OsMAPK6 plays a critical role in regulating rice growth,development,and stress responses.However,the embryonic lethality associated with loss-of-function mutations prevents the generation of homozygous mutant seeds,si...OsMAPK6 plays a critical role in regulating rice growth,development,and stress responses.However,the embryonic lethality associated with loss-of-function mutations prevents the generation of homozygous mutant seeds,significantly hindering functional studies of this gene.Although the weak mutant dsg1 has offered valuable insights into OsMAPK6 function,its extremely low seed-setting rate limits its use for detailed genetic analysis.Here,we employed prime editing to perform precise multi-site modifications at the C-terminus of OsMAPK6,generating a series of osmapk6 mutants with truncated proteins of varying lengths.Among these,the osmapk6(379)and osmapk6(383)mutants exhibited phenotypic defects similar to dsg1,while osmapk6(386)showed a significantly improved seed-setting rate despite persistent developmental defects.Through phenotypic characterization and protein functional analysis,we further clarified how different C-terminal deletion lengths affect plant growth,development,stress responses,and OsMAPK6 protein function.In summary,this study elucidates the importance of the OsMAPK6 C-terminus in rice biology and provides a fertile weak mutant with enhanced seed production,offering a valuable genetic resource for future research on OsMAPK6.展开更多
Grain size is one of the most important agronomic traits controlling grain yield. Development of novel germplasm with large grains would be beneficial for crop improvement. We report the genetic identification and fun...Grain size is one of the most important agronomic traits controlling grain yield. Development of novel germplasm with large grains would be beneficial for crop improvement. We report the genetic identification and functional analysis of the LONG GRAIN 6(LOG6) gene, which is identical to MITOGENACTIVATED PROTEIN KINASE 6(OsMAPK6), affecting grain length of rice. Map-based cloning revealed that the long-grain phenotype of log6-D results from a glutamine(E) to lysine(K) mutation in the conserved TEY motif of OsMAPK6. In near-isogenic lines(NILs), the log6-D allele increased grain length and grain yield of Guichao 2(GC2), Teqing(TQ), and 93–11. Sequence analysis revealed 10 OsMAPK6 haplotypes,with xian(indica) and geng(japonica) harboring different haplotypes. Our findings shed light on the function of MAPKs and offer a novel dominant allele for improving the grain yield of rice.展开更多
Leaf angle is a major agronomic trait that determines plant architecture,which directly affects rice planting density,photosynthetic efficiency,and yield.The plant phytohormones brassinosteroids(BRs)and the MAPK signa...Leaf angle is a major agronomic trait that determines plant architecture,which directly affects rice planting density,photosynthetic efficiency,and yield.The plant phytohormones brassinosteroids(BRs)and the MAPK signaling cascade are known to play crucial roles in regulating leaf angle,but the underlying molecular mechanisms are not fully understood.Here,we report a rice WRKY family transcription factor gene,OsWRKY72,which positively regulates leaf angle by affecting lamina joint development and BR signaling.Phenotypic analysis showed that oswrky72 mutants have smaller leaf angles and exhibit insensitivity to exogenous BRs,whereas OsWRKY72 overexpression lines show enlarged leaf angles and are hypersensitive to exogenous BRs.Histological sections revealed that the change in leaf inclination is due to asymmetric cell proliferation and growth at the lamina joint.Further investigation showed that OsWRKY72 binds directly to the promoter region of BR receptor kinase(OsBRI1),a key gene in theBR signaling pathway,and activates its expression to positively regulate rice BR signaling.In addition,we discovered that OsWRKY72 interacts with and is phosphorylated by OsMAPK6,and this phosphorylation event can enhanceOsWRKY72 activity in promoting OsBRI1 expression.Genetic evidence confirmed that OsMAPK6,OsWRKY72,and OsBRI1 function in a common pathway to regulate leaf angle.Collectively,our findings clarify the critical role of the OsWRKY72 transcription factor in regulating rice leaf angle.These results provide valuable insights into the molecular regulatory networks that govern plant architecture in rice.展开更多
Rice OsLIC encoding a CCCH zinc finger transcription factor plays an important role in immunity.However,the immune signaling pathways that OsLIC-involved and the underlying mechanisms that OsLIC-conferred resistance a...Rice OsLIC encoding a CCCH zinc finger transcription factor plays an important role in immunity.However,the immune signaling pathways that OsLIC-involved and the underlying mechanisms that OsLIC-conferred resistance against pathogens are largely unclear.Here,we show that OsLIC,as a substrate for Os MAPK6,negatively regulates resistance to Xanthomonas oryzae pv.oryzae(Xoo)and X.oryzae pv.oryzicola(Xoc)by directly suppressing OsWRKY30 transcription.Biochemical assays showed that OsLIC bound to OsWRKY30 promoter and suppressed its transcription.Genetic assays confirmed that the osilc knockout mutants and OsWRKY30-overexpressing plants exhibited enhanced resistance to Xoo and Xoc,knocking out OsWRKY30 in the oslic mutants attenuated the resistance against bacterial pathogens.Os MAPK6 physically interacted with and phosphorylated OsLIC leading to decreased OsLIC DNA-binding activity,therefore,overexpression of OsLIC partially suppressed Os MAPK6-mediated rice resistance.In addition,both Os MAPK6-phosphorylated activation of OsLIC and phosphorylation-mimic OsLIC_(5D) had reduced DNA-binding activity towards OsWRKY30 promoter,thereby promoting OsWRKY30 transcription.Collectively,these results reveal that Os MAPK6-mediated phosphorylation of OsLIC positively regulates rice resistance to Xoo and Xoc by modulating OsWRKY30 transcription,suggesting that Os MAPK6-OsLIC-OsWRKY30 module is an immune signaling pathway in response to the bacterial pathogens.展开更多
Grain size and grain weight are important determinants for grain yield.In this study,we identify a novel OsMAPK5–OsWRKY72 module that negatively regulates grain length and grain weight in rice.We found that loss-of-f...Grain size and grain weight are important determinants for grain yield.In this study,we identify a novel OsMAPK5–OsWRKY72 module that negatively regulates grain length and grain weight in rice.We found that loss-of-function of OsMAPK5 leads to larger cell size of the rice spikelet hulls and a significant increase in both grain length and grain weight in an indica variety Minghui 86(MH86).OsMAPK5 interacts with OsMAPKK3/4/5 and OsWRKY72 and phosphorylates OsWRKY72 at T86 and S88.Similar to the osmapk5 MH86 mutants,the oswrky72 knockout MH86 mutants exhibited larger size of spikelet hull cells and increased grain length and grain weight,whereas the OsWRKY72-overexpression MH86 plants showed opposite phenotypes.OsWRKY72 targets the W-box motifs in the promoter of OsARF6,an auxin response factor involved in auxin signaling.Dual-luciferase reporter assays demonstrated that OsWRKY72 activates OsARF6 expression.The activation effect of the phosphorylation-mimicking OsWRKY72T86D/S88D on OsARF6 expression was significantly enhanced,whereas the effects of the OsWRKY72 phosphorylation-null mutants were significantly reduced.In addition,auxin levels in young panicles of the osmapk5 and oswrky72 mutants were significantly higher than that in the wild-type MH86.Collectively,our study uncovered novel connections of the OsMAPKK3/4/5-OsMAPK5-mediated MAPK signaling,OsWRKY72-mediated transcription regulation,and OsARF6-mediated auxin signaling pathways in regulating grain length and grain weight in an indica-type rice,providing promising targets for molecular breeding of rice varieties with high yield and quality.展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.32441024,32572315,and U25A20674)the Youth Innovation Promotion Association of Chinese Academy of Sciences(Grant No.2021229)+1 种基金the Heilongjiang Key Research and Development Program,China(Grant No.2025ZX04B02)the Young Scientist Group Project of Northeast Institute of Geography and Agroecology,Chinese Academy of Sciences(Grant No.2023QNXZ02).
文摘OsMAPK6 plays a critical role in regulating rice growth,development,and stress responses.However,the embryonic lethality associated with loss-of-function mutations prevents the generation of homozygous mutant seeds,significantly hindering functional studies of this gene.Although the weak mutant dsg1 has offered valuable insights into OsMAPK6 function,its extremely low seed-setting rate limits its use for detailed genetic analysis.Here,we employed prime editing to perform precise multi-site modifications at the C-terminus of OsMAPK6,generating a series of osmapk6 mutants with truncated proteins of varying lengths.Among these,the osmapk6(379)and osmapk6(383)mutants exhibited phenotypic defects similar to dsg1,while osmapk6(386)showed a significantly improved seed-setting rate despite persistent developmental defects.Through phenotypic characterization and protein functional analysis,we further clarified how different C-terminal deletion lengths affect plant growth,development,stress responses,and OsMAPK6 protein function.In summary,this study elucidates the importance of the OsMAPK6 C-terminus in rice biology and provides a fertile weak mutant with enhanced seed production,offering a valuable genetic resource for future research on OsMAPK6.
基金supported by the National Natural Science Foundation of China (91935302, 31971870)。
文摘Grain size is one of the most important agronomic traits controlling grain yield. Development of novel germplasm with large grains would be beneficial for crop improvement. We report the genetic identification and functional analysis of the LONG GRAIN 6(LOG6) gene, which is identical to MITOGENACTIVATED PROTEIN KINASE 6(OsMAPK6), affecting grain length of rice. Map-based cloning revealed that the long-grain phenotype of log6-D results from a glutamine(E) to lysine(K) mutation in the conserved TEY motif of OsMAPK6. In near-isogenic lines(NILs), the log6-D allele increased grain length and grain yield of Guichao 2(GC2), Teqing(TQ), and 93–11. Sequence analysis revealed 10 OsMAPK6 haplotypes,with xian(indica) and geng(japonica) harboring different haplotypes. Our findings shed light on the function of MAPKs and offer a novel dominant allele for improving the grain yield of rice.
基金supported by the National Rice Industry Technology Sys-tem of Modern Agriculture for China(grant no.CARS-01-08)the"5511"Collaborative Innovation Project for High-quality Development and Sur-passes of Agriculture between Government of Fujian and Chinese Acad-emy of Agricultural Sciences(grant no.XTCXGC2021001)+4 种基金the Key Pro-gram of Science and Technology in Fujian province,China(grant no.2020NZ08016)the Special Foundation of Non-Profit Research Institutes of Fujian Province(grant no.2020R1023008)the National Natural Science Foundation of China-Youth Science Fund Project(grant no.32201733)the Scientific Research Project of Fujian Academy of Agricultural Sciences(grant no.GJYS202301)the Central Government Guides Local Science and Technology Development Special Project(grant no.2022L3018).
文摘Leaf angle is a major agronomic trait that determines plant architecture,which directly affects rice planting density,photosynthetic efficiency,and yield.The plant phytohormones brassinosteroids(BRs)and the MAPK signaling cascade are known to play crucial roles in regulating leaf angle,but the underlying molecular mechanisms are not fully understood.Here,we report a rice WRKY family transcription factor gene,OsWRKY72,which positively regulates leaf angle by affecting lamina joint development and BR signaling.Phenotypic analysis showed that oswrky72 mutants have smaller leaf angles and exhibit insensitivity to exogenous BRs,whereas OsWRKY72 overexpression lines show enlarged leaf angles and are hypersensitive to exogenous BRs.Histological sections revealed that the change in leaf inclination is due to asymmetric cell proliferation and growth at the lamina joint.Further investigation showed that OsWRKY72 binds directly to the promoter region of BR receptor kinase(OsBRI1),a key gene in theBR signaling pathway,and activates its expression to positively regulate rice BR signaling.In addition,we discovered that OsWRKY72 interacts with and is phosphorylated by OsMAPK6,and this phosphorylation event can enhanceOsWRKY72 activity in promoting OsBRI1 expression.Genetic evidence confirmed that OsMAPK6,OsWRKY72,and OsBRI1 function in a common pathway to regulate leaf angle.Collectively,our findings clarify the critical role of the OsWRKY72 transcription factor in regulating rice leaf angle.These results provide valuable insights into the molecular regulatory networks that govern plant architecture in rice.
基金supported by grants from the National Natural Science Foundation of China(31821005,31871946,32172421)the National Science Foundation of Hubei Province(2020CFA058)。
文摘Rice OsLIC encoding a CCCH zinc finger transcription factor plays an important role in immunity.However,the immune signaling pathways that OsLIC-involved and the underlying mechanisms that OsLIC-conferred resistance against pathogens are largely unclear.Here,we show that OsLIC,as a substrate for Os MAPK6,negatively regulates resistance to Xanthomonas oryzae pv.oryzae(Xoo)and X.oryzae pv.oryzicola(Xoc)by directly suppressing OsWRKY30 transcription.Biochemical assays showed that OsLIC bound to OsWRKY30 promoter and suppressed its transcription.Genetic assays confirmed that the osilc knockout mutants and OsWRKY30-overexpressing plants exhibited enhanced resistance to Xoo and Xoc,knocking out OsWRKY30 in the oslic mutants attenuated the resistance against bacterial pathogens.Os MAPK6 physically interacted with and phosphorylated OsLIC leading to decreased OsLIC DNA-binding activity,therefore,overexpression of OsLIC partially suppressed Os MAPK6-mediated rice resistance.In addition,both Os MAPK6-phosphorylated activation of OsLIC and phosphorylation-mimic OsLIC_(5D) had reduced DNA-binding activity towards OsWRKY30 promoter,thereby promoting OsWRKY30 transcription.Collectively,these results reveal that Os MAPK6-mediated phosphorylation of OsLIC positively regulates rice resistance to Xoo and Xoc by modulating OsWRKY30 transcription,suggesting that Os MAPK6-OsLIC-OsWRKY30 module is an immune signaling pathway in response to the bacterial pathogens.
基金funded by grants from Fujian Provincial Science and Technology Key Project(2022NZ030014)National Natural Science Foundation of China(32071941)+1 种基金Natural Science Foundation of Fujian Province(2023J011418)Minjiang University(MJY22023).
文摘Grain size and grain weight are important determinants for grain yield.In this study,we identify a novel OsMAPK5–OsWRKY72 module that negatively regulates grain length and grain weight in rice.We found that loss-of-function of OsMAPK5 leads to larger cell size of the rice spikelet hulls and a significant increase in both grain length and grain weight in an indica variety Minghui 86(MH86).OsMAPK5 interacts with OsMAPKK3/4/5 and OsWRKY72 and phosphorylates OsWRKY72 at T86 and S88.Similar to the osmapk5 MH86 mutants,the oswrky72 knockout MH86 mutants exhibited larger size of spikelet hull cells and increased grain length and grain weight,whereas the OsWRKY72-overexpression MH86 plants showed opposite phenotypes.OsWRKY72 targets the W-box motifs in the promoter of OsARF6,an auxin response factor involved in auxin signaling.Dual-luciferase reporter assays demonstrated that OsWRKY72 activates OsARF6 expression.The activation effect of the phosphorylation-mimicking OsWRKY72T86D/S88D on OsARF6 expression was significantly enhanced,whereas the effects of the OsWRKY72 phosphorylation-null mutants were significantly reduced.In addition,auxin levels in young panicles of the osmapk5 and oswrky72 mutants were significantly higher than that in the wild-type MH86.Collectively,our study uncovered novel connections of the OsMAPKK3/4/5-OsMAPK5-mediated MAPK signaling,OsWRKY72-mediated transcription regulation,and OsARF6-mediated auxin signaling pathways in regulating grain length and grain weight in an indica-type rice,providing promising targets for molecular breeding of rice varieties with high yield and quality.
