ABSTRACT
Objective To establish a method for the determination of protocatechuic acid, salidroside, and chlorogenic acid in Sargentodoxa cuneata. Methods The separation was performed on a Waters XSELECT CSH C18 (150 mm × 4.6 mm, 5 μm) with methanol-acetonitrile-0.2 % phosphoric acid as the mobile phase in a gradient elution at a flow rate of 0.8 ml/min. The detection wavelength was 260 nm and the column temperature was 35 ℃. Results The linear ranges of protocatechuic acid, salidroside, and chlorogenic acid were 0.0020-0.0120, 0.0600-0.3602, 0.0750-4.5006 mg/ml, respectively. The average recoveries were 98.01% (RSD=0.07%), 98.53 % (RSD=0.12%), and 101.10 % (RSD=1.92%), respectively. Conclusions The method is simple, accurate, and highly reproducible, which could provide the scientific evidence for the quality control of Sargentodoxa cuneata.
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Objective: To study the proper medium for tissue culture of the explants from Hypericum perforatum . Methods: The root, stem with node and leaf of the Hypericum perforatum seedling were used as explants, and cultured in basic Murashige and Skoog (MS) medium supplemented with different kinds and different concentrations of hormones. The callus, buds and roots were induced. The cultivation of the tube seedling were carried out. Results: Different hormones and explants had different effects on the induction of callus and the formation of buds. 2,4 D could promote the formation of the callus of Hypericum perforatum . Among cytokinins, 6 BA had great effect on the formation of buds, but the effects of KT and ZT were insignificant. The combination of NAA 1 mg/L and 6 BA 0.2 mg/L was found most effective in promoting the rooting of Hypericum perforatum . Conclusion: The results indicate that the callus is easy to be induced when MS medium supplemented with 2,4 D. The root of explants is most suitable for the rosette bud rapid induction.
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Objective: To establish the culture system of hairy root of Isatis indigotica and to induce the regeneration plant of hairy root. Methods: Hairy root of Isatis indigotica was obtained from infected cotyledon explants after infection with Agrobacterium rhizogenes strains A4, R1601 and ATCC15834. The better lines were selected. The growth curve was surveyed and extrinsic factors affecting the growth of hairy roots were investigated. The regeneration plant was induced on MS media with different hormones. The transformation of Ri T DNA was examined through high voltage paper electrophoresis. Results: The hairy root was originally obtained from Isatis indigotica . The regeneration plant was induced on MS media with BA. The result of high voltage paper electrophoresis confirmed the transformation of T DNA from Ri plasmid to the hairy root and regeneration plant. Conclusion: The acquisition of hairy root of Isatis indigotica and regeneration plant of it lay a foundation for the production of active component and introduction of foreign gene. [
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Objective Two insect resistance genes Bacillus thuringiensis(Bt) crystal protein gene CryIA(c) and cowpea trypsin inhibitor gene CpTI were introduced into tetraploid Isatis indigotica to enhance the resistance to moths.Methods I.indigotica was transformed with a plasmid,pGBI121S4ABC,containing CryIA(c) Bt and CpTI and the selectable gene(Npt-Ⅱ) driven by the CaMV35S promoter via Agrobacterium tumefaciens LBA4404 mediated transformation.Then PCR and Southern blotting assay were conducted followed by moth bioassay test.Results Co-transgenic rate of the two genes in tetraploid I.indigotica was 16.67%.The integration and expression of introduced genes in T0 regenerated transgenic plants were confirmed by Southern blotting assays.Insect bioassay test demonstrated transgenic lines had significant inhibition to moths,compared with wild type control.Conclusion Co-transformation of Bt and CpTI genes is an effective strategy to enhance the resistance to moths for tetraploid I.indigotica.
ABSTRACT
Object To establish the culture system of hairy root of Eclipta prostrata (L.) L.. Methods Hairy root of E. prostrata was obtained from infected cotyledon explants after infection with Agrobacterium rhizogenes strains A4, R1601 and ATCC15834, elite strains were screened and growth curves determined. The transformation of Ri T DNA was examined through high voltage paper electrophoresis. Results The hairy root was originally obtained from E.prostrata. The result of high voltage paper elctrophoresis confirmed the transformation of T DNA from Ri plasmid to the hairy root. Conclusion The acquisition of hairy root of E. prostrata provided further a foundation for the industrial production of active drug component.