Biocontrol Activity Research Articles

Enhancing saffron (Crocus sativus L.) growth in the Kashmir valley with resilient and widely effective Plant Growth-Promoting Rhizobacteria (PGPR) under field conditions

This study provides a thorough examination of stress-tolerant plant growth-promoting rhizobacteria (PGPR) and their significance in enhancing plant growth and biocontrol activities, and influence on saffron cultivation in the Kashmir Valley. Quantitative analysis of 35 efficient PGPR revealed significant variability in plant growth promotion (PGP) attributes ranging from 60 to 653 µg/ml tri-calcium phosphate (TCP) solubilization, 1548–4311 nM α-ketobutyrate h/mg/protein ACC-deaminase activity, 1.4–74.8 µg/ml IAA-like auxins production, and 94–214 % siderophore production. Molecular fingerprinting disclosed 52 ERIC types that belonged to 25 genera, mainly Bacillus. Bacillus species including B. aryabhattai, B. halotolerans, B. methylotrophicus, B. safensis, B. siamensis and B. simplex dominated the PGPR population, along with Pseudomonas spp., such as P. azotoformans and P. chlororaphis, indicating a selective buildup of PGPR in the saffron rhizosphere. In-vivo and In-vitro dual inoculation test of these widespread PGPR and stress-tolerant also inhibited the mycelial growth of Fusarium species including F. oxysporum, F. oxysporum f. sp. ciceris, and F. proliferatum. The field application of seven selected PGPR strains resulted in significant improvements in saffron productivity, including increased sprouted corms, plant height, stigma length, and stigma fresh and dry weight, which ranged from 0.6-1.8 %, 0.4–0.7 %, 0.3–0.5 %, and 6–19 %, respectively. These strains have the potential to stimulate cormlet growth and ultimately increase saffron production. Therefore, the PGP traits of these stress-tolerant rhizobacteria contributing to nutrients solubilization, auxin biosynthesis, and fungicidal attributes can ameliorate the saffron agricultural practices.

Biochemical Properties of a Novel Cold-Adapted GH19 Chitinase with Three Chitin-Binding Domains from Chitinilyticum aquatile CSC-1 and Its Potential in Biocontrol of Plant Pathogenic Fungi.

GH19 (glycoside hydrolase 19) chitinases play crucial roles in the enzymatic conversion of chitin and biocontrol of phytopathogenic fungi. Herein, a novel multifunctional chitinase of GH19 (CaChi19A), which contains three chitin-binding domains (ChBDs), was successfully cloned from Chitinilyticum aquatile CSC-1 and heterologously expressed in Escherichia coli. We also generated truncated mutants of CaChi19A_ΔI, CaChi19A_ΔIΔII, and CaChi19A_CatD consisting of two ChBDs and a catalytic domain, one ChBD and a catalytic domain, and only a catalytic domain, respectively. CaChi19A, CaChi19A_ΔI, CaChi19A_ΔIΔII, and CaChi19A_CatD exhibited cold adaptation, as their relative enzyme activities at 5 °C were 40.7, 51.6, 66.2, and 82.6%, respectively. Compared with CaChi19A and other variants, CaChi19A_ΔIΔII demonstrated a higher level of stability below 50 °C and retained relatively high activity over a wide pH range of 5-12. Analysis of the hydrolysis products revealed that CaChi19A and CaChi19A_ΔIΔII exhibit exoacting, endoacting, and N-acetyl-β-d-glucosaminidase activities toward colloidal chitin. Furthermore, CaChi19A and CaChi19A_ΔIΔII exhibited inhibitory effects on the hyphal growth of Fusarium oxysporum, Fusarium redolens, Fusarium fujikuroi, Fusarium solani, and Coniothyrium diplodiella, thereby illustrating effective biocontrol activity. These results indicated that CaChi19A and CaChi19A_ΔIΔII show advantages in some applications where low temperatures were demanded in industries as well as the biocontrol of fungal diseases in agriculture.

Exiguobacterium acetylicum Strain SI17: A Potential Biocontrol Agent against Peronophythora litchii Causing Post-Harvest Litchi Downy Blight

Litchi downy blight (LDB) caused by Peronophythora litchii destroys 20–30% of litchi fruit every year and causes significant economic losses. Some Exiguobacterium strains exhibit considerable promise in both agricultural and industrial sectors. E. acetylicum SI17, isolated from the litchi fruit carposphere, demonstrated significant biocontrol activity against LDB through pre-harvest treatment. To elucidate its underlying regulatory mechanisms, the genome of SI17 was sequenced and analyzed, revealing a circular chromosome spanning 3,157,929 bp and containing 3541 protein-coding genes and 101 RNA genes. Notably, 94 genes were implicated in the production of secondary metabolites. Among the 29 Exiguobacterium strains so far sequenced, SI17 possessed the largest genome. In the phylogenomic analysis encompassing the entire genome, SI17 was clustered into Group I. Treating litchi fruit with SI17 before harvesting resulted in a decrease in H2O2 content in the fruit peel and an increase in superoxide dismutase activity, thus enhancing resistance to LDB. Interestingly, SI17 did not display plate antagonism against Peronophythora litchii SC18. It can be inferred that SI17 generates secondary metabolites, which enhance litchi’s resistance to LDB. This study represents the first documentation of an Exiguobacterium strain exhibiting a role in litchi plant disease and showcasing significant potential for the biological control of LDB.

Transcriptomic and metabolomic analyses provide evidence on the biocontrol mechanism responsible for the inhibition of Botrytis cinerea and Alternaria alternata by Bacillus velezensis in kiwifruit

AbstractBotrytis cinerea and Alternaria alternata are major pre‐ and postharvest plant pathogens of kiwifruit and responsible for significant economic losses. Although synthetic fungicides are the main source of controlling these pathogens, their use can lead to pathogen resistance, pollution of the environment, and their residues on agricultural produce can represent a health risk to humans. Bacillus velezensis is an ecofriendly bacterium with biocontrol potential. The objectives of the present study were to determine the efficacy of B. velezensis against B. cinerea and A. alternata, both of which are pre‐ and postharvest pathogens of kiwifruit, and investigate the mechanisms responsible for its biocontrol activity using transcriptomic and metabolomic methodology. Dual cultures of B. velezensis and either B. cinerea or A. alternata versus single cultures were used to conduct a transcriptomic and metabolomic analyses to provide insight into the mechanism by which B. velezensis inhibits B. cinerea and A. alternata. A total of 2499 and 3248 differentially expressed genes (DEGs) were identified in the B. velezensis/B. cinerea and B. velezensis/A. alternata comparisons, respectively. Genes related to sporulation, virulence, and hydrolase activity were downregulated in B. cinerea and A. alternata. Genes associated with MAPK signaling and the TCA cycle were also downregulated. Our study provides new insights into mechanism underlying the inhibition of B. cinerea and A. alternata by B. velezensis. Our results also demonstrate the potential of B. velezensis as a biocontrol agent against these two major pathogens of kiwifruit.

Antifungal metabolites produced by Pseudomonas hunanensis SPT26 effective in biocontrol of fusarium wilt of Lycopersicum esculentum under saline conditions.

In past few years, salinity has become one of the important abiotic stresses in the agricultural fields due to anthropogenic activities. Salinity is leading towards yield losses due to soil infertility and increasing vulnerability of crops to diseases. Fluorescent pseudomonads are a diverse group of soil microorganisms known for promoting plant growth by involving various traits including protecting crops from infection by the phytopathogens. In this investigation, salt tolerant plant growth promoting bacterium Pseudomonas hunanensis SPT26 was selected as an antagonist against Fusarium oxysporum, causal organism of fusarium wilt in tomato. P. hunanensis SPT26 was found capable to produce various antifungal metabolites. Characterization of purified metabolites using Fourier transform infrared spectroscopy (FT-IR) and liquid chromatography-electron spray ionization-mass spectrometry(LC-ESI/MS) showed the production of various antifungal compounds viz., pyrolnitrin, pyochelin and hyroxyphenazine by P. hunanensis SPT26. In the preliminary examination, biocontrol activity of purified antifungal metabolites was checked by dual culture method and results showed 68%, 52% and 65% growth inhibition by pyrolnitrin, 1- hydroxyphenazine and the bacterium (P. hunanensis SPT26) respectively. Images from scanning electron microscopy (SEM) revealed the damage to the mycelia of fungal phytopathogen due to production of antifungal compounds secreted by P. hunanensis SPT26. Application of bioinoculant of P. hunanensis SPT26 and purified metabolites significantly decreased the disease incidence in tomato and increased the plant growth parameters (root and shoot length, antioxidant activity, number of fruits per plant, etc.) under saline conditions. The study reports a novel bioinoculant formulation with the ability to promote plant growth parameters in tomato in presence of phytopathogens even under saline conditions.

Pseudomonas and Acinetobacter spp. capable of metabolizing aromatics displays multifarious plant growth promoting traits: Insights on strategizing consortium-based application to agro-ecosystems

The persistence and slow rate of natural attenuation of genotoxic and endocrine-disrupting aromatic pollutants has impacted the soil heath of agro-ecologies leading to various health and environmental issues. Ecotoxic-xenobiotic compounds degradation property along with plant growth promoting traits of microbes are beneficial for devising strategies for improved agricultural soil health and its clean-up. Present study assessed the multipartite plant growth promoting-, biocontrol- and niche adaptive- traits of five aromatic degrading soil bacteria viz. Pseudomonas bharatica CSV86T, Pseudomonas sp. C5pp, Pseudomonas sp. PP4, Pseudomonas sp. PPD and Acinetobacter sp. ISP4. Solubilization of rock phosphate (P) and feldspar (K), production of plant growth hormone (auxin; IAA), ammonia, siderophores and ACC deaminase by strain CSV86T, C5pp and PP4 were observed. Similarly, high fusaric acid resistance (MIC: 1–1.2 mg mL−1) and tolerance to salinity (5–7.5 %) was observed. In vitro application of CSV86T, C5pp, PP4 and ISP4 as consortium resulted in significant increase (P < 0.05) in the seedling shoot length (15–37 %), root length (65–150 %) and biomass (30–40 %) of wheat, mung bean and fenugreek. In-soil microcosm assay confirmed the plant growth promoting and phytoprotective ability of this consortium against toxicity of aromatics in contaminated soil. Antifungal activity against phytopathogenic fungi Magnaporthe oryzae and Aspergillus spp., ability to produce lytic enzymes and HCN suggests their biocontrol potential. Four aromatic degrading bacteria (as a consortium) exhibiting plant growth promotion and biocontrol activity could be used as an eco-friendly bioremediator-biofertilizer-biocontrol agent (bioformulation) for restoration of agricultural soil with improved crop productivity.

Genome Sequence Resource of Bacillus velezensis Strain HC-5, Revealing Its Potential as a Native Biological Control Agent Against Ginger Bacterial Wilt

The Bacillus velezensis strain HC-5, isolated from rhizosphere soil of ginger, exhibits effective biocontrol activity against ginger bacterial wilt. However, its genomic information has remained unexplored. This study presents the complete genome sequence of B. velezensis HC-5. The genome spans 4,237,244 bp and consists of 4,258 genes, with a total GC content of 45.85%. The HC-5 genome comprises 4,058 coding genes, 86 transfer RNAs, 27 ribosomal RNAs (rRNAs), 19 small RNAs (sRNAs), 70 interspersed repeats, and 74 tandem repeats. Moreover, we identified 10 gene clusters responsible for the biosynthesis of antibiotic compounds and 28 two-component systems associated with the bacterium's environmental adaptability. This genome sequence of B. velezensis HC-5 enhances our understanding of its biocontrol mechanisms and supports its potential development as an effective biocontrol agent for managing bacterial wilt in ginger production. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Biocontrol efficacy of Pseudomonas mediterranea PVCT 3C against Plenodomus tracheiphilus: In vitro and in planta mechanisms at early disease stages

In this study, we investigated the biocontrol activity of the P. mediterranea strain PVCT 3C against Mal secco, a severe disease of citrus caused by the vascular fungus Plenodomus tracheiphilus. In vitro, bacterial diffusible compounds, volatile organic compounds and culture filtrates produced by PVCT 3C reduced the mycelial growth and conidial germination of P. tracheiphilus, also affecting the mycelial pigmentation. The application of bacterial suspensions by leaf-spraying before the inoculation with the pathogen on plants of the highly susceptible species sour orange and lemon led to an overall reduction in incidence and disease index, above all during the early disease stage. PVCT 3C genome was subjected to whole-genome shotgun sequencing to study the molecular mechanisms of action of this strain. In silico annotation of biosynthetic gene clusters for secondary metabolites revealed the presence of numerous clusters encoding antimicrobial compounds (e.g. cyclic lipopeptides, hydrogen cyanide, siderophores) and candidate novel products. During the asymptomatic disease phase (seven days post-inoculation), bacterial treatments interfered with the expression of different fungal genes, as assessed with an NGS and de novo assembly RNA-seq approach. These results suggest that P. mediterranea PVCT 3C or its secondary metabolites may offer a potential effective and sustainable alternative to contain P. tracheiphilus infections via integrated management.

Root Rot Management in Common Bean (Phaseolus vulgaris L.) Through Integrated Biocontrol Strategies using Metabolites from Trichoderma harzianum, Serratia marcescens, and Vermicompost Tea

Common bean (Phaseolus vulgaris L.) is an essential food staple and source of income for small-holder farmers across Africa. However, yields are greatly threatened by fungal diseases like root rot induced by Rhizoctonia solani. This study aimed to evaluate an integrated approach utilizing vermicompost tea (VCT) and antagonistic microbes for effective and sustainable management of R. solani root rot in common beans. Fourteen fungal strains were first isolated from infected common bean plants collected across three Egyptian governorates, with R. solani being the most virulent isolate with 50% dominance. Subsequently, the antagonistic potential of vermicompost tea (VCT), Serratia sp., and Trichoderma sp. was assessed against this destructive pathogen. Combinations of 10% VCT and the biocontrol agent isolates displayed potent inhibition of R. solani growth in vitro, prompting in planta testing. Under greenhouse conditions, integrated applications of 5 or 10% VCT with Serratia marcescens, Trichoderma harzianum, or effective microorganisms (EM1) afforded up to 95% protection against pre- and post-emergence damping-off induced by R. solani in common bean cv. Giza 6. Similarly, under field conditions, combining VCT with EM1 (VCT + EM1) or Trichoderma harzianum (VCT + Trichoderma harzianum) substantially suppressed disease severity by 65.6% and 64.34%, respectively, relative to untreated plants. These treatments also elicited defense enzyme activity and distinctly improved growth parameters including 136.68% and 132.49% increases in pod weight per plant over control plants. GC–MS profiling of Trichoderma harzianum, Serratia marcescens, and vermicompost tea (VCT) extracts revealed unique compounds dominated by cyclic pregnane, fatty acid methyl esters, linoleic acid derivatives, and free fatty acids like oleic, palmitic, and stearic acids with confirmed biocontrol and plant growth-promoting activities. The results verify VCT-mediated delivery of synergistic microbial consortia as a sustainable platform for integrated management of debilitating soil-borne diseases, enhancing productivity and incomes for smallholder bean farmers through regeneration of soil health. Further large-scale validation can pave the adoption of this climate-resilient approach for securing food and nutrition security.

A Comprehensive Approach Combining Short-Chain Polyphosphate and Bacterial Biostimulants for Effective Nutrient Solubilization and Enhanced Wheat Growth.

Phosphorus constitutes a crucial macronutrient for crop growth, yet its availability often limits food production. Efficient phosphorus management is crucial for enhancing crop yields and ensuring food security. This study aimed to enhance the efficiency of a short-chain polyphosphate (PolyP) fertilizer by integrating it with plant growth-promoting bacteria (PGPB) to improve nutrient solubilization and wheat growth. Specifically, the study investigated the effects of various bacterial strains on wheat germination and growth when used in conjunction with PolyP. To achieve this, a greenhouse experiment was conducted in which the wheat rhizosphere was amended with a short-chain PolyP fertilizer. Based on the morphological aspect, eight bacteria, designated P1 to P8, were isolated and further characterized. Plant growth-promoting traits were observed in all bacterial strains, as they presented the ability to produce Indole Acetic Acid (IAA) in significant amounts ranging from 7.5 ± 0.3 µg/mL to 44.1 ± 2 µg/mL, expressed by B. tropicus P4 and P. soyae P1, respectively. They also produced ammonia, hydrogen cyanide (HCN), and siderophores. Their effect against the plant pathogen Fusarium culmorum was also assessed, with P. reinekei P2 demonstrating the highest biocontrol activity as it presented a total inhibitory effect. Additionally, some strains exhibited the ability to solubilize/hydrolyze phosphorus, potassium, and zinc. In vivo, the initial growth potential of wheat seeds indicated that those inoculated with the isolated strains exhibited elevated germination rates and enhanced root growth. Based on their plant growth-promoting traits and performance in the germination assay, three strains were selected for producing the best results, specifically phosphorus hydrolyzation/solubilization, zinc solubilization, IAA production, HCN, and siderophores production. Wheat seeds were inoculated by drenching in a bacterial suspension containing 1010 CFU/mL of log phase culture, and an in planta bioassay was conducted in a growth chamber using three selected strains (Pseudomonas soyae P1, Pseudomonas reinekei P2, and Bacillus tropicus P4), applied either individually or with PolyP on a P-deficient soil (28 mg/kg of P Olsen). Our findings demonstrated that the combination of Pseudomonas soyae P1 and PolyP achieved the highest shoot biomass, averaging 41.99 ± 0.87 g. Notably, applying P. soyae P1 or Bacillus tropicus P4 alone yielded similar results to the use of PolyP alone. At the heading growth stage, the combination of Bacillus tropicus P4 and PolyP significantly increased the Chlorophyll Content Index (CCI) to 37.02 µmol/m2, outperforming both PolyP alone (24.07 µmol/m2) and the control (23.06 µmol/m2). This study presents an innovative approach combining short-chain PolyP with bacterial biostimulants to enhance nutrient availability and plant growth. By identifying and characterizing effective bacterial strains, it offers a sustainable alternative to conventional fertilizers.

The nematode signaling molecule ascr#18 induces prepenetration defenses in wheat against a leaf rust fungus

Leaf rust, caused by the pathogenic biotrophic rust fungus Puccinia triticina (Pt), is one of the most destructive wheat diseases worldwide; its negative impact on crop yields is exacerbated by increasing temperatures due to climate change. Ascarosides are nematode pheromones that induce resistance to microbial pathogens and pests in a wide range of crops, making them valuable components in biocontrol scenarios. We investigated the effect on infection of various wheat (Triticum aestivum) genotypes with the virulent Pt race 77W × R by ascr#18, the major ascaroside secreted into the rhizosphere by plant-parasitic nematodes. Spraying the leaves with ascr#18 24 h before inoculation with fungal uredospores slowed disease development and resulted in a reduction of the number of rust pustules on treated compared to untreated leaves. Dose–response analysis over the nano- and micromolar range revealed a broad optimum concentration down to 0.01nM ascr#18. Microscopic analysis showed very early arrest of the fungus at the appressorial stage, with associated enhanced local accumulation of H2O2 and abortive stoma penetration. Similarly, ascr#18 also induced strong resistance to Pt race PKTTS, confirming its race-unspecific biocontrol activity. The results of this study are consistent with and extend previous research that has shown that ascr#18 activates plant immunity and thus protects plants from pathogens even at very low doses.

Biocontrol potentiality of &lt;i&gt;Burkholderia vietnamiensis&lt;/i&gt; nrv12 against the rice blast fungus &lt;i&gt;magnaporthe oryzae&lt;/i&gt;

Rice blast disease, caused by the pathogenic fungus Magnaporthe oryzae, is a widespread infection leading to serious crop loss worldwide. In order to achieve sustainable agriculture, root-associated bacteria have been applied to manage fungal diseases and promote growth. The present study aimed to evaluate in vitro the growth-promoting ability and in vivo biocontrol activity against M. oryzae of rice rhizosphere bacterium. Out of sixty-eight isolates recovered from the rhizosphere of blast-infected rice plants, isolate NRV12 exhibited the highest antifungal activity against M. oryzae SH, with an inhibition percentage of 72.7±3.44%. By analysis of 16S rRNA sequence associated with morphology, physiological and biochemical tests, the strain was identified as Burkholderia vietnamiensis. In addition, NRV12 produced hydrolytic enzymes (amylase, cellulase, protease), indole acetic acid (IAA), exhibited nitrogen-fixing potential and the ability to solubilize phosphate and zinc. Innoculation with NRV12 significantly promoted in vivo rice seedling growth to 23.3% as compared to the non-bacteria-treated seedlings. Importantly, infected rice seedlings treated with NRV12 led to a 40% disease reduction in rice blast. These findings suggest that NRV12 is a valuable and promising isolate with biocontrol potential against rice blast caused by M. oryzae.

Field isolates of Beauveria bassiana exhibit biological heterogeneity in multitrophic interactions of agricultural importance

Beauveria bassiana (Bb) is a widespread entomopathogenic fungus widely used in agriculture for crop protection. Other than pest control, fungi belonging to the B. bassiana complex represent an important microbial resource in agroecosystems, considering their multiple interactions with other microorganisms as antagonists of phytopathogens, or with plants as endophytic colonizers and growth promoters. Here, we characterised field collected or commercial isolates of B. bassiana relative to the environmental factors that affect their growth. We further compared the metabolome, the entomopathogenic potential and biocontrol activity of the tested isolates respectively on the insect pest Spodoptera littoralis or against the fungal plant pathogen Fusarium oxysporum. Our analysis revealed that the B. bassiana complex is characterised by a high level of inter-isolate heterogeneity in terms of nutritional requirements, establishment of intra- or inter-kingdom interactions, and the nature of metabolites produced. Interestingly, certain B. bassiana isolates demonstrated a preference for low nutrient plant-derived media, which hints at their adaptation towards an endophytic lifestyle over a saprophytic one. In addition, there was a noticeable variation among different B. bassiana isolates in their capacity to kill S. littoralis larvae in a contact infection test, but not in an intrahaemocoelic injection experiment, suggesting a unique level of adaptability specific to the host. On the other hand, most B. bassiana isolates exhibited similar biocontrol efficacy against the soil-dwelling ascomycete F. oxysporum f. sp. lycopersici, a pathogen responsible for vascular wilt disease in tomato plants, effectively averting wilting. Overall, we show that the effectiveness of B. bassiana isolates can greatly vary, emphasising the importance of isolate selection and nutritional adaptability consideration for their use in sustainable agriculture.

Comparative genomic analysis of Bacillus atrophaeus HAB-5 reveals genes associated with antimicrobial and plant growth-promoting activities.

Bacillus atrophaeus HAB-5 is a plant growth-promoting rhizobacterium (PGPR) that exhibits several biotechnological traits, such as enhancing plant growth, colonizing the rhizosphere, and engaging in biocontrol activities. In this study, we conducted whole-genome sequencing of B. atrophaeus HAB-5 using the single-molecule real-time (SMRT) sequencing platform by Pacific Biosciences (PacBio; United States), which has a circular chromosome with a total length of 4,083,597 bp and a G + C content of 44.21%. The comparative genomic analysis of B. atrophaeus HAB-5 with other strains, Bacillus amyloliquefaciens DSM7, B. atrophaeus SRCM101359, Bacillus velezensis FZB42, B. velezensis HAB-2, and Bacillus subtilis 168, revealed that these strains share 2,465 CDSs, while 599 CDSs are exclusive to the B. atrophaeus HAB-5 strain. Many gene clusters in the B. atrophaeus HAB-5 genome are associated with the production of antimicrobial lipopeptides and polypeptides. These gene clusters comprise distinct enzymes that encode three NRPs, two Transat-Pks, one terpene, one lanthipeptide, one T3PKS, one Ripp, and one thiopeptide. In addition to the likely IAA-producing genes (trpA, trpB, trpC, trpD, trpE, trpS, ywkB, miaA, and nadE), there are probable genes that produce volatile chemicals (acoA, acoB, acoR, acuB, and acuC). Moreover, HAB-5 contained genes linked to iron transportation (fbpA, fetB, feuC, feuB, feuA, and fecD), sulfur metabolism (cysC, sat, cysK, cysS, and sulP), phosphorus solubilization (ispH, pstA, pstC, pstS, pstB, gltP, and phoH), and nitrogen fixation (nif3-like, gltP, gltX, glnR, glnA, nadR, nirB, nirD, nasD, narl, narH, narJ, and nark). In conclusion, this study provides a comprehensive genomic analysis of B. atrophaeus HAB-5, pinpointing the genes and genomic regions linked to the antimicrobial properties of the strain. These findings advance our knowledge of the genetic basis of the antimicrobial properties of B. atrophaeus and imply that HAB-5 may employ a variety of commercial biopesticides and biofertilizers as a substitute strategy to increase agricultural output and manage a variety of plant diseases.

Physiological and genomic insights into a psychrotrophic drought-tolerant bacterial consortium for crop improvement in cold, semiarid regions

The agricultural land in the Indian Himalayan region (IHR) is susceptible to various spells of snowfall, which can cause nutrient leaching, low temperatures, and drought conditions. The current study, therefore, sought an indigenous psychrotrophic plant growth-promoting (PGP) bacterial inoculant with the potential to alleviate crop productivity under cold and drought stress. Psychrotrophic bacteria preisolated from the night-soil compost of the Lahaul Valley of northwestern Himalaya were screened for phosphate (P) and potash (K) solubilization, nitrogen fixation, indole acetic acid (IAA) production, siderophore and HCN production) in addition to their tolerance to drought conditions for consortia development. Furthermore, the effects of the selected consortium on the growth and development of wheat (Triticum aestivum L.) and maize (Zea mays L.) were assessed in pot experiments under cold semiarid conditions (50 % field capacity). Among 57 bacteria with P and K solubilization, nitrogen fixation, IAA production, siderophore and HCN production, Pseudomonas protegens LPH60, Pseudomonas atacamensis LSH24, Psychrobacter faecalis LUR13, Serratia proteamaculans LUR44, Pseudomonas mucidolens LUR70, and Glutamicibacter bergerei LUR77 exhibited tolerance to drought stress (-0.73 MPa). The colonization of wheat and maize seeds with these drought-tolerant PGP strains resulted in a germination index >150, indicating no phytotoxicity under drought stress. Remarkably, a particular strain, Pseudomonas sp. LPH60 demonstrated antagonistic activity against three phytopathogens Ustilago maydis, Fusarium oxysporum, and Fusarium graminearum. Treatment with the consortium significantly increased the foliage (100 % and 160 %) and root (200 % and 133 %) biomasses of the wheat and maize plants, respectively. Furthermore, whole-genome sequence comparisons of LPH60 and LUR13 with closely related strains revealed genes associated with plant nutrient uptake, phytohormone synthesis, siderophore production, hydrogen cyanide (HCN) synthesis, volatile organic compound production, trehalose and glycine betaine transport, cold shock response, superoxide dismutase activity, and gene clusters for nonribosomal peptide synthases and polyketide synthetases. With their PGP qualities, biocontrol activity, and ability to withstand environmental challenges, the developed consortium represents a promising cold- and drought-active PGP bioinoculant for cereal crops grown in cold semiarid regions.

Evaluating the Biocontrol Efficacy and Antioxidant Potential of Phellinus caribaeo-quercicola-A First Report Dual-Action Endophyte From Inula racemosa Hook. F.

Phellinus caribaeo-quercicola is a basidiomycetous fungus, isolated as an endophyte in this study from the healthy and symptomless leaves of Inula racemosa Hook. f., an important medicinal herb growing in Kashmir Himalaya. This study combines morphological, molecular and phylogenetic techniques to identify the fungal endophyte, using the ITS sequence of nrDNA. A detached leaf assay was conducted to assess the pathogenicity of the fungal endophyte suggesting its mutually symbiotic relationship with the host. The authors also investigated the antifungal potential of the isolated endophytic strain to ascertain its use as a biocontrol agent. The study shows that P. caribaeo-quercicola INL3-2 strain exhibits biocontrol activity against four key fungal phytopathogens that cause significant agronomic and economic losses: Aspergillus flavus, Aspergillus niger, Fusarium solani, and Fusarium oxysporum. Notably, P. caribaeo-quercicola INL3-2 strain is highly effective against A. flavus, with an inhibition percentage of 57.63%. In addition, this study investigates the antioxidant activity of P. caribaeo-quercicola INL3-2 strain crude extracts using ethyl acetate and methanol as solvents. The results showed that the methanolic fraction of P. caribaeo-quercicola exhibits potential as an antioxidant agent, with an IC50 value of 171.90 ± 1.15 µg/mL. This investigation is first of its kind and marks the initial report of this fungal basidiomycete, P. caribaeo-quercicola, as an endophyte associated with a medicinal plant. The findings of this study highlight the potential of P. caribaeo-quercicola INL3-2 strain as a dual-action agent with both biocontrol and antioxidant properties consistent with the medicinal properties of Inula racemosa. This endophytic fungus could be a promising source of natural compounds for use in agriculture, medicine, and beyond.

Draft genome sequence of Bacillus atrophaeus TL401, a biocontrol bacterium with plant growth-promoting properties.

Bacillus atrophaeus strain TL401 exhibits biocontrol activity against Botrytis cinerea on tomato and plant growth promotion. Here, we present the draft genome sequence of strain ITL401, which includes a circular chromosome with 4,213,034 bp and a guanine-cytosine content of 43.39%.

Transfection of entomopathogenic Metarhizium species with a mycovirus confers hypervirulence against two lepidopteran pests

Although most known viruses infecting fungi pathogenic to higher eukaryotes are asymptomatic or reduce the virulence of their host fungi, those that confer hypervirulence to entomopathogenic fungus still need to be explored. Here, we identified and studied a novel mycovirus in Metarhizium flavoviride, isolated from small brown planthopper (Laodelphax striatellus). Based on molecular analysis, we tentatively designated the mycovirus as Metarhizium flavoviride partitivirus 1 (MfPV1), a species in genus Gammapartitivirus, family Partitiviridae. MfPV1 has two double-stranded RNAs as its genome, 1,775 and 1,575 bp in size respectively, encapsidated in isometric particles. When we transfected commercial strains of Metarhizium anisopliae and Metarhizium pingshaense with MfPV1, conidiation was significantly enhanced (t test; P-value < 0. 01), and the significantly higher mortality rates of the larvae of diamondback moth (Plutella xylostella) and fall armyworm (Spodoptera frugiperda), two important lepidopteran pests were found in virus-transfected strains (ANOVA; P-value < 0.05). Transcriptomic analysis showed that transcript levels of pathogenesis-related genes in MfPV1-infected M. anisopliae were obviously altered, suggesting increased production of metarhizium adhesin-like protein, hydrolyzed protein, and destruxin synthetase. Further studies are required to elucidate the mechanism whereby MfPV1 enhances the expression of pathogenesis-related genes and virulence of Metarhizium to lepidopteran pests. This study presents experimental evidence that the transfection of other entomopathogenic fungal species with a mycovirus can confer significant hypervirulence and provides a good example that mycoviruses could be used as a synergistic agent to enhance the biocontrol activity of entomopathogenic fungi.

Revealing the diversity of Jojoba-associated fungi using amplicon metagenome approach and assessing the in vitro biocontrol activity of its cultivable community.

Jojoba shrubs are wild plants cultivated in arid and semiarid lands and characterized by tolerance to drought, salinity, and high temperatures. Fungi associated with such plants may be attributed to the tolerance of host plants against biotic stress in addition to the promotion of plant growth. Previous studies showed the importance of jojoba as jojoba oil in the agricultural field; however, no prior study discussed the role of jojoba-associated fungi (JAF) in reflecting plant health and the possibility of using JAF in biocontrol. Here, the culture-independent and culture-dependent approaches were performed to study the diversity of the jojoba-associated fungi. Then, the cultivable fungi were evaluated for in-vitro antagonistic activity and in vitro plant growth promotion assays. The metagenome analysis revealed the existence of four fungal phyla: Ascomycota, Aphelidiomycota, Basidiomycota, and Mortierellomycota. The phylum Ascomycota was the most common and had the highest relative abundance in soil, root, branch, and fruit samples (59.7%, 50.7%, 49.8%, and 52.4%, respectively). Alternaria was the most abundant genus in aboveground tissues: branch (43.7%) and fruit (32.1%), while the genus Discosia had the highest abundance in the underground samples: soil (24%) and root (30.7%). For the culture-dependent method, a total of 14 fungi were isolated, identified, and screened for their chitinolytic and antagonist activity against three phytopathogenic fungi (Fusarium oxysporum, Alternaria alternata and Rhizoctonia solani) as well as their in vitro plant growth promotion (PGP) activity. Based on ITS sequence analysis, the selected potent isolates were identified as AspergillusstellatusEJ-JFF3, Aspergillusflavus EJ-JFF4, Stilbocreasp. EJ-JLF1, Fusariumsolani EJ-JRF3, and AmesiaatrobrunneaEJ-JSF4. The endophyte strain A. flavus EJ-JFF4 exhibited the highest chitinolytic activity (9 Enzyme Index) and antagonistic potential against Fusarium oxysporum, Alternaria alternata, and Rhizoctonia solani phytopathogens with inhibitory percentages of 72, 70, and 80 respectively. Also, A. flavus EJ-JFF4 had significant multiple PGP properties, including siderophore production (69.3%), phosphate solubilization (95.4 µg ml-1). The greatest production of Indol-3-Acetic Acid was belonged to A.atrobrunnea EJ-JSF4 (114.5 µg ml-1). The analysis of FUNGuild revealed the abundance of symbiotrophs over other trophic modes, and the guild of endophytes was commonly assigned in all samples. For the first time, this study uncovered fungal diversity associated with jojoba plants using a culture-independent approach and in-vitro assessed the roles of cultivable fungal strains in promoting plant growth and biocontrol. The present study indicated the significance of jojoba shrubs as a potential source of diverse fungi with high biocontrol and PGP activities.

Biocontrol activities of grey mould of grapes with the volatile organic compounds generated by yeast HXMG-1 isolated from grapes

Biocontrol activities of grey mould of grapes with the volatile organic compounds generated by yeast HXMG-1 isolated from grapes