{"title":"Cyclic nucleotide-gated ion channel 20 regulates melatonin-induced calcium signaling and cold tolerance in watermelon","authors":"Jingjing Chang, Yanliang Guo, Jiayue Li, Lingling Liu, Jiahe Liu, Li Yuan, Chunhua Wei, Jianxiang Ma, Yong Zhang, Golam Jalal Ahammed, Feishi Luan, Yunqi Liu, Xian Zhang, Hao Li","doi":"10.1093/plphys/kiae630","DOIUrl":null,"url":null,"abstract":"Melatonin plays a crucial role in regulating plant cold tolerance, but the mechanisms underlying signal transduction remain elusive. In this study, we discovered that overexpression of the melatonin biosynthetic gene caffeic acid O-methyltransferase1 (COMT1) enhanced watermelon (Citrullus lanatus) cold tolerance, accompanied by the accumulation of cytosolic free calcium ([Ca2+]cyt), a stimulation of Ca2+ influx, and upregulation of four Ca2+-permeable channel genes (CNGC2/10/17/20). Conversely, knockout of COMT1 exhibited contrasting effects compared to its overexpression. Knocking out the four CNGC genes revealed that only CNGC20 mediates melatonin-induced Ca2+ influx in response to cold stimuli. CNGC20 deletion impeded watermelon callus redifferentiation, prompting us to employ a virus-induced gene silencing strategy to suppress its expression. Silencing CNGC20 compromised COMT1 overexpression-induced [Ca2+]cyt accumulation, Ca2+ influx, and watermelon cold tolerance. Yeast two-hybrid, bimolecular fluorescence complementation, firefly luciferase complementation imaging, and pull-down assays revealed an interaction between CNGC20 and calmodulin7 (CaM7). Overexpressing CaM7 inhibited melatonin-induced [Ca2+]cyt accumulation, Ca2+ influx, and watermelon cold tolerance. Conversely, silencing CaM7 increased [Ca2+]cyt accumulation, Ca2+ influx, and cold tolerance, whereas COMT1 overexpression failed to further enhance these responses in CaM7-silenced plants, indicating the negative regulation role of CaM7 in melatonin-mediated cold responses. Overall, these findings provide insights into the molecular mechanisms underlying melatonin-enhanced plant cold tolerance via Ca2+ signaling, holding potential for breeding/engineering cold-tolerant cucurbit varieties.","PeriodicalId":20101,"journal":{"name":"Plant Physiology","volume":"77 1","pages":""},"PeriodicalIF":6.5000,"publicationDate":"2024-11-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Plant Physiology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1093/plphys/kiae630","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"PLANT SCIENCES","Score":null,"Total":0}
引用次数: 0
Abstract
Melatonin plays a crucial role in regulating plant cold tolerance, but the mechanisms underlying signal transduction remain elusive. In this study, we discovered that overexpression of the melatonin biosynthetic gene caffeic acid O-methyltransferase1 (COMT1) enhanced watermelon (Citrullus lanatus) cold tolerance, accompanied by the accumulation of cytosolic free calcium ([Ca2+]cyt), a stimulation of Ca2+ influx, and upregulation of four Ca2+-permeable channel genes (CNGC2/10/17/20). Conversely, knockout of COMT1 exhibited contrasting effects compared to its overexpression. Knocking out the four CNGC genes revealed that only CNGC20 mediates melatonin-induced Ca2+ influx in response to cold stimuli. CNGC20 deletion impeded watermelon callus redifferentiation, prompting us to employ a virus-induced gene silencing strategy to suppress its expression. Silencing CNGC20 compromised COMT1 overexpression-induced [Ca2+]cyt accumulation, Ca2+ influx, and watermelon cold tolerance. Yeast two-hybrid, bimolecular fluorescence complementation, firefly luciferase complementation imaging, and pull-down assays revealed an interaction between CNGC20 and calmodulin7 (CaM7). Overexpressing CaM7 inhibited melatonin-induced [Ca2+]cyt accumulation, Ca2+ influx, and watermelon cold tolerance. Conversely, silencing CaM7 increased [Ca2+]cyt accumulation, Ca2+ influx, and cold tolerance, whereas COMT1 overexpression failed to further enhance these responses in CaM7-silenced plants, indicating the negative regulation role of CaM7 in melatonin-mediated cold responses. Overall, these findings provide insights into the molecular mechanisms underlying melatonin-enhanced plant cold tolerance via Ca2+ signaling, holding potential for breeding/engineering cold-tolerant cucurbit varieties.
期刊介绍:
Plant Physiology® is a distinguished and highly respected journal with a rich history dating back to its establishment in 1926. It stands as a leading international publication in the field of plant biology, covering a comprehensive range of topics from the molecular and structural aspects of plant life to systems biology and ecophysiology. Recognized as the most highly cited journal in plant sciences, Plant Physiology® is a testament to its commitment to excellence and the dissemination of groundbreaking research.
As the official publication of the American Society of Plant Biologists, Plant Physiology® upholds rigorous peer-review standards, ensuring that the scientific community receives the highest quality research. The journal releases 12 issues annually, providing a steady stream of new findings and insights to its readership.