Cordyceps militaris(CM)is widely distributed in humid temperate and tropical forests.Commercially available CM is mainly dried fruiting bodies and their further processed products.In the chinese market,these products ...Cordyceps militaris(CM)is widely distributed in humid temperate and tropical forests.Commercially available CM is mainly dried fruiting bodies and their further processed products.In the chinese market,these products are mainly stored at room temperature without vacuum sealing after air drying.In this study,ultra-high performance liquid chromatography-high-resolution mass spectrometry,gas chromatography-mass spectrometry,and microbial community abundance analysis were applied to analyze the changes of major metabolites and microbial communities in CM dried products and explore the changes in quality during storage at room temperature.A total of 856 non-volatile and 431 volatile substances were detected,of which 195 non-volatile and 85 volatile substances showed significant differences.By analyzing the microbial community relative abundance,the highest relative abundance was observed in the second month,which was then followed by a decline.These results demonstrated that the quality,odor,and nutrient content of CM changed considerably after storage from one to two months.These findings can provide new insights for exploring the optimal consumption time for the best flavor and health function of CM dried products in the future.展开更多
Claviceps purpurea produces many pharmacologically important ergot alkaloids(EAS),which are widely used to treat migraine and hypertension and to aid childbirth.Although an EAS biosynthetic cluster of C.purpurea has b...Claviceps purpurea produces many pharmacologically important ergot alkaloids(EAS),which are widely used to treat migraine and hypertension and to aid childbirth.Although an EAS biosynthetic cluster of C.purpurea has been discovered more than 20 years ago,the complete biosynthetic pathway of EAS has not been fully characterized until now.The main obstacle to elucidating this pathway and strain modification is the lack of efficient genome-editing tools for C.purpurea.The conventional gene manipulation method for C.purpurea relies on homologous recombination(HR),although the efficiency of HR in C.purpurea is very low(~1-5%).Consequently,the disruption of target genes is laborious and time-consuming.Although CRISPR/Cas9 genome-editing methods based on in vivo Cas9 expression and gRNA transcription have been reported recently,their gene-disruption efficiency is still very low.Here,we developed an efficient genome-editing system in C.purpurea based on in vitro assembled CRISPR/Cas9 gRNA ribonucleoprotein complexes.As proof of principle,three target genes were efficiently knocked out using this CRISPR/Cas9 ribonucleoprotein complex-mediated HR system,with editing efficiencies ranging from 50%to 100%.Inactivation of the three genes,which are closely related to uridine biosynthesis(ura5),hypha morphology(rac),and EAS production(easA),resulted in a uridine auxotrophic mutant,a mutant with a drastically different phenotype in axenic culture,and a mutant that did not produce EAS,respectively.Our ribonucleoprotein-based genome-editing system has a great advantage over conventional and in vivo CRISPR/Cas9 methods for genome editing in C.purpurea,which will greatly facilitate elucidation of the EAS biosynthetic pathway and other future basic and applied research on C.purpurea.展开更多
Fungi are widely distributed in the environment,and some are beneficial to medicine,industry,agriculture,and food.For example,Penicillium chrysogenum,Acremonium chrysogenum,and Aspergillus terreus can synthesizeβ-lac...Fungi are widely distributed in the environment,and some are beneficial to medicine,industry,agriculture,and food.For example,Penicillium chrysogenum,Acremonium chrysogenum,and Aspergillus terreus can synthesizeβ-lactams,which can be used as drugs[1];Aspergillus niger and Trichoderma reesei are well-known industrial enzyme producers[2].展开更多
基金supported by the National Key R&D Program of china(2023YFF1000800)the Excellent team Plan of Shanghai academy of agricultural Sciences((2022)014).
文摘Cordyceps militaris(CM)is widely distributed in humid temperate and tropical forests.Commercially available CM is mainly dried fruiting bodies and their further processed products.In the chinese market,these products are mainly stored at room temperature without vacuum sealing after air drying.In this study,ultra-high performance liquid chromatography-high-resolution mass spectrometry,gas chromatography-mass spectrometry,and microbial community abundance analysis were applied to analyze the changes of major metabolites and microbial communities in CM dried products and explore the changes in quality during storage at room temperature.A total of 856 non-volatile and 431 volatile substances were detected,of which 195 non-volatile and 85 volatile substances showed significant differences.By analyzing the microbial community relative abundance,the highest relative abundance was observed in the second month,which was then followed by a decline.These results demonstrated that the quality,odor,and nutrient content of CM changed considerably after storage from one to two months.These findings can provide new insights for exploring the optimal consumption time for the best flavor and health function of CM dried products in the future.
基金This work was financially supported by the National Key Research and Development Program of China(Grant No.2018YFA0900500)the National Natural Science Foundation of China(Nos.31921006,31470201,and,31741003)the Strategic Biological Resources Service Network Plan of the Chinese Academy of Sciences(Grant No.KFJ-BRP-009).
文摘Claviceps purpurea produces many pharmacologically important ergot alkaloids(EAS),which are widely used to treat migraine and hypertension and to aid childbirth.Although an EAS biosynthetic cluster of C.purpurea has been discovered more than 20 years ago,the complete biosynthetic pathway of EAS has not been fully characterized until now.The main obstacle to elucidating this pathway and strain modification is the lack of efficient genome-editing tools for C.purpurea.The conventional gene manipulation method for C.purpurea relies on homologous recombination(HR),although the efficiency of HR in C.purpurea is very low(~1-5%).Consequently,the disruption of target genes is laborious and time-consuming.Although CRISPR/Cas9 genome-editing methods based on in vivo Cas9 expression and gRNA transcription have been reported recently,their gene-disruption efficiency is still very low.Here,we developed an efficient genome-editing system in C.purpurea based on in vitro assembled CRISPR/Cas9 gRNA ribonucleoprotein complexes.As proof of principle,three target genes were efficiently knocked out using this CRISPR/Cas9 ribonucleoprotein complex-mediated HR system,with editing efficiencies ranging from 50%to 100%.Inactivation of the three genes,which are closely related to uridine biosynthesis(ura5),hypha morphology(rac),and EAS production(easA),resulted in a uridine auxotrophic mutant,a mutant with a drastically different phenotype in axenic culture,and a mutant that did not produce EAS,respectively.Our ribonucleoprotein-based genome-editing system has a great advantage over conventional and in vivo CRISPR/Cas9 methods for genome editing in C.purpurea,which will greatly facilitate elucidation of the EAS biosynthetic pathway and other future basic and applied research on C.purpurea.
基金This study was funded by the National Natural Science Foundation of China(No.32111530179).
文摘Fungi are widely distributed in the environment,and some are beneficial to medicine,industry,agriculture,and food.For example,Penicillium chrysogenum,Acremonium chrysogenum,and Aspergillus terreus can synthesizeβ-lactams,which can be used as drugs[1];Aspergillus niger and Trichoderma reesei are well-known industrial enzyme producers[2].
