Soybean mosaic virus(SMV),an RNA virus,is the most common and destructive pathogenic virus in soybean fields.The newly developed CRISPR/Cas immune system has provided a novel strategy for improving plant resistance to...Soybean mosaic virus(SMV),an RNA virus,is the most common and destructive pathogenic virus in soybean fields.The newly developed CRISPR/Cas immune system has provided a novel strategy for improving plant resistance to viruses;hence,this study aimed to engineer SMV resistance in soybean using this system.Specifically,multiple sgRNAs were designed to target positive-and/or negative-sense strands of the SMV HC-Pro gene.Subsequently,the corresponding CRISPR/CasRx vectors were constructed and transformed into soybeans.After inoculation with SMV,39.02%,35.77%,and 18.70%of T_(1)plants were confirmed to be highly resistant(HR),resistant(R),and mildly resistant(MR)to SMV,respectively,whereas only 6.50%were identified as susceptible(S).Additionally,qRT-PCR and DAS-ELISA showed that,both at 15 and 30 d post-inoculation(dpi),SMV accumulation significantly decreased or was even undetectable in HR and R plants,followed by MR and S plants.Additionally,the expression level of the CasRx gene varied in almost all T_(1)plants with different resistance level,both at 15 and 30 dpi.Furthermore,when SMV resistance was evaluated in the T_(2)generation,the results were similar to those recorded for the T_(1)generation.These findings provide new insights into the application of the CRISPR/CasRx system for soybean improvement and offer a promising alternative strategy for breeding for resistance to biotic stress that will contribute to the development of SMV-immune soybean germplasm to accelerate progress towards greater soybean crop productivity.展开更多
A novel geminivirus,soybean stay-green associated virus(SoSGV),was previously shown to cause soybean delayed senescence and is associated with the incidence of soybean stay-green syndrome.The modes of SoSGV transmissi...A novel geminivirus,soybean stay-green associated virus(SoSGV),was previously shown to cause soybean delayed senescence and is associated with the incidence of soybean stay-green syndrome.The modes of SoSGV transmission were not yet known.We captured insects belonging to 24 distinct species in a soybean field with the SoSGV outbreak and detected the presence of SoSGV only in leafhoppers and bean bugs(Riptortus pedestris).Caged feeding experiments using captured leafhoppers and bean bugs from soybean fields showed that leafhoppers,but not bean bugs,are vectors transmitting SoSGV.The common brown leafhopper(Orosius orientalis)is identified as the dominant leafhopper species and can establish colonies feeding on soybean plants in experimental conditions.An investigation of SoSGV defective DNA revealed that soybean genomic DNA fragments could be inserted into the SoSGV genome,while sequences from wild soybean,red bean,and cowpea were also identified.We further showed that the common brown leafhopper could transmit SoSGV to wild soybean and red bean plants,emphasizing a vector’s role of the leafhopper in the transmission of SoSGV in the field.展开更多
基金supported by grants from National Natural Science Foundation of China(32001571)R&D Program of Beijing Municipal Education Commission(KM202212448003,KM202312448004)+4 种基金Science and Technology Innovation Project of Beijing Vocational College of Agriculture(XY-YF-22-02)Zhongshan Biological Breeding Laboratory(ZSBBL-KY2023-03)China Agriculture Research System of MOF and MARA(CARS-04)Jiangsu Collaborative Innovation Center for Modern Crop Production(JCICMCP)Collaborative Innovation Center for Modern Crop Production co-sponsored by Province and Ministry(CIC-MCP).
文摘Soybean mosaic virus(SMV),an RNA virus,is the most common and destructive pathogenic virus in soybean fields.The newly developed CRISPR/Cas immune system has provided a novel strategy for improving plant resistance to viruses;hence,this study aimed to engineer SMV resistance in soybean using this system.Specifically,multiple sgRNAs were designed to target positive-and/or negative-sense strands of the SMV HC-Pro gene.Subsequently,the corresponding CRISPR/CasRx vectors were constructed and transformed into soybeans.After inoculation with SMV,39.02%,35.77%,and 18.70%of T_(1)plants were confirmed to be highly resistant(HR),resistant(R),and mildly resistant(MR)to SMV,respectively,whereas only 6.50%were identified as susceptible(S).Additionally,qRT-PCR and DAS-ELISA showed that,both at 15 and 30 d post-inoculation(dpi),SMV accumulation significantly decreased or was even undetectable in HR and R plants,followed by MR and S plants.Additionally,the expression level of the CasRx gene varied in almost all T_(1)plants with different resistance level,both at 15 and 30 dpi.Furthermore,when SMV resistance was evaluated in the T_(2)generation,the results were similar to those recorded for the T_(1)generation.These findings provide new insights into the application of the CRISPR/CasRx system for soybean improvement and offer a promising alternative strategy for breeding for resistance to biotic stress that will contribute to the development of SMV-immune soybean germplasm to accelerate progress towards greater soybean crop productivity.
基金supported by the National Natural Science Foundation of China(31770164)the Jiangsu Province’s Innovation Program(JSSCTD202142)+1 种基金the China Agriculture Research System of MOF and MARA(No.CARS-04)the Jiangsu Collaborative Innovation Center for Modern Crop Production(JCICMCP),and the Collaborative Innovation Center for Modern Crop Production co-sponsored by Province and Ministry(CIC-MCP).
文摘A novel geminivirus,soybean stay-green associated virus(SoSGV),was previously shown to cause soybean delayed senescence and is associated with the incidence of soybean stay-green syndrome.The modes of SoSGV transmission were not yet known.We captured insects belonging to 24 distinct species in a soybean field with the SoSGV outbreak and detected the presence of SoSGV only in leafhoppers and bean bugs(Riptortus pedestris).Caged feeding experiments using captured leafhoppers and bean bugs from soybean fields showed that leafhoppers,but not bean bugs,are vectors transmitting SoSGV.The common brown leafhopper(Orosius orientalis)is identified as the dominant leafhopper species and can establish colonies feeding on soybean plants in experimental conditions.An investigation of SoSGV defective DNA revealed that soybean genomic DNA fragments could be inserted into the SoSGV genome,while sequences from wild soybean,red bean,and cowpea were also identified.We further showed that the common brown leafhopper could transmit SoSGV to wild soybean and red bean plants,emphasizing a vector’s role of the leafhopper in the transmission of SoSGV in the field.
