Biology subjects

Prasad, K.

Publications and source records attributed to Prasad, K..

At least 19 recordsLinked to original sources

bioRxiv2026

Spatial Decoupling of the Endodermal Functions Establishes the Innermost Cortex as the Gravity Sensor in Rice Stems

Gravitropism is a remarkable example of coordinated growth, enabling plants to perceive gravity and generate asymmetric cell expansion that restores organ orientation. In Arabidopsis, gravity sensing in shoots occurs within starch-rich endodermis cells. In rice, however, the stem must accommodate gravitropic bending while preserving integrity of tissues surrounding emerging crown roots, raising the possibility that gravity sensing relies on distinct structural and physiological adaptations. Here, using high-resolution imaging together with auxin-response analyses, we identify a specialized cell layer at the inner boundary of the rice stem cortex, operationally defined by its position adjacent to the lignified endodermis and associated with gravitropic responses. This layer is characterized by circumferentially elongated cells, starch sedimentation, a confined auxin-response domain, and expression of gravity-sensing markers. Following gravistimulation, starch granules realign and the auxin response domain expands asymmetrically within this layer, coinciding with differential growth during stem bending. Our results further indicate that this response depends on polar auxin transport and conserved components of the gravitropic regulatory network. Together, these findings support a model in which gravity sensing and signaling in rice stems are spatially separated from the lignified endodermal barrier. Such tissue organization may represent a developmental adaptation that allows the stem to reconcile barrier integrity with growth flexibility.

plant biology↗

bioRxiv2026

Exploring the impact of intraspecific interaction between sibling colonies of Bacillus cereus MSM-S1 in the absence and presence of antibiotics

Bacillus cereus is a spore-forming pathogen and food contaminant that can survive in hostile environments through different survival strategies. As part of survival, B. cereus may also engage in intra- or interspecific interactions that shape its community structure and stress responses, enabling its persistence under unfavourable conditions. The present study explored intraspecific interactions, at the cell and molecular levels, between two sibling colonies of Bacillus cereus MSM-S1 grown through swarming adjacently on nutrient agar in the absence and presence of gentamicin. While growing alone, gentamicin altered the symmetrical colony morphology of B. cereus MSM-S1, which was associated with cell elongation, decreased flagellation and increased expression of the toxin gene nheA. During interaction, both in the absence and presence of gentamicin, the motility of sibling colonies towards each other was restricted, forming a distinct line/zone of demarcation between the colonies. Microscopic analysis at different time points (day 3, 5 and 7) revealed progressive changes in the swirling arrangement of cells at the interaction zone. In the absence of gentamicin, cells became jammed at the interacting front by day 7, whereas this effect was not observed in the presence of gentamicin on day 7. Cells at the interacting zone underwent gradual sporulation; notably, sporulation was less in the presence of gentamicin. Further analysis suggested that nutrient limitation was the major contributory factor for the formation of the zone of no growth between the interacting colonies. Moreover, the expression of quorum-sensing genes was down-regulated in cells at the interacting zone without gentamicin, but notably, upregulated in the presence of gentamicin. The expression of chemotaxis regulator gene, cheY, was also upregulated in interacting zone cells both in the absence and presence of gentamicin. In addition, increased expression of the global transcriptional regulator codY in the interacting zone under both conditions suggested its involvement in metabolic regulation during sibling colony interaction. The findings highlight modulated quorum sensing and sporulation as adaptive mechanisms in Bacillus cereus MSM-S1 to cope with external stress imposed by nutrient limitations caused by the growth of sibling colonies, whereas, cell elongation and toxin production together is a preferred survival strategy for a growing MSM-S1 colony when stress is imposed by gentamicin as antibiotic but sufficient nutrient is available.

microbiology↗

bioRxiv2026

A complex balance: PLETHORA - JANUS ratiometry explains the PLETHORA dosage regeneration paradox

Molecular gradients are classically thought to encode positional information through concentration thresholds, but how transcription factor dosage is translated into regenerative competence remains unclear. In Arabidopsis roots, PLETHORA (PLT) transcription factors form a gradient that defines a regeneration competence zone and, following injury, rapidly re-establishes their transcriptional domain through an auto-activation. Yet, increasing PLT levels has a paradoxical effect: moderate increases expand the competence zone, whereas further increases impair regeneration. Here, combining mathematical modelling with experiments, we uncover the molecular basis of this non-monotonic response. Our model reveals that PLT activity depends not simply on its abundance, but on the balance between PLT protein complexes and unbound PLT. Following injury, PLT auto-activation rapidly restores the transcriptional domain, whereas increasing PLT dosage progressively shifts the equilibrium towards unbound PLT, reducing the availability of active PLT complexes. We identify and experimentally validate a strategy to extend regeneration competence by maintaining an appropriate ratio of PLT complexes to unbound PLT. Our findings reveal a ratiometric mechanism for decoding transcription factor dosage, in which excess regulator can attenuate rather than enhance activity. Such molecular balancing provides a mechanism by which a single transcription factor gradient can generate distinct, context-dependent outcomes during development and regeneration.

developmental biology↗

bioRxiv2026

Heterotrimeric Gβ subunit orchestrates flavonoid metabolism governing drought-tolerance in groundnut (Arachis hypogaea L.)

Groundnut (Arachis hypogaea) is an important oilseed crop, and its production is severely affected by drought worldwide. Improving the drought-tolerance of groundnut without compromising its agronomic traits is a major challenge for researchers. Here, a meta-transcriptome analysis identified tandemly duplicated G{beta} subunits (AhG{beta}1 and AhG{beta}3) as early signaling genes under water deficit conditions. Heterotrimeric G-protein {beta}-subunit is a major signaling element that can regulate diverse responses in plants. Heterologous complementation studies in Arabidopsis G{beta} mutant (agb1-2) demonstrated evolutionary conservation of AhG{beta} function, while the transgenic overexpression and RNAi knockdown in groundnut established AhG{beta}1 as a positive regulator of plant growth, yield, and drought tolerance. Integrated transcriptomic, transcription factor regulatory network, and metabolomic analyses demonstrated that AhG{beta}1 coordinates drought-responsive transcriptional and metabolic reprogramming characterized by suppression of cell wall-associated processes and activation of phenylpropanoid, flavonoid, and isoflavonoid biosynthesis. This reprogramming resulted in enhanced accumulation of antioxidant flavonoids, including quercetin and rutin, reduced ROS accumulation, and improved drought tolerance. Furthermore, exogenous application of these flavonoids enhanced drought recovery, highlighting their role as protective downstream metabolites. Collectively, our findings establish AhG{beta}1 as a conserved regulator linking early G-protein signaling with transcriptional and metabolic reprogramming to maintain redox homeostasis during drought stress and identify promising molecular and metabolic targets for improving drought resilience in groundnut.

plant biology↗

bioRxiv2026

Clustered structural variant hotspots enable oncogenic addiction and plasticity in osteosarcoma

The genomic landscape of osteosarcoma, the most common bone cancer worldwide, is among the most structurally complex of all human malignancies. The identification of recurrent and functionally consequential patterns has thus remained a challenge. Across 236 whole-genome sequencing osteosarcoma samples, we uncovered five genomic hotspots of clustered structural variation collectively altered in 58% of tumors. Four were associated with amplification of oncogenes ( MYC , CCND3 , CCNE1 , CDK4) , while the fifth mapped largely upstream of TP53 . Hotspot events showed coordinated patterns of co-occurrence and mutual exclusivity with each other and with tumor suppressor alterations, suggesting genomic context-specific selection. We found localized transcriptional dysregulation at hotspot event loci, and single cells harboring these events converged on a neural crest-like program, linking these structural alterations to a less differentiated cell state. These events were also detectable non-invasively through liquid biopsies and displayed ongoing structural evolution throughout disease progression, a finding with potential clinical utility. Our results provide novel insight into how complex rearrangements shape oncogenesis in osteosarcoma, with broader relevance to other cancers characterized by complex genomes.

cancer biology↗

bioRxiv2026

Guide RNA truncation as a tunable approach for modulating base editing outcomes

Base editing enables programmable nucleotide conversion but can be limited by bystander editing and unintended genomic alterations. Here, we investigated guide RNA (gRNA) truncation as a strategy to modulate base editing outcomes. On editing across multiple genomic loci using ABE8e in HUDEP-2 cells, we found that truncated gRNAs retained measurable editing activity at lengths as short as 15 nucleotides, although activity depended strongly on target sequence, gRNA length, and delivery method. Importantly, truncation altered editing efficiencies at individual nucleotides within the editing window, enabling selective reduction of bystander editing while preserving on-target conversion at several loci, including a therapeutically relevant HbE target. Truncated gRNAs also eliminated detectable large deletions at homologous globin loci, consistent with loss of Cas9 nickase activity at short gRNAs lengths. Similar length dependent effects were observed with cytosine base editing, PAM relaxed ABE8e, and in primary hematopoietic stem and progenitor cells (HSPCs), although the optimal gRNA length varied between contexts. Together, these findings establish gRNA length as an additional parameter for tuning nucleotide level base editing outcomes and reducing bystander editing.

bioengineering↗

bioRxiv2026

A Spatial Proteomic Atlas of Tertiary Lymphoid Structures in Non-Small Cell Lung Cancer Identifies a Novel Predictive Class of Lymphoid Aggregates

Tertiary lymphoid structures (TLS) predict benefit from immune checkpoint inhibitors (CPIs), yet mature, germinal-center-rich TLS are infrequent in solid tumors by histological review. Here, using 38-plex MIBI spatial proteomics across 165 lymphoid structures from 14 NSCLC resections, we establish a continuum of TLS maturity using high dimensional compositional, spatial, and molecular features. We demonstrate that histologically-defined lymphoid aggregates (LA) comprise a heterogeneous class of structures, which span this continuum of maturity. We identify a subset of lymphoid aggregates that harbor follicular dendritic cell networks, T follicular helper cells, and activated B cell states characteristic of mature TLS, yet are not readily distinguished from other LA structures in our histological review. We developed a novel digital pathology classifier to identify mature LAs in CPI trials, and demonstrate in a retrospective analysis of Atezolizumab in advanced NSCLC that the inclusion of mature LAs greatly expands the biomarker-eligible population while maintaining strong predicted benefit. Together, these data redefine the biological spectrum of tumor-associated lymphoid aggregates and provide a framework for implementing maturity-informed TLS biomarker strategies.

cancer biology↗

bioRxiv2026

Manipulation of rhizosphere microbiome by Microbacterium sp. GB16_1_BI to promote plant growth.

Plant growth promoting microbes enhance developmental progression of the host by influencing its nutrient availability or by deploying secondary metabolites responsible for manipulating the hormonal crosstalk. Microbacterium bengalense sp. nov. GB16_1_BI (Accession number: SRX9280401), a newly identified ammonium releasing Actinomycetota, could enhance plant growth by manipulating rhizosphere bacteria. Amplicon sequencing of the 16S rRNA V3-V4 region from the rhizosphere of the black rice (Chakhao Poireiton) showed that GB16_1_BI could inhibit most bacteria. However, GB16_1_BI inoculation encouraged the growth of rare bacteria specific to waterlogged rice rhizosphere. Analysis of the OTUs using PICRUSt2 (Phylogenetic investigation of communities by reconstruction of unobserved states) showed increased abundance in the marker genes for nitrogen cycling (nifH, nrfA and nrt) but not for nifD or nifK which was also reflected in the ANOSIM analysis in the OTUs of the N-fixing bacteria. Marker genes for methane metabolism (comA, comB, cofG and cofH) were also more abundant in the inoculated plants than the control; however, ANOSIM studies did not support this observation in the OTUs of methane cycling bacteria. Both Methylosinus and Methylocystis, the two most abundant methanotrophic OTUs, are also known to be nitrogen fixers. Hence, GB16_1_BI could influence plant growth predominantly by manipulating nitrogen cycling microbes. The genome sequence as well as untargeted metabolome analyses of GB16_1_BI showed abundance of secondary metabolites with probable antimicrobial activity. GB16_1_BI could utilize varied carbohydrates and amino acid as energy source and form persister-like cells may help it to survive in the soil in absence of the host plant.

microbiology↗

bioRxiv2026

Reshaping Organellar Translation and tRNA Metabolism: The Consequences of Photosynthesis Loss and Massive Horizontal Gene Transfer

The transition to holoparasitism in plants precipitates the loss of photosynthesis, fundamentally altering the selective landscape acting on organellar genomes. These changes raise questions about the mechanisms by which the essential, coevolved machinery of translation responds to extreme genomic erosion and metabolic dependency. Integrating comparative genomics, tRNA sequencing, and subcellular localization assays, we elucidate the extensive rewiring of organellar translation systems and the tRNA-dependent tetrapyrrole biosynthesis pathway in the holoparasitic angiosperm family Balanophoraceae, which exhibits extreme reduction of tRNA content in plastid and mitochondrial genomes. We identified a rare evolutionary event: the putative intracellular transfer of the plastid initiator tRNA (tRNA-iMet) to the nucleus, which compensates for its loss from the plastid genome. We also demonstrate that the unusual UAG-to-Trp reassignment in the Balanophora plastid genetic code is driven by the loss of release factor pRF1 and the recruitment of a mutated nuclear tRNA-Trp. Furthermore, we reveal that the retention of organellar nuclear-encoded aminoacyl-tRNA synthetases is dictated by the presence/absence of cognate organellar tRNAs, which appear to be functional regardless of their foreign (horizontal transfer from the host plant) or native origins. Finally, we uncover a striking evolutionary asymmetry in nuclear-encoded ribosomal proteins: while plastid subunits exhibit elevated substitution rates consistent with relaxed selection and compensatory coevolution, mitochondrial subunits display high sequence conservation, likely maintaining compatibility with the extensive horizontal gene transfer observed in this lineage. Collectively, these findings represent some of the most extreme changes ever identified in the anciently conserved machinery of plant organellar translation.

evolutionary biology↗

bioRxiv2024

Salt-induced reduction of hyperswarming motility in Bacillus cereus MHS is associated with reduced flagellation, reduced nanotube formation and reduction in expression of quorum sensing regulator

Bacteria have been known to thrive in challenging environmental niches through diverse phenomena. Swarming is one such favourable adaptation that could help bacteria survive extreme conditions. Therefore, targeting swarming is crucial for improving our understanding of bacterial motility and preventing related infections. Bacillus cereus, which causes food poisoning, has been shown to perform swarming, and salts like NaCl can act as a food preservative to control bacterial growth. In order to explore the possible alterations in the swarming of Bacillus cereus in the presence of salt, the present study encompasses the effect of NaCl on the swarming characteristics of a natural bacterial isolate, Bacillus cereus MHS, with a hyperswarming phenotype. Here we report that the presence of an increased amount of NaCl in growth media could induce a reduction in swarming motility and swarming pattern of MHS on Luria agar plates and the observed reduction in swarming was associated with the reduced flagellation and reduction in bacterial nanotube formation. Gene expression studies supported the phenotypic and ultrastructure observations as the expressions of bfla and ymdb genes, involved in formations of flagella and nanotubes respectively, were found to be reduced in the swarming MHS cells in the presence of increased NaCl in swarm media. It was also observed that the salt-induced reduction in swarming of MHS is associated with the reduced expression of the quorum sensing regulator gene plcR. This study first time reports the bacterial nanotubes in a Bacillus cereus strain and indicates a possible link between the bacterial nanotube formation and hyperswarming phenotype in Bacillus cereus MHS.

microbiology↗

bioRxiv2023

Molecular principles of CRISPR-Cas13 mismatch intolerance enable selective silencing of point-mutated oncogenic RNA with single-base precision

Single nucleotide variants (SNVs) are extremely prevalent in human cancers. For instance, KRAS mutations occur in over 90% of pancreatic cancers and [~]40% of colorectal cancers. Virtually all KRAS mutations are SNVs, most of which remain clinically unactionable. The programmable RNA nuclease CRISPR-Cas13 has been deployed to specifically target RNAs such as overexpressed oncogenes and fusion transcripts. However, silencing oncogenic SNVs with single-base precision remains extremely challenging due to the intrinsic mismatch tolerance of Cas13. Here, we developed a comprehensive mutagenesis analysis of target-spacer interactions at single-nucleotide resolution, which revealed key spacer nucleotide positions intolerant to mismatches. We show that introducing synthetic mismatches at these precise positions enables de nova design of CRISPR RNA (crRNA) with strong preferential silencing of SNV transcripts. We demonstrate that our top-performing crRNAs possess prominent SNV-selectivity with dose-dependent silencing activity against all KRAS G12 variants at both the RNA and protein levels with minimal off-target silencing of wildtype KRAS. We applied these design principles to effectively silence oncogenic NRAS G12D and BRAF V600E transcripts, underscoring the adaptability of this platform to silence various SNVs. These findings demonstrate that the CRISPR Cas13 system can be reprogrammed to target mutant transcripts with single-base precision, showcasing the tremendous potential of this tool in personalized transcriptome editing. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=176 SRC="FIGDIR/small/557083v1_ufig1.gif" ALT="Figure 1"> View larger version (58K): org.highwire.dtl.DTLVardef@20a7b3org.highwire.dtl.DTLVardef@1e1f79corg.highwire.dtl.DTLVardef@4724c1org.highwire.dtl.DTLVardef@be3063_HPS_FORMAT_FIGEXP M_FIG C_FIG

molecular biology↗

bioRxiv2023

Gametophytic epigenetic regulators MEA and DME synergistically suppress ectopic shoot formation in Arabidopsis

The gametophyte to sporophyte transition facilitates the alternation of generations in a plant life cycle. The epigenetic regulators DEMETER(DME) and MEDEA(MEA) synergistically control central cell proliferation and differentiation, ensuring proper gametophyte to sporo-phyte transition in Arabidopsis. Mutant alleles of DME and MEA are female gametophyte lethal, eluding the recovery of recessive homozygotes to examine their role in the sporophyte. Here, we exploited the paternal transmission of these mutant alleles coupled with CENH3-haploid inducer to generate mea-1;dme-2 sporophytes. Strikingly, the simultaneous loss of function of MEA and DME leads to the emergence of ectopic shoot meristems at the apical pole of the plant body axis. DME and MEA are expressed in the developing shoot apex and regulate the expression of various shoot-promoting factors. Chromatin immunoprecipitation(ChIP), DNA methylation and gene expression analysis revealed several shoot regulators as potential targets of MEA and DME. Knockdown of upregulated shoot-promoting factors STM, CUC2, and PLT5 rescued the ectopic shoot phenotypes. Our findings reveal a previously unrecognized synergistic role of MEA and DME in restricting the meristematic activity at the shoot apex during sporophytic development to orchestrate the plant architecture.

plant biology↗

bioRxiv2022

Relating enhancer genetic variation across mammals to complex phenotypes using machine learning

Protein-coding differences between mammals often fail to explain phenotypic diversity, suggesting involvement of enhancers, often rapidly evolving regions that regulate gene expression. Identifying associations between enhancers and phenotypes is challenging because enhancer activity is context-dependent and may be conserved without much sequence conservation. We developed TACIT (Tissue-Aware Conservation Inference Toolkit) to associate open chromatin regions (OCRs) with phenotypes using predictions in hundreds of mammalian genomes from machine learning models trained to learn tissue-specific regulatory codes. Applying TACIT for motor cortex and parvalbumin-positive interneurons to neurological phenotypes revealed dozens of new OCR-phenotype associations. Many associated OCRs were near relevant genes, including brain size-associated OCRs near genes mutated in microcephaly or macrocephaly. Our work creates a forward genomics foundation for identifying candidate enhancers associated with phenotype evolution. One Sentence SummaryA new machine learning-based approach associates enhancers with the evolution of brain size and behavior across mammals.

genomics↗

bioRxiv2022

Precise modelling and correction of a spectrum of β-thalassemic mutations in human erythroid cells by base editors

{beta}-thalassemia and HbE result from mutations in the {beta}-globin locus that impedes the production of functional {beta}-hemoglobin and represents one of the most common genetic disorders worldwide. Recent advances in genome engineering have opened up new therapeutic opportunities to directly correct these pathogenic mutations using base editors that install transition mutations (A>G and C>T) in the target region with minimal generation of indels. Herein, for the first time, we demonstrate the usage of base editor in the correction of point mutations spanning multiple regions of the HBB gene, including promoter, intron and exon. To this end, we have engineered human erythroid cells harbouring the diverse HBB mutations, thus eliminating the requirement of patient CD34+ HSPCs with desired mutations for the primary screening by base editors. We further performed precise creation and correction of individual HBB point mutations in human erythroid cells using base editors, which were effectively corrected in the HBB-engineered erythroid model. Intriguingly, most bystander effects produced by the base editor at the target site were reported to exhibit normal hemoglobin variants. Overall, our study provides the proof-of-concept for the precise, efficient and scarless creation and correction of various pathogenic mutations at the coding and non-coding regions of HBB gene in human erythroid cells using base editors and establishes a novel therapeutic platform for the treatment of {beta}-thalassemia/HbE patients. This study can be further explored in correcting the other monogenic disorders caused due to single base substitutions.

genomics↗

bioRxiv2021

A Greener Recyclable Process for Extraction of Microbial Melanin using Ionic Liquid

Conventional techniques for the extraction of the melanin encounter several disadvantages such as poor extraction efficiency, excessive use of acid and alkali, and lengthy time duration. To extract melanin in a more sustainable manner, herein suitability of an aqueous solution of quaternary ammonium hydroxide known as tetrabutylammonium hydroxide (40% w/w TBAOH in water) to extract and purify the pigment from the endophyte Streptomyces hyderabadensis 7VPT5-5R was investigated. The solvent could extract 5.54{+/-}0.03 g/L of pure melanin against 3.31{+/-} 0.02 g/L of pure melanin by conventional method from bacterial culture resulting 66% increase in the yield of the pigment. The quaternary ammonium based solvent was further recycled for 5 consecutive cycles and production of melanin with yield ranging from 5.54{+/-}0.03 g/L to 5.47{+/-} 0.02 g/L was obtained. The yield of the pigment was 38-40% higher in the recycled solvent in comparison to the melanin yield extracted by conventional method. Considering the efficiency of the solvent for effective extraction and purification of melanin and its recyclability potential, it can be considered as a sustainable solvent for the extraction of the pigments from bacterial culture. Furthermore, the developed method for extraction of melanin from microorganisms as a sustainable source would help in reducing the cost of melanin leading to its efficient utilization in diverse fields.

microbiology↗

bioRxiv2021

Whole-genome duplication shapes the aneuploidy landscape of human cancers

Aneuploidy - a hallmark of cancer - has tissue-specific recurrence patterns suggesting it plays a driving role in cancer initiation and progression. However, the contribution of aneuploidy to tumorigenesis depends on the cellular and genomic context in which it arises. Whole-genome duplication (WGD) is a common macro-evolutionary event that occurs in >25% of human tumors during the early stages of tumorigenesis. Although tumors that have undergone WGD are reported to be more permissive to aneuploidy than tumors that have not, it remains unknown whether WGD affects aneuploidy recurrence patterns in human cancers. Here we analyzed clinical tumor samples from 449 WGD- tumors and 157 WGD+ tumors across 22 tumor types. We found distinct recurrence patterns of aneuploidy in WGD- and WGD+ tumors. The relative prevalence of recurrent aneuploidies decreased in WGD+ tumors, in line with increased aneuploidy tolerance. Moreover, the genetic interactions between chromosome arms differed between WGD- and WGD+ tumors, giving rise to distinct co-occurrence and mutual exclusivity aneuploidy patterns. The proportion of whole-chromosome aneuploidy vs. arm-level aneuploidy was significantly higher in WGD+ tumors, indicating distinct dominant mechanisms for aneuploidy formation in WGD- and WGD+ tumors. Human cancer cell lines successfully reproduced these WGD/aneuploidy interactions, confirming the relevance of studying this phenomenon in culture. Lastly, we induced WGD in human colon cancer cell lines, and followed aneuploidy formation in the isogenic WGD+/WGD-cells under standard or selective conditions. These experiments validated key findings from the clinical tumor analysis, and revealed a causal link between WGD and altered aneuploidy landscapes. We conclude that WGD shapes the aneuploidy landscape of human tumors, and propose that the interaction between WGD and aneuploidy is a major contributor to tumor evolution.

cancer biology↗

bioRxiv2020

Regulation of touch dependant de novo root regeneration in Arabidopsis

The versatile regeneration capability of leaves enable even a detached Arabidopsis leaf to yield two kinds of regenerative responses namely, wound healing at the cut end in form of callus formation or de novo root regeneration (DNRR). Using various experimental approaches, we show that the factor favouring DNRR over callus formation seems to be a mechanical cue, possibly touch, at the cut end of the detached leaf. Here, we show that the forced expression of a PLETHORA transcription factor can bypass the need for touch to initiate DNRR. Our findings provide a genetic frame-work for touch dependant DNRR and suggest that a core PLT transcription regulatory module acts in response to mechano-sensing stimuli.

plant biology↗

bioRxiv2020

Age, wound size and position of injury - dependent vascular regeneration assay in growing leaves

BackgroundRecurring damage to aerial organs of plants necessitates their prompt repair, particularly their vasculature. While vascular regeneration assay in aerial plant parts such as stem and inflorescence stalk are well established, those on leaf vasculature remained unexplored. Recently we established a new vascular regeneration assay in growing leaf and discovered the underlying molecular mechanism. ResultsHere we describe the detailed stepwise method of incision and the regeneration assay used for studying the leaf vascular regeneration. By using a combination of micro-surgical perturbations, brightfield microscopy and other experimental approaches, our new findings show that the regeneration efficiency decreases with aging of the leaf, and increases with the nearness of the wound towards the proximal end of the leaf. ConclusionThis easy-to-master vascular regeneration assay is an efficient and rapid method to study the mechanism of vascular regeneration in growing leaves. It can be readily adapted for other plant species and can be combined with cellular and molecular biology techniques.

plant biology↗