Source-sink coordination serves as the foundation for improving crop yield.Current research primarily focuses on individual factors,such as increasing the source or expanding the sink,which often leads to disrupted so...Source-sink coordination serves as the foundation for improving crop yield.Current research primarily focuses on individual factors,such as increasing the source or expanding the sink,which often leads to disrupted source-sink balance,causing trade-offs among photosynthesis,yield,and stress response.To address these limitations,we present an integrated synthetic biological framework that synergistically enhances photosynthetic efficiency(source capacity),sink optimization,and abiotic stress tolerance.We developed an editing-overexpression coupling(EOC)vector system enabling simultaneous overexpression of four photosynthesis-enhancing genes(Cyt c6,PsbA,FBPase,OsMGT3),knockout of three yield-limiting genes(GS3,Gn1a,OsAAP5),and self-excision of selection markers,gene-editing modules,and fragment deletion cassettes.Field evaluations of CFMP-gga transgenic lines revealed significant physiological improvements,including 13%–17%increase in photosynthetic rates,improved chlorophyll fluorescence parameters,and increased stomatal conductance.These enhancements translated into remarkable agronomic gains,including 18.7%–22.3%higher grain yield,23.1%–26.1%increased biomass,and improved panicle architecture(increased grain size and grain number per panicle).The engineered lines maintained superior thermotolerance(under 42°C stress)and alkali tolerance(at pH 10)compared to wild-type controls.This study provides a strategy for enhancing crop yield by demonstrating that coordinated multi-gene regulation of source-sink dynamics,coupled with stress resilience engineering,achieves concurrent improvements.展开更多
The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of t...The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sg RNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sg RNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering.展开更多
基金the National Key Research and Development Program of China(2020YFA0907600)National Natural Science Foundation of China(31100869)+1 种基金Central Public-interest Scientific Institutions Basal Research Fund for Zhang Zhiguo(Y2025YY06)the Fundamental Research Funds for Central Nonprofit Scientific Institutions for Lu Tiegang,and Cui Xuean.
文摘Source-sink coordination serves as the foundation for improving crop yield.Current research primarily focuses on individual factors,such as increasing the source or expanding the sink,which often leads to disrupted source-sink balance,causing trade-offs among photosynthesis,yield,and stress response.To address these limitations,we present an integrated synthetic biological framework that synergistically enhances photosynthetic efficiency(source capacity),sink optimization,and abiotic stress tolerance.We developed an editing-overexpression coupling(EOC)vector system enabling simultaneous overexpression of four photosynthesis-enhancing genes(Cyt c6,PsbA,FBPase,OsMGT3),knockout of three yield-limiting genes(GS3,Gn1a,OsAAP5),and self-excision of selection markers,gene-editing modules,and fragment deletion cassettes.Field evaluations of CFMP-gga transgenic lines revealed significant physiological improvements,including 13%–17%increase in photosynthetic rates,improved chlorophyll fluorescence parameters,and increased stomatal conductance.These enhancements translated into remarkable agronomic gains,including 18.7%–22.3%higher grain yield,23.1%–26.1%increased biomass,and improved panicle architecture(increased grain size and grain number per panicle).The engineered lines maintained superior thermotolerance(under 42°C stress)and alkali tolerance(at pH 10)compared to wild-type controls.This study provides a strategy for enhancing crop yield by demonstrating that coordinated multi-gene regulation of source-sink dynamics,coupled with stress resilience engineering,achieves concurrent improvements.
基金supported by the National Natural Science Foundation of China(32001532 and 31860411)the National Key Research and Development Program of China,(2022YFF1000020)+1 种基金Hunan Seed Industry Innovation Project(2021NK1012)the Yunnan Tobacco Company Project(2020530000241009)。
文摘The development and maturation of the CRISPR/Cas genome editing system provides a valuable tool for plant functional genomics and genetic improvement.Currently available genome-editing tools have a limited number of targets,restricting their application in genetic research.In this study,we developed a novel CRISPR/Cas9 plant ultra-multiplex genome editing system consisting of two template vectors,eight donor vectors,four destination vectors,and one primer-design software package.By combining the advantages of Golden Gate cloning to assemble multiple repetitive fragments and Gateway recombination to assemble large fragments and by changing the structure of the amplicons used to assemble sg RNA expression cassettes,the plant ultra-multiplex genome editing system can assemble a single binary vector targeting more than 40 genomic loci.A rice knockout vector containing 49 sg RNA expression cassettes was assembled and a high co-editing efficiency was observed.This plant ultra-multiplex genome editing system advances synthetic biology and plant genetic engineering.
