two rhizosphere-competent strains, Pantoea sp. MR1 and Ochrobactrum sp. SSR, for solubilization of different organic and inorganic P sources in vitro. In a pot experiment we further revealed the impact of the two strains on wheat seedling performance in soil amended with either phytate, rock phosphate or K2HPO4 as solely P source. To directly link P-solubilizing activity to the strain-specific genetic potential, we designed novel primers for glucose dehydrogenase (gcd), phosphatase (pho) and phytase (phy) genes, which are related to the organic and inorganic P solubilization potential. Quantitative tracing of these functional genes in the inoculated soils of the conducted pot experiment further allowed to compare strain abundances in the soil in dependency on the present P source. We observed strain- and P source-dependent patterns of the P solubilization in vitro as well as in the pot experiment, whereby P release, particularly from phytate, was linked to the strain abundance. We further revealed that the activity of microbial phosphatases is determined by the interplay between functional gene abundance, available soil P, and substrate availability. Moreover, positive impacts of microbial seed inoculation on wheat root architecture and aboveground growth parameters were observed. Our results suggest that screening for rhizosphere-competent strains with gcd, pho and phy genes may help to identify new microbial taxa that are able to solubilize and mineralize inorganic as well as organic bound P. Subsequently, the targeted use of corresponding strains may improve P availability in agricultural soils and consequently reduce fertilizer application.

Rasul M ，Yasmin S ，Yahya M ，Breitkreuz C ，Tarkka M ，Reitz T ... -  《-》

Novel Glucose-1-Phosphatase with High Phytase Activity and Unusual Metal Ion Activation from Soil Bacterium Pantoea sp. Strain 3.5.1.

Phosphorus is an important macronutrient, but its availability in soil is limited. Many soil microorganisms improve the bioavailability of phosphate by releasing it from various organic compounds, including phytate. To investigate the diversity of phytate-hydrolyzing bacteria in soil, we sampled soils of various ecological habitats, including forest, private homesteads, large agricultural complexes, and urban landscapes. Bacterial isolate Pantoea sp. strain 3.5.1 with the highest level of phytase activity was isolated from forest soil and investigated further. The Pantoea sp. 3.5.1 agpP gene encoding a novel glucose-1-phosphatase with high phytase activity was identified, and the corresponding protein was purified to apparent homogeneity, sequenced by mass spectroscopy, and biochemically characterized. The AgpP enzyme exhibits maximum activity and stability at pH 4.5 and at 37°C. The enzyme belongs to a group of histidine acid phosphatases and has the lowest Km values toward phytate, glucose-6-phosphate, and glucose-1-phosphate. Unexpectedly, stimulation of enzymatic activity by several divalent metal ions was observed for the AgpP enzyme. High-performance liquid chromatography (HPLC) and high-performance ion chromatography (HPIC) analyses of phytate hydrolysis products identify dl-myo-inositol 1,2,4,5,6-pentakisphosphate as the final product of the reaction, indicating that the Pantoea sp. AgpP glucose-1-phosphatase can be classified as a 3-phytase. The identification of the Pantoea sp. AgpP phytase and its unusual regulation by metal ions highlight the remarkable diversity of phosphorus metabolism regulation in soil bacteria. Furthermore, our data indicate that natural forest soils harbor rich reservoirs of novel phytate-hydrolyzing enzymes with unique biochemical features.

Suleimanova AD ，Beinhauer A ，Valeeva LR ，Chastukhina IB ，Balaban NP ，Shakirov EV ，Greiner R ，Sharipova MR ... -  《-》

Inoculation of pqqE gene inhabiting Pantoea and Pseudomonas strains improves the growth and grain yield of wheat with a reduced amount of chemical fertilizer.

The present study was performed to examine the role of pqqE inhabiting rhizobacteria in organic acid production and relationship of the organic acids with phosphate solubilization by the bacteria in vitro as well as in vivo. The pqqE gene was PCR amplified and sequenced in genomic DNA of Pantoea sp. WP-5 and Pseudomonas sp. NN-4. Nucleotide sequence obtained from WP-5 and NN-4 showed maximum sequence similarity (88 and 89%, respectively) with the pqqE gene of Pseudomonas fluorescens strain CMR12a (KM251420). Deduced amino acid sequence from pqqE gene of Pseudomonas sp. NN-4 and Pantoea sp. WP-5 showed 75 and 93% similarity, respectively, with protein pyrroloquinoline quinone. Phosphate solubilization and acid production assay were quantified on spectrophotometer and high-profile liquid chromatograph, respectively, by each bacterial strain. Both strains produced organic acids such as acetic, citric, gluconic, succinic and malic acid and lowered the pH of Pikovskaya broth medium under laboratory conditions. Phosphate solubilization by Pantoea sp. WP-5 was 311 ± 4 and 204 ± 3 µg ml-1 in the culture medium supplemented with glucose and sucrose as carbon source, respectively. Pseudomonas sp. NN-4 solubilized 176 ± 3 and 298 ± 5 µg ml-1 phosphate in Pikovskaya broth medium under similar conditions. In field experiments conducted during two consecutive years, the concentration of acetic acid and gluconic acid was higher in root exudates of plants treated with Pantoea sp. WP-5 at 30% reduced doses of nitrogen (N)- and phosphorus (P)-based chemical fertilizers as compared to non-inoculated plants. Values of chlorophyll contents, crop growth rate, leaf area index, straw yield and P contents were recorded higher in plants inoculated with Pantoea sp. WP-5 and Pseudomonas sp. NN-4 as compared to non-inoculated control. Grain yield was increased by 10-12% due to inoculation with Pantoea sp. WP-5 and Pseudomonas sp. NN-4 over non-inoculated control in the field experiments. These results lead to the conclusions that the rhizobacteria inhabiting pqqE gene produced organic acids and solubilized the phosphate in vitro. On inoculation to wheat plants in field experiments, these strains produced the organic acids, solubilized the phosphate, and improved the P uptake and productivity of wheat. The Pantoea sp. WP-5 and Pseudomonas sp. NN-4 are the potential candidates for inoculation to wheat as phosphate solubilizer even with reduced chemical fertilizer dose. The inoculation of the strains may enhance grain yield and net income of the farmer even with less chemical fertilizer application. This practice will be helpfull inminimizing environmental pollution.

Tahir M ，Naeem MA ，Shahid M ，Khalid U ，Farooq ABU ，Ahmad N ，Ahmad I ，Arshad M ，Waqar A ... -  《-》

Phosphate solubilizing bacteria with glucose dehydrogenase gene for phosphorus uptake and beneficial effects on wheat.

Suleman M ，Yasmin S ，Rasul M ，Yahya M ，Atta BM ，Mirza MS ... -  《PLoS One》

Assessment of two carrier materials for phosphate solubilizing biofertilizers and their effect on growth of wheat (Triticum aestivum L.).

Biofertilizers are usually carrier-based inoculants containing beneficial microorganisms. Incorporation of microorganisms in carrier material enables easy-handling, long-term storage and high effectiveness of biofertilizers. Objective of the present study was to assess enriched biogas sludge and soil as biofertilizer carriers on growth and yield of wheat. Six phosphate solubilizing strains were used in this study. Three phosphate solubilizing strains, 77-NS2 (Bacillus endophyticus), 77-CS-S1 (Bacillus sphaericus) and 77-NS5 (Enterobacter aerogenes) were isolated from the rhizosphere of sugarcane, two strains, PSB5 (Bacillus safensis) and PSB12 (Bacillus megaterium) from the rhizosphere of wheat and one halophilic phosphate solubilizing strain AT2RP3 (Virgibacillus sp.) from the rhizosphere of Atriplex amnicola, were used as bioinoculants. Phosphate solubilization ability of these strains was checked in vitro in Pikovskaya medium, containing rock phosphate (RP) as insoluble P source, individually supplemented with three different carbon sources, i.e., glucose, sucrose and maltose. Maximum phosphate solubilization; 305.6μg/ml, 217.2μg/ml and 148.1μg/ml was observed in Bacillus strain PSB12 in Pikovskaya medium containing sucrose, maltose and glucose respectively. A field experiment and pot experiments in climate control room were conducted to study the effects of biogas sludge and enriched soil based phosphorous biofertilizers on growth of wheat. Bacillus strain PSB12 significantly increased root and shoot dry weights and lengths using biogas sludge as carrier material in climate control room experiments. While in field conditions, significant increase in root and shoot dry weights, lengths and seed weights was seen by PSB12 and PSB5 (Bacillus) and Enterobacter strain 77-NS5 using biogas sludge as carrier. PSB12 also significantly increased both root and shoot dry weights and lengths in field conditions when used as enriched soil based inoculum. These results indicated that bacterial isolates having plant beneficial traits such as P solubilization are more promising candidates as biofertilizer when used with carrier materials.

Mukhtar S ，Shahid I ，Mehnaz S ，Malik KA ... -  《-》