VIGS(Virus-induced gene silencing),a method for posttranscriptional gene silencing,is an effective technique for investigating the activities of genes in plants.Since there is no report for available VIGS system in St...VIGS(Virus-induced gene silencing),a method for posttranscriptional gene silencing,is an effective technique for investigating the activities of genes in plants.Since there is no report for available VIGS system in Styrax japonicus,the application of a VIGS approach that results in a gene knockdown to study gene function is limited.In this study,we compared the characteristics that could affect the viability of VIGS in S.japonicus,including the acetosyringone(AS)concentration,the Agrobacterium’s optical density and the inoculation method.The stable reference genes of S.japonicus were selected to validate the gene’s knockdown by quantitative PCR.As a result,we successfully constructed 2 VIGS systems based on TRV virus:vacuum with AS concentration of 200μmol·L^(-1)and OD600of 0.5,and friction-osmosis with AS concentration of 200μmol·L^(-1)and OD600of 1.0,which silencing efficiency was 83.33%and 74.19%,respectively.The successfully applied VIGS method provides a rapid and effective reverse gene functional analysis approach in S.japonicus to identify unknown gene functions.展开更多
The sea cucumber Stichopus japonicus is one of the"Eight Treasures of Seafood"and contains a number of bioactive components involved in multiple physiological and pharmacological functions.Proteins and pepti...The sea cucumber Stichopus japonicus is one of the"Eight Treasures of Seafood"and contains a number of bioactive components involved in multiple physiological and pharmacological functions.Proteins and peptides are generally considered to be responsible for these beneficial properties.In this study,a total of 3478 proteins and 17390 peptides were identified in Stichopus japonicus by proteomics methods.Among them,4 proteins were involved in 8 metabolic pathways,especially oxidative phosphorylation and cell senescence.Subsequently,lifespan assay and oxidative stress test were performed to investigate the peptides prepared from sea cucumber protein hydrolyzate using the aging model of Caenorhabditis elegans.The results of the anti-aging experiment demonstrated that high-dose peptides significantly prolonged the lifespan of nematodes(30.50%),and improved their capacity to inhibit oxidative stress.The results provide evidence supporting the development of bioactive proteins and peptides derived from Stichopus japonicus as functional foods and lay the foundation for the research of an anti-aging drug.展开更多
基金supported by the Subject of Key R&D Plan of Shandong Province(Major Scientific and Technological Innovation Project)“Mining and Accurate Identification of Forest Tree Germplasm Resources”(Grant Nos.2021LZGC02303)Science&Technology Specific Projects in Agricultural High-tech Industrial Demonstration Area of the Yellow River Delta(Grant No.022SZX16)。
文摘VIGS(Virus-induced gene silencing),a method for posttranscriptional gene silencing,is an effective technique for investigating the activities of genes in plants.Since there is no report for available VIGS system in Styrax japonicus,the application of a VIGS approach that results in a gene knockdown to study gene function is limited.In this study,we compared the characteristics that could affect the viability of VIGS in S.japonicus,including the acetosyringone(AS)concentration,the Agrobacterium’s optical density and the inoculation method.The stable reference genes of S.japonicus were selected to validate the gene’s knockdown by quantitative PCR.As a result,we successfully constructed 2 VIGS systems based on TRV virus:vacuum with AS concentration of 200μmol·L^(-1)and OD600of 0.5,and friction-osmosis with AS concentration of 200μmol·L^(-1)and OD600of 1.0,which silencing efficiency was 83.33%and 74.19%,respectively.The successfully applied VIGS method provides a rapid and effective reverse gene functional analysis approach in S.japonicus to identify unknown gene functions.
基金Supported by Scientific Research Program of Higher Education Institutions in Hainan Province(Hnky2019-110)Hainan Provincial Natural Science Foundation of China(424QN258)+1 种基金Hainan Provincial Key Point Research and Invention Program(ZDYF2022SHFZ309)Special Scientific Research Project of Hainan Academician Innovation Platform(YSPTZX202132).
文摘The sea cucumber Stichopus japonicus is one of the"Eight Treasures of Seafood"and contains a number of bioactive components involved in multiple physiological and pharmacological functions.Proteins and peptides are generally considered to be responsible for these beneficial properties.In this study,a total of 3478 proteins and 17390 peptides were identified in Stichopus japonicus by proteomics methods.Among them,4 proteins were involved in 8 metabolic pathways,especially oxidative phosphorylation and cell senescence.Subsequently,lifespan assay and oxidative stress test were performed to investigate the peptides prepared from sea cucumber protein hydrolyzate using the aging model of Caenorhabditis elegans.The results of the anti-aging experiment demonstrated that high-dose peptides significantly prolonged the lifespan of nematodes(30.50%),and improved their capacity to inhibit oxidative stress.The results provide evidence supporting the development of bioactive proteins and peptides derived from Stichopus japonicus as functional foods and lay the foundation for the research of an anti-aging drug.
