Genome editing approaches,particularly the CRISPR/Cas9 technology,are becoming state-of-the-art for trait development in numerous breeding programs.Significant advances in improving plant traits are enabled by this in...Genome editing approaches,particularly the CRISPR/Cas9 technology,are becoming state-of-the-art for trait development in numerous breeding programs.Significant advances in improving plant traits are enabled by this influential tool,especially for disease resistance,compared to traditional breeding.One of the potyviruses,the turnip mosaic virus(TuMV),is the most widespread and damaging virus that infects Brassica spp.worldwide.We generated the targeted mutation at the eIF(iso)4E gene in the TuMV-susceptible cultivar“Seoul”using CRISPR/Cas9 to develop TuMV-resistant Chinese cabbage.We detected several heritable indel mutations in the edited T0 plants and developed T1 through generational progression.It was indicated in the sequence analysis of the eIF(iso)4E-edited T1 plants that the mutations were transferred to succeeding generations.These edited T1 plants conferred resistance to TuMV.It was shown with ELISA analysis the lack of accumulation of viral particles.Furthermore,we found a strong negative correlation(r=-0.938)between TuMV resistance and the genome editing frequency of eIF(iso)4E.Consequently,it was revealed in this study that CRISPR/Cas9 technique can expedite the breeding process to improve traits in Chinese cabbage plants.展开更多
Turnip mosaic virus(TuMV)constitutes one of the primary diseases affecting Brassica rapa,severely impacting its production and resulting in crop failures in various regions worldwide.Recent research has demonstrated t...Turnip mosaic virus(TuMV)constitutes one of the primary diseases affecting Brassica rapa,severely impacting its production and resulting in crop failures in various regions worldwide.Recent research has demonstrated the significance of plant translation initiation factors,specifically the eIF4E and eIF4G family genes,as essential recessive disease resistance genes.In our study,we conducted evolutionary and gene expression studies,leading us to identify e IF(iso)4E.c as a potential TuMV-resistant gene.Leveraging CRISPR/Cas9 technology,we obtained mutant B.rapa plants with edited eIF(iso)4E.c gene.We confirmed eIF(iso)4E.c confers resistance against TuMV through phenotypic observations and virus content evaluations.Furthermore,we employed ribosome profiling assays on eif(iso)4e.c mutant seedlings to unravel the translation landscape in response to TuMV.Interestingly,we observed a moderate correlation between the fold changes in gene expression at the transcriptional and translational levels(R^(2)=0.729).Comparative analysis of ribosome profiling and RNA-seq data revealed that plant-pathogen interaction,and MAPK signaling pathway-plant pathways were involved in eIF(iso)4E.c-mediated TuMV resistance.Further analysis revealed that sequence features,coding sequence length,and normalized minimal free energy,influenced the translation efficiency of genes.Our study highlights that the loss of e IF(iso)4E.c can result in a highly intricate translation mechanism,acting synergistically with transcription to confer resistance against TuMV.展开更多
基金This work was carried out with the support of“Cooperative Research Program for Agriculture Science and Technology Development(Project No.PJ01652201)”Rural Development Administration,Republic of Korea.
文摘Genome editing approaches,particularly the CRISPR/Cas9 technology,are becoming state-of-the-art for trait development in numerous breeding programs.Significant advances in improving plant traits are enabled by this influential tool,especially for disease resistance,compared to traditional breeding.One of the potyviruses,the turnip mosaic virus(TuMV),is the most widespread and damaging virus that infects Brassica spp.worldwide.We generated the targeted mutation at the eIF(iso)4E gene in the TuMV-susceptible cultivar“Seoul”using CRISPR/Cas9 to develop TuMV-resistant Chinese cabbage.We detected several heritable indel mutations in the edited T0 plants and developed T1 through generational progression.It was indicated in the sequence analysis of the eIF(iso)4E-edited T1 plants that the mutations were transferred to succeeding generations.These edited T1 plants conferred resistance to TuMV.It was shown with ELISA analysis the lack of accumulation of viral particles.Furthermore,we found a strong negative correlation(r=-0.938)between TuMV resistance and the genome editing frequency of eIF(iso)4E.Consequently,it was revealed in this study that CRISPR/Cas9 technique can expedite the breeding process to improve traits in Chinese cabbage plants.
基金supported by grants from the Scientist Training Program of BAAFS (Grant No.JKZX202406)the Innovation and Capacity-Building Project of BAAFS (Grant No.KJCX20230221)+2 种基金Collaborative innovation program of the Beijing Vegetable Research Center (Grant No.XTCX202302)the National Natural Science Foundation of China (Grant No.32072567)the China Agriculture Research System of MOF and MARA (Grant No.CARS-A03)。
文摘Turnip mosaic virus(TuMV)constitutes one of the primary diseases affecting Brassica rapa,severely impacting its production and resulting in crop failures in various regions worldwide.Recent research has demonstrated the significance of plant translation initiation factors,specifically the eIF4E and eIF4G family genes,as essential recessive disease resistance genes.In our study,we conducted evolutionary and gene expression studies,leading us to identify e IF(iso)4E.c as a potential TuMV-resistant gene.Leveraging CRISPR/Cas9 technology,we obtained mutant B.rapa plants with edited eIF(iso)4E.c gene.We confirmed eIF(iso)4E.c confers resistance against TuMV through phenotypic observations and virus content evaluations.Furthermore,we employed ribosome profiling assays on eif(iso)4e.c mutant seedlings to unravel the translation landscape in response to TuMV.Interestingly,we observed a moderate correlation between the fold changes in gene expression at the transcriptional and translational levels(R^(2)=0.729).Comparative analysis of ribosome profiling and RNA-seq data revealed that plant-pathogen interaction,and MAPK signaling pathway-plant pathways were involved in eIF(iso)4E.c-mediated TuMV resistance.Further analysis revealed that sequence features,coding sequence length,and normalized minimal free energy,influenced the translation efficiency of genes.Our study highlights that the loss of e IF(iso)4E.c can result in a highly intricate translation mechanism,acting synergistically with transcription to confer resistance against TuMV.
