Biology subjects

Rajabal, V.

Publications and source records attributed to Rajabal, V..

9 recordsLinked to original sources

bioRxiv2026

Drought and rewetting drive divergent microbial dynamics across soil compartments and reveal Trinickia as a key genus for soybean drought resilience

Drought conditions are projected to intensify under climate change, and engineering more resilient plant microbiomes is a promising route to supporting crop productivity. However, effectively harnessing these microbiomes depends on identifying keystone taxa and functional traits that promote plant drought tolerance. Here, we combine deep shotgun metagenomics with absolute-abundance calibration to track genome-resolved microbial dynamics across a prolonged soybean drought and rewetting cycle. Leveraging 728 high-purity metagenome-assembled genomes that capture ~83% of prokaryotic reads on average, we show that drought drives significantly greater change in community structure in the rhizosphere than in bulk soil. This was accompanied by declining taxonomic, yet rising functional diversity. While rhizosphere communities rapidly recovered within 24 hours of rewetting, bulk soil communities instead underwent a progressive post-rewetting disturbance, consistent with a Birch-effect-driven successional shift. Rhizosphere drought enrichment was dominated by a single, taxonomically underexplored genus, Trinickia, whose genomes encode a coordinated suite of osmotic-stress-tolerance, bio-fertilisation, and phytohormone/polyamine biosynthesis traits consistent with plant growth promotion. Supporting this genomic inference, Trinickia abundance strongly correlated with increased root biomass during active drought, independent of plant developmental stage. Together, these findings identify the rhizosphere as a dynamically responsive compartment during drought and rewetting, and highlight Trinickia species as genomically and phenotypically supported candidates for microbiome-based strategies to improve soybean drought resilience.

microbiology↗

bioRxiv2026

Diverse conjugative and mobilisable elements underpin key adaptive traits in Xanthomonas

Horizontal gene transfer mediated by mobile genetic elements (MGEs) is a major driver of bacterial evolution and ecological adaptation. In the plant-associated genus Xanthomonas, multiple MGEs have been implicated in virulence, host specialisation, and environmental persistence, yet MGE diversity and evolutionary dynamics across the genus remain poorly understood. Here, we performed a comparative analysis of conjugative and mobilisable plasmids, integrative and conjugative elements (ICEs), integrative and mobilisable elements (IMEs), and their cargo genes across 516 complete genomes of three major Xanthomonas species: X. campestris, X. cissicola, and X. oryzae. We identified pronounced interspecific differences, with X. cissicola and X. campestris harbouring large and diverse MGE repertoires, comprising 28.3% and 26.7% of their respective pangenomes, whereas X. oryzae contained far fewer MGEs, making up only 3.6% of the identified pangenome. These differences were associated with host defence systems, including CRISPR-Cas and restriction-modification systems, and with variation in CRISPR spacer diversity. IMEs were the most abundant MGEs across all species, encoding diverse defence systems and accessory genes. ICEs exhibited signatures of horizontal transfer within and between species, and across genera. Notably, nearly identical ICEs carrying heavy-metal resistance genes were identified in Xanthomonas and Pseudomonas aeruginosa, indicating recent transfer between genera. MGEs collectively carried genes involved in virulence, interbacterial interactions, defence against phages, and plant cell wall degradation, with several elements associated with specific pathovars. Together, our findings establish MGEs as key drivers of genome plasticity and adaptive evolution in Xanthomonas, shaped by a dynamic interplay with host defence systems.

genomics↗

bioRxiv2026

Discovery of novel antimicrobial resistance genes in food and fertiliser using a high-throughput gene capture and functional screening platform

Integrons are genetic elements that drive bacterial adaptation by capturing and expressing mobile gene cassettes. They play a key role in dissemination of antimicrobial resistance (AMR) genes, particularly in Gram-negative bacteria. In addition to known AMR determinants, integron gene cassettes carry a vast reservoir of novel genes whose functions are largely uncharacterised, making it diNicult to assess their full contribution to the resistome. Contributing to this are limitations in current sequence-based prediction methods which often lack the ability to identify unknown AMR or other adaptive genes with novel mechanisms. To address this, we developed an integron gene cassette capture system, a functional screening platform that captures environmental gene cassettes for direct phenotypic testing. Using this system, we recovered previously unknown AMR determinants while also providing insights into the prevalence of known clinical AMR genes in a range of environmental samples, including food items. Here we provide experimental data on multiple novel bleomycin resistance genes and a stress response gene conferring gentamicin and tobramycin resistance. Our sequence analysis of the captured library also highlighted the diversity of the environmental cassette pool, with 656 unique cassettes recovered, the majority of which encoded proteins with unknown functions. The cassette capture system is a powerful tool for accessing hidden elements of the resistome and discovering novel adaptive genes that may go undetected using current sequence-based approaches. Environmental implicationAntimicrobial resistance (AMR) genes are hazardous biological contaminants, yet the vast majority of environmental integron gene cassettes remain functionally uncharacterised. This study addresses this sequence-to-function gap by deploying a novel functional capture platform directly on realistic environmental matrices, including agricultural fertilisers, coastal seawater, and commercial food products. By characterising these cassettes, we uncovered hidden reservoirs of both novel and clinically established AMR genes circulating in critical exposure pathways. This work reveals the true hazardous potential of the mobile environmental resistome, validating a proactive One Health surveillance tool for monitoring emerging biological threats.

microbiology↗

bioRxiv2026

Revegetation following grazing cessation drives sequential shifts in soil microbial functions and life-history strategies over decadal time scales

Revegetation is a key strategy for restoring degraded lands globally. While this process can reshape belowground microbial communities, the extent to which such changes restore soil ecosystem functions, and whether different microbial traits recover synchronously or on distinct timescales remains less clear. Understanding this is essential for evaluating restoration success, as different microbial traits underpin distinct ecosystem services, from carbon storage to plant growth promotion, and the sequence in which these are restored can inform restoration targets and provide meaningful indicators of long-term ecological recovery. To address this, we applied deep metagenomic sequencing to characterise soil microbial responses following revegetation (spanning 1-31 years prior to sampling) on grazing agricultural lands. We find that revegetation following grazing cessation drove significant and sequential shifts in dominant microbial functions and life-history strategies. Functional changes occurred in distinct phases: an early, rapid restructuring of core soil health processes, detectable as early as three years, including enrichment of nutrient retention and carbon fixation pathways, followed by a more gradual development of plant growth-promoting traits as the plant community matures. Genome-resolved analyses of nearly 500 metagenome-assembled genomes revealed a fundamental shift in dominant microbial life history strategies: from a resource-scavenging and stress tolerance profile in grazing soils to strategies that prioritise biosynthesis and growth yields in revegetated soils. These lifestyle shifts have important implications for enhancing the microbial biomass and carbon sink potential of soils. Together, these findings show that microbial functional shifts following revegetation are temporally structured - informing expectations for when key restoration targets may be achieved, and providing a practical framework for monitoring ecosystem recovery.

microbiology↗

bioRxiv2025

EcoFoldDB: Protein structure-guided functional profiling of ecologically relevant microbial traits at the metagenome scale

Microbial communities are fundamental to planetary health and ecosystem processes. High-throughput metagenomic sequencing has provided unprecedented insights into the structure and function of these communities. However, functionally profiling metagenomes remains constrained due to the limited sensitivity of existing sequence homology-based methods to annotate evolutionarily divergent genes. Protein structure, more conserved than sequence, and intrinsically tied to molecular function, offers a solution. Capitalising on recent breakthroughs in structural bioinformatics, we present EcoFoldDB, a database of protein structures curated for ecologically relevant microbial traits, and its companion pipeline, EcoFoldDB-annotate, which leverages Foldseek with the ProstT5 protein language model for rapid structural homology searching directly from sequence data. EcoFoldDB-annotate outperforms state-of-the-art sequence-based methods in annotating metagenomic proteins, in terms of sensitivity and precision. To demonstrate its utility and scalability, we performed structure-guided functional profiling of 32 million proteins encoded by 8,000 high-quality metagenome-assembled genomes from the global soil microbiome. EcoFoldDB-annotate could resolve the phylogenetic partitioning of important nitrogen cycling pathways, from taxonomically restricted nitrifiers to more widespread denitrifiers, as well as identifying novel, uncultivated bacterial taxa enriched in plant growth-promoting traits. We anticipate that EcoFoldDB will enable researchers to extract ecological insights from environmental genomes and metagenomes, and accelerate discoveries in microbial ecology.

microbiology↗

bioRxiv2024

Seed banking alters native seed microbiome composition and function relative to natural populations

O_LISeed banks are a vital resource for preserving plant species diversity globally. However, seedling establishment and survival rates from banked seeds can be poor. Despite a growing appreciation for the role of seed microbiota in supporting seed quality and plant health, our understanding of the effects of conventional seed banking processes on seed microbiomes remains limited. C_LIO_LIWe investigated the composition and functional potential of the epiphytic seed microbiome of a native plant species using both 16S rRNA gene sequencing and culture-based approaches. C_LIO_LIComparing the bacterial community composition of freshly collected seeds and those sourced from seed banking organisations, we found stored seeds hosted significantly less diverse bacterial populations, with substantial reductions in both low-abundance taxa and some core community members identified in unstored seeds. Bacteria with key plant growth promoting traits including IAA production, ACC deaminase activity, phosphate solubilisation, siderophore activity, and nitrogen fixation were identified in seed epiphytic communities, but these beneficial traits were less prevalent in stored seed compared to fresh seeds. C_LIO_LIOverall, these results suggest that epiphytic seed microbiomes may undergo significant changes during the storage process, selecting for bacteria tolerant to storage conditions, and potentially reducing the population of plant-growth promoting bacteria on seeds. C_LI

microbiology↗

bioRxiv2023

Uncovering bacterial hosts of class 1 integrons in an urban coastal aquatic environment with a single-cell fusion-PCR technology

Horizontal gene transfer (HGT) is a key driver of bacterial evolution via transmission of genetic materials across taxa. Class 1 integrons are genetic elements that correlate strongly with anthropogenic pollution and contribute to the spread of antimicrobial resistance (AMR) genes via HGT. Despite their significance to human health, there is a shortage of robust, culture-free surveillance technologies for identifying uncultivated environmental taxa that harbour class 1 integrons. We developed a modified version of epicPCR (emulsion, paired isolation and concatenation polymerase chain reaction) that links class 1 integrons amplified from single bacterial cells to taxonomic markers from the same cells in emulsified aqueous droplets. Using this single-cell genomic approach and Nanopore sequencing, we successfully assigned class 1 integron gene cassette arrays containing mostly AMR genes to their hosts in coastal water samples that were affected by pollution. Our work presents the first application of epicPCR for targeting variable, multi-gene loci of interest. We also identified the Rhizobacter genus as novel hosts of class 1 integrons. These findings establish epicPCR as a powerful tool for linking taxa to class 1 integrons in environmental bacterial communities and offer the potential to direct mitigation efforts towards hotspots of class 1 integron-mediated dissemination of AMR. SynopsisWe present a novel single-cell genomic surveillance technology for identifying environmental bacterial hosts of a class of mobile genetic elements that are linked to anthropogenic pollution and contribute to the dissemination of antimicrobial resistance.

microbiology↗

bioRxiv2022

Discovery of integrons in Archaea: platforms for cross-domain gene transfer

Horizontal gene transfer between different domains of life is increasingly being recognised as an important driver of evolution, with the potential to provide the recipient with new gene functionality and assist niche adaptation1-3. However, the molecular mechanisms underlying the integration of exogenous genes from foreign domains are mostly unknown. Integrons are a family of genetic elements that facilitate this process within Bacteria via site-specific DNA recombination4-7. Integrons, however, have not been reported outside Bacteria, and thus their potential role in cross-domain gene transfer has not been investigated. Here we show that integrons are also present among diverse phyla within the domain Archaea. Further, we provide experimental evidence that integron-mediated recombination can facilitate the recruitment of archaeal genes by bacteria. Our findings establish a new mechanism that can facilitate horizontal gene transfer between the two domains of prokaryotes, which has important implications for prokaryotic evolution in both clinical and environmental contexts.

microbiology↗

bioRxiv2021

Methods for the targeted sequencing and analysis of integrons and their gene cassettes from complex microbial communities

Integrons are bacterial genetic elements that can integrate mobile gene cassettes. They are mostly known for spreading antibiotic resistance cassettes among human pathogens. However, beyond clinical settings, gene cassettes encode an extraordinarily diverse range of functions important for bacterial adaptation. The recovery and sequencing of cassettes has promising applications, including: surveillance of clinically important genes, particularly antibiotic resistance determinants; investigating the functional diversity of integron-carrying bacteria; and novel enzyme discovery. Although gene cassettes can be directly recovered using PCR, there are no standardised methods for their amplification and, importantly, for validating sequences as genuine integron gene cassettes. Here, we present reproducible methods for the PCR amplification, sequence processing, and validation of gene cassette amplicons from complex communities. We describe two different PCR assays that either amplify cassettes together with integron integrases, or gene cassettes together within cassette arrays. We compare the use of Nanopore and Illumina sequencing, and present bioinformatic pipelines that filter sequences to ensure that they represent amplicons from genuine integrons. Using a diverse set of environmental DNAs, we show that our approach can consistently recover thousands of unique cassettes per sample and up to hundreds of different integron integrases. Recovered cassettes confer a wide range of functions, including antibiotic resistance, with as many as 300 resistance cassettes found in a single sample. In particular, we show that class 1 integrons appear to be collecting and concentrating antibiotic resistance genes out of the broader diversity of cassette functions. The methods described here can be applied to any environmental or clinical microbiome sample.

microbiology↗