Bioactive Metabolites of Serratia sp. NhPB1 Isolated from Pitcher of Nepenthes and its Application to Control Pythium aphanidermatum.

Plant-associated bacteria have already been considered as the store house of bioactive compounds that confer the plant growth promotion and disease protection. Hence, the unique plant parts have already been expected to harbor diverse microbial communities with multi-beneficial properties. Based on this, the current study has been designed to identify the potential of Serratia sp. NhPB1 isolated from the pitcher of Nepenthes plant for its activity against the infamous pathogen Pythium aphanidermatum. The in vitro antifungal, plant growth promoting and enzymatic activities of the isolate indicated its promises for agricultural application. The isolate NhPB1 was also demonstrated to have positive effect on Solanum lycopersicum and Capsicum annuum, due to its plant beneficial metabolites. From the results of LC-MS/MS analysis, the isolate has also been revealed to have the ability to synthesize bioactive compounds including salicylic acid, cyclodipeptides, acyl homoserine lactone, indole-3-acetic acid, and serrawettin W1. These identified compounds and their known biological properties make the isolate characterized in the study to have significant promises as an eco-friendly solution for the improvement of agricultural productivity.

Multifarious Plant Probiotic Features of Bacillus sp. W11 Isolated from Vermicast and Its Promises for Biocontrol Activity Against Phytopathogens.

Organic agricultural practice using vermisources is considered as one of the common sustainable strategies in agriculture. Apart from their nutrient content, beneficial microbes associated with natural vermicast serve as an efficient bioinoculant for improving agricultural productivity. Hence, studies on the identification of suitable microbial inoculants with multi-trait plant beneficial properties from these hotspots are highly promising as an ecofriendly substitute against harmful chemical fertilizers. The current study has been designed to isolate bacterial strains with various plant probiotic and biocontrol properties against various fungal phytopathogens. Among the various bacteria obtained in the study, the isolate W11 was found to have remarkable inhibitory activity against a broad range of phytopathogens like Fusarium oxysporum, Pythium myriotylum, and Rhizoctonia solani, in addition to its multiple plant growth-promoting properties. The W11 was further identified as Bacillus sp. by 16S rDNA sequence-based analysis. The W11 treatment was also found to enhance the plant growth parameters of Vigna unguiculata. In addition, the priming of Bacillus sp. W11 on potato tuber has confirmed to protect it from Fusarium wilt caused by Fusarium oxysporum. This highlights the possible protective effect of W11 during the post-harvest storage of potatoes. Also, the metabolite analysis of W11 extract by LC-QTOF-MS/MS analysis has revealed the presence of lipopeptide surfactin derivatives with m/z of 1008, 1022, and 1036 (M + H)+. All the results obtained in the study thus indicate the remarkable agricultural promises of Bacillus sp. W11 isolated from vermicast. Even though vermicast has been studied for its beneficial agricultural applications, the isolation of plant beneficial bacteria from it and detailed characterization of its beneficial effects make the study important.

Phenazine 1-carboxylic acid Producing Seed Harbored Endophytic Bacteria from Cultivated Rice Variety of Kerala and Its Broad Range Antagonism to Diverse Plant Pathogens.

Endophytic microorganisms residing within the diverse parts of plants play a significant role in the plant growth and defense response. In the case of the vertically transmitted seed-borne endophytes, they form the promising initiator of the juvenile plant microbiome by supporting the growth and establishment of the seedlings. Hence, the current study emphasizes the isolation and screening of plant beneficial traits of seed endophytes from the cultivated rice variety Jyothi of Kerala, India. Among the 14 bacterial endophytes obtained in the study, the isolate S3 was found to have promising activity against the phytopathogens such as Fusarium oxysporum, Pythium aphanidermatum, Pythium myriotylum, Phytophthora infestans, Rhizoctonia solani, Colletotrichum acutatum, and Sclerotium rolfsii. The isolate S3 was further identified as Paenibacillus polymyxa by the 16S rRNA-based sequence analysis. Furthermore, the isolate was confirmed for its capability for hydrogen cyanide (HCN) production, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, biofilm formation, and nitrogen fixation. The P. polymyxa S3 was also found to have the potential to provide post-harvest protection to the rice kernels from Sclerotium rolfsii. By the LC-MS/MS analysis, the organism was confirmed for the production of phenazine 1-carboxylic acid which could be the prime chemical basis of its antifungal activity. The in vivo plant growth evaluation has also demonstrated the root length enhancement effect of P. polymyxa S3 in Vigna unguiculata. Here, the root length of P. polymyxa S3-treated plant was enhanced to 12.44 ± 0.58223 cm when compared with distilled water control (10.261 ± 0.38151 cm) and the observed change was statistically significant as per the analysis of variance at P value less than 0.05. Based on all these properties, the isolated P. polymyxa S3 could be considered as a promising agent to be used for the development of competent plant probiotic formulations.

Effect of zinc oxide nanoparticle supplementation on the enhanced production of surfactin and iturin lipopeptides of endophytic Bacillus sp. Fcl1 and its ameliorated antifungal activity.

BACKGROUND: Lipopeptides from the Bacillus spp. possess an excellent spectrum of antimicrobial properties which make them suitable candidates to be explored for the food, agricultural, pharmaceutical and biotechnological applications. As the low yield of the lipopeptides limits their applications, methods to enhance their production are highly significant. RESULTS: In this study, extracts prepared from endophytic Bacillus sp. Fcl1 cultured in the presence of various supplements were screened for antifungal activity against Pythium aphanidermatum. From the results, the supplementation of carbon sources and zinc oxide nanoparticles (ZnONPs) was found to have an enhancement effect on the antifungal activity of Bacillus sp. Fcl1. Among these, the highest antifungal activity (73.2%) could be observed for the Fcl1 sample cultured with 5 mg L-1 of ZnONP supplementation. The growth of Fcl1 in the presence of ZnONPs also indicated its compatibility with the nano-supplement in the concentration range used. By liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) analysis, the synthesis of increased numbers of lipopeptide surfactin derivatives could be identified from the extracts of Fcl1 prepared from the carbon sources and ZnONP-supplemented cultures. In addition to the surfactin derivatives, the presence of another lipopeptide iturin was also detected from the extracts of Fcl1 cultured with ZnONPs. CONCLUSION: ZnONP supplementation was found to enhance antifungal activity and lipopeptide production in the Bacillus sp. Fcl1. The use of nanoparticles to enhance the antifungal mechanisms of Fcl1 as observed in the study provides novel insights to explore its applications for sustainable agricultural productivity.

Impact of Supplements on Enhanced Activity of Bacillus amyloliquefaciens BmB1 Against Pythium aphanidermatum Through Lipopeptide Modulation.

The present study has been designed to improve the activity of endophytic Bacillus amyloliquefaciens BmB1 against Pythium aphanidermatum through the culture supplementation with carbon sources, nitrogen sources and zinc oxide nanoparticles (ZnONPs). From the results of the study, supplementation with glucose (45 g/L), yeast extract (7.5 g/L) and ZnONPs (5 mg/L) were found to enhance the antifungal activity of B. amyloliquefaciens BmB1. This was also confirmed by comparative statistical analysis with experimental control. Further LC-Q-TOF-MS analysis of extracts of B. amyloliquefaciens BmB1 cultured with supplements showed a remarkable modulation of its lipopeptide profile. The blend of lipopeptides enhanced during the culture supplementation of B. amyloliquefaciens BmB1 as evidenced by the mass spectrometric analysis can consider to be the basis of its increased activity against P. aphanidermatum. As Bacillus spp. are well known for their biocontrol activities, the results of the study offer ways to improve its agricultural applications.

Plant Beneficial Features and Application of Paraburkholderia sp. NhPBG1 Isolated from Pitcher of Nepenthes hamblack.

Pitchers are the unique structures of carnivorous plants used for the trapping of insects and other small invertebrates. The digestion of captured prey here is assisted by the bacteria, which have been associated with pitchers. These bacterial communities can therefore expect to have a variety of plant beneficial functions. In this study, the bacterial isolate NhPBG1 from the pitcher of Nepenthes hamblack was screened for activity against Pythium aphanidermatum, Rhizoctonia solani, Fusarium oxysporum, and Colletotrichum accutatum and was found to have the inhibitory activity towards all the tested phytopathogens. Interestingly, the isolate was found to have hyper-inhibitory effect against P. aphanidermatum. Further to this, the isolate was also shown to be positive for plant beneficial traits such as indole-3-acetic acid (IAA) and ammonia production, phosphate, potassium and zinc solubilization, nitrogen fixation, and 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity. BLAST analysis of the 16S rDNA sequence of NhPBG1 has identified it as Paraburkholderia sp. Also, the Zingiber officinale rhizome pre-treated with NhPBG1 was found to get protected from P. aphanidermatum induced infection, whereas the control showed symptoms of infection. This was further confirmed by the microscopic evaluation of the presence of fungal mycelia in the tissues of control. However, the mycelial invasion could not be detected in the NhPBG1 treated rhizome. The metabolite profiling of NhPBG1 by GC-MS has identified variety of general metabolites, while the antifungal compounds pyocyanin and 1-hydroxyphenazine could be identified by the LC-MS/MS analysis.

Distribution of antibiotic resistance and virulence factors among the bacteria isolated from diseased Etroplus suratensis.

Considering the emerging concern with the antimicrobial resistance (AMR) evolution, the study has been designed to identify the antibiotic resistance and virulence properties of culturable bacteria isolated from the diseased fish Etroplus suratensis. This has resulted in the purification of 18 morphologically distinct bacterial isolates which were identified by both biochemical and molecular methods. Antibiotic resistance analysis showed the resistance of these isolates to multiple antibiotics and remarkable evolution of AMR. Further screening for virulence factors confirmed five isolates to be positive for haemolytic activity, eight with caseinase, four with DNase, one with gelatinase and three with biofilm-forming properties. In addition to these, the isolates were subjected to PCR-based screening to detect the presence of genes coding for aerolysin and haemolysin. Results showed the presence aerolysin gene in the isolates ESS3.2, ESS3.8, ESI3.3 and ESS3.6, while haemolysin gene was observed to be present in ESG3.1 and ESI3.2. The observed results hence indicate the need for frequent monitoring of these properties among bacterial isolates from diverse environment especially those associated with edible fish.

Assessment of Plant-Probiotic Performance of Novel Endophytic Bacillus sp. in Talc-Based Formulation.

Endophytic bacteria are considered to have a plethora of plant growth promoting and anti-phytopathogenic traits to live within the plants. Hence, they have immense promises for plant probiotic development. In the current study, plant probiotic endophytic Bacillus sp. CaB5 which has been previously isolated from Capsicum annuum was investigated for its performance in talc-based formulation. For this, CaB5 was made into formulation with sterile talc, calcium carbonate, and carboxymethyl cellulose. The viability analysis of the formulation by standard plate count and fluorescence methods has confirmed the stable microbial count up to 45 days. Plant probiotic performance of the prepared formulation was analyzed on cowpea (Vigna unguiculata) and lady's finger (Abelmoschus esculentus). The results showed the formulation treatment to have enhancement effect on seed germination as well as plant growth in both selected plants. The results highlight the potential of CaB5-based formulation for field application to enhance growth of economically important plants.