Inflorescence architecture is determined by inflorescence length,branch angles and the density of siliques,which affects planting density,lodging resistance and mechanical operation in rapeseed.However,the molecular m...Inflorescence architecture is determined by inflorescence length,branch angles and the density of siliques,which affects planting density,lodging resistance and mechanical operation in rapeseed.However,the molecular mechanisms controlling inflorescence architecture are poorly understood,restricting the progress of breeding varieties with ideal plant architecture in oilseed rape.In this study,we have identified and characterized a rapeseed inflorescence development mutant,reduced inflorescence length(ril),which exhibits determinate and shortened inflorescences,reduced plant height,compact branches,and increased silique density.Through BSA-seq and map-based cloning,we find that RIL encodes a cyclic nucleotide-gated channel 20(BnaA01.CNGC20).A substitution of proline at the 304th position to leucine(P304L)was identified in the conserved transmembrane domain of BnaA01.CNGC20.This P304L substitution neither affects the subcellular localization and self-assembly of BnaA01.CNGC20,nor disrupts the interactions with BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1(BAK1),which interacts with CNGC20 and phosphorylates it to regulate Ca^(2+)channel stability.However,the P304L substitution increases channel activity and Ca^(2+)influx,which in turn induces immune responses such as cell death,H2O2 accumulation,upregulation of pathogenesis-related genes,and pattern-triggered immunity.The enhanced immunity improves the resistance to Leptosphaeria biglobosa and Sclerotinia sclerotiorum.Transcriptome analysis further revealed that CNGC20 plays dual roles in regulating plant growth and immunity via the brassinosteroid and auxin signaling pathways.The findings in this study provide deeper insights into the intricate relationship between cytosolic Ca^(2+)level and plant development and immunity,as well as the trade-off between immunity and the performance of yield-related traits in the heterozygous plants(+/ril),which may serve as a guide for balancing yield and disease resistance in oilseed rape breeding.展开更多
环核苷酸门控离子通道(Cyclic nucleotide-gated ion channels,CNGC)是非选择性的阳离子通道,受细胞内信使小分子环核苷酸(cAMP和cGMP)以及Ca^(2+)/CaM调控。哺乳动物CNGC功能的变构调节机制受到CaM结合影响,哺乳动物CNGC在胞质N和/或C...环核苷酸门控离子通道(Cyclic nucleotide-gated ion channels,CNGC)是非选择性的阳离子通道,受细胞内信使小分子环核苷酸(cAMP和cGMP)以及Ca^(2+)/CaM调控。哺乳动物CNGC功能的变构调节机制受到CaM结合影响,哺乳动物CNGC在胞质N和/或C末端具有CaMBD。在植物方面,研究大多集中于与植物CNGC的环核苷酸结合结构域重叠的C端CaM结合结构域(CaMBD)。然而近期对模式植物拟南芥CNGC12的研究提供了单个植物CNGC同种型具有多个CaMBD的证据。重点总结了动植物钙调蛋白多个结合位点调控环核苷酸门控离子通道的研究进展。展开更多
Members of the cyclic nucleotide-gated channel(CNGC)proteins are reportedly involved in a variety of biotic and abiotic responses and stomatal movement.However,it is unknown if and how a single member could regulate m...Members of the cyclic nucleotide-gated channel(CNGC)proteins are reportedly involved in a variety of biotic and abiotic responses and stomatal movement.However,it is unknown if and how a single member could regulate multiple responses.Here we characterized three closely related CNGC genes in rice,OsCNGC14,OsCNGC15 and OsCNGC16,to determine whether they function in multiple abiotic stresses.The loss-of-function mutants of each of these three genes had reduced calcium ion(Ca^(2+))influx and slower stomatal closure in response to heat,chilling,drought and the stress hormone abscisic acid(ABA).These mutants also had reduced tolerance to heat,chilling and drought compared with the wildtype.Conversely,overexpression of OsCNGC16 led to a more rapid stomatal closure response to stresses and enhanced tolerance to heat,chilling and drought.The tight association of stomatal closure and stress tolerance strongly suggests that tolerance to multiple abiotic stresses conferred by these Os CNGC genes results at least partially from their regulation of stomatal movement.In addition,physical interactions were observed among the three Os CNGC proteins but not with a distantly related CNGC,suggesting the formation of hetero-oligomers among themselves.This study unveils the crucial role of OsCNGC14,15 and 16 proteins in stomatal response and tolerance to multiple stresses,suggesting a mechanism of tolerance to multiple stresses that involves calcium influx and stomatal movement regulation.展开更多
Extracellular adenosine triphosphate(eATP)functions as a damage-associated molecular pattern in plant immunity.P2K1,a purinergic receptor with a cytoplasmic serine/threonine kinase domain,initiates ATP-responsive sign...Extracellular adenosine triphosphate(eATP)functions as a damage-associated molecular pattern in plant immunity.P2K1,a purinergic receptor with a cytoplasmic serine/threonine kinase domain,initiates ATP-responsive signaling cascades characterized by a rapid spike in cytosolic Ca^(2+),which acts as a critical second messenger.In this study,we identified the cyclic nucleotide-gated channel complex CNGC2–CNGC4 as essential for eATP-induced calcium signaling and bacterial resistance in plants.A biochemical link between eATP perception and CNGC2–CNGC4 function was established by demonstrating the physical association between the channel complex and the eATP receptor P2K1 at the plasma membrane.Furthermore,we discovered that P2K1 phosphorylates the CNGC2 subunit of the CNGC2–CNGC4 channel in response to eATP,establishing a phosphorylation-dependent mechanism that connects eATP perception to calcium influx.Through AlphaFold-Multimer prediction,electrophysiological assay,and genetic analysis,we identified serine residues S705 and S718 in CNGC2 as the key phosphorylation sites mediating P2K1-dependent channel activation and eATP-triggered immunity.Notably,P2K1 selectively phosphorylates CNGC2,in contrast to BIK1 that phosphorylates CNGC4 during pathogen-associated molecular pattern-triggered immunity.Together,these findings indicate that the CNGC2-CNGC4 channel complex serves as a core component of calcium-dependent plant immunity,with distinct kinases phosphorylating different subunits in response to specific immune elicitors.展开更多
Cyclic nucleotide-gatedion channels(CNGCs)are key components in pattern-triggeredimmunity(PTl)signaling.Tight control of CNGC homeostasis is crucial for maintaining a balance between plant growth and immunity.Neverthe...Cyclic nucleotide-gatedion channels(CNGCs)are key components in pattern-triggeredimmunity(PTl)signaling.Tight control of CNGC homeostasis is crucial for maintaining a balance between plant growth and immunity.Nevertheless,the mechanisms for fine-tuning CNGC homeostasis remain largely unknown.Here,we report that Arabidopsis thaliana CNGC3 is a functionalcalcium channel to mediate patterninduced Ca2+influx,PTl,and resistance to Sclerotinia sclerotiorum.We identified a CNGC interactor,Skp1-interacting protein 31(SKIP31).In the absence of a pathogen,SKIP31ubiquitinates CNGC3 at Lys8 and Lys33 of the K-X-V-R motif for degradation to repress plant immunity.When a pathogen attacks,activated receptor-like cytoplasmic kinase(RLCK)BOTRYTIS-INDUCED KINASE1(BIK1)phosphorylates SKIP31 to inhibit its ubiquitin ligase activity and interaction with the CNGC3 N-terminal region,thereby suppressing CNGC3 protein degradation to promote immunity.Phosphorylation within the F box of SKlP31 at Ser88 and Ser93 and at the C-terminal Ser261 prevents its interaction with Skp1 and CNGC3,respectively.These phosphorylation sites are conserved in SKlP31 of different plant species,and SKlP31 interacts with all examined CNGCs,suggesting a pivotal role of SKIP31 phosphorylation in regulating CNGC stability and plant immunity.Moreover,biochemical assays revealed that BIK1 directly phosphorylates the CNGC3 cytoplasmic C-terminal region at four Ser residues to enhance its Ca²+channel activity,demonstrating dual roles of BlK1 in both promoting CNGC channel activity and stabilizing the channel protein.Collectively,our work unveils an SCF ubiquitin ligase-RLCK control system that fine-tunes the homeostasis of CNGCs for orchestrating plant immunity.展开更多
基金supported by the National Natural Science Foundation of China (U22A20477 and 32201791)the China Postdoctoral Science Foundation (2020M682440)the Postdoctoral Fellowship Program of CPSF (GZB20230825).
文摘Inflorescence architecture is determined by inflorescence length,branch angles and the density of siliques,which affects planting density,lodging resistance and mechanical operation in rapeseed.However,the molecular mechanisms controlling inflorescence architecture are poorly understood,restricting the progress of breeding varieties with ideal plant architecture in oilseed rape.In this study,we have identified and characterized a rapeseed inflorescence development mutant,reduced inflorescence length(ril),which exhibits determinate and shortened inflorescences,reduced plant height,compact branches,and increased silique density.Through BSA-seq and map-based cloning,we find that RIL encodes a cyclic nucleotide-gated channel 20(BnaA01.CNGC20).A substitution of proline at the 304th position to leucine(P304L)was identified in the conserved transmembrane domain of BnaA01.CNGC20.This P304L substitution neither affects the subcellular localization and self-assembly of BnaA01.CNGC20,nor disrupts the interactions with BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1(BAK1),which interacts with CNGC20 and phosphorylates it to regulate Ca^(2+)channel stability.However,the P304L substitution increases channel activity and Ca^(2+)influx,which in turn induces immune responses such as cell death,H2O2 accumulation,upregulation of pathogenesis-related genes,and pattern-triggered immunity.The enhanced immunity improves the resistance to Leptosphaeria biglobosa and Sclerotinia sclerotiorum.Transcriptome analysis further revealed that CNGC20 plays dual roles in regulating plant growth and immunity via the brassinosteroid and auxin signaling pathways.The findings in this study provide deeper insights into the intricate relationship between cytosolic Ca^(2+)level and plant development and immunity,as well as the trade-off between immunity and the performance of yield-related traits in the heterozygous plants(+/ril),which may serve as a guide for balancing yield and disease resistance in oilseed rape breeding.
文摘环核苷酸门控离子通道(Cyclic nucleotide-gated ion channels,CNGC)是非选择性的阳离子通道,受细胞内信使小分子环核苷酸(cAMP和cGMP)以及Ca^(2+)/CaM调控。哺乳动物CNGC功能的变构调节机制受到CaM结合影响,哺乳动物CNGC在胞质N和/或C末端具有CaMBD。在植物方面,研究大多集中于与植物CNGC的环核苷酸结合结构域重叠的C端CaM结合结构域(CaMBD)。然而近期对模式植物拟南芥CNGC12的研究提供了单个植物CNGC同种型具有多个CaMBD的证据。重点总结了动植物钙调蛋白多个结合位点调控环核苷酸门控离子通道的研究进展。
基金supported by grants from the National Natural Science Foundation of China(32370309)the STI 2030-Major Projects(2023ZD040710X)+2 种基金the National Natural Science Foundation of China(31700223)Jiangsu Agricultural Science and Technology Innovation Fund(CX(23)1033)Jiangsu Collaborative Innovation Center for Modern Crop Production and Cyrus Tang Innovation Center for Crop Seed Industry。
文摘Members of the cyclic nucleotide-gated channel(CNGC)proteins are reportedly involved in a variety of biotic and abiotic responses and stomatal movement.However,it is unknown if and how a single member could regulate multiple responses.Here we characterized three closely related CNGC genes in rice,OsCNGC14,OsCNGC15 and OsCNGC16,to determine whether they function in multiple abiotic stresses.The loss-of-function mutants of each of these three genes had reduced calcium ion(Ca^(2+))influx and slower stomatal closure in response to heat,chilling,drought and the stress hormone abscisic acid(ABA).These mutants also had reduced tolerance to heat,chilling and drought compared with the wildtype.Conversely,overexpression of OsCNGC16 led to a more rapid stomatal closure response to stresses and enhanced tolerance to heat,chilling and drought.The tight association of stomatal closure and stress tolerance strongly suggests that tolerance to multiple abiotic stresses conferred by these Os CNGC genes results at least partially from their regulation of stomatal movement.In addition,physical interactions were observed among the three Os CNGC proteins but not with a distantly related CNGC,suggesting the formation of hetero-oligomers among themselves.This study unveils the crucial role of OsCNGC14,15 and 16 proteins in stomatal response and tolerance to multiple stresses,suggesting a mechanism of tolerance to multiple stresses that involves calcium influx and stomatal movement regulation.
基金supported by the Key Program of the National Natural Science Foundation of China(no.31930010 to L.L.)the National Key Research and Development Program of China(2021YFA1300700 to W.T.)the National Natural Science Foundation of China(nos.32270326 to W.T.,32270265 to L.L.,32170263 to D.K.,32222011 to L.L.).
文摘Extracellular adenosine triphosphate(eATP)functions as a damage-associated molecular pattern in plant immunity.P2K1,a purinergic receptor with a cytoplasmic serine/threonine kinase domain,initiates ATP-responsive signaling cascades characterized by a rapid spike in cytosolic Ca^(2+),which acts as a critical second messenger.In this study,we identified the cyclic nucleotide-gated channel complex CNGC2–CNGC4 as essential for eATP-induced calcium signaling and bacterial resistance in plants.A biochemical link between eATP perception and CNGC2–CNGC4 function was established by demonstrating the physical association between the channel complex and the eATP receptor P2K1 at the plasma membrane.Furthermore,we discovered that P2K1 phosphorylates the CNGC2 subunit of the CNGC2–CNGC4 channel in response to eATP,establishing a phosphorylation-dependent mechanism that connects eATP perception to calcium influx.Through AlphaFold-Multimer prediction,electrophysiological assay,and genetic analysis,we identified serine residues S705 and S718 in CNGC2 as the key phosphorylation sites mediating P2K1-dependent channel activation and eATP-triggered immunity.Notably,P2K1 selectively phosphorylates CNGC2,in contrast to BIK1 that phosphorylates CNGC4 during pathogen-associated molecular pattern-triggered immunity.Together,these findings indicate that the CNGC2-CNGC4 channel complex serves as a core component of calcium-dependent plant immunity,with distinct kinases phosphorylating different subunits in response to specific immune elicitors.
基金supported by grants from the Zhejiang Province Core Technology Innovation Project in Agriculture,the Hainan Provincial Natural Science Foundation of China(324CXTD430)the National Natural Science Foundation of China(31871947)the Zhejiang Provincial Natural Science Foundation of China(LZ18C140002).
文摘Cyclic nucleotide-gatedion channels(CNGCs)are key components in pattern-triggeredimmunity(PTl)signaling.Tight control of CNGC homeostasis is crucial for maintaining a balance between plant growth and immunity.Nevertheless,the mechanisms for fine-tuning CNGC homeostasis remain largely unknown.Here,we report that Arabidopsis thaliana CNGC3 is a functionalcalcium channel to mediate patterninduced Ca2+influx,PTl,and resistance to Sclerotinia sclerotiorum.We identified a CNGC interactor,Skp1-interacting protein 31(SKIP31).In the absence of a pathogen,SKIP31ubiquitinates CNGC3 at Lys8 and Lys33 of the K-X-V-R motif for degradation to repress plant immunity.When a pathogen attacks,activated receptor-like cytoplasmic kinase(RLCK)BOTRYTIS-INDUCED KINASE1(BIK1)phosphorylates SKIP31 to inhibit its ubiquitin ligase activity and interaction with the CNGC3 N-terminal region,thereby suppressing CNGC3 protein degradation to promote immunity.Phosphorylation within the F box of SKlP31 at Ser88 and Ser93 and at the C-terminal Ser261 prevents its interaction with Skp1 and CNGC3,respectively.These phosphorylation sites are conserved in SKlP31 of different plant species,and SKlP31 interacts with all examined CNGCs,suggesting a pivotal role of SKIP31 phosphorylation in regulating CNGC stability and plant immunity.Moreover,biochemical assays revealed that BIK1 directly phosphorylates the CNGC3 cytoplasmic C-terminal region at four Ser residues to enhance its Ca²+channel activity,demonstrating dual roles of BlK1 in both promoting CNGC channel activity and stabilizing the channel protein.Collectively,our work unveils an SCF ubiquitin ligase-RLCK control system that fine-tunes the homeostasis of CNGCs for orchestrating plant immunity.
