Enzyme And Secondary Metabolites In Different Development Stages Of Thyme Changed And Responsed By Inducer

Thyme is lamiaceae of perennial fragrant plant. The mainly chemical contentnaphtha and flavonoids.The naphtha including volatile material taht divided intoaromatic compounds,fatty acid derivatives,single sesquiterpanes and half terpenoids,etc. Exogenous inducer materials can dramatically improving the secondarymetabolites of thyme.The experimental material was cultivation during difference phase of thyme, thatmeasured lipoxygenase and phenylalanine transaminase activity; mRNA expressionof secondary metabolites of key enzymes lipoxygenase was tested by RT-PCR; Useorganic solvent extraction ssential oil of thyme. The qualitative analysis andquantitative analysis ware tested by the GC-MS; The study were research jasmonicacid methyl ester, salicylic acid and chitosan the effect development stages of thymesecondary metabolites in the process of key enzymes LOX and PAL and mRNAexpression of LOX and PAL. Secondary metabolites to release influence on thecharacter; Study through adjusting culture medium carbon sources, nitrogen sourcesand amino acid. Plant resistance induction system in that important regulatory effect,confirmed that exogenous induction and precursor induction will adjust thymesecondary metabolites process enzymes and gene expression and increase theaccumulation of product.The mainly result showed that thyme cultured for20d and30d and induced byMeJA of concentration of1.0mmol/L groups that LOX enzymes singfiancntlyincreased that the activity is655.4%as CK,the concentration of150mmol/L SA after12h as503.8%as CK. The concentration200mmol/L CTS was treated24h after for429.1%as CK. The thyme of culture for30d was treated MeJA by0.5mmol/L for48h after LOX688.1%as CK and was treated with150mg/L SA that induction activityof LOX12h after for383.8%as CK, the200mmol/L CTS induction24h after theactivity of LOX for360.8%of CK.40d thyme exogenous factors in processing ofLOX active effect was not obvious. The RT-PCR results show that differentconcentration of exogenous induction that they can change significantly mRNA expresstion level of thyme LOX. The thyme cultured for30d after and treated withinduced material, LOX mRNA expression is significantly higher than control group.When the thymen was cultured for20d and30d that1.0mmol MeJA,100mg/L SA and200mg/L groups of PAL activity ware higher than control groups,thatwas respectively610.3%,788.1%,592.5%,402.4%,541.2%and400.1%; And the40dafter,that0.5mmol/L MeJA,100mg/L SA and200mg/L chitosan droups havehighest activity of the PAL that respectively390.2%,377.1%and413.4%. The thymecultured for30d that PAL gene expression quantity achieved peak. Exogenousinduced material can adjust the thyme secondary metabolism related enzymes PALgene expression.The different stage secondary metabolites accumulation of thyme was tested byGC-MS. Results showed that the extract of detection including46compounds and11kinds of compounds of content than the1%. The results showed that three elicitorwere significantly increased aromatic compound that Benzene1-methyl-3-(1-methylet-hyl)-1,4-Cyclohexadiene and1-methyl-4-(1-methylethyl)-as a representative ofoxidation terpenoids and terpenes compounds effect is very significantly.Thyme culture for30d of the medium content sugar concentration is4%andNO3/NH4+2:1ratio that the LOX acyivity high others groups. The0.6mmol/Lphenylalanine and0.4mmol/L groups highest increasing activity of enzymes LOX,pyruvic acid no effect LOX, Thyme of the PAL activity of the sucrose density is2%,NO3-/NH4+2:1ratio for the highest, The0.6mmol/L phenylalanine,0.8mmol/Lglutamic acid and0.4mmol/L tyrosine groups active PAL higher than control, thatrespectively126.3%,150.6%and153.9%as the control.Conclusion: different inducer material that can promote thyme secondarymetabolism contents also can induce the key enzyme mRNA expression levels.Different materials and induction was existed material optiomal of concentration andbest induced time.