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<p>Rapid Alkalinization Factor, one of the small secretary peptides, functions to increase the extracellular pH rapidly and regulates root phenomics during adverse conditions. Emerging studies confirm that RALFs participate in liquid-liquid phase separation (LLPS) via pectin and the receptors to regulate various signaling mechanisms. Although, several RALFs have been analyzed across the plant kingdom, their functional roles in signaling during developmental and stress responses remains largely unknown. The present study focuses on determining the intrinsically disordered regions in rice RALFs and the expressional analysis of the disordered ones in PEG based drought model followed by correlational analysis. In the present study, in-silico characterization of Rice RALFs in terms of motif analysis using MEME, sub-cellular localization using Plant mPlock, gene ontology using Shiny GO, cis-regulatory element analysis using PlantCare, transcription factor binding site anaysis using Plantreg Map and prediction of intrinsically disordered regions in RALFs using AIUPRED and MEGAPRED tools were carried out to shortlist the RALFs that could be involved in drought. Based on the presence of stress-responsive cis-regulatory elements, enriched GO-terms and site of action, total of 18 RALFs (RALF3/4/5/6/7/8/9/13/22/23/24/31/32/33/ 34/35/37/39) were shortlisted. Further, RALF3/4/13/22/23/24/32/34/35/37/39 were determined to be structurally disordered and hence considered for the further studies. The differential expression of disordered RALFs along with ordered RALF9/33 in in-vitro PEG based drought model were analyzed using qRT-PCR. RALFs showed distinct expression patterns in root and shoot tissues suggesting their major role in mediating drought responses and furthering our understanding on the downstream responses. Further, correlating drought RALF candidates with responses such as cell-wall integrity, lateral root development and ROS accumulation highlights the novel insights into IDRs. providing an preliminary framework for RALF functions.</p> Graphical Abstract <p>Red Arrows indicate the existing information with respect to IDRs and RALFs. The Black arrows indicates the conventional signalling flow with respect to RALFs and stress. The Balck dashed arrow indicates the un - validated link between the drought responsive RALFs and the drought specific responses. </p> <p></p>

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Functional Insights into Intrinsically Disordered Drought Responsive RALFs in Oryza sativa

  • 期刊论文

  • 发表日期：

    2026年3月21日

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  • Durvasula Sumana Pratyusha,
  • Dronamraju VL Sarada

摘要

Rapid Alkalinization Factor, one of the small secretary peptides, functions to increase the extracellular pH rapidly and regulates root phenomics during adverse conditions. Emerging studies confirm that RALFs participate in liquid-liquid phase separation (LLPS) via pectin and the receptors to regulate various signaling mechanisms. Although, several RALFs have been analyzed across the plant kingdom, their functional roles in signaling during developmental and stress responses remains largely unknown. The present study focuses on determining the intrinsically disordered regions in rice RALFs and the expressional analysis of the disordered ones in PEG based drought model followed by correlational analysis. In the present study, in-silico characterization of Rice RALFs in terms of motif analysis using MEME, sub-cellular localization using Plant mPlock, gene ontology using Shiny GO, cis-regulatory element analysis using PlantCare, transcription factor binding site anaysis using Plantreg Map and prediction of intrinsically disordered regions in RALFs using AIUPRED and MEGAPRED tools were carried out to shortlist the RALFs that could be involved in drought. Based on the presence of stress-responsive cis-regulatory elements, enriched GO-terms and site of action, total of 18 RALFs (RALF3/4/5/6/7/8/9/13/22/23/24/31/32/33/ 34/35/37/39) were shortlisted. Further, RALF3/4/13/22/23/24/32/34/35/37/39 were determined to be structurally disordered and hence considered for the further studies. The differential expression of disordered RALFs along with ordered RALF9/33 in in-vitro PEG based drought model were analyzed using qRT-PCR. RALFs showed distinct expression patterns in root and shoot tissues suggesting their major role in mediating drought responses and furthering our understanding on the downstream responses. Further, correlating drought RALF candidates with responses such as cell-wall integrity, lateral root development and ROS accumulation highlights the novel insights into IDRs. providing an preliminary framework for RALF functions.

Graphical Abstract

Red Arrows indicate the existing information with respect to IDRs and RALFs. The Black arrows indicates the conventional signalling flow with respect to RALFs and stress. The Balck dashed arrow indicates the un - validated link between the drought responsive RALFs and the drought specific responses.