The imperative aspect of the CRISPR/Cas9 system is a short stretch of 20 nucleotides of gRNA that control the overall specificity.Due to the small size,the chance of its multiple occurrences in the genome increases;how...The imperative aspect of the CRISPR/Cas9 system is a short stretch of 20 nucleotides of gRNA that control the overall specificity.Due to the small size,the chance of its multiple occurrences in the genome increases;however,a few mismatches are tolerated by the Cas9 endonuclease activity.An accurate and careful in silico-based off-target prediction while target selection is preferred to address the issue.These predictions are based on a comprehensive set of selectable parameters.Therefore,we investigated the possible off-target prediction and their screening in StERF3 gene-edited potato plants while developing StERF3-loss-of-function mutants using CRISPR/Cas9 approach.The 201 off-targets for the selected targets of the StERF3 gene were predicted,and 79 werefiltered as potential off-targets.Of these 79,twenty-five off-targets showed scores with defined cut-off values<0.5 and were analyzed in Sterf3-edited potato plants compared to wild-type plants.No off-targeting was found to have occurred in edited plants.展开更多
Stripe rust is a continuous threat to wheat crop all over the world.It causes considerable yield losses in wheat crop every year.Continuous deployment of adult plant resistance(APR)genes in newly developing wheat cult...Stripe rust is a continuous threat to wheat crop all over the world.It causes considerable yield losses in wheat crop every year.Continuous deployment of adult plant resistance(APR)genes in newly developing wheat cultivars is the most judicious strategy to combat this disease.Herein,we dissected the genetics underpinning stripe rust resistance in Pakistani wheat germplasm.An association panel of 94 spring wheat genotypes was phenotyped for two years to score the infestation of stripe rust on each accession and was scanned with 203 polymorphic SSRs.Based on D’measure,linkage disequilibrium(LD)exhibited between loci distant up to 45 c M.Marker-trait associations(MTAs)were determined using mixed linear model(MLM).Total 31 quantitative trait loci(QTLs)were observed on all 21 wheat chromosomes.Twelve QTLs were newly discovered as well as 19 QTLs and 35 previously reported Yr genes were validated in Pakistani wheat germplasm.The major QTLs were QYr.uaf.2 AL and QYr.uaf.3 BS(PVE,11.9%).Dissection of genes from the newly observed QTLs can provide new APR genes to improve genetic resources for APR resistance in wheat crop.展开更多
文摘The imperative aspect of the CRISPR/Cas9 system is a short stretch of 20 nucleotides of gRNA that control the overall specificity.Due to the small size,the chance of its multiple occurrences in the genome increases;however,a few mismatches are tolerated by the Cas9 endonuclease activity.An accurate and careful in silico-based off-target prediction while target selection is preferred to address the issue.These predictions are based on a comprehensive set of selectable parameters.Therefore,we investigated the possible off-target prediction and their screening in StERF3 gene-edited potato plants while developing StERF3-loss-of-function mutants using CRISPR/Cas9 approach.The 201 off-targets for the selected targets of the StERF3 gene were predicted,and 79 werefiltered as potential off-targets.Of these 79,twenty-five off-targets showed scores with defined cut-off values<0.5 and were analyzed in Sterf3-edited potato plants compared to wild-type plants.No off-targeting was found to have occurred in edited plants.
基金Higher Education Commission(HEC),Islamabad,Pakistan for providing for funding(21179/SRGP/R&D/HEC/2014)the International Research Support Initiative Program(IRSIP)Scholarship for this research work。
文摘Stripe rust is a continuous threat to wheat crop all over the world.It causes considerable yield losses in wheat crop every year.Continuous deployment of adult plant resistance(APR)genes in newly developing wheat cultivars is the most judicious strategy to combat this disease.Herein,we dissected the genetics underpinning stripe rust resistance in Pakistani wheat germplasm.An association panel of 94 spring wheat genotypes was phenotyped for two years to score the infestation of stripe rust on each accession and was scanned with 203 polymorphic SSRs.Based on D’measure,linkage disequilibrium(LD)exhibited between loci distant up to 45 c M.Marker-trait associations(MTAs)were determined using mixed linear model(MLM).Total 31 quantitative trait loci(QTLs)were observed on all 21 wheat chromosomes.Twelve QTLs were newly discovered as well as 19 QTLs and 35 previously reported Yr genes were validated in Pakistani wheat germplasm.The major QTLs were QYr.uaf.2 AL and QYr.uaf.3 BS(PVE,11.9%).Dissection of genes from the newly observed QTLs can provide new APR genes to improve genetic resources for APR resistance in wheat crop.