Jack Melia Plant Endophytic Fungi In The Related Research And Sophora Sffpt Gene Plant Dual Construction Of Expression Vector

Diversity and in vitro antitumor activity of endophytic fungi from mangrove plants XylocarpusIn recent years, the studies on the secondary metabolites from mangrove endophytic fungi are significantly increasing. As for its unique growth environment, mangrove endophytic fungi often have special metabolic pathways and genetic background. The types of its secondary metabolites and products are also quite abundant.In this study, Xylocarpus plant materials were collected from Sanya, Hainan and southern Thailand Trang Province Coast. The biologically pure endophytic fungal strains from stems, leaves and seed coats of Xylocarpus plants were acquired by repeated purification, and identified with Internal Transcribed Spacer (ITS) rDNA molecular methods. In vitro antitumor activity was tested by MTT method.A total of26entophytic fungal strains were isolated and identified, which belonging to15genera,11families,9orders and3classes in Ascomycota, and one genera in Basidiomycota. There were differences in genus and species levels among three plant materials from different habitats and species. The strains of Phomopsis and Colletotrichum existed in all three plant materials and they were dominant species.The EtoAc extracts of many strains showed strong in vitro antitumor activity according to bioassay. Aspergillus, Botryosphaeria, Gibberella and Penicillium strains showed good inhibitory effect on the tumor cell lines HCT-116, Bel-7402, BGC823, A549and A2780. These results provide a basis for further selection of the active strains and separation of the extract and isolation of the secondary metabolites.Trichoderma sp. Xy24(0106) with good in vitro antitumor activity was scaled up in10L level, and3compounds were isolated and one of them was identified as Trichodimerol by various spectroscopic data analyses.The diversity and the strains with antitumor activity of endophytic fungi from Xylocarpus plants were observed, and it is of value for further investigation. Construction of plant expression vector fused the SfFPT geneFlavonoids have a variety of biological properties, such as cardiovascular activity, antibacterial and antiviral activity, antitumor activity, etc. The prenylation of flavonoids is not only the contributor to the diversity of plant secondary metabolites but also to their bioactivity, bioavailability and druggability improvement.In the previous work of our laboratory, function characterization of the flavonoids prenyltransferases(SfFPT) from Sophora flavescent was carried out by preparation of the recombinant yeast microsomal fraction. Functional identification and catalytic efficiency research showed that SfFPT was a kind of plant flavonoids prenyltransferases with broad substrate spectrum.The purpose of this investigation is to construct plant binary expression vectors and transform the SfFPT gene into tobacco, aiming to do preparation for the expression and functional identification of SfFPT in plants. The main results are as follows:(1) Primers were designed to introduce restriction sites and the SfFPT gene was inserted into the position between CaMV35S promoter and NOS terminator of the expression vector pCAMBIA1305.1and pCAMBIA1302by DNA recombinant technology. Three plant binary expression vectors pC05-Sf-GUS、pC05-Sf-NR、pC02-Sf-GFP have been constructed.(2) The three recombinant vectors were introduced into Agrobacterium tumefaciens LBA4404, Agrobacterium rhizogenes K.599using CaCl2method.(3) We try to investigate the conditions and methods of transferring SfFPT gene into tobacco using leaf dish transformation and laid the foundation for the next step to cultivate transgenic tobacco and study the functional identification of SfFPT in tobacco.