Objective To clone and sequence cDNA encoding 3-hydroxy-3-methylglutaryl-coenzyme A reductase(HMGR) from Atractylodes lancea.Methods The cDNA,encoding HMGR in A.lancea,was amplified by RACE strategy with the cDNA of the total RNA of young leaves as the template.The partial fragments of HMGR were cloned and sequenced.Results The analysis results revealed that the conserved fragments were 458 bp.At the same time,the two fragments had been obtained 84.28% identification in nucleotide acid and 92.11% identification in corresponding amino acid,named as HMGRcr1 and HMGRcr2,respectively.It was deduced that they may be members of the HMGR gene family in A.lancea.Sequencing analysis showed that HMGRcr1 and HMGRcr2 had high identity with HMGR from other plants.Conclusion The cDNA encoding HMGR from A.lancea is cloned and reported for the first time.The work will provided a foundation for exploring the mechanism of terpenes biosynthesis and application to the other medicinal plants.

Objective To elucidate the constituents from the root of Euphorbia pekinensis and to look for new products with antitumor activity.Methods The chromatography on silica gel was used and the structures were identified by IR,NMR,and MS spectral analyses and their physicochemical properties were comparied.Results Eight compounds were isolated and identified as octadecanol(Ⅰ),octadecanyl-3-methoxy-4-hydroxybenzeneacrylate(Ⅱ),?-sitosterol(Ⅲ),triacontanoic acid(Ⅳ),2,2′-dimethoxy-3,3′-dihydroxy-5,5′-oxo-6,6′-biphenylformic anhydride(Ⅴ),euphol(Ⅵ),tirucallol(Ⅶ),and euphpekinensin(Ⅷ).Conclusion The compounds Ⅰ,Ⅳ,Ⅵ,and Ⅶ are isolated from the roots of E.pekinensis for the first time.

Objective To establish the suspension cell line of Atractylodes lancea and to study the effect of two endophytic fungal elicitors on its essential oil production.Methods The essential oil was extracted by using ultrasonic wave after suspension cell was treated with endophytic fungal elicitors.Then,the determination of four compounds(atractylone,hinesol,?-eudesmol,and atractylodin) was carried out by gas chromatography.Results By testing in various conditions,the suspension cell line with a rapid growth rate was established.Its highest biomass(6.95 g/L) was obtained on day 21.?-Eudesmol was the only detection in the control suspension cell,and its highest content(17.469 ?g/g) was also reached on day 21.The effect of crude elicitors of two endophytic fungi(belong to Cunninghamella sp.and Gilmaniella sp.respectively,named AL4 and AL12) on the cell growth and the production of essentia1 oil were investigated.Overall AL4 elicitor got better effect.When suspension cell of 14-day-old cultures was exposed to AL4 elicitor(carbohydrate 20 mg/L medium) for 7 d,the biomass increased 3.31% over the control,and the four compounds(atractylone: 14.715 ?g/g,hinesol: 28.395 ?g/g,?-eudesmol: 38.794 ?g/g,and atractylodin: 8.310 ?g/g) were all detected.Among them,the content of ?-eudesmol reached 2.22 times as much as the control.Conclusion The cell growth and the accumulation of essential oil of A.lancea could also be promoted by adding crude elicitors of the endophytic fungi AL4 and AL12.

Objective To study the producing method for Euphorbia pekinensis promoted by endophytic fungi.Methods The tissue culture plantlets were innoculated with endophytic fungi,then it was transplanted to the pods and cultured for one year.The biomass and content of terpenoids(isoeuphpekinensin and euphol)were determined.Results The growth of the E.pekinensis was promoted by endophytic fungi E4(Fusarium sp.Ⅰ)and E5(Fusarium sp.Ⅱ).The fresh weight of the root increased 49.45% and 59.74% for E4 and E5 treated,respectively.The plants dry weight coefficient increased 2.99% and 6.48% for E4 and E5 treated,respectively.The contents of isoeuphpekinensin and euphol in the plants were increased for tissue cultured in both bottles and pods.For the plants cultured in pods for one year,the isoeuphpekinensin and euphol increased 92.79% and 40%,respectively treated by E4.The isoeuphpekinensin and euphol increassed 105.32% and 241.38%,respectively treated by E5.Conclusion The endophytic fungi E4 and E5 could promote the growth of E.pekinensis and accumulated terpenoids.

Purpose　The aim is to compare the sterols in Hericium crinaceus ethanol extract and water extract from solid fermented hyphae and to study the pharmaceutical chemical basis of the different medicinal effects. Methods　The nonsaponifiable lipids were isolated by saponification.The sterols were then detected by TLC and RP-HPLC.Results　The content of sterols in ethanol extract was found to be higher than that in water extract.And one type of sterols from Hericium erinaceus hyphae was identified as ergosterol.Conclusion　Due to ergosterol′s multifunction in biological activities,it may well be one of the active components of hyphae.And higher content of lipids, especially sterols may be one of the reasons for the better medicinal effect of ethanol extract than water extract.

Crude elicitor of one endophytic fungi (belong to Cunninghamella sp., named AL4) induced multiple responses in Atractylodes lancea suspension cells, including rapid generation of nitric oxide (NO) and hydrogen peroxide (H2O2), sequentially followed by enhancement of essential oil production. Adding NO-specific scavenger 2-4-carboxyphenyl-4,4,5, 5-tetramethylimidazol ine-1-oxyl-3-oxide (cPTIO) and H2O2 scavenger catalase (CAT) could block elicitor-induced NO and H2O2 generation respectively, but could all partly block elicitor-induced essential oil biosynthesis. Adding NO-donor sodium nitroprusside (SNP) and H2O2 could all promote essential oil accumulation in A. lancea cells, but the effect of both was different. These results strongly suggested that NO and H2O2 may all act as signaling molecule to mediate AL4 elicitor promoting essential oil accumulation in suspension cells of A. lancea. Furthermore, adding cPTIO and CAT contemporarily could not completely inhibit essential oil accumulation induced by AL4 elicitor. This result suggested that AL4 elicitor could also promote essential oil accumulation in suspension cells of A. lancea by other means.
Atractylodes ; cytology ; metabolism ; Benzoates ; pharmacology ; Catalase ; pharmacology ; Cells, Cultured ; Cunninghamella ; physiology ; Hydrogen Peroxide ; metabolism ; Imidazoles ; pharmacology ; Nitric Oxide ; metabolism ; Oils, Volatile ; analysis ; metabolism

The effect of the endophytic fungi Botrytis sp. (C1) or Chaetomium globosum (C4) on the drought resistance of Chrysanthemum morifolium was studied. Ch. morifolium plantlets were inoculated with C1, C4 and cultured in the pots for 60 days, then the plantlets were stressed by 0%, 10%, 20%, 30%, 40% PEG6000 respectively in order to simulate different drought conditions. Biomass, the activities of SOD, POD, PAL, the contents of MDA and soluble protein of each group were determined. The results showed that endophytic fungi groups grew better than the control (without inoculation endophytic fungi). With the increasing of the concentration of PEG6000, the biomass of Ch. morifolium of each groups decreased, while the biomass of fungi groups was significantly higher than that of control, moreover C4 group higher than C1 group. With the concentration of PEG increasing, the content of MDA of each group increased too, while POD activity and soluble protein content of all treatments increased at first and then decreased. SOD activity and PAL activity of the control were increased with the increase of PEG concentration, but SOD activity of the two fungi groups were stable. After been stressed by different concentrations of PEG, MDA content of two fungi groups were always lower than the control, while SOD activity, POD activity, PAL activity and soluble protein content were higher. In conclusion, endophytic fungi can increase the drought resistance of Ch. morifolium.
Biomass ; Botrytis ; drug effects ; growth & development ; isolation & purification ; Chaetomium ; drug effects ; growth & development ; isolation & purification ; Chrysanthemum ; drug effects ; metabolism ; microbiology ; Droughts ; Peroxidases ; metabolism ; Polyethylene Glycols ; pharmacology ; Stress, Physiological ; drug effects ; Superoxide Dismutase ; metabolism