The complex of calcineurin B-like protein(CBL)and CBL-interacting protein kinase(CIPK)serves as key components in calcium-sensing,orchestrating various signals crucial for plant growth,development,and responses to bio...The complex of calcineurin B-like protein(CBL)and CBL-interacting protein kinase(CIPK)serves as key components in calcium-sensing,orchestrating various signals crucial for plant growth,development,and responses to biotic and abiotic stresses.However,the mechanism underlying the response of this module to cold stress and its role in flower development in wintersweet(Chimonanthus praecox)remains unclear.Through expression pattern analysis,calcium ion(Ca^(2+))concentration assays,correlation analysis,and linear regression analysis,we found that the[Ca^(2+)],along with CpCBL8 and CpCIPK9 expression levels in wintersweet flower buds(FBs),significantly decreased during the initial flowering stage when the chilling requirement reached 570 chill units(CU).Notably,there was a significant positive correlation between[Ca^(2+)]and CpCBL8 expression.Ca^(2+)increased the expression of Cp CBL8 and CpCIPK9 in FBs,causing a significant delay in the flowering of wintersweet.Furthermore,the function of CpCBL8 was studied using heterologous transformation.Overexpression of CpCBL8 significantly delayed flowering time and significantly reduced cold tolerance and altered the expression pattern of endogenous genes related to low-temperature stress and flower development in transgenic Arabidopsis thaliana.Additionally,transcriptome analysis of chilling-induced dormancy breaking and flower bud enlargement revealed that CpCBL8 and CpCIPK9 were negatively regulated by cold,and the expression pattern of endogenous genes related to flower development and cold stress in wintersweet were similar to that of in A.thaliana.Moreover,protein-protein interaction(PPI)analysis revealed that CpCBL8 and CpCIPK9 interacted in the plasma membrane and nucleus.On the basis of these findings,we speculated that the CpCBL8-CpCIPK9 module plays a crucial role in regulating responses to cold stress and flower development in wintersweet.This study elucidated molecular mechanisms through which the downregulation of the Ca^(2+)-induced CpCBL8-CpCIPK9 module results in dormancy breaking and enhances cold tolerance.This study provides valuable insights for the cultivation of new varieties of wintersweet with increased ornamental value and enhanced cold stress tolerance.展开更多
Plant calcineurin B-like （CBL） proteins have been proposed as important Ca2＋ sensors and specifically interact with CBL-interacting protein kinases （CIPKs） in plant-specific calcium signaling. Here, we identified...Plant calcineurin B-like （CBL） proteins have been proposed as important Ca2＋ sensors and specifically interact with CBL-interacting protein kinases （CIPKs） in plant-specific calcium signaling. Here, we identified and isolated 15 CIPK genes in a japonica rice variety Nipponbare based on the predicted sequences of rice CIPK gene family. Gene structure analysis showed that these 15 genes were divided into intron-less and intron-rich groups, and OsCIPK3 and OsCIPK24 exhibited alternative splicing in their mature process. The phylogenetic analyses indicated that rice CIPKs shared an ancestor with Arabidopsis and poplar CIPKs. Analyses of gene expression showed that these OsCIPK genes were differentially induced by biotic stresses such as bacterial blight and abiotic stresses （heavy metal such as Hg2＋, high salinity, cold and ABA）. Interestingly, five OsCIPK genes, OsCIPK1, 2, 10, 11 and 12, were transcriptionally up-regulated after bacterial blight infection whereas four OsCIPK genes, OsCIPK2, 10, 11 and 14, were induced by all treatments, indicating that some of OsCIPK genes are involved in multiple stress response pathways in plants. Our finding suggests that CIPKs play a key role in both biotic and abiotic stress responses.展开更多
Calcium(Ca^(2+))plays a pivotal role in various signal transduction pathways.Calcineurin B-like proteins(CBLs)are a unique group of Ca^(2+)sensors that decode Ca^(2+)signals by activating the plant specific protein ki...Calcium(Ca^(2+))plays a pivotal role in various signal transduction pathways.Calcineurin B-like proteins(CBLs)are a unique group of Ca^(2+)sensors that decode Ca^(2+)signals by activating the plant specific protein kinase known as the CBL-interacting protein kinase(CIPK).In plants,the CBL-CIPK signaling network regulates multiple signals in response to different extracellular cues including abiotic stress.However,the genome wide annotation and expression patterns of CBLs and CIPKs in woody cutting flower plants are still unclear.In this study,a total number of 7 CBLs(RcCBLs)and 17 CIPKs(RcCIPKs)genes,divided into four and five subfamilies,respectively,were identified from the rose genome.All RcCBLs possess a classic elongation factor-hand(EF-hand)domain,while all RcCIPKs possess both the classic kinase and NAF domains.Most RcCBLs were predicted to be plasma membrane localized,whereas most RcCIPKs were predicted to be cytoplasmic localized.Synteny analysis showed that one RcCBL gene pair and five RcCIPK gene pairs have gone through whole genome duplication events.Promoter cis-element prediction assays indicated that RcCBLs and RcCIPKs could function in different abiotic stress responses in rose plants.Further quantitative real-time PCR analysis demonstrated that RcCBLs and RcCIPKs were expressed in different organs with overlapped but distinct patterns in response to various abiotic stresses.The findings in this work will provide fundamental information and gene resources for further functional research on RcCBLs and RcCIPKs.展开更多
Manganese(Mn)is an essential micronutrient for all living organisms.However,excess Mn supply that can occur in acid or waterlogged soils has toxic effects on plant physiology and development.Although a variety of Mn t...Manganese(Mn)is an essential micronutrient for all living organisms.However,excess Mn supply that can occur in acid or waterlogged soils has toxic effects on plant physiology and development.Although a variety of Mn transporter families have been characterized,we have only a rudimentary understanding of how these transporters are regulated to uphold and adjust Mn homeostasis in plants.Here,we demonstrate that two calcineurin-B-like proteins,CBL2/3,and their interacting kinases,CIPK3/9/26,are key regulators of plant Mn homeostasis.Arabidopsis mutants lacking CBL2 and 3 or their interacting protein kinases CIPK3/9/26 exhibit remarkably high Mn tolerance.Intriguingly,CIPK3/9/26 interact with and phosphorylate the tonoplast-localized Mn and iron(Fe)transporter MTP8 primarily at Ser35,which is conserved among MTP8 proteins from various species.Mn transport complementation assays in yeast combined with multiple physiological assays indicate that CBL-CIPK-mediated phosphorylation of MTP8 negatively regulates its transport activity from the cytoplasm to the vacuole.Moreover,we show that sequential phosphorylation of MTP8,initially at Ser31/32 by the calcium-dependent protein kinase CPK5 and subsequently at Ser35 by CIPK26,provides an activation/deactivation fine-tuning mechanism for differential regulation of Mn transport.Collectively,our findings define a two-tiered calcium-controlled mechanism for dynamic regulation of Mn homeostasis under conditions of fluctuating Mn supply.展开更多
基金funded by the Natural Science Foundation of Chongqing(CSTB2023NSCQ-MSX0236)Fundamental Research Funds for the Central Universities(SWU-XDJH202308)Earmarked Funds for the China Agriculture Research System(CARS-26)。
文摘The complex of calcineurin B-like protein(CBL)and CBL-interacting protein kinase(CIPK)serves as key components in calcium-sensing,orchestrating various signals crucial for plant growth,development,and responses to biotic and abiotic stresses.However,the mechanism underlying the response of this module to cold stress and its role in flower development in wintersweet(Chimonanthus praecox)remains unclear.Through expression pattern analysis,calcium ion(Ca^(2+))concentration assays,correlation analysis,and linear regression analysis,we found that the[Ca^(2+)],along with CpCBL8 and CpCIPK9 expression levels in wintersweet flower buds(FBs),significantly decreased during the initial flowering stage when the chilling requirement reached 570 chill units(CU).Notably,there was a significant positive correlation between[Ca^(2+)]and CpCBL8 expression.Ca^(2+)increased the expression of Cp CBL8 and CpCIPK9 in FBs,causing a significant delay in the flowering of wintersweet.Furthermore,the function of CpCBL8 was studied using heterologous transformation.Overexpression of CpCBL8 significantly delayed flowering time and significantly reduced cold tolerance and altered the expression pattern of endogenous genes related to low-temperature stress and flower development in transgenic Arabidopsis thaliana.Additionally,transcriptome analysis of chilling-induced dormancy breaking and flower bud enlargement revealed that CpCBL8 and CpCIPK9 were negatively regulated by cold,and the expression pattern of endogenous genes related to flower development and cold stress in wintersweet were similar to that of in A.thaliana.Moreover,protein-protein interaction(PPI)analysis revealed that CpCBL8 and CpCIPK9 interacted in the plasma membrane and nucleus.On the basis of these findings,we speculated that the CpCBL8-CpCIPK9 module plays a crucial role in regulating responses to cold stress and flower development in wintersweet.This study elucidated molecular mechanisms through which the downregulation of the Ca^(2+)-induced CpCBL8-CpCIPK9 module results in dormancy breaking and enhances cold tolerance.This study provides valuable insights for the cultivation of new varieties of wintersweet with increased ornamental value and enhanced cold stress tolerance.
基金supported by the National Natural Science Foundation of China (Grants Nos.30800677 and 30771329)the Natural Science Foundation of Zhejiang Province, China (Grant No. Y3080359)+1 种基金National Key Programs for Transgenic Crops (Grant Nos. 2008ZX08009-001 and 2009ZX08009-076B)Zhejiang Normal University Innovative Research Team Program, China
文摘Plant calcineurin B-like （CBL） proteins have been proposed as important Ca2＋ sensors and specifically interact with CBL-interacting protein kinases （CIPKs） in plant-specific calcium signaling. Here, we identified and isolated 15 CIPK genes in a japonica rice variety Nipponbare based on the predicted sequences of rice CIPK gene family. Gene structure analysis showed that these 15 genes were divided into intron-less and intron-rich groups, and OsCIPK3 and OsCIPK24 exhibited alternative splicing in their mature process. The phylogenetic analyses indicated that rice CIPKs shared an ancestor with Arabidopsis and poplar CIPKs. Analyses of gene expression showed that these OsCIPK genes were differentially induced by biotic stresses such as bacterial blight and abiotic stresses （heavy metal such as Hg2＋, high salinity, cold and ABA）. Interestingly, five OsCIPK genes, OsCIPK1, 2, 10, 11 and 12, were transcriptionally up-regulated after bacterial blight infection whereas four OsCIPK genes, OsCIPK2, 10, 11 and 14, were induced by all treatments, indicating that some of OsCIPK genes are involved in multiple stress response pathways in plants. Our finding suggests that CIPKs play a key role in both biotic and abiotic stress responses.
基金funded by the following grants:the Natural Science Foundation of Shandong Province of China(ZR2021MC169)the Cooperation Project of University and Local Enterprise in Yantai of Shandong Province(2021XDRHXMPT09).
文摘Calcium(Ca^(2+))plays a pivotal role in various signal transduction pathways.Calcineurin B-like proteins(CBLs)are a unique group of Ca^(2+)sensors that decode Ca^(2+)signals by activating the plant specific protein kinase known as the CBL-interacting protein kinase(CIPK).In plants,the CBL-CIPK signaling network regulates multiple signals in response to different extracellular cues including abiotic stress.However,the genome wide annotation and expression patterns of CBLs and CIPKs in woody cutting flower plants are still unclear.In this study,a total number of 7 CBLs(RcCBLs)and 17 CIPKs(RcCIPKs)genes,divided into four and five subfamilies,respectively,were identified from the rose genome.All RcCBLs possess a classic elongation factor-hand(EF-hand)domain,while all RcCIPKs possess both the classic kinase and NAF domains.Most RcCBLs were predicted to be plasma membrane localized,whereas most RcCIPKs were predicted to be cytoplasmic localized.Synteny analysis showed that one RcCBL gene pair and five RcCIPK gene pairs have gone through whole genome duplication events.Promoter cis-element prediction assays indicated that RcCBLs and RcCIPKs could function in different abiotic stress responses in rose plants.Further quantitative real-time PCR analysis demonstrated that RcCBLs and RcCIPKs were expressed in different organs with overlapped but distinct patterns in response to various abiotic stresses.The findings in this work will provide fundamental information and gene resources for further functional research on RcCBLs and RcCIPKs.
基金This research was funded by a grant from the National Natural Science Foundation of China(31770289 to C.W.)Northwest A&F University(Z111021604 to C.W.)+3 种基金the National Natural Science Foundation of China(31900236 to Z.Z.)supported in part by the open funds of China Postdoctoral Science Foundation(2018M643740 to Z.Z.)the Natural Science Basic Research Plan in Shaanxi Province of China(program no.2019JQ-150)the State Key Laboratory of Plant Physiology and Biochemistry(SKLPPBKF2101 to C.W.).
文摘Manganese(Mn)is an essential micronutrient for all living organisms.However,excess Mn supply that can occur in acid or waterlogged soils has toxic effects on plant physiology and development.Although a variety of Mn transporter families have been characterized,we have only a rudimentary understanding of how these transporters are regulated to uphold and adjust Mn homeostasis in plants.Here,we demonstrate that two calcineurin-B-like proteins,CBL2/3,and their interacting kinases,CIPK3/9/26,are key regulators of plant Mn homeostasis.Arabidopsis mutants lacking CBL2 and 3 or their interacting protein kinases CIPK3/9/26 exhibit remarkably high Mn tolerance.Intriguingly,CIPK3/9/26 interact with and phosphorylate the tonoplast-localized Mn and iron(Fe)transporter MTP8 primarily at Ser35,which is conserved among MTP8 proteins from various species.Mn transport complementation assays in yeast combined with multiple physiological assays indicate that CBL-CIPK-mediated phosphorylation of MTP8 negatively regulates its transport activity from the cytoplasm to the vacuole.Moreover,we show that sequential phosphorylation of MTP8,initially at Ser31/32 by the calcium-dependent protein kinase CPK5 and subsequently at Ser35 by CIPK26,provides an activation/deactivation fine-tuning mechanism for differential regulation of Mn transport.Collectively,our findings define a two-tiered calcium-controlled mechanism for dynamic regulation of Mn homeostasis under conditions of fluctuating Mn supply.
