The Validation Of Glycosyltransferase Gene Ugtpgx And The Studing On Induction In Adventitious Roots Of Panax Quinquefolium.l

Panax quinquefolium L.is famous and used herbs,which have a wide range of applications in the medicine,food,cosmetics and other fields.It has an agreat market demand in domestic and international.In this study,taking advantage of the established systems for cultivating adventitious roots of P.quinquefolium,we conducted studies about using fungal elicitors to enhance the secondary metabolite content.The induced mechanism of methyl jasmonate(MJ)elicitation(5 mg/L)and the glycosyltransferase gene in adventitious roots of Panax quinquefolium L were validated.Validation the experiments about UGTPgX gene function,the first is construction an expression vector UGTPgX-pYES2-NTA.The builded expression vector UGTPgX-pYES2-NTActransform into Saccharomvces cerevisiae INVSc1.Screening positive transformation,get the mutant strain YWJ1.To galactose induced expression of YWJ1 strains,then the product of iquid-mass chromatography analysis,validate the UGTPgX genotype with the darma sihuan terpene compounds in C-20 hydroxyl function of glycosylation.UGTPgX can catalytic synthesis Rb1 to Rd,Rg1 to Rh1.Fungal elicitors(Candida tropicalis,Aspergillus niger and a mixture of these cultures)were added to Panax quinquefolium adventitious roots to determine their ability to stimulate ginsenoside production.Addition of the elicitors resulted in the accumulation of ginsenosides over and above that in the control group,with peak ginsenosides content reaching 11.47 mg/g,8.03 mg/g and 9.59 mg/g when C.tropicalis,A.niger and a mixed elicitor preparation was used,respectively.The polysaccharide content reached a peak of 152.81 mg/g,213.01 mg/g-1 and 212.36 mg/gwhen C.tropicalis(800 mg/L),A.niger(200 mg/L)and the mixed elicitors(100 mg/L),respectively,were added.RT-PCR was used to analyze the expression of different genes from the elicitor treated groups of P.quinquefolium.Fungal elicitor treatment up-regulated squalene synthase(SS),squalene epoxidase(SE),dammarenediol synthase(DDS),β-amyrin synthase(β-AS),protopanaxadiol synthase(PPDS)and protopanaxatriol synthase(PPTS).Elicitors can by adjusting of some of the key enzyme genes in Panax quinquefolium L adventitious roots to regulate the content of secondary metabolites of American ginseng.But by 5 mg/L MJ as inducer of the experimental results prove the elicitor induced not only causeing the change of genes,MJ can stimulate the NO signaling pathway,JA and SA to trigger the synthesis of secondary metabolites of plant cells.