Apresentação efectuada na 51th GA Meeting, Kiel, Germany, 2003.Fundação para a Ciência e a Tecnologia (FCT

Conceição, Luís F. R.

Dias, Alberto Carlos Pires

Ribeiro, Catarina

Universidade do Minho: RepositoriUM

Changes of metabolism in Hypericum perforatum cells after elicitation with Colletotrichum gloeosporioides51th GA Meeting, Kiel September 2003L.F. Conceição, C.L. Ribeiro, A.C.P. DiasUniversidade do MinhoDep. BiologiaBraga - Portugal• important medicinal plant• antidepressive, antiviral, anticancer activities• economic importanceHypericum perforatumHigh demand of biomass ecological problems (wild harvesting) phytochemical variabilitySolution: cultivationbiomass consistence anthracnose (Collethotrichum gloeosporioides)Major problemColletotrichum gloeosporioides• common plant pathogen• host-pathogen model• major pathogen of H. perforatum L.In vitro cultures of H. perforatum L.• easy to manipulate• closed systemPathogen vs hostExperimentalH. perforatumsuspension culturesElicitation (6th day of culture)HPLC-DAD-MS analysis of phenolsCell viabillity (tripan blue)AOS measurement (DCFH-DA)Methyl jasmonate (MeJ) or salicylic acid (SA) Induction (5th day of culture)Autoclaved C. gloeosporioidesbiomass suspension AnalysisMeJ/fungi elicitation – culture changesMeJ 100mM0 2 4 6 8 10 12 14051015Control (A)MeJ (B)MeJ + fungi (D)fungi (C)time (day)dwb (mg/ml)A B C D- addiction of MeJ (100 mM)- addiction of fungiMeJ/fungi elicitation – HPLC profilescontrol MeJ 100 mMfungi fungi + MeJ1,3,6,7 xanthones derivatives - quercetin derivatives - luteolin derivatives -MeJ/fungi elicitation – phenolic changesXanthone accumulation- addiction of MeJ (100 mM)- addiction of fungi5.0 7.5 10.0 12.502468controlMeJfungiMeJ+fungidayxant onas (mg/g dwb)MeJ/fungi elicitation – oxidative burst0 100 200 300 4000.00.51.01.52.02.53.03.5time (min)AOS (net)DCFH-DA (dichlorodihydrofluorescin diacetate)fluorescence(An+1 - An)0 100 200 300 400 500 600 7000510controlMeJfungiMeJ+fungitime (min)total AOSConclusionsH. perforatum cultures vs C. gloeosporioidesInduction of an oxidative burst – incompatible interaction ?Induction of the xanthone accumulation – phytoalexins ?Methyl jasmonate(ISR)Salicylic acid(SAR)(+) (no effect)Acknowledgements(POCTI Project, granted by FCT)SA 25 mM0 2 4 6 8 10 12 14051015Control (A)SA (B)SA + fungi (D)fungi (C)time (day)dwb (mg/mL)SA 100 mM0 2 4 6 8 10 12 14051015Control (A)SA (B)SA + fungi (D)fungi (C)time (day)dwb (mg/mL)A B C DSA/fungi elicitation – culture changesA B CD5.0 7.5 10.0 12.501002003000255075100dayflavonols (g/gdwb) flavones (g/gdwb)ConclusionsFlavonoids accumulationluteolin derivatives – dot linequercetin derivatives – straight line

Changes of metabolism in Hypericum perforatum cells after elicitation with Colletotrichum gloeosporioides

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Changes of metabolism in Hypericum perforatum cells after elicitation with Colletotrichum gloeosporioides

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