Bioinformatics Analysis Of DCs In Jilin Panax Ginseng And Research On The Transformation Of Large-Insert DNA (B9) Of Panax Ginseng To Ganoderma Lucidum

Panax ginseng is a medicinal herb of Araliaceae.Ginsenosides are the mainly effective components of Panax ginseng which become a focus in the study of pharmacological activity.Ganoderma lucidum is a kind of ancient medicinal fungus.Ganoderma Triterpene as one of the effective components in Ganoderma lucidum has been identified with many pharmacological activities by mass researches.Both Ginsenosides and Ganoderma Triterpene are Terpenoids,and the anabolic pathways of these two components have part of the same in the mevalonate pathway(MVA).The aim of this study is to isolate and analysis Jilin ginseng decarboxylase unigenes,to investigate its origin and evolution,the structure and function,activity and expression,and to analysis its role in the synthesis of ginsenosides during the growth and development process in Jilin ginseng.This study will provide resources and theoretical basis on the ginseng decarboxylase system study.As the mainly transformation method of multiple genes,the transformation of large fragment DNA are more applied to the study of the regulating body traits and functions by multiple genes.Study on the transformation of the large fragment which carrying ginseng decarboxylase genes related to skeleton synthesis of the skeleton into Ganoderma lucidum,can enrich the germplasm resource of Ganoderma lucidum and lay the foundation of the produce of ginsensides which realizing Ganoderma lucidum as biological reactor.The main results are as follows:1.A total of 166 decarboxylase unigenes were isolated from the database of Jilin ginseng unigenes.2.Analysis of the pathway of Jilin Panax ginseng DC unigenes,33 metabolic pathways have navigated and the results showed that there are 4 unigenes involved in terpenoid backbone biosynthesis process and all the four unigenes are diphosphomevalonate decarboxylase.3.The phylogenic analysis of Jilin Panax ginseng DC unigenes revealed that all the decarboxylase was highly conservative In the process of evolution;the evolution of Jilin Panax ginseng PgMVD gene and Panax notoginseng PnMVD is similar.4.Heatmap of Jilin Panax ginseng DC Unigenes on 14 tissues,four different years-old roots and42 farm varieties revealed that the expression of DC Unigenes has a significantly differences at different tissues,developmental stages and growth environment.The network analysis showed that the activity of most DC Unigenes is significantly correlated.5.Analysis of the structure and function of Jilin ginseng Pg MVD protein showed that Pg MVD4 is a hydrophilic protein and the molecular weight is 46,618.4Da.It contains four kinds ofsecondary structures including Alpha helix,Beta turn,Extended strand and Random coil.Six GO-Terms were annotated on biological process,molecular function and cellular component which including isoprenoid biosynthesis(GO:0008299),isopentenyl pyrophosphate metabolism(GO:0019287),mevalonate pyrophosphate decarboxylase activity(GO:0004163),ATP binding activity(GO:0005524),carboxylase(GO:0016831),cytoplasmic(GO:0005829).6.The large fragment DNA which carrying ginseng decarboxylase genes related to skeleton synthesis of the skeleton was transformed into the Ganoderma lucidum protoplast by electroporation.The transformation condition was that the age of protoplast for cultured was 6days,the kanamycin resistance concentration was 12mg/ml,the voltage was 700 V,the ratio of DNA to protoplast was 1:4,a total of 2 transformants were obtained.