Differential Root Exudation and Architecture for Improved Growth of Wheat Mediated by Phosphate Solubilizing Bacteria.

Phosphorous (P) deficiency is a major challenge faced by global agriculture. Phosphate-solubilizing bacteria (PSB) provide a sustainable approach to supply available phosphates to plants with improved crop productivity through synergistic interaction with plant roots. The present study demonstrates an insight into this synergistic P-solubilizing mechanism of PSB isolated from rhizosphere soils of major wheat-growing agro-ecological zones of Pakistan. Seven isolates were the efficient P solubilizers based on in vitro P-solubilizing activity (233-365 μg ml-1) with a concomitant decrease in pH (up to 3.5) by the production of organic acids, predominantly acetic acid (∼182 μg ml-1) and gluconic acid (∼117 μg ml-1). Amplification and phylogenetic analysis of gcd, pqqE, and phy genes of Enterobacter sp. ZW32, Ochrobactrum sp. SSR, and Pantoea sp. S1 showed the potential of these PSB to release orthophosphate from recalcitrant forms of phosphorus. Principal component analysis indicates the inoculation response of PSB consortia on the differential composition of root exudation (amino acids, sugars, and organic acids) with subsequently modified root architecture of three wheat varieties grown hydroponically. Rhizoscanning showed a significant increase in root parameters, i.e., root tips, diameter, and surface area of PSB-inoculated plants as compared to uninoculated controls. Efficiency of PSB consortia was validated by significant increase in plant P and oxidative stress management under P-deficient conditions. Reactive oxygen species (ROS)-induced oxidative damages mainly indicated by elevated levels of malondialdehyde (MDA) and H2O2 contents were significantly reduced in inoculated plants by the production of antioxidant enzymes, i.e., superoxide dismutase, catalase, and peroxidase. Furthermore, the inoculation response of these PSB on respective wheat varieties grown in native soils under greenhouse conditions was positively correlated with improved plant growth and soil P contents. Additionally, grain yield (8%) and seed P (14%) were significantly increased in inoculated wheat plants with 20% reduced application of diammonium phosphate (DAP) fertilizer under net house conditions. Thus, PSB capable of such synergistic strategies can confer P biofortification in wheat by modulating root morphophysiology and root exudation and can alleviate oxidative stress under P deficit conditions.

Yahya M ，Islam EU ，Rasul M ，Farooq I ，Mahreen N ，Tawab A ，Irfan M ，Rajput L ，Amin I ，Yasmin S ... -  《Frontiers in Microbiology》

Integrated analysis of potential microbial consortia, soil nutritional status, and agro-climatic datasets to modulate P nutrient uptake and yield effectiveness of wheat under climate change resilience.

Climate change has a devastating effect on wheat production; therefore, crop production might decline by 2030. Phosphorus (P) nutrient deficiency is another main limiting factor of reduced yield. Hence, there is a dire need to judiciously consider wheat yield, so that human requirements and nutrition balance can be sustained efficiently. Despite the great significance of biostimulants in sustainable agriculture, there is still a lack of integrated technology encompassing the successful competitiveness of inoculated phosphate-solubilizing bacteria (PSB) in agricultural systems in the context of climatic conditions/meteorological factors and soil nutritional status. Therefore, the present study reveals the modulation of an integrated P nutrient management approach to develop potential PSB consortia for recommended wheat varieties by considering the respective soil health and agro-climatic conditions. The designed consortia were found to maintain adequate viability for up to 9 months, verified through field emission scanning electron microscopy and viable count. Furthermore, a significant increase in grain yield (5%-8%) and seed P (4%) content was observed in consortia-inoculated wheat plants with 20% reduced Diammonium phosphate (DAP) application under net house conditions. Fluorescence in situ hybridization analysis of roots and amplification of the gcd gene of Ochrobactrum sp. SSR indicated the survival and rhizosphere competency of the inoculated PSB. Categorical principal component analysis (CAT-PCA) showed a positive correlation of inoculated field-grown wheat varieties in native soils to grain yield, soil P content, and precipitation for sites belonging to irrigated plains and seed P content, soil organic matter, and number of tillers for sites belonging to Northern dry mountains. However, the impact of inoculation at sites belonging to the Indus delta was found significantly correlated to soil potassium (K) content, electrical conductivity (EC), and temperature. Additionally, a significant increase in grain yield (15%) and seed P (14%) content was observed in inoculated wheat plants. Thus, the present study demonstrates for the first time the need to integrate soil biological health and agro-climatic conditions for consistent performance of augmented PSB and enhanced P nutrient uptake to curtail soil pollution caused by the extensive use of agrochemicals. This study provides innovative insights and identifies key questions for future research on PSB to promote its successful implementation in agriculture.

Yahya M ，Rasul M ，Hussain SZ ，Dilawar A ，Ullah M ，Rajput L ，Afzal A ，Asif M ，Wubet T ，Yasmin S ... -  《Frontiers in Plant Science》

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》

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 ... -  《-》

Designing Synergistic Biostimulants Formulation Containing Autochthonous Phosphate-Solubilizing Bacteria for Sustainable Wheat Production.

Applying phosphate-solubilizing bacteria (PSB) as biofertilizers has enormous potential for sustainable agriculture. Despite this, there is still a lack of information regarding the expression of key genes related to phosphate-solubilization (PS) and efficient formulation strategies. In this study, we investigated rock PS by Ochrobactrum sp. SSR (DSM 109610) by relating it to bacterial gene expression and searching for an efficient formulation. The quantitative PCR (qPCR) primers were designed for PS marker genes glucose dehydrogenase (gcd), pyrroloquinoline quinone biosynthesis protein C (pqqC), and phosphatase (pho). The SSR-inoculated soil supplemented with rock phosphate (RP) showed a 6-fold higher expression of pqqC and pho compared to inoculated soil without RP. Additionally, an increase in plant phosphorous (P) (2%), available soil P (4.7%), and alkaline phosphatase (6%) activity was observed in PSB-inoculated plants supplemented with RP. The root architecture improved by SSR, with higher root length, diameter, and volume. Ochrobactrum sp. SSR was further used to design bioformulations with two well-characterized PS, Enterobacter spp. DSM 109592 and DSM 109593, using the four organic amendments, biochar, compost, filter mud (FM), and humic acid. All four carrier materials maintained adequate survival and inoculum shelf life of the bacterium, as indicated by the field emission scanning electron microscopy analysis. The FM-based bioformulation was most efficacious and enhanced not only wheat grain yield (4-9%) but also seed P (9%). Moreover, FM-based bioformulation enhanced soil available P (8.5-11%) and phosphatase activity (4-5%). Positive correlations were observed between the PSB solubilization in the presence of different insoluble P sources, and soil available P, soil phosphatase activity, seed P content, and grain yield of the field grown inoculated wheat variety Faisalabad-2008, when di-ammonium phosphate fertilizer application was reduced by 20%. This study reports for the first time the marker gene expression of an inoculated PSB strain and provides a valuable groundwork to design field scale formulations that can maintain inoculum dynamics and increase its shelf life. This may constitute a step-change in the sustainable cultivation of wheat under the P-deficient soil conditions.

Yahya M ，Rasul M ，Sarwar Y ，Suleman M ，Tariq M ，Hussain SZ ，Sajid ZI ，Imran A ，Amin I ，Reitz T ，Tarkka MT ，Yasmin S ... -  《Frontiers in Microbiology》