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Biology subjects

Bhat, S. S.

Publications and source records attributed to Bhat, S. S..

8 recordsLinked to original sources

Leaves in Transition: Single nuclei RNA sequencing provides insights into sorghums juvenile and adult phases

The transition from juvenile to adult phase (JA) is a key developmental process in plants, driven by conserved pathways that shape growth and stress responses. In monocots such as sorghum, this transition influences traits linked to environmental resilience, yet the regulation of micro-anatomical features such as trichomes and bulliform cells remains poorly understood. Here, we use single-nucleus RNA sequencing (snRNA-seq) to generate high-resolution gene expression maps of juvenile and adult sorghum leaves. Contrary to the traditional developmental model, we find that trichomes are present in both juvenile and adult leaves, with stage-dependent differences in gene expression. We also observe that bulliform cells, typically considered adult-specific, are present in juvenile leaves. At the transcriptomic level, cell populations cluster more strongly by developmental stage than by cell type, with juvenile cells grouping with other juvenile cells and adult cells with adult counterparts, indicating a dominant effect of developmental state on gene expression. In addition, we identify enrichment of dhurrin biosynthetic genes in trichome-associated cells as well as elevated expression of aquaporin genes in these cells, pointing to a potential role for trichomes in coordinating defense and water-related processes. Together, these findings refine current models of sorghum development and suggest that key epidermal cells and their associated functions may be established earlier than previously known. Collectively, this study provides a single nucleus resolved framework for understanding how developmental stage shapes gene expression in sorghum leaves.

plant biology↗

Chronic Dietary Exposure to Methylparaben and Ethyl paraben Induces Developmental, Biochemical, and Behavioural Toxicity in Drosophila melanogaster

Parabens, particularly methylparaben (MP) and ethylparaben (EP), are extensively used preservatives in cosmetics, foods, and pharmaceuticals. Although considered safe at low concentrations, recent evidence questions their biological inertness under chronic exposure. This study evaluated the developmental, biochemical, and behavioral effects of continuous dietary MP and EP exposure in Drosophila melanogaster, an established in vivo model for toxicological screening. Flies were chronically exposed to MP (0.5-2%) or EP (0.5-1.5%) throughout development and adulthood. Developmental timing, lifespan, oxidative-stress markers (MDA, FRAP, total protein), and locomotor performance (negative geotaxis in adults, crawling in larvae) were quantified. Paraben exposure significantly delayed development ([~]15% increase in eclosion time), reduced median lifespan (up to 50% decrease at 2% MP), and elevated oxidative damage ({uparrow}MDA, {downarrow}FRAP) in a dose-dependent manner. Protein content declined more rapidly with age, suggesting oxidative degradation or proteolysis. Both adult climbing and larval crawling performances were impaired, linking biochemical stress to neuromuscular dysfunction. MP produced stronger oxidative and behavioral effects than EP. Feeding controls confirmed that observed deficits were not due to nutritional differences. Chronic MP and EP exposure induces systemic toxicity in D. melanogaster, integrating endocrine disruption and redox imbalance as plausible mechanisms. Given conserved stress and hormonal pathways, these findings reinforce the need to re-evaluate low-dose paraben safety limits and highlight Drosophila as a rapid, ethically viable platform for screening environmental preservatives and safer substitutes.

neuroscience↗

Metagenomics to Metabolomics: Integrating Genomic Insights for Mulberry Crop Protection and Enhancement

Indian Mulberry (Morus indica) is vital in sericulture, with their leaves serving as the primary food source for silkworms. The soil microbiome surrounding mulberry trees plays a pivotal role in nutrient cycling and ecosystem functioning. This study utilizes metagenomic analysis to explore the taxonomic diversity and functional potential of microbial communities in mulberry soil, particularly focusing on the rhizosphere. Key bacterial species such as Pseudomonas, Frankia, Azosipirulum are identified, highlighting their importance in mulberry health and disease dynamics. The abundance distribution of these bacterial populations reveals significant trends, offering insights into mulberry agroecosystem microbial ecology. Understanding garden soil-derived microbial consortia provides a foundation for exploring their role in nutrient cycling and plant health. The study reveals the intricate web of interactions between mulberry and their surrounding soil microbiota and identification of metabolites. Leveraging high-throughput sequencing and bioinformatics, the research identifies potential metabolites as biofertilizers and biopesticides, aiming to improve agricultural sustainability. The findings underscore the critical role of soil microbes in maintaining soil fertility, supporting plant health, and enhancing ecosystem resilience. Despite limitations and gaps, the study contributes to advancing eco-friendly agricultural practices and promoting soil health in mulberry cultivation. Ultimately, the research transcends the laboratory, resonating with stakeholders as it unravels the genetic blueprints of soil life and sows seeds of sustainable progress.

genomics↗

Interplay between m6A modification and overall transcripts quantity: Impacts on mRNA composition in plant stress granules

Stress granules (SGs) are cytoplasmic structures that emerge in response to unfavorable environmental conditions. They contain a rich pool of RNA, including non-translated mRNA. The mechanisms governing transcripts accumulation in SGs is only partially understood. Despite the recognized role of m6A in plant transcriptome regulation, its impact on SGs composition and assembly remains elusive. We examined the formation of SGs, the presence of m6A, and the transcription-level-dependent localization of selected mRNAs within these granules during hypoxia in the roots of Lupinus angustifolius and Arabidopsis thaliana. In lupine, SGs exhibit a distinctive bi-zonal structure, comprising of a ring and a central area with differences in ultrastructure and composition. Following the transcriptome analysis, mRNAs were selected for examination of their localization in SGs and m6A levels. Transcripts from genes responsive to hypoxia (ADH1 and HUP7) exhibited significant lower levels of m6A compared to housekeeping genes but only ADH1 was not present in SGs. HUP7 mRNA with low quantity of m6A, is present both in the SGs and cytoplasm probably due to extremely high expression level. It was also shown that the amount of m6A in SGs was higher than in the cytoplasm only in the first hours of hypoxia and then decreased. In mutants of A. thaliana with reduced level of m6A, formation and quantity of SGs were studied. In this line, ECT2 was not observed and poly(A) RNA levels were slightly reduced in SGs. Additionally the number of SGs was lower than that of the wild type. In summary, our findings demonstrate the limited impact of m6A modification on SGs assembly. However the interplay between m6A modification and the overall transcript quantity in the cytoplasm plays a regulatory role in mRNA partitioning into SGs.

cell biology↗

MTA influences RNA Polymerase II transcription dynamics and regulates the cold response in Arabidopsis.

RNA methylation at the N6 position of adenosine residues (m6A) has been demonstrated to be a vital epigenetic mark in eukaryotes. In plants, it is involved in a myriad of developmental and stress responsive pathways, such as cold stress. However, the role of MTA (m6A writer) itself in such processes remains obscured. Here, we show that the cold induced changes in abundance of differentially methylated transcripts are at-least partly due to the differences in the rates of transcription of such transcripts. RNAPII stalling was observed only on RNA methylation motifs that were decorated with m6A, and those that had a specific nucleotide composition. We note that the downregulation of genes in a mta mutant is a result of lower transcription of these genes. Moreover, 5 ends of transcripts from a subset of genes are disproportionately more accumulated in mta as compared to wild type under cold stress. In the mta mutant, genes corresponding to these transcripts had significantly reduced RNAPII occupancy on their 3 ends but not on 5 ends indicating that mta serves as an important factor for RNAPII elongation in a subset of genes. Taken together, our data suggest a novel direct role for MTA as a gene specific influencer of RNAPII elongation dynamics and thus introduces a new path by which the m6A methylation machinery may affect differential gene expression during stress in plants.

plant biology↗

SERRATE interacts with the nuclear exosome targeting (NEXT) complex to degrade primary miRNA precursors in Arabidopsis

SERRATE/ARS2 is a conserved RNA effector protein involved in transcription, processing and export of different types of RNAs. In Arabidopsis, the best-studied function of SERRATE (SE) is to promote miRNA processing. Here, we report that SE interacts with the Nuclear Exosome Targeting (NEXT) complex, comprising the RNA helicase HEN2, the RNA binding protein RBM7 and one of the two zinc-knuckle proteins ZCCHC8A/ZCCHC8B. The identification of common targets of SE and HEN2 by RNA-seq supports the idea that SE cooperates with NEXT for RNA surveillance by the nuclear exosome. Among the RNA targets accumulating in absence of SE or NEXT are miRNA precursors. Loss of NEXT components results in the accumulation of pri-miRNAs without affecting levels of miRNAs, indicating that NEXT is, unlike SE, not required for miRNA processing. As compared to se-2, se-2 hen2-2 double mutants showed increased accumulation of pri-miRNAs, but partially restored levels of mature miRNAs and attenuated developmental defects. We propose that the slow degradation of pri-miRNAs caused by loss of HEN2 compensates for the poor miRNA processing efficiency in se-2 mutants, and that SE regulates miRNA biogenesis through its double contribution in promoting miRNA processing but also pri-miRNA degradation through the recruitment of the NEXT complex.

plant biology↗

DRB1 as a mediator between transcription and microRNA processing

DRB1 (HYL1) is a double-stranded RNA binding protein involved in miRNA processing in plants. It is a core component of the Microprocessor complex and enhances the efficiency and precision of miRNA processing by DCL1 protein. In this work, we report a novel function of DRB1 protein in the transcription of MIR genes. DRB1 co-localizes with RNA Polymerase II and affects its distribution along MIR genes. Moreover, proteomic experiments revealed that DRB1 protein interacts with many transcription factors. Finally, we show that the action of DRB1 is not limited to MIR genes as it impacts expression of many other genes, majority of which are involved in plant response to light. These discoveries add DRB1 as another player of gene regulation at transcriptional level, independent of its role in miRNA biogenesis.

plant biology↗

mRNA adenosine methylase (MTA) deposits m6A on pri-miRNAs to modulate miRNA biogenesis in Arabidopsis thaliana

m6A, one of the most abundant mRNA modifications, has been associated with various metabolic processes in plants. Here we show that m6A also plays a role in miRNA biogenesis in Arabidopsis thaliana. Significant reductions in plant m6A/MTA levels results in lower accumulation of miRNAs whereas pri-miRNA levels tend to be higher in such plants. m6A-IP Seq and MTA-GFP RIP were used to show that many pri-miRNAs are m6A methylated and are bound by MTA, further demonstrating that pri-miRNAs can also be substrates for m6A methylation by MTA. We report that MTA interacts with RNA Pol II, supporting the assumption that m6A methylation is a co-transcriptional process, and also identify TGH, a known miRNA biogenesis related protein, as a novel protein that interacts with MTA. Finally, reduced levels of miR393b may partially explain the strong auxin insensitivity seen in Arabidopsis plants with reduced m6A levels.

plant biology↗