Inoculation Of Crops Research Articles

Antibiotic resistance in plant growth promoting bacteria: A comprehensive review and future perspectives to mitigate potential gene invasion risks.

Plant growth-promoting bacteria (PGPB) are endowed with several attributes that can be beneficial for host plants. They opened myriad doors toward green technology approach to reduce the use of chemical inputs, improve soil fertility, and promote plants’ health. However, many of these PGPB harbor antibiotic resistance genes (ARGs). Less attention has been given to multi-resistant bacterial bioinoculants which may transfer their ARGs to native soil microbial communities and other environmental reservoirs including animals, waters, and humans. Therefore, large-scale inoculation of crops by ARGs-harboring bacteria could worsen the evolution and dissemination of antibiotic resistance and aggravate the negative impacts on such ecosystem and ultimately public health. Their introduction into the soil could serve as ARGs invasion which may inter into the food chain. In this review, we underscore the antibiotic resistance of plant-associated bacteria, criticize the lack of consideration for this phenomenon in the screening and application processes, and provide some recommendations as well as a regulation framework relating to the development of bacteria-based biofertilizers to aid maximizing their value and applications in crop improvement while reducing the risks of ARGs invasion.

Effects of Rhizosphere Bacteria on Strawberry Plants (Fragaria × ananassa Duch.) under Water Deficit.

Due to the observed climate warming, water deficiency in soil is currently one of the most important stressors limiting the size and quality of plant crops. Drought stress causes a number of morphological, physiological, and biochemical changes in plants, limiting their growth, development, and yield. Innovative methods of inducing resistance and protecting plants against stressors include the inoculation of crops with beneficial microorganisms isolated from the rhizosphere of the plant species to which they are to be applied. The aim of the present study was to evaluate 12 different strains of rhizosphere bacteria of the genera Pantoea, Bacillus, Azotobacter, and Pseudomonas by using them to inoculate strawberry plants and assessing their impact on mitigating the negative effects of drought stress. Bacterial populations were assessed by estimates of their size based on bacterial counts in the growth substrate and with bioassays for plant growth-promoting traits. The physiological condition of strawberry plants was determined based on the parameters of chlorophyll fluorescence. The usefulness of the test methods used to assess the influence of plant inoculation with rhizosphere bacteria on the response of plants growing under water deficit was also evaluated. A two-factor experiment was performed in a complete randomization design. The first experimental factor was the inoculation of plant roots with rhizosphere bacteria. The second experimental factor was the different moisture content of the growth substrate. The water potential was maintained at −10 to −15 kPa under control conditions, and at −40 to −45 kPa under the conditions of water deficit in the substrate. The tests on strawberry plants showed that the highest sensitivity to water deficiency, and thus the greatest usefulness for characterizing water stress, was demonstrated by the following indices of chlorophyll “a” fluorescence: FM, FV, FV/FM, PI, and Area. Based on the assessment of the condition of the photosynthetic apparatus and the analysis of chlorophyll “a” fluorescence indices, including hierarchical cluster analysis, the following strains of rhizosphere bacteria were found to have favorable effects on strawberry plants under water deficit: the Bacillus sp. strains DLGB2 and DKB26 and the Pantoea sp. strains DKB63, DKB70, DKB68, DKB64, and DKB65. In the tests, these strains of Bacillus sp. exhibited a common trait—the ability to produce siderophores, while those of Pantoea sp. were notable for phosphate mobilization and ACCD activity.

Prospects for Using Phosphate-Solubilizing Microorganisms as Natural Fertilizers in Agriculture.

Phosphates are known to be essential for plant growth and development, with phosphorus compounds being involved in various physiological and biochemical reactions. Phosphates are known as one of the most important factors limiting crop yields. The problem of phosphorus deficiency in the soil has traditionally been solved by applying phosphate fertilizers. However, chemical phosphate fertilizers are considered ineffective compared to the organic fertilizers manure and compost. Therefore, increasing the bioavailability of phosphates for plants is one of the primary goals of sustainable agriculture. Phosphate-solubilizing soil microorganisms can make soil-insoluble phosphate bioavailable for plants through solubilization and mineralization. These microorganisms are currently in the focus of interest due to their advantages, such as environmental friendliness, low cost, and high biological efficiency. In this regard, the solubilization of phosphates by soil microorganisms holds strong potential in research, and inoculation of soils or crops with phosphate-solubilizing bacteria is a promising strategy to improve plant phosphate uptake. In this review, we analyze all the species of phosphate-solubilizing bacteria described in the literature to date. We discuss key mechanisms of solubilization of mineral phosphates and mineralization of organic phosphate-containing compounds: organic acids secreted by bacteria for the mobilization of insoluble inorganic phosphates, and the enzymes hydrolyzing phosphorus-containing organic compounds. We demonstrate that phosphate-solubilizing microorganisms have enormous potency as biofertilizers since they increase phosphorus bioavailability for the plant, promote sustainable agriculture, improve soil fertility, and raise crop yields. The use of phosphate-solubilizing microbes is regarded as a new frontier in increasing plant productivity.

The Role of Synthetic Microbial Communities (SynCom) in Sustainable Agriculture

Modern agriculture faces several challenges due to climate change, limited resources, and land degradation. Plant-associated soil microbes harbor beneficial plant growth-promoting (PGP) traits that can be used to address some of these challenges. These microbes are often formulated as inoculants for many crops. However, inconsistent productivity can be a problem since the performance of individual inoculants/microbes vary with environmental conditions. Over the past decade, the ability to utilize Next Generation Sequencing (NGS) approaches with soil microbes has led to an explosion of information regarding plant associated microbiomes. Although this type of work has been predominantly sequence-based and often descriptive in nature, increasingly it is moving towards microbiome functionality. The synthetic microbial communities (SynCom) approach is an emerging technique that involves co-culturing multiple taxa under well-defined conditions to mimic the structure and function of a microbiome. The SynCom approach hopes to increase microbial community stability through synergistic interactions between its members. This review will focus on plant-soil-microbiome interactions and how they have the potential to improve crop production. Current approaches in the formulation of synthetic microbial communities will be discussed, and its practical application in agriculture will be considered.

The Mechanisms of Sodium Chloride Stress Mitigation by Salt-Tolerant Plant Growth Promoting Rhizobacteria in Wheat

We investigated the growth-promoting mechanism of salt-tolerant plant growth promoting rhizobacteria (ST-PGPR) in wheat under sodium chloride (NaCl) stress by measuring the growth and physiological and biochemical responses of wheat plants inoculated with ST-PGPR under 0–400 mM NaCl. The results showed that ST-PGPR plays a significant role in the growth of wheat under NaCl stress. Under 300 mM NaCl, wheat plants inoculated with the three ST-PGPR strains increased in plant height, root length, dry weight, and fresh weight by 71.21%, 89.19%, 140.94%, and 36.31%, respectively, compared to the control group. The proline and soluble sugar contents of wheat inoculated with Bacillus thuringiensis increased by 38.8% and 21.4%, respectively. The average content of antioxidant enzymes increased by 13.89%, and compared with the control, in wheat inoculated with the three species of ST-PGPR, the average content of ethylene decreased 2.16-fold. In addition, a mathematical model based on the “interaction equation” revealed that the best results of mixed inoculation were due to the complementary strengths of the strains. The analysis of experimental phenomena and data revealed the mechanisms by which Brevibacterium frigoritolerans, Bacillus thuringiensis, and Bacillus velezensis alleviate NaCl stress in wheat: (1) by lowering of osmotic stress, oxidative stress, and ethylene stress in wheat and (2) by using root secretions to provide substances needed for wheat. This study provides a new approach for the comprehensive understanding and evaluation of ST-PGPR as a biological inoculant for crops under salt stress.

Exploiting Beneficial Pseudomonas spp. for Cannabis Production.

Among the oldest domesticated crops, cannabis plants (Cannabis sativa L., marijuana and hemp) have been used to produce food, fiber, and drugs for thousands of years. With the ongoing legalization of cannabis in several jurisdictions worldwide, a new high-value market is emerging for the supply of marijuana and hemp products. This creates unprecedented challenges to achieve better yields and environmental sustainability, while lowering production costs. In this review, we discuss the opportunities and challenges pertaining to the use of beneficial Pseudomonas spp. bacteria as crop inoculants to improve productivity. The prevalence and diversity of naturally occurring Pseudomonas strains within the cannabis microbiome is overviewed, followed by their potential mechanisms involved in plant growth promotion and tolerance to abiotic and biotic stresses. Emphasis is placed on specific aspects relevant for hemp and marijuana crops in various production systems. Finally, factors likely to influence inoculant efficacy are provided, along with strategies to identify promising strains, overcome commercialization bottlenecks, and design adapted formulations. This work aims at supporting the development of the cannabis industry in a sustainable way, by exploiting the many beneficial attributes of Pseudomonas spp.

From Lab to Farm: Elucidating the Beneficial Roles of Photosynthetic Bacteria in Sustainable Agriculture.

Photosynthetic bacteria (PSB) possess versatile metabolic abilities and are widely applied in environmental bioremediation, bioenergy production and agriculture. In this review, we summarize examples of purple non-sulfur bacteria (PNSB) through biofertilization, biostimulation and biocontrol mechanisms to promote plant growth. They include improvement of nutrient acquisition, production of phytohormones, induction of immune system responses, interaction with resident microbial community. It has also been reported that PNSB can produce an endogenous 5-aminolevulinic acid (5-ALA) to alleviate abiotic stress in plants. Under biotic stress, these bacteria can trigger induced systemic resistance (ISR) of plants against pathogens. The nutrient elements in soil are significantly increased by PNSB inoculation, thus improving fertility. We share experiences of researching and developing an elite PNSB inoculant (Rhodopseudomonas palustris PS3), including strategies for screening and verifying beneficial bacteria as well as the establishment of optimal fermentation and formulation processes for commercialization. The effectiveness of PS3 inoculants for various crops under field conditions, including conventional and organic farming, is presented. We also discuss the underlying plant growth-promoting mechanisms of this bacterium from both microbial and plant viewpoints. This review improves our understanding of the application of PNSB in sustainable crop production and could inspire the development of diverse inoculants to overcome the changes in agricultural environments created by climate change.

Assessing the effect of P-solubilizing bacteria and mycorrhizal fungi on tomato yield and quality under different crop rotations

Little is known about the effects of inoculation with beneficial microorganisms under diversified crop rotations on agronomic performance of the high-value vegetable crop tomato (Solanum lycopersicum L.). A greenhouse experiment was conducted to evaluate the interactive effect of crop precession with two native species, faba bean (Leguminosae) and barley (Graminaceae), and inoculations with commercial arbuscular mycorrhizal fungi (AMF), and phosphate solubilizing Streptomyces roseocinereus MS1B15 on tomato plant growth, yield, fruit quality and the contents of various mineral nutrients. The overall results revealed that crop inoculation with S. roseocinereus MS1B15 significantly increased tomato growth and yield while crop precession showed no differences among tested treatments. Plant nutrient assimilation was significantly increased by both inoculations and was differentially modulated under faba bean and barley rotations. Furthermore, fruit quality demonstrated significant variations between treatments: whereas the inoculations significantly increased tomato vitamin C, crop rotations increased antioxidant compounds and organic acids. Overall, faba bean rotation combined with S. roseocinereus MS1B15 inoculation showed the best benefits for tomato plants, increasing most of the biometric variables evaluated in this study. Such combinations could be recommended as a sustainable and feasible model for tomato production.

Effects of Microbial Consortia, Applied as Fertilizer Coating, on Soil and Rhizosphere Microbial Communities and Potato Yield

The use of biological inputs in crop production systems, as complements to synthetic inputs, is gaining popularity in the agricultural industry due to increasing consumer demand for more environmentally friendly agriculture. An approach to meeting this demand is the inoculation of field crops with beneficial microbes to promote plant growth and resistance to biotic and abiotic stresses. However, the scientific literature reports inconsistent results following applications of bio-inoculant to fields. The effects of inoculation with beneficial microbes on bulk soil and rhizospheric microbial communities is often overlooked as precise monitoring of soil microbial communities is difficult. The aim of this research was to use Illumina high throughput sequencing (HTS) to shed light on bulk soil and rhizospheric microbial community responses to two commercial microbial inoculants coated onto fertilizer granules, applied to potato fields. Bulk soil samples were collected 4 days before seeding (May 27th), 7 days after seeding (June 7th), at potato shoot emergence (June 21st) and at mid-flowering (July 26th). Rhizospheric soil was collected at the mid-flowering stage. The Illumina MiSeq HTS results indicated that the bulk soil microbial community composition, especially prokaryotes, changed significantly across potato growth stages. Microbial inoculation did not affect bulk soil or rhizospheric microbial communities sampled at the mid-flowering stage. However, a detailed analysis of the HTS results showed that bulk soil and rhizospheric microbial community richness and composition were different for the first treatment block compared to the other three blocks. The spatial heterogeneity of the soil microbial community between blocks of plots was associated with potato tuber yield changes, indicating links between crop productivity and soil microbial community composition. Understanding these links could help in production of high-quality microbial inoculants to promote potato productivity.

The Long-Term Effects of Using Phosphate-Solubilizing Bacteria and Photosynthetic Bacteria as Biofertilizers on Peanut Yield and Soil Bacteria Community.

Microbial inoculation is a promising strategy to improve crop yields and reduce the use of chemical fertilizers, thereby creating environment-friendly agriculture. In this study, the long-term (5 years) effects of a phosphate-solubilizing bacterium Burkholderia cepacia ISOP5, a purple non-sulfur bacterium Rhodopseudomonas palustris ISP-1, and a mixed inoculation of these two bacteria (MB) on peanut yield, soil microbial community structure, and microbial metabolic functions were evaluated in a field experiment. After 5 years of inoculation, total peanut yield with B. cepacia ISOP5, R. palustris ISP-1, and MB treatments increased by 8.1%, 12.5%, and 19.5%, respectively. The treatments also significantly promoted the absorption of N and increased the protein content in peanut seeds. Nutrient content also increased to some extent in the bacteria-inoculum-treated soil. However, bacterial community diversity and richness were not significantly affected by bacterial inoculums, and only minor changes occurred in the bacterial community composition. Functional prediction revealed that bacterial inoculums reduced the relative abundance of those genes associated with P uptake and transport as well as increased the abundance of genes associated with inorganic P solubilization and organic P mineralization. Bacterial inoculums also increased the total relative abundance of genes associated with N metabolism. In addition to developing sustainable and eco-friendly agricultural practice, crop inoculation with B. cepacia ISOP5 and R. palustris ISP-1 would improve soil fertility, enhance microbial metabolic activity, and increase crop yield.

The Phosphate Inhibition Paradigm: Host and Fungal Genotypes Determine Arbuscular Mycorrhizal Fungal Colonization and Responsiveness to Inoculation in Cassava With Increasing Phosphorus Supply.

A vast majority of terrestrial plants are dependent on arbuscular mycorrhizal fungi (AMF) for their nutrient acquisition. AMF act as an extension of the root system helping phosphate uptake. In agriculture, harnessing the symbiosis can potentially increase plant growth. Application of the AMF Rhizophagus irregularis has been demonstrated to increase the yields of various crops. However, there is a paradigm that AMF colonization of roots, as well as the plant benefits afforded by inoculation with AMF, decreases with increasing phosphorus (P) supply in the soil. The paradigm suggests that when fertilized with sufficient P, inoculation of crops would not be beneficial. However, the majority of experiments demonstrating the paradigm were conducted in sterile conditions without a background AMF or soil microbial community. Interestingly, intraspecific variation in R. irregularis can greatly alter the yield of cassava even at a full application of the recommended P dose. Cassava is a globally important crop, feeding 800 million people worldwide, and a crop that is highly dependent on AMF for P uptake. In this study, field trials were conducted at three locations in Kenya and Tanzania using different AMF and cassava varieties under different P fertilization levels to test if the paradigm occurs in tropical field conditions. We found that AMF colonization and inoculation responsiveness of cassava does not always decrease with an increased P supply as expected by the paradigm. The obtained results demonstrate that maximizing the inoculation responsiveness of cassava is not necessarily only in conditions of low P availability, but that this is dependent on cassava and fungal genotypes. Thus, the modeling of plant symbiosis with AMF under different P levels in nature should be considered with caution.

Biological Nitrogen Fixation and Denitrification in Rhizosphere of Potato Plants in Response to the Fertilization and Inoculation

The study aim was to evaluate the potential nitrogen fixation and denitrification in the rhizosphere soil of potato plants, crop yield and output quality in response to the different fertilization systems and the inoculation with Azospirillum brasilense 410. Field stationary experiment was conducted between 2016 and 2019 with potato in a crop rotation system on leached chernozem soil. Farmyard manure, 40 t/ha, applied prior to potatoes planting promotes nitrogen fixation (0.8–2.0 times compared to control). However, it has also affected denitrification (in 1.4–2.2 times higher compared to control). The lowest rate of mineral fertilizers used in the experiment, N40P40K40, was shown as most environmentally feasible. Under its use the increase of soil nitrogenase activity and low denitrification levels were observed. Same trends were also noted for the medium fertilizer rate, N80P80K80. The highest doses of mineral fertilizers, N120P120K120, substantially affected the denitrification process and reduced the nitrogen fixation activity (in 1.9–2.2 times). The combination of manure with the medium fertilizers rate has also resulted in high denitrification levels, while the soil nitrogen fixation activity has restored only at flowering stage. Crop inoculation with A. brasilense combined with the manure application, has not affected studied processes. However, crop inoculation after the green manure intercropping has shown the growth of nitrogenase activity. Used on the mineral fertilizers background inoculation has activated nitrogen fixation and has ensured the decrease of denitrification levels, subject to the fertilization background. High fertilizer rates have hampered the inoculation efficiency. Inoculation has promoted crop yields on unfertilized and mineral backgrounds or following green manure. Crop inoculation following organic and the organo-mineral backgrounds had no significant effect, probably due to the competition for A. brasilense from microorganisms that have created a competitive environment for A. brasilense. Despite its environmental expediency, inoculation combined with the low fertilizer doses underperforms the action of inoculation combined with the medium fertilizer rates showing the latter as the compromise between the environmental requirements and crop productivity. The use of inoculation has promoted the accumulation of starch and ascorbic acid and has contributed to the reduction of nitrate contents in the tubers of inoculated plants.

Weeds as Pathogen Hosts and Disease Risk for Crops in the Wake of a Reduced Use of Herbicides: Evidence from Yam (Dioscorea alata) Fields and Colletotrichum Pathogens in the Tropics.

The transition toward sustainable agriculture requires rethinking cropping systems in the light of less intensive and chemically reliant practices. Weed management is one of the target practices to evolve cropping systems with decreased impact on the environment. While softened management will lead to increased weeds/crops coexistence, it is of importance to assess the relative benefits and drawbacks of new practices. Among the potential drawbacks of weeds/crops coexistence, disease risk may increase if weeds are hosting pathogens. In this study, we assessed the potential of weeds for hosting pathogenic generalist fungi known to translate into disease in crops. We first describe prevalence in fields after harvest and relate prevalence to species characteristics and communities. Then, we directly test the idea that weeds serve as inoculums sources during cropping with a natural experiment. This study highlights variation in host skill among feral weeds for Colletotrichum species, including potential congeneric sub-specialization on different weeds within communities. Last, prevalence within fields was more correlated to focal crop inoculation rates compared to local weed load, but there was a significant correlation effect with prevalence on weeds in the vicinity of fields, suggesting that weeds are mediating disease levels at the local scale, too. Results pointed to the importance of weed host skill in disease risk yet open the door to the potential control of pathogens via targeted weed management.

Comparative Study of the Rhizosphere and Root Endosphere Microbiomes of Cholistan Desert Plants.

Microbial communities associated with the rhizosphere and roots of desert halophytes play an important role in plants’ growth and development. Very limited information has been available on the microbial diversity of arid environments of Pakistan. Hence in the current study, the microbial diversity of rhizosphere and root endosphere of desert halophytes, Zygophyllum simplex, Haloxylon salicoricum, Aerva javanica, and Capparis decidua was evaluated. The rhizosphere and root endosphere samples of desert halophytes collected from the three geographic sites of Cholistan desert, Punjab, Pakistan were analyzed by using 16S rRNA based Illumina sequencing. The results showed that Proteobacteria were more abundant in the rhizospheric soils while Actinobacteria were more dominant in the root endosphere of halophytes. Bacteroidetes, Firmicutes, and Deinococcus-Thermus were identified from all rhizospheric soils and roots across the three sites, with variable percentage. Bacillus, Kocuria, Pseudomonas, Halomonas, and Flavobacterium were commonly identified from the rhizosphere and root endosphere of halophytes across all the three sites. At the genus level, microbial diversity from Haloxylon showed the greatest variations between the rhizosphere and root endosphere from the site 2. This study revealed that microbial diversity analysis can be used to study how changes in abiotic factors such as soil moisture content and salinity affect the microbial communities associated with the rhizospheric soils and root endosphere of halophytes across the three sites. This study will also help in the discovery of potential inoculants for crops growing in arid and semi-arid regions of Pakistan.

Development of a taxon-discriminating molecular marker to trace and quantify a mycorrhizal inoculum in roots and soils of agroecosystems.

Crop inoculation with Glomus cubense isolate (INCAM-4, DAOM-241198) promotes yield in banana, cassava, forages, and others. Yield improvements range from 20 to 80% depending on crops, nutrient supply, and edaphoclimatic conditions. However, it is difficult to connect yield effects with G. cubense abundance in roots due to the lack of an adequate methodology to trace this taxon in the field. It is necessary to establish an accurate evaluation framework of its contribution to root colonization separated from native arbuscular mycorrhizal fungi (AMF). A taxon-discriminating primer set was designed based on the ITS nrDNA marker and two molecular approaches were optimized and validated (endpoint PCR and quantitative real-time PCR) to trace and quantify the G. cubense isolate in root and soil samples under greenhouse and environmental conditions. The detection limit and specificity assays were performed by both approaches. Different 18 AMF taxa were used for endpoint PCR specificity assay, showing that primers specifically amplified the INCAM-4 isolate yielding a 370bp-PCR product. In the greenhouse, Urochloa brizantha plants inoculated with three isolates (Rhizophagus irregularis, R. clarus, and G. cubense) and environmental root and soil samples were successfully traced and quantified by qPCR. The AMF root colonization reached 41-70% and the spore number 4-128 per g of soil. This study demonstrates for the first time the feasibility to trace and quantify the G. cubense isolate using a taxon-discriminating ITS marker in roots and soils. The validated approaches reveal their potential to be used for the quality control of other mycorrhizal inoculants and their relative quantification in agroecosystems.

The Impact of Nitrogen Fixers and Phosphate Solubilizing Microbes on Sorghum (Sorghum bicolor) Yield

Objective: The inoculants, mainly nitrogen fixers and phosphate solubilizing microbes, have an influence on plant growth attributes. The current study was conducted to assess the fertilizing activity of A. chroocaccum and B. megaterium strains on growth, yield parameters and nutrient uptake of Sorghum bicolor. Methods: The isolation and identification of the nitrogen-fixing bacterium- A. chroocaccum and phosphate solubilizing microbe- B. megaterium and the growth and yield parameters of Sorghum bicolor as well as NPK uptake levels were studied. Results: A. chroocaccum and B. megaterium exhibited the proliferation of microbial population in soil by synergistic interaction with plants, and the application increased the availability of NPK in the soil after harvest of the crop. These beneficial inoculants are also known to help in the uptake of some other nutrients. The maximum beneficial aspects of the plant were noticed in the plants inoculated with 100% Recommended Dose of Fertilizer (RDF)+ A. chroocaccum+ B. megaterium, and the least was noticed in the control plants at all growth stages and at the time of harvest. Conclusion: From this study, it can be concluded that the beneficial effect of A. chroocaccum and B. megaterium is observed in sorghum crop with improved yield and nutrient uptake. Therefore, this species can be used extensively for future inoculation of the sorghum crop for better growth and development and for good returns in an ecological way.

Inorganic Phosphate Solubilization by Rhizosphere Bacterium Paenibacillus sonchi: Gene Expression and Physiological Functions.

Due to the importance of phosphorus (P) in agriculture, crop inoculation with phosphate-solubilizing bacteria is a relevant subject of study. Paenibacillus sonchi genomovar Riograndensis SBR5 is a promising candidate for crop inoculation, as it can fix nitrogen and excrete ammonium at a remarkably high rate. However, its trait of phosphate solubilization (PS) has not yet been studied in detail. Here, differential gene expression and functional analyses were performed to characterize PS in this bacterium. SBR5 was cultivated with two distinct P sources: NaH2PO4 as soluble phosphate source (SPi) and hydroxyapatite as insoluble phosphate source (IPi). Total RNA of SBR5 cultivated in those two conditions was isolated and sequenced, and bacterial growth and product formation were monitored. In the IPi medium, the expression of 68 genes was upregulated, whereas 100 genes were downregulated. Among those, genes involved in carbon metabolism, including those coding for subunits of 2-oxoglutarate dehydrogenase, were identified. Quantitation of organic acids showed that the production of tricarboxylic acid cycle-derived organic acids was reduced in IPi condition, whereas acetate and gluconate were overproduced. Increased concentrations of proline, trehalose, and glycine betaine revealed active osmoprotection during growth in IPi. The cultivation with hydroxyapatite also caused the reduction in the motility of SBR5 cells as a response to Pi depletion at the beginning of its growth. SBR5 was able to solubilize hydroxyapatite, which suggests that this organism is a promising phosphate-solubilizing bacterium. Our findings are the initial step in the elucidation of the PS process in P. sonchi SBR5 and will be a valuable groundwork for further studies of this organism as a plant growth-promoting rhizobacterium.

Engineering rhizobacteria for sustainable agriculture.

Exploitation of plant growth promoting (PGP) rhizobacteria (PGPR) as crop inoculants could propel sustainable intensification of agriculture to feed our rapidly growing population. However, field performance of PGPR is typically inconsistent due to suboptimal rhizosphere colonisation and persistence in foreign soils, promiscuous host-specificity, and in some cases, the existence of undesirable genetic regulation that has evolved to repress PGP traits. While the genetics underlying these problems remain largely unresolved, molecular mechanisms of PGP have been elucidated in rigorous detail. Engineering and subsequent transfer of PGP traits into selected efficacious rhizobacterial isolates or entire bacterial rhizosphere communities now offers a powerful strategy to generate improved PGPR that are tailored for agricultural use. Through harnessing of synthetic plant-to-bacteria signalling, attempts are currently underway to establish exclusive coupling of plant-bacteria interactions in the field, which will be crucial to optimise efficacy and establish biocontainment of engineered PGPR. This review explores the many ecological and biotechnical facets of this research.

BENEFITS OF JOINT MANAGEMENT OF GREEN MANURE AND MYCORRHIZAL INOCULANTS IN CROP PRODUCTION

<p><strong>Background.</strong> Nutrient supply is a decisive component of agricultural systems, but in Cuba only 25 % of cultivated areas can be fertilized, influencing their productivity negatively. From the benefits of the use of mycorrhizal inoculants and green manures and the mycorrhizal dependence of <em>Canavalia ensiformis</em>, the hypothesis of integrating both was evaluated. <strong>Objective</strong>. To establish nutrient supply schemes that guarantee high yields with lower amounts of fertilizers, as well as new routes for mycorrhizal inoculation of crops. <strong>Methodology.</strong> The canavalia response to the inoculation of three generalist AMF strains was evaluated in eight edaphic conditions, as well as eight integration experiments of the green fertilizers inoculated with the efficient strain in the nutrient supply schemes of cassava, banana, corn, tobacco, mulberry and king-grass. <strong>Results.</strong> Canavalia responded positively to inoculation, with average increases in biomass and NPK contents of 50 and 70 % respectively, as well as mycorrhizal propagules. The effectiveness of the applied strains depended on the edaphic conditions and presumably the soil pH. The canavalia inoculated with the efficient AMF strain and used as the preceding and/or intercropped green manure was successfully integrated into the nutrient supply systems of the different crops, guaranteeing high yields, effective mycorrhizal function and decreasing between 25 and 75 % the amounts of fertilizers applied to them. <strong>Implications.</strong> The benefits and feasibility of the joint management of green manures and mycorrhizal inoculants in agricultural production are demonstrated, as well as being a successful way to achieve effective mycorrhization in crops whose direct inoculation is prohibitive due to the high amounts of biofertilizer or laboriousness. <strong>Conclusion.</strong> The integration of AMF inoculants and green manure increased the benefits of both practices and it is a promising way of managing the arbuscular mycorrhizal symbiosis in sustainable crop production.</p>

Pseudomonas fluorescens and Azospirillum brasilense Increase Yield and Fruit Quality of Tomato Under Field Conditions

Crop inoculation with plant growth-promoting rhizobacteria (PGPR) is a sustainable alternative to diminish the excessive use of chemical fertilizers in agriculture. However, there is little information about PGPR inoculation effects under field conditions and even less on industrial tomato production. We aimed to study the effects of a sole inoculation at seedling stage with Pseudomonas fluorescens Rt6M10, Azospirillum brasilense Az39, and their combination on growth and yield of two industrial tomato varieties UCO 14 (UCO) and Harris Moran 3861 (HM). We compared these PGPR inoculation treatments with chemically fertilized and non-fertilized (control) seedlings under field conditions. We found that inoculation with Rt6M10, Az39, and their combination increased seedling root dry weight by 62%, 41%, and 23%, respectively and shoot dry weight by 29%, 23%, and 2%, respectively compared with non-inoculated control, improving tolerance to transplant stress. Inoculation with Rt6M10, Az39, and their combination increased stem diameter by 15%, 16%, and 13%, respectively, while Rt6M10 and the combination treatments increased leaf chlorophyll and carotenoid levels compared with non-inoculated plants. Az39 increased fruit number (35%) and fruit weight (38%) per plant in HM, whereas in UCO variety, the increase was the highest (48% and 49%, respectively). Seedling inoculation increase fruit firmness and equatorial and polar fruit diameter by 24%, 10%, and 12%, respectively in HM and by 21%, 14%, and 14%, respectively in UCO. Overall, bio-inoculation with Rt6M10 and/or Az39 was beneficial for tomato seedlings at transplanting and supported fruit yield and quality (total soluble solid content, pH, and titratable acidity) equally or better than chemically fertilized seedlings.