Drought-tolerant rhizobacteria with predicted functional traits enhanced wheat growth and P uptake under moderate drought and low P-availability.

This study aims to investigate the effect of isolated drought-tolerant rhizobacteria, spanning various groups, such as nitrogen-fixing bacteria (NFB), phosphate solubilizing bacteria (PSB), and other plant growth promoting rhizobacteria (PGPR), on the growth of wheat (Triticum durum) plants, focusing on various morphological and physiological responses under moderate drought and low-P availability. Among 343 rhizobacterial morphotypes, 16 exhibited tolerance to NaCl and PEG-6000. These included 8 PSB, 4 NFB, and 4 osmotolerant-PGPR groups, distributed across 14 different genera. Biochemical characterization showcased diverse PGP capabilities, particularly in P solubilization. The dynamic responses of drought-tolerant PSB to salt and PEG-6000-induced drought stress involved variations in organic acid (OA) secretion, with specific acids, including palmitic, lactic, and stearic, playing crucial roles in enhancing available P fractions. Inoculation with rhizobacteria significantly increased both shoot (SDW) and root (RDW) dry weights of wheat plants, as well as rhizosphere available P. PSB11 (Arthrobacter oryzae) emerged as the most effective strain, plausibly due to its positive impact on root morphological traits (length, surface, and volume). Other isolates, PSB10 (Priestia flexa), PSB13 (Bacillus haynesii), and particularly PGPR2 (Arthrobacter pascens) significantly increased shoot P content (up to 68.91 %), with a 2-fold increase in chlorophyll content. The correlation analysis highlighted positive associations between SDW, shoot P content, chlorophyll content index (CCI), and leaf area. Additionally, a negative correlation emerged between microbial biomass P and root morphophysiological parameters. This pattern could be explained by reduced competition between plants and rhizobacteria for accessible P, as indicated by low microbial biomass P and strong plant growth. Our investigation reveals the potential of drought-tolerant rhizobacteria in enhancing wheat resilience to moderate drought and low-P conditions. This is demonstrated through exceptional performance in influencing root architecture, P utilization efficiency, and overall plant physiological parameters. Beyond these outcomes, the innovative isolation procedure employed, targeting rhizobacteria from diverse groups, opens new avenues for targeted isolation techniques. This unique approach contributes to the novelty of our study, offering promising prospects for targeted bioinoculants in mitigating the challenges of drought and P deficiency in wheat cultivation.

Agronomic advantage of bacterial biological nitrogen fixation on wheat plant growth under contrasting nitrogen and phosphorus regimes.

Introduction: Given their remarkable capacity to convert atmospheric nitrogen into plant-accessible ammonia, nitrogen-fixing microbial species hold promise as a sustainable alternative to chemical nitrogen fertilizers, particularly in economically significant crops like wheat. This study aimed to identify strains with optimal attributes for promoting wheat growth sustainably, with a primary emphasis on reducing reliance on chemical nitrogen fertilizers. Methods: We isolated free nitrogen-fixing strains from diverse rhizospheric soils across Morocco. Subsequently, we conducted a rigorous screening process to evaluate their plant growth-promoting traits, including nitrogen fixation, phosphate solubilization, phytohormone production and their ability to enhance wheat plant growth under controlled conditions. Two specific strains, Rhodotorula mucilaginosa NF 516 and Arthrobacter sp. NF 528, were selected for in-depth evaluation, with the focus on their ability to reduce the need for chemical nitrogen supply, particularly when used in conjunction with TSP fertilizer and natural rock phosphate. These two sources of phosphate were chosen to assess their agricultural effectiveness on wheat plants. Results and discussion: Twenty-two nitrogen-fixing strains (nif-H+) were isolated from various Moroccan rhizospheric soils, representing Bacillus sp., Pseudomonas sp., Arthrobacter sp., Burkholderia sp. and a yeast-like microorganism. These strains were carefully selected based on their potential to promote plant growth. The findings revealed that the application of Rhodotorula mucilaginosa NF 516 and Arthrobacter sp. NF 528 individually or in combination, significantly improved wheat plant growth and enhanced nutrients (N and P) uptake under reduced nitrogen regimes. Notably, their effectiveness was evident in response to both natural rock phosphate and TSP, demonstrating their important role in wheat production under conditions of low nitrogen and complex phosphorus inputs. This research underscores the significant role of nitrogen-fixing microorganisms, particularly Rhodotorula mucilaginosa NF 516 and Arthrobacter sp. NF 528, in wheat production under conditions of low nitrogen and complex phosphorus inputs. It showcases their potential to reduce chemical nitrogen fertilization requirements by up to 50% without compromising wheat plant yields. Our study emphasizes the importance of bacterial biological nitrogen fixation in meeting the remaining nitrogen requirements beyond this reduction. This underscores the vital role of microbial contributions in providing essential nitrogen for optimal plant growth and highlights the significance of biological nitrogen fixation in sustainable agriculture practices.

Bioinoculants as a means of increasing crop tolerance to drought and phosphorus deficiency in legume-cereal intercropping systems.

Ensuring plant resilience to drought and phosphorus (P) stresses is crucial to support global food security. The phytobiome, shaped by selective pressures, harbors stress-adapted microorganisms that confer host benefits like enhanced growth and stress tolerance. Intercropping systems also offer benefits through facilitative interactions, improving plant growth in water- and P-deficient soils. Application of microbial consortia can boost the benefits of intercropping, although questions remain about the establishment, persistence, and legacy effects within resident soil microbiomes. Understanding microbe- and plant-microbe dynamics in drought-prone soils is key. This review highlights the beneficial effects of rhizobacterial consortia-based inoculants in legume-cereal intercropping systems, discusses challenges, proposes a roadmap for development of P-solubilizing drought-adapted consortia, and identifies research gaps in crop-microbe interactions.

Integrated use of polyphosphate and P-solubilizing bacteria enhanced P use efficiency and growth performance of durum wheat.

Coupling phosphate-solubilizing bacteria (PSB) with P fertilizers, including polyphosphates (PolyP), was reported as eco-efficient approach to enhance P use efficiency. Although PSB have been recently reported to hydrolyze PolyP, the plant growth promoting mechanisms of PolyP-PSB co-application were not yet uncovered. This study aims to evaluate the effect of a PSB consortium (PSBCs) on growth, P use efficiency (PUE), and wheat yield parameters under PolyP (PolyB) application. Co-application of PolyB-PSBCs significantly enhanced wheat growth at 75 days after sowing (DAS) compared to 30 DAS. A significant increase in shoot dry biomass (47%), shoot inorganic P content (222%), PUE (91%), and root P absorption efficiency (RPAE, 99%) was noted compared to unfertilized plants. Similarly, the PolyB-PSBCs co-application enhanced morphological root traits at 30 DAS, while acid phosphatase activities (root and rhizosphere), RPAE, and PUE were significantly increased at 75 DAS. The improved wheat P acquisition could be attributed to a lower investment in root biomass production, and significant induction of acid phosphatase activity in roots and rhizosphere soil under PolyB-PSBCs co-application. Consequently, the PolyB-PSBCs co-application significantly improved aboveground performance, which is reflected by increased shoot nutrient contents (P 300%, K 65%), dry weight (54%), and number (50%) of spikes. Altogether, this study provides relevant evidence that co-application of PolyP-PSBCs can be an integrated and environmentally preferred P fertilization approach owing to the dual effects of PolyP and PSBCs on wheat PUE.

Microbial Catabolic Activity: Methods, Pertinence, and Potential Interest for Improving Microbial Inoculant Efficiency.

Microbial catabolic activity (MCA) defined as the degrading activity of microorganisms toward various organic compounds for their growth and energy is commonly used to assess soil microbial function potential. For its measure, several methods are available including multi-substrate-induced respiration (MSIR) measurement which allow to estimate functional diversity using selected carbon substrates targeting specific biochemical pathways. In this review, the techniques used to measure soil MCA are described and compared with respect to their accuracy and practical use. Particularly the efficiency of MSIR-based approaches as soil microbial function indicators was discussed by (i) showing their sensitivity to different agricultural practices including tillage, amendments, and cropping systems and (ii) by investigating their relationship with soil enzyme activities and some soil chemical properties (pH, soil organic carbon, cation exchange capacity). We highlighted the potential of these MSIR-based MCA measurements to improve microbial inoculant composition and to determine their potential effects on soil microbial functions. Finally, we have proposed ideas for improving MCA measurement notably through the use of molecular tools and stable isotope probing which can be combined with classic MSIR methods. Graphical abstract describing the interrelation between the different parts and the concepts developed in the review.

The current status and challenges of biomass biorefineries in Africa: A critical review and future perspectives for bioeconomy development.

Africa benefits from diverse biomasses that are rich in high-added value materials and precursors for energy, food, agricultural, cosmetic and medicinal applications. Many African countries are interested in valorizing biomasses to develop efficient and integrated biorefinery processes and their use for local and regional economic development. Thus, this report critically reviews the current status of African biomass richness, its diversity, and potential applications. Moreover, particular attention is given to bioenergy production, mainly by biological and thermochemical conversion processes. This also includes biomass valorization in agriculture, particularly for the production of plant-based biostimulants, which are a potential emerging agri-input sector worldwide. This study points out that even though several processes for biofuel, biogas, biofertilizer and biostimulant production have already been established in Africa, their development on a larger scale remains limited. This study also reports the different socioeconomic and political aspects of biomass applications, along with their challenges, opportunities, and future research perspectives, to promote concrete technologies transferable into an industrial level.

Screening of potential phosphate solubilizing bacteria inoculants should consider the contrast in phosphorus bio-solubilization rate along with plant growth promotion and phosphorus use efficiency.

AIMS: Although phosphate solubilizing bacteria (PSB) have been globally reported to improve soil phosphorus (P) availability and plant growth, technical gaps such as the lack of an ideal screening approach, is yet to be addressed. The potential of non-halo-forming PSB remains underestimated because of the currently adopted screening protocols that exclusively consider halo-forming and PSB with high phosphorus solubilization (PS) capacities. Yet, caution should be taken to properly assess PSB with contrasting PS rates regardless of the presence or absence of the solubilization halo. METHODS AND RESULTS: This study sought to examine the PS rate and plant growth promotion ability of 12 PSB categorized as high PSB (H-PSB), medium PSB (M-PSB), and low PSB (L-PSB) based on their PS rates of rock phosphate (RP). The non-halo-forming PSB Arthrobacter pascens was categorized as H-PSB, which might have been eliminated during the classical screening process. In addition, induction of organic acids and phosphatase activity in rhizosphere soils by H-, M-, and L-PSB was proportional to increased wheat P content by 143.22, 154.21, and 77.76 mg P g-1 compared to uninoculated plants (18.1 mg P g-1). CONCLUSIONS: Isolates considered as M- and L-PSB could positively influence wheat above-ground physiology and root traits as high as H-PSB. In addition, non-halo-forming PSB revealed significant PS rates along with positive effects on plant growth as high as halo-forming PSB.

Polyphosphate application influences morpho-physiological root traits involved in P acquisition and durum wheat growth performance.

BACKGROUND: Among phosphate (P) fertilizers, polyphosphates (PolyPs) have shown promising results in terms of crop yield and plant P nutrition. However, compared to conventional P inputs, very little is known on the impact of PolyPs fertilizers on below- and above-ground plant functional traits involved in P acquisition. This study aims to evaluate agro-physiological responses of durum wheat variety ´Karim´ under different PolyPs applications. Three PolyPs fertilizers (PolyA, PolyB, and PolyC) versus one orthophosphate (OrthoP) were applied at three doses; 30 (D30), 60 (D60), and 90 (D90) kg P/ha under controlled conditions. The PolyPs (especially PolyB and PolyC) application at D60 significantly increased morphophysiological root traits (e.g., RL: 42 and 130%; RSA:40 and 60%), shoot inorganic P (Pi) content (159 and 88%), and root P acquisition efficiency (471 and 296%) under PolyB and PolyC, respectively compared to unfertilized plants. Above-ground physiological parameters, mainly nutrient acquisition, chlorophyll content and chlorophyll fluorescence parameters were also improved under PolyB and PolyA application at D60. A significant and positive correlation between shoot Pi content and rhizosphere soil acid phosphatase activity was observed, which reveal the key role of these enzymes in PolyPs (A and B) use efficiency. Furthermore, increased P uptake/RL ratio along with shoot Pi indicates more efficient P allocation to shoots with less investment in root biomass production under PolyPs (especially A and B). CONCLUSIONS: Under our experimental conditions, these findings report positive impacts of PolyPs on wheat growth performance, particularly on photosynthesis and nutrient acquisition at D60, along with modulation of root morpho-physiological traits likely responsible of P acquisition efficiency.

Phosphate solubilizing bacteria can significantly contribute to enhance P availability from polyphosphates and their use efficiency in wheat.

Rhizosphere microbes significantly enhance phosphorus (P) availability from a variety of unavailable P pools in agricultural soils. However, little is known about the contribution of root-associated microorganisms, notably P solubilizing bacteria (PSB), to enhance the use of polyphosphate (PolyP) fertilizers as well as the key mechanisms involved. This study assesses the ability of four PSB (Bacillus siamensis, Rahnella aceris, Pantoea hericii, Bacillus paramycoides) and their consortium (Cs) to enhance the release rate of available P from two types of PolyP ("PolyB" and "PolyC") with a focus on the key role of phosphatase enzyme activities and organic acids production. Wheat growth performance and P acquisition efficiency were evaluated in response to co-application of PSB and PolyP. Results showed that inoculation with PSB, notably Cs, significantly enhanced available P from PolyC, PolyB and tri-calcium P. Increased available P in response to inoculation with PSB significantly correlated with medium acidification, organic acids production (notably glycolic acid) and induced activities of acid phosphatase and pyrophosphatase. In planta, the co-application of PSB-PolyP improved wheat plant biomass, root growth and P acquisition, with best results obtained from Cs-PolyP co-application as compared to uninoculated and unfertilized plants. At seedling stage, the co-application of Cs-PolyP (PolyB and PolyC) enhanced root hairs length (125 % and 131 %), root length (26 % and 37 %) and root inorganic P (Pi) content (160 % and 182 %), respectively compared to uninoculated plants. Similarly, at tillering stage, plant biomass (35 % and 47 %), Pi content (43 % and 253 %), P translocation (215 % and 315 %) and soil phosphatases (213 % and 219 %) significantly improved under PolyB and PolyC application, respectively. Findings from this study demonstrate the key role of PSB to enhance the use of PolyP through production of organic acids and phosphatases, exhibiting differential traits patterns between the two PolyP. Improved wheat growth and root P acquisition in response to PSB-PolyP co-application can be attributed to induced rhizosphere processes leading to enhanced available P taken up by roots.

A phospho-compost biological-based approach increases phosphate rock agronomic efficiency in faba bean as compared to chemical and physical treatments.

Under arid and semi-arid conditions, direct application of phosphate rock (PR) as a source of phosphorus (P) for crop production is likely influenced by agricultural practices and soil properties. Different approaches could be used to improve the agronomic efficiency of low-grade PR over a wider range of soils and crops. In this study, biological, physical, and chemical treatments of low-grade Moroccan PR were investigated and compared through agronomic trials on faba bean grown under alkaline soil conditions. The physical treatment was based on blending PR with triple superphosphate (TSP) at 75:25 and 50:50 ratios, the biological treatments involved co-application of PR with compost at 50:50 ratio and phospho-compost elaborated from PR (20%), sewage sludge (46%), and wheat residues (34%), while the chemical treatment was obtained by a 30% acidulation of PR by phosphoric acid. Control treatments consisting of zero P application (control), PR alone, and TSP alone were considered to assess the effectiveness of the abovementioned techniques to improve PR agronomic efficiency. A pot experiment was conducted in sandy soil (Jorf Lasfar, central Morocco) for 60 days in a completely randomized design considering eight treatments. All treatments, except the control, were amended with 52 mg kg-1 of P from different PR-based fertilizers before sowing. At the flowering stage (60-day-old plants), results indicated that all PR treatments significantly improved plant growth, root nodulation, and nutrient uptake compared to the control. The relative agronomic efficiency of pretreated PR was significantly higher with phospho-compost treatment (86%) than the partially acidulated PR (78%) or the PR/TSP blend 50:50 (64%). Likewise, P uptake, P use efficiency, number of root nodules, and N uptake all were improved under PR treatments. Our finding revealed that the biological technique based on phospho-compost yielded better compared to chemical and physical treatments.

Phosphate bacterial solubilization: A key rhizosphere driving force enabling higher P use efficiency and crop productivity.

Background: Increasing crop production to feed a growing population has driven the use of mineral fertilizers to ensure nutrients availability and fertility of agricultural soils. After nitrogen, phosphorus (P) is the second most important nutrient for plant growth and productivity. However, P availability in most agricultural soils is often limited because P strongly binds to soil particles and divalent cations forming insoluble P-complexes. Therefore, there is a constant need to sustainably improve soil P availability. This may include, among other strategies, the application of microbial resources specialized in P cycling, such as phosphate solubilizing bacteria (PSB). This P-mediating bacterial component can improve soil biological fertility and crop production, and should be integrated in well-established formulations to enhance availability and efficiency in use of P. This is of importance to P fertilization, including both organic and mineral P such as rock phosphate (RP) aiming to improve its agronomic efficiency within an integrated crop nutrition system where agronomic profitability of P and PSB can synergistically occur. Aim of Review: The purpose of this review is to discuss critically the important contribution of PSB to crop P nutrition in concert with P fertilizers, with a specific focus on RP. We also highlight the need for PSB bioformulations being a sustainable approach to enhance P fertilizer use efficiency and crop production. Key Scientific Concepts of Review: We first recognize the important contribution of PSB to sustain crop production, which requires a rational approach for both screening and evaluation of PSB enabling an accurate assessment of the bacterial effects both alone and in intertwined interaction with plant roots. Furthermore, we propose new research ideas about the development of microbial bioformulations based on PSB with a particular focus on strains exhibiting synergetic effects with RP.

Benefits of phosphate solubilizing bacteria on belowground crop performance for improved crop acquisition of phosphorus.

Although research on plant growth promoting bacteria began in the 1950s, basic and applied research on bacteria improving use of phosphorus (P) continues to be a priority among many agricultural research institutions. Ultimately, identifying agriculturally beneficial microbes, notably P solubilizing bacteria (PSB), that enhance the efficient use of P supports more sustainable cropping systems and the judicious use of mineral nutrients. In parallel, there is more attention on improving crop root P acquisition of existing soil P pools as well as by increasing the proportion of fertilizer P that is taken up by crops. Today, new lines of research are emerging to investigate the co-optimization of PSB-fertilizer-crop root processes for improved P efficiency and agricultural performance. In this review, we compile and summarize available findings on the beneficial effects of PSB on crop production with a focus on crop P acquisition via root system responses at the structural, functional and transcriptional levels. We discuss the current state of knowledge on the mechanisms of PSB-mediated P availability, both soil- and root-associated, as well as crop uptake via P solubilization, mineralization and mobilization, mainly through the production of organic acids and P-hydrolyzing enzymes, and effects on phytohormone signaling for crop root developement. The systematic changes caused by PSB on crop roots are discussed and contextualized within promising functional trait-based frameworks. We also detail agronomic profitability of P (mineral and organic) and PSB co-application, in amended soils and inoculated crops, establishing the connection between the influence of PSB on agroecosystem production and the impact of P fertilization on microbial diversity and crop functional traits for P acquisition.

Defining the wheat microbiome: Towards microbiome-facilitated crop production.

Wheat is one of the world's most important crops, but its production relies heavily on agrochemical inputs which can be harmful to the environment when used excessively. It is well known that a multitude of microbes interact with eukaryotic organisms, including plants, and the sum of microbes and their functions associated with a given host is termed the microbiome. Plant-microbe interactions can be beneficial, neutral or harmful to the host plant. Over the last decade, with the development of next generation DNA sequencing technology, our understanding of the plant microbiome structure has dramatically increased. Considering that defining the wheat microbiome is key to leverage crop production in a sustainable way, here we describe how different factors drive microbiome assembly in wheat, including crop management, edaphic-environmental conditions and host selection. In addition, we highlight the benefits to take a multidisciplinary approach to define and explore the wheat core microbiome to generate solutions based on microbial (synthetic) communities or single inoculants. Advances in plant microbiome research will facilitate the development of microbial strategies to guarantee a sustainable intensification of crop production.

Nitrogen Fixing Azotobacter Species as Potential Soil Biological Enhancers for Crop Nutrition and Yield Stability.

Biological nitrogen fixation (BNF) refers to a microbial mediated process based upon an enzymatic "Nitrogenase" conversion of atmospheric nitrogen (N2) into ammonium readily absorbable by roots. N2-fixing microorganisms collectively termed as "diazotrophs" are able to fix biologically N2 in association with plant roots. Specifically, the symbiotic rhizobacteria induce structural and physiological modifications of bacterial cells and plant roots into specialized structures called nodules. Other N2-fixing bacteria are free-living fixers that are highly diverse and globally widespread in cropland. They represent key natural source of nitrogen (N) in natural and agricultural ecosystems lacking symbiotic N fixation (SNF). In this review, the importance of Azotobacter species was highlighted as both important free-living N2-fixing bacteria and potential bacterial biofertilizer with proven efficacy for plant nutrition and biological soil fertility. In addition, we described Azotobacter beneficial plant promoting traits (e.g., nutrient use efficiency, protection against phytopathogens, phytohormone biosynthesis, etc.). We shed light also on the agronomic features of Azotobacter that are likely an effective component of integrated plant nutrition strategy, which contributes positively to sustainable agricultural production. We pointed out Azotobacter based-biofertilizers, which possess unique characteristics such as cyst formation conferring resistance to environmental stresses. Such beneficial traits can be explored profoundly for the utmost aim to research and develop specific formulations based on inoculant Azotobacter cysts. Furthermore, Azotobacter species still need to be wisely exploited in order to address specific agricultural challenges (e.g., nutrient deficiencies, biotic and abiotic constraints) taking into consideration several variables including their biological functions, synergies and multi-trophic interactions, and biogeography and abundance distribution.

Phosphate Solubilizing Rhizobacteria Could Have a Stronger Influence on Wheat Root Traits and Aboveground Physiology Than Rhizosphere P Solubilization.

Limited P availability in several agricultural areas is one of the key challenges facing current agriculture. Exploiting P-solubilizing bacteria (PSB) has been an emerging bio-solution for a higher rhizosphere P-availability, meanwhile the above- and below-ground interactions that PSB would trigger remain unclear over plant growing stages. We hypothesized that PSB effects on plant growth may be greater on root traits that positively links with aboveground physiology more than the commonly believed rhizosphere P bio-solubilization. In this study, five contrasting PSB (Pseudomonas spp.) isolates (low "PSB1", moderate "PSB2 and PSB4" and high "PSB3 and PSB5" P-solubilizing capacity "PSC") were used to investigate above- and below-ground responses in wheat fertilized with rock P (RP) under controlled conditions. Our findings show that all PSB isolates increased wheat root traits, particularly PSB5 which increased root biomass and PSB3 that had greater effect on root diameter in 7-, 15- and 42-day old plants. The length, surface and volume of roots significantly increased along with higher rhizosphere available P in 15- and 42-day old plants inoculated with PSB4 and PSB2. Shoot biomass significantly increased with both PSB2 and PSB5. Root and shoot physiology significantly improved with PSB1 (lowest PSC) and PSB4 (moderate PSC), notably shoot total P (78.38%) and root phosphatase activity (390%). Moreover, nutrients acquisition and chlorophyll content increased in inoculated plants and was stimulated (PSB2, PSB4) more than rhizosphere P-solubilization, which was also revealed by the significant above- and below-ground inter-correlations, mainly chlorophyll and both total (R = 0.75, p = 0.001**) and intracellular (R = 0.7, p = 0.000114*) P contents. These findings demonstrate the necessity to timely monitor the plant-rhizosphere continuum responses, which may be a relevant approach to accurately evaluate PSB through considering below- and above-ground relationships; thus enabling unbiased interpretations prior to field applications.

Soil Microbial Resources for Improving Fertilizers Efficiency in an Integrated Plant Nutrient Management System.

Tomorrow's agriculture, challenged by increasing global demand for food, scarcity of arable lands, and resources alongside multiple environment pressures, needs to be managed smartly through sustainable and eco-efficient approaches. Modern agriculture has to be more productive, sustainable, and environmentally friendly. While macronutrients such as nitrogen (N), phosphorus (P), potassium (K), and sulfur (S) supplied by mineral fertilizers are vital to crop production, agriculturally beneficial microorganisms may also contribute directly (i.e., biological N2 fixation, P solubilization, and phytohormone production, etc.) or indirectly (i.e., antimicrobial compounds biosynthesis and elicitation of induced systemic resistance, etc.) to crop improvement and fertilizers efficiency. Microbial-based bioformulations that increase plant performance are greatly needed, and in particular bioformulations that exhibit complementary and synergistic effects with mineral fertilization. Such an integrated soil fertility management strategy has been demonstrated through several controlled and non-controlled experiments, but more efforts have to be made in order to thoroughly understand the multiple functions of beneficial microorganisms within the soil microbial community itself and in interaction with plants and mineral resources. In fact, the combined usage of microbial [i.e., beneficial microorganisms: N2-fixing (NF), P-solubilizing, and P mobilizing, etc.] and mineral resources is an emerging research area that aims to design and develop efficient microbial formulations which are highly compatible with mineral inputs, with positive impacts on both crops and environment. This novel approach is likely to be of a global interest, especially in most N- and P-deficient agro-ecosystems. In this review, we report on the importance of NF bacteria and P solubilizing/mobilizing microbes as well as their interactions with mineral P fertilization in improving crop productivity and fertilizers efficiency. In addition, we shed light on the interactive and synergistic effects that may occur within multi-trophic interactions involving those two microbial groups and positive consequences on plant mineral uptake, crop productivity, and resiliency to environmental constraints. Improving use of mineral nutrients is a must to securing higher yield and productivity in a sustainable manner, therefore continuously designing, developing and testing innovative integrated plant nutrient management systems based on relevant biological resources (crops and microorganisms) is highly required.

Expression of a phosphate-starvation inducible fructose-1,6-bisphosphatase gene in common bean nodules correlates with phosphorus use efficiency.

While increased P-hydrolysing acid phosphatases (APase) activity in bean nodules is well documented under phosphorus (P) limitation, gene expression and subcellular localization patterns within the N2-fixing nodule tissues are poorly understood. The aim of this research was to track the enzyme activity along with the intra-nodular localization of fructose-1,6-bisphosphatase (FBPase), and its contribution to P use efficiency (PUE) under symbiotic nitrogen fixation (SNF) in Phaseolus vulgaris. The FBPase transcript were localized in situ using RT-PCR and the protein activity was measured in nodules of two contrasting recombinant inbred lines (RILs) of P. vulgaris, namely RILs 115 (P-efficient) and 147 (P-inefficient), that were grown under sufficient versus deficient P supply. Under P-deficiency, higher FBPase transcript fluorescence was found in the inner cortex as compared to the infected zone of RIL115. In addition, both the specific FBPase and total APase enzyme activities significantly increased in both RILs, but to a more significant extent in RIL115 as compared to RIL147. Furthermore, the increased FBPase activity in nodules of RIL115 positively correlated with higher use efficiency of both the rhizobial symbiosis (23%) and P for SNF (14% calculated as the ratio of N2 fixed per nodule total P content). It is concluded that the abundant tissue-specific localized FBPase transcript along with induced enzymatic activity provides evidence of a specific tolerance mechanism where N2-fixing nodules overexpress under P-deficiency conditions. Such a mechanism would maximise the intra-nodular inorganic P fraction necessary to compensate for large amount of P needed during the SNF process.

Localization of phytase transcripts in germinating seeds of the common bean (Phaseolus vulgaris L.).

MAIN CONCLUSION: The work provides the first-time evidence of tissue-specific expression of a phytase gene in the germinating seeds of Phaseolus vulgaris. Phytase enzyme plays a major role in germinating seeds. It is also active during N2 fixation within nodules of legumes. The effect of phosphorus (P) deficiency on phytase gene expression and localization in N2-fixing root nodules has been recently studied in hydroaeroponic culture of Phaseolus vulgaris. In this study, phytase gene transcripts within the germinating seed tissues of the P-inefficient P. vulgaris recombinant inbred line RIL147 were in situ localized with a similar RT-PCR recipe as that used for nodules. Our results show that the phytase gene expression was mainly localized in the outer layers, vascular cells and parenchyma of germinating seeds whereas it was localized in the inner and middle cortex of nodules. Image analysis quantified higher fluorescence intensity of the phytase transcript signal in the seed embryo than in radicles, cotyledons or the nodule cortex. Furthermore, the phytase activity was 22-fold higher in cotyledons (43 nmol min(-1) g(-1) dry weight) than in nodules (2 nmol min(-1) g(-1) dry weight). The K m and V m values of phytase activity in cotyledons were also significantly higher than in nodules. Interestingly, the amplified sequence of cDNA phytase exhibited highest homology with the Glycine max purple acid phosphatase (NM_001289274) 90 % for germinating seed as compared to nodule phytase cDNA displaying 94 % homology with the Glycine max phytase (GQ422774.1). It is concluded that phytase enzymes are likely to vary from seeds to nodules and that phytase enzymes play key roles in the use of organic P or N2 fixation, as it is well known for germination.

Localization of the Bacillus subtilis beta-propeller phytase transcripts in nodulated roots of Phaseolus vulgaris supplied with phytate.

Soil organic phosphorus (Po) such as phytate, which comprises up to 80 % of total Po, must be hydrolyzed by specific enzymes called phytases to be used by plants. In contrast to plants, bacteria, such as Bacillus subtilis, have the ability to use phytate as the sole source of P due to the excretion of a beta-propeller phytase (BPP). In order to assess whether the B. subtilis BPP could make P available from phytate for the benefit of a nodulated legume, the P-sensitive recombinant inbred line RIL147 of Phaseolus vulgaris was grown under hydroaeroponic conditions with either 12.5 µM phytate (C6H18O24P6) or 75 µmol Pi (K2HPO4), and inoculated with Rhizobium tropici CIAT899 alone, or co-inoculated with both B. subtilis DSM 10 and CIAT899. The in situ RT-PCR of BPP genes displayed the most intense fluorescent BPP signal on root tips. Some BPP signal was found inside the root cortex and the endorhizosphere of the root tip, suggesting endophytic bacteria expressing BPP. However, the co-inoculation with B. subtilis was associated with a decrease in plant P content, nodulation and the subsequent plant growth. Such a competitive effect of B. subtilis on P acquisition from phytate in symbiotic nitrogen fixation might be circumvented if the rate of inoculation were reasoned in order to avoid the inhibition of nodulation by excess B. subtilis proliferation. It is concluded that B. subtilis BPP gene is expressed in P. vulgaris rhizosphere.