A micropeptide regulates seed desiccation DOI Creative Commons

Rong Chun Lu,

Zilin Zhang,

Songping Hu

et al.

Frontiers in Plant Science, Journal Year: 2025, Volume and Issue: 16

Published: March 26, 2025

Seeds serve as the major means of reproduction for most plant species and form foundation both agriculture natural ecosystems (Waterworth et al., 2024). are also key genetic resources to deal with increasing human population climate fluctuations (Leprince 2017).Seed development can be categorized into three stages: maturation, dormancy, germination (Figure 1A; Zinsmeister 2020;Powell, 2022;Nadarajan 2023;Waterworth 2024).In maturation phase, seeds acquire desiccation tolerance, followed by developmental processes that expands longevity dormancy stage. Maturation drying reduces seed moisture content 5% -15% fresh weight 2020). Dormancy under optimal conditions, such low temperature humidity, prolongs viability, while suboptimal conditions lead aging Powell 2023). Seed is modulated a complex interplay genetic, biochemical, molecular determinants intricately connected environmental signals light, temperature, nitrate availability, phytohormones including abscisic acid (ABA) gibberellin (GA) (Chahtane 2017;Matilla, 2024;Rachappanavar, 2025).The difference between dormant non-dormant could attributed number gene expression changes (Meimoun 2014), physiological, developmental, morphological features grains on spike, pericarp color, transparency, hairiness, waxiness, permeability water, α-amylase activity, concentrations growth regulators ABA GA within embryo (Sohn al. 2021). initiated water uptake (imbibition), resulting in activation multiple cellular actions, completed emergence young roots shoots 1A;Carrera-Castaño 2020;Waterworth 2024).The acquisition tolerance at late stage provides critical survival mechanism crops, enabling them adaptive adverse extreme temperatures drought 2017;Zinsmeister The majority crop plants generate classified orthodox seeds, which possess ability withstand (below 7%) harsh freezing (-10 ℃) long time (Nadarajan maize, kernel suitable mechanized harvesting from 15% -25%, however, some regions China, maize varieties have high grain harvest, ranging 30% -40% (Xiang 2012;Kebebe 2015;Dai 2017;Li 2018aLi , 2018b;;Zhou 2018). Kernel dehydration rate (KDR), defined loss two adjacent periods after pollination (Zhang 2024a), determinant quality exerts significant impact efficiency mechanical (Li 2018c).Besides, removal free leads phase transition cytoplasm mobility fluid glassy state (Buitink Leprince, 2008), metabolic quiescence increased (Zinsmeister To date, elements implicated modulation reactive oxygen (ROS) signaling, lipid peroxidation cell membrane, preservation DNA RNA integrity, methylation status, biosynthesis storage proteins (SSPs), ABA, auxin, GA, brassinosteroids (BRs) been documented crucial 2023;Pirredda 2024).Abiotic factors thermal salinity stress significantly longevity, availability emerging predominant In several quantitative trait loci (QTLs) characterized pivotal players regulation KDR 2020(Li 2021a;;Zhang 2023;Jin Collectively, higher level harvesting, preventing breakage, mildew, reducing costs associated harvest (de Jager 2004;Xiang 2015;Wang 2022;Xia Thus, comprehensive understanding mechanisms governing necessary crucial.Seed linked multitude physiological modifications, accumulation macromolecules (proteins, lipids, carbohydrates), enhanced membrane defense mechanisms, governed hormone signaling pathways ethylene (Angelovici 2010;Bewley 2013;Kijak Ratajczak, 2020;Oliver 2020;Smolikova onset occurs when enter 2017;Smolikova Numerous components Late Embryogenesis Abundant (LEA) proteins, small heat shock (sHSPs), non-reducing oligosaccharides, antioxidants, (ROS), well (GA), identified 2010;Kijak addition, many transcription ABA-INSENSITIVE 3 (ABI3), FUSCA (FUS3) LEAFY COTYLEDONS 2 (LEC2) discovered defines balance finally initiate (Smolikova However, regulatory mediated peptides remain largely elusive.Micropeptides, referred microproteins or short open reading frame (sORF)-encoded peptides, essential products derived larger polypeptide MicroRNAs (miRNAs), non-coding (lncRNA), circular (circRNA), typically an arbitrary length less than 100 -150 amino acids (Hashimoto 2008;Makarewich Olson, 2017;Sousa Farkas, 2018;Vitorino 2021;Pan 2022;Sruthi 2022;Bhar Roy, 2023;Gautam A growing evidence show roles micropeptides various but not limited (Sharma 2020;Erokhina 2021;Badola 2022), adventitious root formation (Chen 2020), nodule (Couzigou 2016), cold response anthocyanin (Vale 2024), responses cadmium arsenic stressors (Kumar 2023;Lu immunity (Zhou 2022). Recently, microRPG1 (micropeptide RPG ORF1) peptide governs (KDR) has identified, offering novel perspectives regulate micropeptide providing valuable insights future breeding cereal crops 1B; Lyu, 2024;Yu 2024).Maize (Zea mays) one important world-wide, annual global production over 1147 million tons (Yang Yan, Mechanized kernels viable solution reduce labor enhance efficiency. yet achieved China due absence appropriate corn cultivars 2018a;Wang 2018;Liu requires sufficiently (15% -25%) (Liu This poses challenge exhibit during usually 40% (Dai 2018a;Zhou Consequently, enhancing minimizing become aim modern (Sala 2006;Qu this end, prominent locus (QTL) KDR, designated Dehydration Rate 1 (qKDR1), recombinant inbred line population, originated crossbreeding lines K22 DAN340, known their variant KDRs (Pan 2016;Xiao 2016;Yu 2024).qKDR1 located chromosome 1, specifically 1417 base pair (bp) intergenic noncoding region genome (Yu Targeted deletion sequence via CRISPR-Cas9 yields varying KDRs, demonstrating 1417-bp segment qKDR1 variability, its knockout impaired KDR. investigate transient transcriptional activity assays were conducted protoplasts. findings reveal functions silencer, 369-bp repressive element. Subsequent RNA-sequencing analysis performed ascertain potential targets qKDR1, leading identification target gene, Regulated Peptide Gene (RPG). situated 10 kilobases upstream exhibits levels kernels, declines later stages maturation. where knocked out, markedly elevated. these results indicate acts repressor expression. Furthermore, public chromatin immunoprecipitation sequencing (ChIP-seq) datasets uncovered MYB-related factors, ZmMYBST1 ZmMYBR43, bind locus. Both ZmMYBR43 patterns similar RPG, they inhibit activity. Additionally, CRISPR-Cas9-generated double mutants demonstrate reduced These suggest interact downregulate expression, thereby modulating KDR.Ribosome profiling (Ribo-seq) reveals mRNA ribosome bound frames, ORF1, ORF2, ORF3. Mutations ORF1 accelerated whereas mutations other ORFs no obvious effect Overexpressing resulted decelerated decreased different environments. endogenous verified (IP) mass spectrometry (MS). encodes functional (microRPG1). ZmEIL1 ZmEIL3, downstream RNA-seq assay. ZmEIL3 upregulated downregulated overexpression lines, respectively. Consistently, contrast, application facilitates rate. Hence, microRPG1peptide represses further decelerates 1B).Although function established Arabidopsis precise warrant exploration. First, binding affinities sites elucidated. Secondly, inhibits potentially through native promoter investigation. It proposed localized plasma nucleus, cytoplasm, indicating unidentified receptors may exist recognize peptide, initiating cascades, nucleus finely tolerance. advanced CRISPR screening platform powerful methodology generating single receptor-like kinases (RLKs) simultaneously (Gaillochet 2021), will facilitate uncharacterized signal modulate it plausible effects independently any specific receptors.Additionally, interactions (Kijak necessitate scrutiny. Importantly, single-cell transcriptomic facilitated involved (Liew 2024;Yao technique unveil resolution establish unprecedented networks govern Notably, develop sustain 1A). question remains unresolved how initiates peptide. interacts yet-to-be-identified require elucidation 2020;Farrant 2022;Waterworth 2024).Despite fact exclusively found genera Zea influence Desiccation maternal array programmed suggesting process, CLAVATA3/EMBRYO SURROUNDING REGION 9 (CLE9) 2019) andCLE25/26 (Takahashi 2018;Endo Fukuda, C-TERMINALLY ENCODED PEPTIDE 5 (CEP5) (Smith RAPID ALKALINIZATION FACTOR (RALF) (Jing along droughtresponsive (Xie 2022;Ji 2025;Zhang 2025), examinations. imaging (MSI) employed elucidate spatial distribution structurally diverse hormones 2024) compounds (Garcí a-Rojas 2024;Yin 2024;Zou even (Croslow 2024;Zhang 2024b). successfully identify mammalian cells (David 2018;Bottomley MSI instrumental discovering existing knowledge underlying addition miPEPs horticultural (Ji 2025); biological uncharacterized. CRISPR-Cas system generation miPEP 2021b), Moreover, CRISPR-mediated tools, interference (CRISPRi), (CRISPRa), CRISPRoff, CROP-seq, CRISP-seq, CRISPR-based epigenetic Perturb-seq coupled transcriptomics utilized growth, agronomic traits, resolution. tools enable construction peptides.In summary, discovery contributes improvement adapt harvesting. According Worldostats (https:://worldostats.com/corn-maize-production-by-country-2025/) FAO (Food Agriculture Organization United Nations) staggering 1.16 billion tones per year. top producing countries USA (348.8 tons), (277.2 tons) Brazil (109.4 accounting half production. would lower prevent increase worldwide food supplement future. possible introduce rice, wheat millet artificially exogenous manipulate beneficial future.Identifying subsequently promote period. reduction levels.DT maintained intricate dormancy. Optimal humidity prolong favorable contribute senescence. Water imbibition triggers activities processes, culminating germination. Cutting-edge technologies rehydrate desiccation,

Language: Английский

A micropeptide regulates seed desiccation DOI Creative Commons

Rong Chun Lu,

Zilin Zhang,

Songping Hu

et al.

Frontiers in Plant Science, Journal Year: 2025, Volume and Issue: 16

Published: March 26, 2025

Seeds serve as the major means of reproduction for most plant species and form foundation both agriculture natural ecosystems (Waterworth et al., 2024). are also key genetic resources to deal with increasing human population climate fluctuations (Leprince 2017).Seed development can be categorized into three stages: maturation, dormancy, germination (Figure 1A; Zinsmeister 2020;Powell, 2022;Nadarajan 2023;Waterworth 2024).In maturation phase, seeds acquire desiccation tolerance, followed by developmental processes that expands longevity dormancy stage. Maturation drying reduces seed moisture content 5% -15% fresh weight 2020). Dormancy under optimal conditions, such low temperature humidity, prolongs viability, while suboptimal conditions lead aging Powell 2023). Seed is modulated a complex interplay genetic, biochemical, molecular determinants intricately connected environmental signals light, temperature, nitrate availability, phytohormones including abscisic acid (ABA) gibberellin (GA) (Chahtane 2017;Matilla, 2024;Rachappanavar, 2025).The difference between dormant non-dormant could attributed number gene expression changes (Meimoun 2014), physiological, developmental, morphological features grains on spike, pericarp color, transparency, hairiness, waxiness, permeability water, α-amylase activity, concentrations growth regulators ABA GA within embryo (Sohn al. 2021). initiated water uptake (imbibition), resulting in activation multiple cellular actions, completed emergence young roots shoots 1A;Carrera-Castaño 2020;Waterworth 2024).The acquisition tolerance at late stage provides critical survival mechanism crops, enabling them adaptive adverse extreme temperatures drought 2017;Zinsmeister The majority crop plants generate classified orthodox seeds, which possess ability withstand (below 7%) harsh freezing (-10 ℃) long time (Nadarajan maize, kernel suitable mechanized harvesting from 15% -25%, however, some regions China, maize varieties have high grain harvest, ranging 30% -40% (Xiang 2012;Kebebe 2015;Dai 2017;Li 2018aLi , 2018b;;Zhou 2018). Kernel dehydration rate (KDR), defined loss two adjacent periods after pollination (Zhang 2024a), determinant quality exerts significant impact efficiency mechanical (Li 2018c).Besides, removal free leads phase transition cytoplasm mobility fluid glassy state (Buitink Leprince, 2008), metabolic quiescence increased (Zinsmeister To date, elements implicated modulation reactive oxygen (ROS) signaling, lipid peroxidation cell membrane, preservation DNA RNA integrity, methylation status, biosynthesis storage proteins (SSPs), ABA, auxin, GA, brassinosteroids (BRs) been documented crucial 2023;Pirredda 2024).Abiotic factors thermal salinity stress significantly longevity, availability emerging predominant In several quantitative trait loci (QTLs) characterized pivotal players regulation KDR 2020(Li 2021a;;Zhang 2023;Jin Collectively, higher level harvesting, preventing breakage, mildew, reducing costs associated harvest (de Jager 2004;Xiang 2015;Wang 2022;Xia Thus, comprehensive understanding mechanisms governing necessary crucial.Seed linked multitude physiological modifications, accumulation macromolecules (proteins, lipids, carbohydrates), enhanced membrane defense mechanisms, governed hormone signaling pathways ethylene (Angelovici 2010;Bewley 2013;Kijak Ratajczak, 2020;Oliver 2020;Smolikova onset occurs when enter 2017;Smolikova Numerous components Late Embryogenesis Abundant (LEA) proteins, small heat shock (sHSPs), non-reducing oligosaccharides, antioxidants, (ROS), well (GA), identified 2010;Kijak addition, many transcription ABA-INSENSITIVE 3 (ABI3), FUSCA (FUS3) LEAFY COTYLEDONS 2 (LEC2) discovered defines balance finally initiate (Smolikova However, regulatory mediated peptides remain largely elusive.Micropeptides, referred microproteins or short open reading frame (sORF)-encoded peptides, essential products derived larger polypeptide MicroRNAs (miRNAs), non-coding (lncRNA), circular (circRNA), typically an arbitrary length less than 100 -150 amino acids (Hashimoto 2008;Makarewich Olson, 2017;Sousa Farkas, 2018;Vitorino 2021;Pan 2022;Sruthi 2022;Bhar Roy, 2023;Gautam A growing evidence show roles micropeptides various but not limited (Sharma 2020;Erokhina 2021;Badola 2022), adventitious root formation (Chen 2020), nodule (Couzigou 2016), cold response anthocyanin (Vale 2024), responses cadmium arsenic stressors (Kumar 2023;Lu immunity (Zhou 2022). Recently, microRPG1 (micropeptide RPG ORF1) peptide governs (KDR) has identified, offering novel perspectives regulate micropeptide providing valuable insights future breeding cereal crops 1B; Lyu, 2024;Yu 2024).Maize (Zea mays) one important world-wide, annual global production over 1147 million tons (Yang Yan, Mechanized kernels viable solution reduce labor enhance efficiency. yet achieved China due absence appropriate corn cultivars 2018a;Wang 2018;Liu requires sufficiently (15% -25%) (Liu This poses challenge exhibit during usually 40% (Dai 2018a;Zhou Consequently, enhancing minimizing become aim modern (Sala 2006;Qu this end, prominent locus (QTL) KDR, designated Dehydration Rate 1 (qKDR1), recombinant inbred line population, originated crossbreeding lines K22 DAN340, known their variant KDRs (Pan 2016;Xiao 2016;Yu 2024).qKDR1 located chromosome 1, specifically 1417 base pair (bp) intergenic noncoding region genome (Yu Targeted deletion sequence via CRISPR-Cas9 yields varying KDRs, demonstrating 1417-bp segment qKDR1 variability, its knockout impaired KDR. investigate transient transcriptional activity assays were conducted protoplasts. findings reveal functions silencer, 369-bp repressive element. Subsequent RNA-sequencing analysis performed ascertain potential targets qKDR1, leading identification target gene, Regulated Peptide Gene (RPG). situated 10 kilobases upstream exhibits levels kernels, declines later stages maturation. where knocked out, markedly elevated. these results indicate acts repressor expression. Furthermore, public chromatin immunoprecipitation sequencing (ChIP-seq) datasets uncovered MYB-related factors, ZmMYBST1 ZmMYBR43, bind locus. Both ZmMYBR43 patterns similar RPG, they inhibit activity. Additionally, CRISPR-Cas9-generated double mutants demonstrate reduced These suggest interact downregulate expression, thereby modulating KDR.Ribosome profiling (Ribo-seq) reveals mRNA ribosome bound frames, ORF1, ORF2, ORF3. Mutations ORF1 accelerated whereas mutations other ORFs no obvious effect Overexpressing resulted decelerated decreased different environments. endogenous verified (IP) mass spectrometry (MS). encodes functional (microRPG1). ZmEIL1 ZmEIL3, downstream RNA-seq assay. ZmEIL3 upregulated downregulated overexpression lines, respectively. Consistently, contrast, application facilitates rate. Hence, microRPG1peptide represses further decelerates 1B).Although function established Arabidopsis precise warrant exploration. First, binding affinities sites elucidated. Secondly, inhibits potentially through native promoter investigation. It proposed localized plasma nucleus, cytoplasm, indicating unidentified receptors may exist recognize peptide, initiating cascades, nucleus finely tolerance. advanced CRISPR screening platform powerful methodology generating single receptor-like kinases (RLKs) simultaneously (Gaillochet 2021), will facilitate uncharacterized signal modulate it plausible effects independently any specific receptors.Additionally, interactions (Kijak necessitate scrutiny. Importantly, single-cell transcriptomic facilitated involved (Liew 2024;Yao technique unveil resolution establish unprecedented networks govern Notably, develop sustain 1A). question remains unresolved how initiates peptide. interacts yet-to-be-identified require elucidation 2020;Farrant 2022;Waterworth 2024).Despite fact exclusively found genera Zea influence Desiccation maternal array programmed suggesting process, CLAVATA3/EMBRYO SURROUNDING REGION 9 (CLE9) 2019) andCLE25/26 (Takahashi 2018;Endo Fukuda, C-TERMINALLY ENCODED PEPTIDE 5 (CEP5) (Smith RAPID ALKALINIZATION FACTOR (RALF) (Jing along droughtresponsive (Xie 2022;Ji 2025;Zhang 2025), examinations. imaging (MSI) employed elucidate spatial distribution structurally diverse hormones 2024) compounds (Garcí a-Rojas 2024;Yin 2024;Zou even (Croslow 2024;Zhang 2024b). successfully identify mammalian cells (David 2018;Bottomley MSI instrumental discovering existing knowledge underlying addition miPEPs horticultural (Ji 2025); biological uncharacterized. CRISPR-Cas system generation miPEP 2021b), Moreover, CRISPR-mediated tools, interference (CRISPRi), (CRISPRa), CRISPRoff, CROP-seq, CRISP-seq, CRISPR-based epigenetic Perturb-seq coupled transcriptomics utilized growth, agronomic traits, resolution. tools enable construction peptides.In summary, discovery contributes improvement adapt harvesting. According Worldostats (https:://worldostats.com/corn-maize-production-by-country-2025/) FAO (Food Agriculture Organization United Nations) staggering 1.16 billion tones per year. top producing countries USA (348.8 tons), (277.2 tons) Brazil (109.4 accounting half production. would lower prevent increase worldwide food supplement future. possible introduce rice, wheat millet artificially exogenous manipulate beneficial future.Identifying subsequently promote period. reduction levels.DT maintained intricate dormancy. Optimal humidity prolong favorable contribute senescence. Water imbibition triggers activities processes, culminating germination. Cutting-edge technologies rehydrate desiccation,

Language: Английский

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