Seed endophytic bacterial profiling from wheat varieties of contrasting heat sensitivity

Wheat yield can be limited by many biotic and abiotic factors. Heat stress at the grain filling stage is a factor that reduces wheat production tremendously. The potential role of endophytic microorganisms in mitigating plant stress through various biomolecules like enzymes and growth hormones and also by improving plant nutrition has led to a more in-depth exploration of the plant microbiome for such functions. Hence, we devised this study to investigate the abundance and diversity of wheat seed endophytic bacteria (WSEB) from heatS (heat susceptible, GW322) and heatT (heat tolerant, HD3298 and HD3271) varieties by culturable and unculturable approaches. The results evidenced that the culturable diversity was higher in the heatS variety than in the heatT variety and Bacillus was found to be dominant among the 10 different bacterial genera identified. Though the WSEB population was higher in the heatS variety, a greater number of isolates from the heatT variety showed tolerance to higher temperatures (up to 55°C) along with PGP activities such as indole acetic acid (IAA) production and nutrient acquisition. Additionally, the metagenomic analysis of seed microbiota unveiled higher bacterial diversity, with a predominance of the phyla Proteobacteria covering >50% of OTUs, followed by Firmicutes and Actinobacteria. There were considerable variations in the abundance and diversity between heat sensitivity contrasting varieties, where notably more thermophilic bacterial OTUs were observed in the heatT samples, which could be attributed to conferring tolerance against heat stress. Furthermore, exploring the functional characteristics of culturable and unculturable microbiomes would provide more comprehensive information on improving plant growth and productivity for sustainable agriculture

Protocol for Quantitative Estimation of Hydrogen Cyanide Production from Bacteria

Hydrogen cyanide (HCN) is a volatile, nitrogen-containing secondary metabolite produced by various bacterial species, primarily during the idiophase of growth under nutrient-limiting or competitive conditions. It plays a significant ecological role as a biocontrol agent by inhibiting the respiratory enzymes of plant pathogens and modulating microbial competition in the rhizosphere. Although protocols for detecting HCN production have existed for over a century, they have largely remained qualitative and are rarely optimized for quantitative assessment. This is mainly due to the volatile nature of HCN, unidentified stable reference standards, and the absence of a robust, universally accepted protocol that ensures consistency across diverse microbial types. In this study, we present a simplified and efficient colorimetric method to quantify HCN production in both Gram-positive and Gram-negative bacteria. Qualitatively, HCN production was observed by a color change due to the isopurpurate complex. This compound was then eluted and quantified by measuring absorbance at 625 nm. The method uses potassium ferrocyanide as a standard, whose slow dissociation constant enables a stable and controlled release of cyanide ions for calibration, unlike highly dissociative salts like KCN that introduce early volatilization errors. This protocol demonstrated high sensitivity, capable of detecting HCN at concentrations as low as ppm levels, with strong correlation to the standard curve (R2 > 0.99). Achieving such sensitivity with other conventional methods, such as gas detection tubes or electrochemical sensors, often requires more sophisticated instrumentation and strict handling conditions. In contrast, this approach offers a cost-effective, reproducible, and user-friendly alternative. While a universally adopted method is still lacking due to standardization challenges and HCN volatility, the proposed protocol marks a significant advancement toward accurate and accessible quantitative assessment in microbiological and agricultural applications

Determination of nodule coverage parameters using multibeam normal incidence echo characteristics: A study in the Indian Ocean

A study of the echo peak amplitudes from known nodule areas is initiated to observe the acoustic response for varying nodule abundances and number densities. A statistical study of the peak amplitudes from different nodule areas confirms that the coefficient of variation is the highest for medium nodule abundance and number density. Echo fluctuation study based on the Rician probability density function (PDF) establishes that the non‐nodule sediment bottom contributes to less scattering, i.e., it is a microtopographic type, whereas scattering is dominant in the nodule‐bearing areas. The spectral studies are conducted on depth data of different areas. This study ensures that the signal scattering in the nodule bottom area is due to the nodules lying on the seabed rather than the large / small‐scale topographic variations. The study based on Poisson PDF for nodule area confirms this fact again. Agreement between the nodule distribution and the Poisson distribution parameter is clearly seen. Such a relation is not observed in the case of Rician density functions

Prospecting catabolic diversity of microbial strains for developing microbial consortia and their synergistic effect on Lentil (Lens esculenta) growth, yield and iron biofortification

Not AvailableCarbon profiling of heterotrophic microbial inoculants is worthwhile strategy for formulating consortium-based biofertilizers. Consortium-based biofertilizers are better than single strain-based biofertilizers for sustaining agricultural productivity and enhancing micronutrient concentration in grains. Currently, we investigated catabolic diversity among microbes using different carbon sources and certain enzyme activities. A field experiment was also carried to evaluate the synergistic effect of selected lentil Rhizobia and plant growth promoting rhizobacteria strains on lentil growth, yield, nitrogen fixation, and Fe-content in seeds. On the basis of carbon profiling Bacillus sp. RB1 and Pseudomonas sp. RP1 were selected for synergistic study with lentil Rhizobium-Rhizobium leguminosarum subsp. viciae RR1. Co-inoculation of Rhizobium with Bacillus sp. RB1 and Pseudomonas sp. RP1 significantly enhanced the plant height, number of pods per plant, seed yield, number of nodules per plant, nitrogenase activity and Fe biofortification in seed over the single Rhizobium inoculation or dual combination of Rhizobium + RB1 or RP1. The response of single Rhizobium inoculation or co-inoculation of Rhizobium with RB1 and/or RP1 at 50% RDF was almost similar or higher than full dose of recommended N:P:K with respect to lentil yield and Fe biofortification in seed. This deciphered grouping of microbial strains for formulation of microbial consortia-based biofertilizers and revealed the promise of consortium of Rhizobium and plant growth promoting rhizobacteria in improving the biological yield and enhancing the Fe content of lentil seed.Not Availabl

Community-forming traits play role in effective colonization of plant-growth-promoting bacteria and improved plant growth

Community-forming traits (CFts) play an important role in the effective colonization of plant-growth-promoting bacterial communities that influence host plants positively by modulating their adaptive functions. In this study, by considering plant-growth-promoting traits (PGPts) and community-forming traits (CFts), three communities were constructed, viz., SM1 (PGPts), SM2 (CFts), and SM3 (PGPts+CFts). Each category isolates were picked up on the basis of their catabolic diversity of different carbon sources. Results revealed a distinctive pattern in the colonization of the communities possessed with CF traits. It was observed that the community with CFts colonized inside the plant in groups or in large aggregations, whereas the community with only PGPts colonized as separate individual and small colonies inside the plant root and leaf. The effect of SM3 in the microcosm experiment was more significant than the uninoculated control by 22.12%, 27.19%, and 9.11% improvement in germination percentage, chlorophyll content, and plant biomass, respectively. The significant difference shown by the microbial community SM3 clearly demonstrates the integrated effect of CFts and PGPts on effective colonization vis-à-vis positive influence on the host plant. Further detailed characterization of the interaction will take this technology ahead in sustainable agriculture

Microbial Community and Function-Based Synthetic Bioinoculants: A Perspective for Sustainable Agriculture

Interactions among the plant microbiome and its host are dynamic, both spatially and temporally, leading to beneficial or pathogenic relationships in the rhizosphere, phyllosphere, and endosphere. These interactions range from cellular to molecular and genomic levels, exemplified by many complementing and coevolutionary relationships. The host plants acquire many metabolic and developmental traits such as alteration in their exudation pattern, acquisition of systemic tolerance, and coordination of signaling metabolites to interact with the microbial partners including bacteria, fungi, archaea, protists, and viruses. The microbiome responds by gaining or losing its traits to various molecular signals from the host plants and the environment. Such adaptive traits in the host and microbial partners make way for their coexistence, living together on, around, or inside the plants. The beneficial plant microbiome interactions have been exploited using traditional culturable approaches by isolating microbes with target functions, clearly contributing toward the host plants’ growth, fitness, and stress resilience. The new knowledge gained on the unculturable members of the plant microbiome using metagenome research has clearly indicated the predominance of particular phyla/genera with presumptive functions. Practically, the culturable approach gives beneficial microbes in hand for direct use, whereas the unculturable approach gives the perfect theoretical information about the taxonomy and metabolic potential of well-colonized major microbial groups associated with the plants. To capitalize on such beneficial, endemic, and functionally diverse microbiome, the strategic approach of concomitant use of culture-dependent and culture-independent techniques would help in designing novel “biologicals” for various crops. The designed biologicals (or bioinoculants) should ensure the community’s persistence due to their genomic and functional abilities. Here, we discuss the current paradigm on plant-microbiome-induced adaptive functions for the host and the strategies for synthesizing novel bioinoculants based on functions or phylum predominance of microbial communities using culturable and unculturable approaches. The effective crop-specific inclusive microbial community bioinoculants may lead to reduction in the cost of cultivation and improvement in soil and plant health for sustainable agriculture.</jats:p

DataSheet_1_Seed endophytic bacterial profiling from wheat varieties of contrasting heat sensitivity.docx

Wheat yield can be limited by many biotic and abiotic factors. Heat stress at the grain filling stage is a factor that reduces wheat production tremendously. The potential role of endophytic microorganisms in mitigating plant stress through various biomolecules like enzymes and growth hormones and also by improving plant nutrition has led to a more in-depth exploration of the plant microbiome for such functions. Hence, we devised this study to investigate the abundance and diversity of wheat seed endophytic bacteria (WSEB) from heatS (heat susceptible, GW322) and heatT (heat tolerant, HD3298 and HD3271) varieties by culturable and unculturable approaches. The results evidenced that the culturable diversity was higher in the heatS variety than in the heatT variety and Bacillus was found to be dominant among the 10 different bacterial genera identified. Though the WSEB population was higher in the heatS variety, a greater number of isolates from the heatT variety showed tolerance to higher temperatures (up to 55°C) along with PGP activities such as indole acetic acid (IAA) production and nutrient acquisition. Additionally, the metagenomic analysis of seed microbiota unveiled higher bacterial diversity, with a predominance of the phyla Proteobacteria covering >50% of OTUs, followed by Firmicutes and Actinobacteria. There were considerable variations in the abundance and diversity between heat sensitivity contrasting varieties, where notably more thermophilic bacterial OTUs were observed in the heatT samples, which could be attributed to conferring tolerance against heat stress. Furthermore, exploring the functional characteristics of culturable and unculturable microbiomes would provide more comprehensive information on improving plant growth and productivity for sustainable agriculture.</p

DataSheet_1_Community-forming traits play role in effective colonization of plant-growth-promoting bacteria and improved plant growth.docx

Community-forming traits (CFts) play an important role in the effective colonization of plant-growth-promoting bacterial communities that influence host plants positively by modulating their adaptive functions. In this study, by considering plant-growth-promoting traits (PGPts) and community-forming traits (CFts), three communities were constructed, viz., SM1 (PGPts), SM2 (CFts), and SM3 (PGPts+CFts). Each category isolates were picked up on the basis of their catabolic diversity of different carbon sources. Results revealed a distinctive pattern in the colonization of the communities possessed with CF traits. It was observed that the community with CFts colonized inside the plant in groups or in large aggregations, whereas the community with only PGPts colonized as separate individual and small colonies inside the plant root and leaf. The effect of SM3 in the microcosm experiment was more significant than the uninoculated control by 22.12%, 27.19%, and 9.11% improvement in germination percentage, chlorophyll content, and plant biomass, respectively. The significant difference shown by the microbial community SM3 clearly demonstrates the integrated effect of CFts and PGPts on effective colonization vis-à-vis positive influence on the host plant. Further detailed characterization of the interaction will take this technology ahead in sustainable agriculture.</p