RESUMO
To research the correlation between accumulation of triterpenoids and expression of key enzymes genes in triterpenoid biosynthesis of Alisma orientale,the study utilized UPLC-MS/MS method to detect eight triterpenoids content in the tuber of A. orientale from different growth stages,including alisol A,alisol A 24 acetate,alisol B,alisol B 23 acetate,alisol C 23 acetate,alisol F,alisol F 24 acetate and alisol G,and then the Real time quantitative PCR was used to analyze the expression of key enzymes genes HMGR and FPPS in triterpenoid biosynthesis. Correlation analysis showed that there was a significant positive relation between the total growth of these eight triterpenoids and the average relative expression of HMGR and FPPS(HMGR: r = 0. 998,P<0. 01; FPPS: r = 0. 957,P<0. 05),respectively. Therefore,the study preliminarily determined that HMGR and FPPS genes could regulate the biosynthesis of triterpenoids in A. orientale,which laid a foundation for further research on the biosynthesis and regulation mechanism of triterpenoids in A. orientale.
Assuntos
Alisma/química , Alisma/genética , Geraniltranstransferase/genética , Triterpenos/análise , Cromatografia Líquida , Hidroximetilglutaril-CoA-Redutases NADP-Dependentes/genética , Compostos Fitoquímicos/análise , Extratos Vegetais , Proteínas de Plantas/genética , Tubérculos/química , Espectrometria de Massas em TandemRESUMO
To engineer endophytic Enterobacter cloacae as a biocontrol agent against banana fusarium wilt, a promoter-probe plasmid pUCK was constructed to identify a strong promoter to express disease resistance genes. Using a kanamycin resistance gene for selection, 10 fragments with strong promoter activity were identified from the genome of the E. cloacae KKWB-10 strain. The regions of these 10 fragments that were the primary contributors to the promoter function were identified, and their promoter activities were further evaluated using green fluorescent protein (GFP) as a reporter gene. Fragment 132aâ³ drove the highest level of GFP activity when the bacteria bearing the fragments were cultured in Luria-Bertani and banana stem extract media. The GFP-expressing strain harboring fragment 132aâ³ (K-pUCK7-132aâ³-GT) was then inoculated into banana plantlets (about 1 × 10(7) CFU per plant) to verify the activity of fragment 132aâ³ in planta. Ten days after inoculation, tissue sections of these banana plantlets were observed by laser confocal scanning microscope. Green fluorescence was observed in the tissues of banana plantlets inoculated with K-pUCK7-132aâ³-GT but not in uninoculated controls. These results suggest that fragment 132aâ³ possesses strong promoter activity when its host strain colonizes the banana plants and can be used to engineer endophytic E. cloacae KKWB-10 for biocontrol.
Assuntos
Enterobacter cloacae/genética , Musa/microbiologia , Regiões Promotoras Genéticas , Antibacterianos/farmacologia , Fusão Gênica Artificial , DNA Bacteriano/química , DNA Bacteriano/genética , Endófitos/genética , Genes Reporter , Proteínas de Fluorescência Verde/análise , Proteínas de Fluorescência Verde/genética , Canamicina/farmacologia , Dados de Sequência Molecular , Seleção Genética , Análise de Sequência de DNAAssuntos
Artrite Gotosa/patologia , Artrite Gotosa/cirurgia , Articulações do Pé/patologia , Articulações do Pé/cirurgia , Articulação da Mão/patologia , Articulação da Mão/cirurgia , Artrite Gotosa/diagnóstico por imagem , Articulações do Pé/diagnóstico por imagem , Articulação da Mão/diagnóstico por imagem , Humanos , Masculino , Pessoa de Meia-Idade , RadiografiaRESUMO
A bisdesmosidic steroidal saponins library, composed of 16 novel kryptogenin glycosides, was set up via six random glycosylation procedures, wherein two compounds showed their antitumor activity against HeLa cell in the preliminary pharmacological research.
