Harnessing chickpea bacterial endophytes for improved plant health and fitness.

Endophytic bacteria live asymptomatically inside the tissues of host plants without inflicting any damage. Endophytes can confer several beneficial traits to plants, which can contribute to their growth, development, and overall health. They have been found to stimulate plant growth by enhancing nutrient uptake and availability. They can produce plant growth-promoting substances such as auxins, cytokinins, and gibberellins, which regulate various aspects of plant growth and development. Endophytes can also improve root system architecture, leading to increased nutrient and water absorption. Some endophytes possess the ability to solubilize nutrients, such as phosphorus and potassium, making them more available for plant uptake, and fixing atmospheric nitrogen. Chickpea (Cicer arietinum) is a major legume crop that has mutualistic interactions with endophytes. These endophytes can benefit the chickpea plant in various ways, including higher growth, improved nutrient uptake, increased tolerance to abiotic and biotic stressors, and disease suppression. They can produce enzymes and metabolites that scavenge harmful reactive oxygen species, thus reducing oxidative stress. Moreover, several studies reported that endophytes produce antimicrobial compounds, lytic enzymes, and volatile organic compounds that inhibit the growth of fungal pathogens and trigger systemic defense responses in plants, leading to increased resistance against a broad range of pathogens. They can activate plant defense pathways, including the production of defense-related enzymes, phytoalexins, and pathogenesis-related proteins, thereby providing long-lasting protection. It is important to note that the diversity and function of chickpea-associated endophytes can vary depending on factors such as variety, geographical location, and environmental conditions. The mechanisms behind the plant-beneficial interactions are still being intensively explored. In this review, new biotechnologies in agricultural production and ecosystem stability were presented. Thus, harnessing chickpea endophytes could be exploited in developing drought-resistant cultivars that can maintain productivity in arid and semi-arid environments, crucial for meeting the global demand for chickpeas.

Insight into endophytic microbial diversity in two halophytes and plant beneficial attributes of Bacillus swezeyi.

This study utilized high-throughput sequencing to investigate endophytic bacteria diversity in halophytic plants Anabasis truncate (AT) and Anabasis eriopoda (AE) from the Aral Sea region. Following sequence processing, 356 Amplicon Sequence Variants (ASVs) were discovered. The abundance and variety of endophytic bacteria were higher in AT. Bacillota, Pseudomonadota, Actinomycetota, and Bacteroidota constituted the dominant in AE, whereas Pseudomonadota, Actinomycetota, Bacteroidota, and Chloroflexota constituted the dominant in AT. Biomarkers were identified through LEFSe analysis, showing host-specific patterns. PCoA indicated distinct bacterial community structures. Phylogenetic analysis revealed diverse endophytic bacteria, including potential novel taxa. PICRUSt2 predicted diverse functions for endophytic bacteria in halophytes, indicating recruitment of beneficial bacterial taxa to adapt to extreme hypersaline conditions, including plant growth-promoting, biocontrol, and halophilic/tolerant bacteria. Moreover, the evolutionary relationship, metabolic capabilities, and plant beneficial potentials of the Bacillus swezeyi strains, previously isolated from the above two halophytes, were analyzed using comparative genomic and physiological analysis. The B. swezeyi strains displayed versatile environmental adaptability, as shown by their ability to use a wide range of carbon sources and their salt tolerances. B. swezeyi possessed a wide range of enzymatic capabilities, including but not limited to proteases, cellulases, and chitinases. Comparative genomic analysis revealed that despite some variations, they shared genetic similarities and metabolic capabilities among the B. swezeyi strains. B. swezeyi strains also displayed outstanding plant-growth-promoting and antagonistic potentials, offering potential solutions to the global food crisis. This study enhances our understanding of microbial diversity in halophytes on saline-alkali land in the West Aral Sea, shedding light on the halophyte microbiome and its collaboration with hosts in highly hypersaline environments. This study also provides a scientific basis for developing high-quality microbial fertilizers and implementing sustainable agricultural practices.

Plant beneficial traits of endophytic bacteria associated with fennel (Foeniculum vulgare Mill.).

In this study, we used 16S rRNA gene sequence analysis to describe the diversity of cultivable endophytic bacteria associated with fennel (Foeniculum vulgare Mill.) and determined their plant-beneficial traits. The bacterial isolates from the roots of fennel belonged to four phyla: Firmicutes (BRN1 and BRN3), Proteobacteria (BRN5, BRN6, and BRN7), Gammaproteobacteria (BRN2), and Actinobacteria (BRN4). The bacterial isolates from the shoot of fennel represented the phyla Proteobacteria (BSN1, BSN2, BSN3, BSN5, BSN6, BSN7, and BSN8), Firmicutes (BSN4, BRN1, and BRN3), and Actinobacteria (BRN4). The bacterial species Bacillus megaterium, Bacillus aryabhattai, and Brevibacterium frigoritolerans were found both in the roots and shoots of fennel. The bacterial isolates were found to produce siderophores, HCN, and indole-3-acetic acid (IAA), as well as hydrolytic enzymes such as chitinase, protease, glucanase, and lipase. Seven bacterial isolates showed antagonistic activity against Fusarium culmorum, Fusarium solani, and Rhizoctonia. solani. Our findings show that medicinal plants with antibacterial activity may serve as a source for the selection of microorganisms that exhibit antagonistic activity against plant fungal infections and may be considered as a viable option for the management of fungal diseases. They can also serve as an active part of biopreparation, improving plant growth.

Molecular genotyping analysis suggested high clonality and simple marker loci in Puccinia striiformis f. sp. tritici population in Uzbekistan, Central Asian region.

Puccinia striiformis f. sp. tritici (Pst) is a destructive pathogen that causes wheat stripe rust worldwide. Understanding the population structure and dynamic of pathogen spread is critical to fight against this disease. Limited information is available for the population genetic structure of Pst in Uzbekistan, Central Asia. In this study, we carried out surveillance from 9 different regions (Andijan, Fergana, Jizzakh, Kashkadarya, Namangan, Samarkand, Sirdaryo, Surkhandarya and Tashkent) of Uzbekistan to fill this gap. A total of 255 isolates were collected, which were genotyped using 17 polymorphic simple sequence repeats (SSR) markers. The DAPC analysis results showed no population subdivision in these sample-collected regions except Surkhandarya. Multilocus genotype (MLG) analysis, FST, and Nei's genetic distance results indicated a clonal population (rBarD ≤ 0.12) and merely three MLGs accounting for 70% of the overall population. MLG-34 was predominant in all Uzbekistan regions, followed by MLG-36 and MLG-42. Low genotypic diversity was observed in Andijan, Fergana, Jizzakh, Kashkadarya, Namangan, Sirdaryo, and Tashkent (0.56 to 0.76), compared with Samarkand (0.82) and Surkhandarya (0.97). No virulence against Yr5, Yr15, YrSp, and Yr26 was found, while resistant was overcome against Yr1, Yr2, Yr6, Yr9, Yr17, and Yr44 genes (Virulence frequency =≥75%). Comparative study results of Uzbekistan with previous Himalayan population were showed divergence from China and Pakistan populations. Further studies need to be conducted in a worldwide context to understand migration patterns; for that purpose, collaborative work is essential due to the Pst long-distance migration capability.

Antimicrobial Action Mechanisms of Natural Compounds Isolated from Endophytic Microorganisms.

Infectious diseases are a significant challenge to global healthcare, especially in the face of increasing antibiotic resistance. This urgent issue requires the continuous exploration and development of new antimicrobial drugs. In this regard, the secondary metabolites derived from endophytic microorganisms stand out as promising sources for finding antimicrobials. Endophytic microorganisms, residing within the internal tissues of plants, have demonstrated the capacity to produce diverse bioactive compounds with substantial pharmacological potential. Therefore, numerous new antimicrobial compounds have been isolated from endophytes, particularly from endophytic fungi and actinomycetes. However, only a limited number of these compounds have been subjected to comprehensive studies regarding their mechanisms of action against bacterial cells. Furthermore, the investigation of their effects on antibiotic-resistant bacteria and the identification of biosynthetic gene clusters responsible for synthesizing these secondary metabolites have been conducted for only a subset of these promising compounds. Through a comprehensive analysis of current research findings, this review describes the mechanisms of action of antimicrobial drugs and secondary metabolites isolated from endophytes, antibacterial activities of the natural compounds derived from endophytes against antibiotic-resistant bacteria, and biosynthetic gene clusters of endophytic fungi responsible for the synthesis of bioactive secondary metabolites.

Shared governance in the plant holobiont and implications for one health.

The holobiont Holobiont theory is more than 80 years old, while the importance of microbial communities for plant holobionts was already identified by Lorenz Hiltner more than a century ago. Both concepts are strongly supported by results from the new field of microbiome research. Here, we present ecological and genetic features of the plant holobiont that underpin principles of a shared governance between hosts and microbes and summarize the relevance of plant holobionts in the context of global change. Moreover, we uncover knowledge gaps that arise when integrating plant holobionts in the broader perspective of the holobiome as well as one and planetary health concepts. Action is needed to consider interacting holobionts at the holobiome scale, for prediction and control of microbiome function to improve human and environmental health outcomes.

Antimicrobial and Cytotoxic Activities of the Secondary Metabolites of Endophytic Fungi Isolated from the Medicinal Plant Hyssopus officinalis.

According to the World Health Organization, it is estimated that by 2050, drug-resistant infections could cause up to 10 million deaths annually. Therefore, finding a new generation of antibiotics is crucial. Natural compounds from endophytic fungi are considered a potential source of new-generation antibiotics. The antimicrobial and cytotoxic effects of ethyl acetate extracts of nine endophytic fungal isolates obtained from Hyssopus officinalis were investigated for bioassay-guided isolation of the natural compounds. An extract of isolate VII showed the highest antimicrobial activities against Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus (30.12 ± 0.20 mm and 35.21 ± 0.20 mm) and Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa (30.41 ± 0.23 mm and 25.12 ± 0.25 mm) among the tested extracts of isolates. Molecular identification of isolate VII confirmed it as Chaetomium elatum based on sequencing of its ITS genes, and it was discovered that this was the first time C. elatum had been isolated from H. officinalis. This isolate was cultured at a large scale for the isolation and identification of the active compound. Penicillic acid was isolated for the first time from C. elatum and its chemical structure was established by NMR spectroscopy. The penicillic acid showed strong antibacterial activities against Bacillus subtilis and Staphylococcus aureus with 20.68 mm and 25.51 mm inhibition zones, respectively. In addition, MIC and MBC values and antibiofilm activities of penicillic acid were determined. It was found that penicillic acid reduced the level of biofilms in proportion to antibacterial activity.

The potential of biochar as a microbial carrier for agricultural and environmental applications.

Biochar can be an effective carrier for microbial inoculants because of its favourable properties promoting microbial life. In this review, we assess the effectiveness of biochar as a microbial carrier for agricultural and environmental applications. Biochar is enriched with organic carbon, contains nitrogen, phosphorus, and potassium as nutrients, and has a high porosity and moisture-holding capacity. The large number of active hydroxyl, carboxyl, sulfonic acid group, amino, imino, and acylamino hydroxyl and carboxyl functional groups are effective for microbial cell adhesion and proliferation. The use of biochar as a carrier of microbial inoculum has been shown to enhance the persistence, survival and colonization of inoculated microbes in soil and plant roots, which play a crucial role in soil biochemical processes, nutrient and carbon cycling, and soil contamination remediation. Moreover, biochar-based microbial inoculants including probiotics effectively promote plant growth and remediate soil contaminated with organic pollutants. These findings suggest that biochar can serve as a promising substitute for non-renewable substrates, such as peat, to formulate and deliver microbial inoculants. The future research directions in relation to improving the carrier material performance and expanding the potential applications of this emerging biochar-based microbial immobilization technology have been proposed.

Viral Community Structure and Potential Functions in the Dried-Out Aral Sea Basin Change along a Desiccation Gradient.

The dried-out Aral Sea basin represents an extreme environment due to a man-made ecological disaster. Studies conducted in this unique environment revealed high levels of pollution and a specifically adapted microbiota; however, viral populations remained entirely unexplored. By employing an in-depth analysis based on the sequencing of metagenomic DNA recovered from rhizosphere samples of Suaeda acuminata (C. A. Mey.) Moq. along a desiccation gradient of 5, 10, and 40 years, we detected a diverse viral community comprising 674 viral populations (viral operational taxonomic units [vOTUs]) dominated by Caudovirales. Targeted analyses highlighted that viral populations in this habitat are subjected to certain dynamics that are driven mainly by the gradient of desiccation, the corresponding salinity, and the rhizosphere bacterial populations. In silico predictions linked the viruses to dominant prokaryotic taxa in the Aral Sea basin, such as Gammaproteobacteria, Actinomycetia, and Bacilli. The lysogenic lifestyle was predicted to be predominant in areas that dried out 5 years ago, representing the early revegetation phase. Metabolic prediction of viral auxiliary metabolic genes (AMGs) suggests that viruses may play a role in the biogeochemical cycles, stress resilience, and competitiveness of their hosts due to the presence of genes that are involved in biofilm formation. Overall, our study provides important insights into viral ecology in an extreme environment and expands our knowledge related to virus occurrence in terrestrial systems. IMPORTANCE Environmental viruses have added a wealth of knowledge to ecological studies with the emergence of metagenomic technology and approaches. They are also becoming recognized as important genetic repositories that underpin the functioning of terrestrial ecosystems but have remain moslty unexplored. Using shotgun metagenome sequencing and bioinformatic tools, we found that the viral community structure was affected during natural revegetation in the dried-up Aral Sea area, a model habitat for investigating natural ecological restoration but still understudied. In this study, we highlight the importance of viruses, elements that are overlooked, for their potential contribution to terrestrial ecosystems, i.e., nutrient cycles, stress resilience, and host competitiveness, during natural revegetation.

Function-Based Rhizosphere Assembly along a Gradient of Desiccation in the Former Aral Sea.

The desiccation of the Aral Sea represents one of the largest human-made environmental regional disasters. The salt- and toxin-enriched dried-out basin provides a natural laboratory for studying ecosystem functioning and rhizosphere assembly under extreme anthropogenic conditions. Here, we investigated the prokaryotic rhizosphere communities of the native pioneer plant Suaeda acuminata (C.A.Mey.) Moq. in comparison to bulk soil across a gradient of desiccation (5, 10, and 40 years) by metagenome and amplicon sequencing combined with quantitative PCR (qPCR) analyses. The rhizosphere effect was evident due to significantly higher bacterial abundances but less diversity in the rhizosphere compared to bulk soil. Interestingly, in the highest salinity (5 years of desiccation), rhizosphere functions were mainly provided by archaeal communities. Along the desiccation gradient, we observed a significant change in the rhizosphere microbiota, which was reflected by (i) a decreasing archaeon-bacterium ratio, (ii) replacement of halophilic archaea by specific plant-associated bacteria, i.e., Alphaproteobacteria and Actinobacteria, and (iii) an adaptation of specific, potentially plant-beneficial biosynthetic pathways. In general, both bacteria and archaea were found to be involved in carbon cycling and fixation, as well as methane and nitrogen metabolism. Analysis of metagenome-assembled genomes (MAGs) showed specific signatures for production of osmoprotectants, assimilatory nitrate reduction, and transport system induction. Our results provide evidence that rhizosphere assembly by cofiltering specific taxa with distinct traits is a mechanism which allows plants to thrive under extreme conditions. Overall, our findings highlight a function-based rhizosphere assembly, the importance of plant-microbe interactions in salinated soils, and their exploitation potential for ecosystem restoration approaches. IMPORTANCE The desertification of the Aral Sea basin in Uzbekistan and Kazakhstan represents one of the most serious anthropogenic environmental disasters of the last century. Since the 1960s, the world's fourth-largest inland body of water has been constantly shrinking, which has resulted in an extreme increase of salinity accompanied by accumulation of many hazardous and carcinogenic substances, as well as heavy metals, in the dried-out basin. Here, we investigated bacterial and archaeal communities in the rhizosphere of pioneer plants by combining classic molecular methods with amplicon sequencing as well as metagenomics for functional insights. By implementing a desiccation gradient, we observed (i) remarkable differences in the archaeon-bacterium ratio of plant rhizosphere samples, (ii) replacement of archaeal indicator taxa during succession, and (iii) the presence of specific, potentially plant-beneficial biosynthetic pathways in archaea present during the early stages. In addition, our results provide hitherto-undescribed insights into the functional redundancy between plant-associated archaea and bacteria.

The Metabolic Potential of Endophytic Actinobacteria Associated with Medicinal Plant Thymus roseus as a Plant-Growth Stimulator.

Bio-fertilizer practice considers not only economical but also environmentally friendly, sustainable agriculture. Endophytes can play important beneficiary roles in plant development, directly, indirectly, or synergistically. In this study, the majority of our endophytic actinobacteria were able to possess direct plant growth-promoting (PGP) traits, including auxin (88%), ammonia (96%), siderophore production (94%), and phosphate solubilization (24%), along with cell-wall degrading enzymes such as protease (75%), cellulase (81%), lipase (81%), and chitinase (18%). About 45% of tested strains have an inhibitory effect on the phytopathogen Fusarium oxysporum, followed by 26% for Verticillium dahlia. Overall, our results showed that strains XIEG63 and XIEG55 were the potent strains with various PGP traits that caused a higher significant increase (p ≤ 0.05) in length and biomass in the aerial part and roots of tomato and cotton, compared to the uninoculated plants. Our data showed that the greatest inhibition percentages of two phytopathogens were achieved due to treatment with strains XIEG05, XIEG07, XIEG45, and XIEG51. The GC-MS analysis showed that most of the compounds were mainly alkanes, fatty acid esters, phenols, alkenes, and aromatic chemicals and have been reported to have antifungal activity. Our investigation emphasizes that endophytic actinobacteria associated with medicinal plants might help reduce the use of chemical fertilization and potentially lead to increased agricultural productivity and sustainability.

The diversity of bacterial endophytes from Iris pseudacorus L. and their plant beneficial traits.

This study reports the diversity of cultivable endophytic bacteria associated with yellow iris (Iris pseudacorus L.) by using 16S rRNA gene analysis and their plant beneficial traits. The 16S rRNA sequence similarities of endophytic bacteria isolated from the leaves and roots of yellow iris showed that the isolates belonged to the genera Staphylococcus, Streptomyces, Variovorax, Pantoea, Paenibacillus, Bacillus, Janthinobacterium, Enterobacter, Brevibacterium, Agrobacterium, Rhizobium, Xanthomonas translucens, and Pseudomonas. The endophytic bacteria Pseudomonas gessardii HRT18, Brevibacterium frigoritolerans HRT8, Streptomyces atratus HRT13, and Bacillus toyonensis HST13 exhibited antimicrobial activity against five plant pathogenic fungi Fusarium, Rhizoctonia, Botrytis, Pythium, and Alternaria. They also demonstrated the capability to produce chitinase, protease, glucanase, lipase, HCN, and indole-3-acetic acid (IAA). Thirteen isolates (46%) produced IAA, and the most active IAA producers were Bacillus cereus, Agrobacterium tumefaciens, Agrobacterium vitis, Bacillus megaterium, and Bacillus aryabhattai. The IAA producing bacterial isolates stimulated root and shoot growth of garden cress. Our findings suggest that medicinal plants could be a promising source for isolating plant-beneficial bacteria that can be used to enhance the growth and protect plants against soil-borne pathogens.

A Novel Bacillus safensis-Based Formulation along with Mycorrhiza Inoculation for Controlling Alternaria alternata and Simultaneously Improving Growth, Nutrient Uptake, and Steviol Glycosides in Stevia rebaudiana under Field Conditions.

The excess use of chemicals by farmers in the agroecosystems degrades soil quality, disturbs soil ecology, and increases soil salinity and health hazards in humans. Stevia rebaudiana is an important medicinal and aromatic crop whose leaves contain steviol glycosides (SGs). The Bacillus safensis NAIMCC-B-02323 strain STJP from the rhizosphere of S. rebaudiana producing salicylic acid (16.80 µg/mL), chitinase (75.58 U/mL), ß-1,3-glucanase (220.36 U/mL), and cellulase (170 U/mL) was taken as a plant growth-promoting rhizobacteria (PGPR). The cell-free supernatant (CFS) from strain STJP showed significant biocontrol activity against Alternaria alternata (80%), suggesting the protective role of extracellular metabolite(s) against phytopathogens. Paneer whey-based bioformulation (P-WBF) was developed to exploit B. safensis STJP to enhance the growth, nutrient uptake, soil properties, stevioside content, and SGs biosynthesis in S. rebaudiana under an A. alternata-infested field. The combined treatment of P-WBF and mycorrhiza (Glomus fasciculatum ABTEC) significantly enhanced plant growth parameters after 90 days, in comparison with control. The symbiotic action (P-WBF and mycorrhiza) displayed much better results in terms of chlorophyll a and b (improved by 132.85% and 39.80%, respectively), protein (by 278.75%), flavonoid (by 86.99%), carbohydrate (by 103.84%), antioxidant (by 75.11%), and stevioside (by 120.62%) contents in plants as compared to the untreated set. Further, the augmentation of potassium (by 132.39%), phosphorous (by 94.22%), and zinc (by 111.11%) uptake in plant tissues and soil was also observed by the application of P-WBF and mycorrhiza. The expression of UGT74G1 and UGT85C2 genes related to SG biosynthesis was upregulated (2.7- and 3.2-fold, respectively) in plants treated with P-WBF and mycorrhiza as further confirmed by the accumulation of SGs. The results suggest that the application of P-WBF and mycorrhiza not only provides an ecofriendly and sustainable solution to improve stevioside content in S. rebaudiana by a nutrient-linked mechanism but also paves the way to enhanced production of stevioside.

The Integrated Effect of Microbial Inoculants and Biochar Types on Soil Biological Properties, and Plant Growth of Lettuce (Lactuca sativa L.).

Numerous reports confirm the positive effect of biochar application on soil properties and plant development. However, the interaction between root-associated beneficial microbes and different types of biochar is not well understood. The objective of this study was to evaluate the plant growth of lettuce after the application of three types of biochar in loamy, sandy soil individually and in combination with plant-beneficial microbes. Furthermore, total microbial activity in rhizosphere soil of lettuce was measured by means of fluorescein diacetate (FDA) hydrolase and enzyme activities linked to carbon, nitrogen, and phosphorus cycling. We used three types of biochar: (i) pyrolysis char from cherry wood (CWBC), (ii) pyrolysis char from wood (WBC), and (iii) pyrolysis char from maize (MBC) at 2% concentration. Our results showed that pyrolysis biochars positively affected plant interaction with microbial inoculants. Plant dry biomass grown on soil amended with MBC in combination with Klebsiella sp. BS13 and Klebsiella sp. BS13 + Talaromyces purpureogenus BS16aPP inoculants was significantly increased by 5.8% and 18%, respectively, compared to the control plants. Comprehensively, interaction analysis showed that the biochar effect on soil enzyme activities involved in N and P cycling depends on the type of microbial inoculant. Microbial strains exhibited plant growth-promoting traits, including the production of indole 3-acetic-acid and hydrogen cyanide and phosphate-solubilizing ability. The effect of microbial inoculant also depends on the biochar type. In summary, these findings provide new insights into the understanding of the interactions between biochar and microbial inoculants, which may affect lettuce growth and development.

Characterization of cadmium-tolerant endophytic fungi isolated from soybean (Glycine max) and barley (Hordeum vulgare).

Cadmium stress disrupts plant-microbial interactions and reduces plant growth and development. In plants, the tolerance to stress can be increased by inoculation with endophytic microorganisms. The aim of this study was to investigate the distribution of endophytic fungi in various plant organs of barley and soybean and evaluate their Cd removal ability. Two hundred fifty-three fungal strains were isolated from various organs of barley (Hordeum vulgare cv Arna) and soybean (Glycine max cv Almaty). The colonization rate ranged from 13.6% to 57.3% and was significantly higher in the roots. Ten genera were identified: Fusarium, Penicillium, Aspergillus, Metarhizium, Beauveria, Trichoderma, Rhodotorula, Cryptococcus, Aureobasidium and Metschnikowia. Twenty-three fungal strains have a Cd tolerance index from 0.24 to 1.12. Five strains (Beauveria bassiana T7, Beauveria bassiana T15, Rhodotorula mucilaginosa MK1, Rhodotorula mucilaginosa RH2, Metschnikowia pulcherrima MP2) with the highest level of Cd tolerance have minimum inhibitory concentrations from 290 to 2400 µg/ml. These fungi were able to remove Cd up to 59%. The bioaccumulation capacity ranged from 2.3 to 11.9 mg/g. Selected fungal strains could be considered as biological agents for their potential application in the bioremediation of contaminated sites.