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
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.
