Glucosinolates(GSLs)are a class of sulfur-and nitrogen-containing and amino acid-derived important secondary metabolites,which mainly present in plants of Brassicaceae family,including Brassica crops,such as broccoli,...Glucosinolates(GSLs)are a class of sulfur-and nitrogen-containing and amino acid-derived important secondary metabolites,which mainly present in plants of Brassicaceae family,including Brassica crops,such as broccoli,cabbage,and oilseed rape.The bioactive GSL meta bolites confer benefits to plant defense,human health,and the unique flavor of some Brassica crops.However,certain GSL profiles have adverse effects and are known as anti-nutritional factors.This has attracted mounting attempts to increase beneficial GSLs and reduce detrimental ones in the most commonly consumed Brassica crops.We provide a comprehensive overview of metabolic engineering applied in Brassica crops to achieve this purpose,including modulation of GSL biosynthesis,ablation of GSL hydrolysis,inhibition of GSL transport processes,and redirection of meta bolic flux to GSL.Moreover,advances in omics approaches,i.e,genomics,transcriptome,and metabolome,applied in the elucidation of GSL metabolism in Brassica crops,as well as promising and potential genome-editing technologies are also discussed.展开更多
Clubroot disease is a severe threat to Brassica crops globally,particularly in western Canada.Genetic resistance,achieved through pyramiding clubroot resistance(CR)genes with different modes of action,is the most impo...Clubroot disease is a severe threat to Brassica crops globally,particularly in western Canada.Genetic resistance,achieved through pyramiding clubroot resistance(CR)genes with different modes of action,is the most important strategy for managing the disease.However,studies on the CR gene functions are quite limited.In this study,we have conducted investigations into the temporal,structural,and interacting features of a newly cloned CR gene,Rcr1,using CRISPR/Cas9 technology.For temporal functionality,we developed a novel CRISPR/Cas9-based binary vector,pHHIGR-Hsp18.2,to deliver Rcr1 into a susceptible canola line(DH12075)and observed that early expression of Rcr1 is critical for conferring resistance.For structural functionality,several independent mutations in specific domains of Rcr1 resulted in loss-offunction,highlighting their importance for CR phenotype.In the study of the interacting features of Rcr1,a cysteine protease gene and its homologous allele in canola were successfully disrupted via CRISPR/Cas9 as an interacting component with Rcr1 protein,resulting in the conversion from clubroot resistant to susceptible in plants carrying intact Rcr1.These results indicated an indispensable role of these two cysteine proteases in Rcr1-mediated resistance response.This study,the first of its kind,provides valuable insights into the functionality of Rcr1.Further,the new vector p HHIGR-Hsp18.2 demonstrated an inducible feature on the removal of add-on traits,which should be useful for functional genomics and other similar research in brassica crops.展开更多
基金Zhejiang Provincial Ten-thousand Program for Leading Talents of Science and Technology Innovation(2018R52026)National Natural Science Foundation of China(Nos.31800252,32072586 and 31500247)Natural Science Foundation of Zhejiang Province(No.LY21C020002).
文摘Glucosinolates(GSLs)are a class of sulfur-and nitrogen-containing and amino acid-derived important secondary metabolites,which mainly present in plants of Brassicaceae family,including Brassica crops,such as broccoli,cabbage,and oilseed rape.The bioactive GSL meta bolites confer benefits to plant defense,human health,and the unique flavor of some Brassica crops.However,certain GSL profiles have adverse effects and are known as anti-nutritional factors.This has attracted mounting attempts to increase beneficial GSLs and reduce detrimental ones in the most commonly consumed Brassica crops.We provide a comprehensive overview of metabolic engineering applied in Brassica crops to achieve this purpose,including modulation of GSL biosynthesis,ablation of GSL hydrolysis,inhibition of GSL transport processes,and redirection of meta bolic flux to GSL.Moreover,advances in omics approaches,i.e,genomics,transcriptome,and metabolome,applied in the elucidation of GSL metabolism in Brassica crops,as well as promising and potential genome-editing technologies are also discussed.
基金supported by the Genomics Initiative of Agriculture and Agri-Food Canada。
文摘Clubroot disease is a severe threat to Brassica crops globally,particularly in western Canada.Genetic resistance,achieved through pyramiding clubroot resistance(CR)genes with different modes of action,is the most important strategy for managing the disease.However,studies on the CR gene functions are quite limited.In this study,we have conducted investigations into the temporal,structural,and interacting features of a newly cloned CR gene,Rcr1,using CRISPR/Cas9 technology.For temporal functionality,we developed a novel CRISPR/Cas9-based binary vector,pHHIGR-Hsp18.2,to deliver Rcr1 into a susceptible canola line(DH12075)and observed that early expression of Rcr1 is critical for conferring resistance.For structural functionality,several independent mutations in specific domains of Rcr1 resulted in loss-offunction,highlighting their importance for CR phenotype.In the study of the interacting features of Rcr1,a cysteine protease gene and its homologous allele in canola were successfully disrupted via CRISPR/Cas9 as an interacting component with Rcr1 protein,resulting in the conversion from clubroot resistant to susceptible in plants carrying intact Rcr1.These results indicated an indispensable role of these two cysteine proteases in Rcr1-mediated resistance response.This study,the first of its kind,provides valuable insights into the functionality of Rcr1.Further,the new vector p HHIGR-Hsp18.2 demonstrated an inducible feature on the removal of add-on traits,which should be useful for functional genomics and other similar research in brassica crops.
