ChicoryextractsfromCichoriumintybusL.aspotentialantifungals
D.Mares1,C.Romagnoli2,B.Tosi1,E.Andreotti1,G.Chillemi3&F.Poli
4
1DepartmentofNaturalandCulturalResources,UniversityofFerrara,Ferrara,Italy;2
Department
ofPaleobiologyandBotanicalGarden,UniversityofModenaandReggioEmilia,Modena,Italy;
3
Veneto
Agricoltura,SezioneRicercaeSperimentazione,Padova,Italy;4DepartmentofEvolutionaryand
ExperimentalBiology,UniversityofBologna,Bologna,Italy
Received4October2004;acceptedinrevisedform23November2004
Abstract
InthisworkextractsfromrootsofthecommonvegetableCichoriumintybusL.,highlyappreciatedforits
bittertaste,werestudiedtoinvestigatetheirpossiblebiologicalactivityonfungifromavarietyofecological
environments:someareparasitesonplants(phytopathogens)orofanimalsandhumans(zoophilicand
anthropophilicdermatophytes),othersliveonthesoilandonlyseldomparasitizeanimals(geophilic
dermatophytes).Theextractswereineffectiveongeophilicspeciesandontestedphytopathogens,withthe
exceptionofPythiumultimum,whereastheyinhibitedthegrowthofzoophilicandanthropophilic
dermatophytes,inparticularTrichophytontonsuransvar.sulfureum,whosetreatmentcausedmorphological
anomalies,hereobservedbyscanningelectronmicroscopy.Thisbehaviourisdiscussedonthebasisofthe
presenceinthechicoryextractofthetwomainsesquiterpenelactones,8-deoxylactucinand11b,13-dihy-
drolactucin.
Keywords:antifungalactivity,Cichoriumintybus,dermatophytes,phytopathogens
Introduction
Aninterestindrugsfromnaturalsourceshas
recentlybeenrevived,withconsequentrenewed
attentiondevotedtoplantseitherascosmeticsor
assourcesoftherapeuticallyeffectivemedicines.
InthisworkweconsideredCichoriumintybus
L.(Asteraceae),aplantcommonlyknownaschic-
oryorassuccory,oftenfoundonourtablesasa
vegetable,andhighlyappreciatedforitsbittertaste.
Plantswithverydifferentmorphologicaland
chromaticcharacteristicsoftheepigeanpartwere
knownaschicory;amongthem,C.intybus
L.var.‘RossodiChioggia’,withredandvariegated
leaves,isaspecialityoftheVenetoregion(North-
easternItaly).Thisspecieswasachievedbyalong
processofselection,madebyhorticulturists,in
ordertoobtainaplantformingaheart,havingthe
centralleavestightlyimbricatedandendowedwith
aredvariegationonthewholesurfaceoftheleaf.
Chicorycultivationwasmentionedinan
ancientEgyptianmedicaldocument(1550B.C.)
reportingthattwotypesofthisplantwereappre-
ciatedinparticular:onefortheleavesusedas
salad,theotherasadepurantanddigestive.
Chicoryisstillusedtodayforitsbitter-tonic,
diuretic,digestive,lightlylaxativeandcholagogic
properties,duetoitscontentofmucilages,resins
andbittersubstances[1].Leavesareusedasinfu-
sionsforanemiaanddigestivedisorders:rootsare
employedasinfusiontopurifytheorganismand
tostimulatetheappetite;flowershavecholagogic
activity,buttheyarealsoutilizedexternally,such
asinskinwashing,coolingandsoftening[2].
ThemaincomponentsofC.intybusaresesqui-
terpenelactones,suchaslactucin,8-deoxylactucin,
lactupicrinand11b-dihydro-terpenederivatives[3],
whichareresponsibleforthebittertasteofthis
plant.Othercomponentsarecoumarins,suchas
cichoriin,esculin,umbelliferone,scopoletinand
Mycopathologia(2005)160:85–92ÓSpringer2005
DOI:10.1007/s11046-004-6635-2
6,7-dihydroxycoumarin[4],flavonederivatives
(cichoricacid,chlorogenicacid,apigenin,quercitin)
[5,6],carbohydrates(glucose,fructoseandinulin)
[7]andvitamins[8].
Thecomponentsvarywidely,notonlyfrom
onecultivartoanother,butalsowithinasingle
cultivar,dependingonwhichperiodoftheyearthe
cropisharvested[9]andalsoaccordingtothe
methodusedforextractpreparation[10,11].
Thefunctionofthesecondarymetabolitesof
chicorycouldperhapsbeconnectedwithapassive
defenceoftheplantagainstpathogens:infacta
studybyMondeetal.[12]showedthattheinoc-
ulationofthebacteriumPseudomonascichoriiinto
theplantleadstotheformationofaphytoalexin,
theguaianolidecichoralexin,withanobvious
defencefunction.Itisnotknownifthesedefence
mechanismsarealsoeffectiveagainstphytopath-
ogenicfungi,which,asarule,attackthechicory;
toourknowledge,theantifungalactivityof
Cichoriumintybusextractsonsoil-bornepatho-
genshasneverbeenstudiedbefore.
Thepurposeofthepresentworkwastoeval-
uatetheefficacyofchicoryextractsonthegrowth
ofsomegroupsofpathogenicfungi:somephyto-
pathogenicfungiandsomefungiparasiticonman
andotheranimals.Thelatters,knownasderma-
tophytes,arekeratinophilicandkeratinolytic,i.e.
theyareabletodigestandusekeratinandkeratin-
likesubstratescausingdiseasesknownastineas.
Dermatophytesbelongtothreeecologicalgroups:
geophiles(suchasMicrosporumgypseumand
Nannizziacajetani),thatmainlyliveinthesoil,
zoophiles(suchasTrichophytonmentagrophytes),
essentiallypathogenicforanimals,andanthropo-
philes(suchasEpidermophytonfloccosum,Trich-
ophytonrubrumandTrichophytontonsurans)
whichgenerallyonlyinfectman.
Materialsandmethods
Plantmaterial
TheplantmaterialwasthebiennialCichorium
intybusL.var.‘RossodiChioggia’,cv.medium-
initial,kindlyprovidedbythe‘CentroSperimen-
taleOrtofloricolodiVenetoAgricoltura,Podi
Tramontana’(Rosolina,Rovigo,Italy).Plants
belongingtoselectionline‘ea1°p5a/38-7-7-6’(C
2
)
wereplantedinthenurseryand,after30days,the
plantulesweretransplantedtotheopenfield,
followingtheestablishedcultivationtechnique
[9,13].After60days,forthestudyonantifungal
activity,samplesweretaken,harvesting,asplant
portion,theroots,astheyhaveanhighercontentof
guaianolidesthanthatoftheheadandouterleaves
[7,10].
Preparationofchicoryextracts
Tohavethebestresultswemodifiedtheprocedure
describedbyPyrek[14];ourextractionmethodisthe
following:200gofdriedandpulverizedrootswere
putinasolid–liquidSoxhletextractorwith1000ml
ofacetoneat50°Cfor24h;then,theacetonewas
replacedwithfurther1000mlofpureacetoneand
theextractioncontinuedfor24h,obtainingan
exhaustiveextraction.Atthispointtheacetone
extracts(2000ml)wereblendedandconcentrated
withaBuchiRotavaporsetat40°C;thissolution
wasthenpurifiedfor24hwithhexane(1000ml)
withaliquid–liquidSoxhletextractortoeliminate
lipophiliccompounds.Theaqueousphase,con-
taininghydrophilicsubstancessuchasguaianolides,
wasthenseparatedfromhexaneandreducedto
100mlintheRotavaporkeptat70°C.Finallythe
extractwassubjectedtochromatography.
Sesquiterpenelactones
Themainsesquiterpenelactonespresentinthefirst-
year-growthsamplesofC.intybusvar.‘Rossodi
Chioggia’weretheguaianolides8-deoxylactucin
and11b,13-dihydrolactucin.Thesewereisolated
fromtherootsinaccordancewithprocedures
describedabove.Thechemical–physicalcharacter-
istics(UV,MS,NMR,TLC,HPLC)arecompara-
bletothosereportedinliterature[14–17].The
qualitativeandquantitativedeterminationofthe
twoguaianolideswasperformedbyanautomated
multipledevelopmenthigh-performancethin-layer
chromatographicanalysis(AMD/HPTLC)(Camag,
Switzerland)asdescribedbyPolietal.[7].The
concentrationsof8-deoxylactucinand11b,13-dihy-
drolactucinweredeterminedin10samplesofthe
product;themeanamountof8-deoxylactucinand
11b,13-dihydrolactucincontainedintheextracts,
afterfiltrationonMillipore0.22lm,wererespec-
tively240.75(standarddeviation3.1)and50.06
(standarddeviation1.5)lg/ml.Thesemeanamounts
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