Flower palate ultrastructure of the carnivorous plant Genlisea hispidula Stapf with remarks on the structure and function of the palate in the subgenus Genlisea (Lentibulariaceae)

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Protoplasma(2018)255:1139–

1146https://doi.org/10.1007/s00709-018- 1220-6

ORIGINALARTICLE

FlowerpalateultrastructureofthecarnivorousplantGenliseahispidula Stapfwithremarksonthestructureandfunctionofthepalateinthe subgenusGenlisea(Lentibulariaceae)

BartoszJ . Płachno¹^&PiotrŚwiątek²^&MałgorzataS tpiczyńska³^&VitorF ernandesO liveiraM iranda⁴

Received:1December2017/Accepted:29January2018/Publishedonline:14February2018

#TheAuthor(s)2018.Thisarticleisanopenaccesspublication

Abstract

InthegenusGenliseaaswellasinitssistergenusUtricularia,thepalateprobablyplaysakeyroleinprovidingthecolour,mechanical andolfactorystimulitoattractinsectpollinatorsandtoguidethemtothegenerativestructuresandthenectaryspur.However,informat ionaboutthemicro-

morphologyofthepalateofGenliseaisscarce.ThisstudyaimstoexaminethestructureofthepalateinGenliseahispidulaindetailaswell asthepalatefromotherfivespeciesfromthesubgenusGenlisea.Inparticular,itsaimistoascertainwhetherthesepalatesfunctionasana reafortheosmophoresintheflowerorwhethertheyproducenectar.Weshowedthatthepalateinallofthespeciesthatwereexaminedw astheglandulartypeandthatithadcapitate,glandulartrichomes,whichhadasimilargeneralarchitectureacrossthespeciesthatwere examined.Nonectarsecretionwasobservedonthepalates.Theultrastructureofthepalatetrichomesshowedthatthepalateglandulart richomesmostprobablyfunctionasscentglandsthatproduceanolfactorystimulusforflowerpollinators.

KeywordsBladderwort.Carnivorousplant.Corkscrewplants.Floralmicro-

morphology.Genlisea.Glands.Lentibulariaceae.Lamiales.Osmophore.Pollination.Trichome.Utricularia.Ultrastructure

Introduction

ThegenusGenliseaA.St.-Hil.includesapproximately30–

31speciesthatareclassifiedintotwosubgenera—

GenliseaandTayloria.Genliseaaresmallherbaceousrootless plantsthat

HandlingEditor:Hanns-HeinzKassemeyer

ElectronicsupplementarymaterialTheonlineversionofthisarticle(http s://doi.org/10.1007/s00709-018-1220-6)containssupplementary m

a te r ia l, w h i ch i s a v a i l a b le t o a u t h o r i z ed u s e r s.

*BartoszJ.Płachnobartosz.pl achno@uj.edu.pl

1 DepartmentofPlantCytologyandEmbryology,JagiellonianUnivers ityinKraków,9GronostajowaStr,30-387Kraków,Poland

2 DepartmentofAnimalHistologyandEmbryology,UniversityofSi lesiainKatowice,9BankowaStr,40-007Katowice,Poland 3 BotanicGarden,FacultyofBiology,UniversityofWarsaw,Al.

Ujazdowskie4,00-478Warsaw,Poland

4 FaculdadedeCiênciasAgráriaseVeterinárias,Jaboticabal,Departa mentodeBiologiaAplicadaàAgropecuária,UniversidadeEstadual Paulista(UNESP),SãoPaulo,Brazil

formspecifictrapstocatchsmallsoil/waterorganisms(e.g.Re ut1993;Płachnoetal.2007;Fleischmann2012a;Fleischma nnetal.2011,2017,andreferencestherein).

Genliseahavezygomorphicflowersinwhichthechasm ogamousandbilabiatecorollaismadeupoffivefusedpetalswith aspur.Theupperlipiscreatedbythefusionoftwopetalsandthel owerlipbythefusionofthreepetals.Thelowerlipisflat,extende dandformsapalateatthebaseandlaterathroatthathidesthesexu alorgansandextendstothespur(Fromm-

Trinta1979;Fleischmannetal.2011;Fleischmann2012a).

AccordingtoFleischmann(2012a,2012b),thespeciesofG.sub genusGenliseaarecharacterisedbymaskedflowers—Bsnap- dragonblossoms^.Thepalateformsagibbousmask,whichisfir mlyappressedtotheupperlipand blocksaccesstotheentrance to thethroat(corollatube)andnectarspur(Fig.1a–

c).Thereareonlyrareobser-

vationsofthepollinatorsoftheG.subgenusGenlisea;smallbees (familyHalictidae)arethepollinatorsoftheAfrican G.stapfiiandlargerbees(familyMegachilidae)arevisitorstoth eBrazilianG.aurea(Fleischmann2012,2012a).

ThespeciesoftheG.subgenusTayloriahaveadifferentcoro lladesignthantheoneinthesubgenusGenlisea(a

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114

0 B.J.Płachnoetal.

Fig.1G eneralfloralmorphology andpalatemicromorphologyofGe nliseahispidula.aGeneralfloralm orphologyof

G.hispidula,palate(arrow),spur(s );bar=1mm.b,cMicromorpholog yofthepalate,notethenumerousgl andulartri-

chomes;bar=500and200μm.dMi cromorphologyofthepalateinSE M;bar=1mm.e,fAsectionofthepa latewithglandulartri-

chomesandpapillae;bar=100and 40μm.gPartofthesectionthrought hepalatewithaglandu-

lartrichomeandpapillae,notethes ubcuticularspacesoftheheadcells :pedestalcell(arrow),basalcell(B c),head(star);lightmi-

croscopy(LM).Semi-thinsec- tion,stainedwithmethyleneblue, bar=20μm.hPartofthesectionthro ughthefreshpalateshowingaccu mulationsofanthocyaninsinthehe adcellsofthetrichome:pedestalce ll(arrow),basalcell(Bc),head(sta r);bar=20μm

flowerwitha longslendercorollatubewitha narrowedentran cean ddisc-likespreadin gcoroll alobes)

(Fleischmann2012b).Fleischmann(2012a,2012b)sug- gestedthattheflowersofthemembersofthesubgenusTayloria shouldbepollinatedbyinsectswithalongpro-

boscissuchasbutterflies,mothsorbydipteranssuchasbombyli idflies.However,Arangurenetal.

(2017)foundthatG.violaceaflowersweremainlypollinatedb ythebeesLasioglossumsp.(Halictidae)andCeratinasp.

(Apidae)andalsothefliesToxomerus(Syrphidae).Theseflow erswerealsovisitedbysomespeciesofDiptera,Lepidopteraan dHymenoptera.

InUtricularia(thegenusmostrelatedtoGenlisea),therear elargedifferencesbetweenthespeciesof

pollinators.Forexample,speciesthathavesmallflowerss ucha sU.alboc aerule a,U.p urpurasc ensan d

U.reticulata(Hobbhahnetal.2006)werevisitedbyvari- ousinsects:bees,butterflies,mothsandflies,andsomespecies ofHymenoptera,whichwereeffectivepollinators.InUtricula riabremii,smallHymenoptera(Mymaridae,Braconidae)we reobservedasflowervisitors(Płachnoetal.2017a).Thelargef loweredUtriculariaspecies,e.g.

U.reniformis(Clivatietal.2014),U.cornigeraand

U.nelumbifolia(Płachnoetal.2017b),havelarge,strongpollin atorsthathaveeasyaccesstotheirsexualorgansandnectar.Cliv atietal.(2014)observedlargebees(thecar-

penterbeeXylocopasp.andthebumblebeeBombussp.)aspoll inatorsofU.reniformisflowers.

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InUtriculariaflowers,thepalatesarem orphologicallyvery diverse(Taylor1989)andprobablyplayakeyroleinproviding thecolour,mechanicalandolfactorystimulitoat-

tractinsectpollinatorsandtoguidethemtotheirgenerativestru cturesandthespurwithnectar(Płachnoetal.2016,2017a,b).Th epalatesinGenliseaalsoprovidethecoloursig-

nalsforpollinators(Fleischmann2012);however,information aboutthemicro-

morphologyofGenliseapalatesisscarce.Fleischmann(201 2a)observedglandulartrichomesonthepalatesurfaceofGen liseasubglabraandinterpretedthemasnectarglands.

ThisstudyaimstoexaminethestructureofthepalateinGenl iseahispidulaindetailaswellasthepalatefromotherfivespecie sfromthesubgenusGenlisea.Inparticular,itaimstoascertainw hetherthesepalatesfunctionasaunguentarius(Płachnoetal.20 17a;=areaofosmophoresintheflower;Endress1994)orwheth ertheyproducenectartoattractflowervisitors.Itshouldbestresse dthatisinterestingtocomparetheglandulartrichomesfromth eGenliseapalatetotheUtriculariapalateduetotheirclosesyst ematicrelationship.

Materialandmethods

ThespeciesthatwereusedinthisstudyincludeG.hispidulaStapf ,G . subglabraS tapf,G . roraimensisN . E . Br.a n d G . africana Oliv,whichwereobtainedfromthecollectionsofBotanickáz ahradahl.m.Prahy,CzechRepublic;BotanickázahradaLibere c,CzechRepublic,thecollectionofMr.KamilPasek(bestcarniv orousplants.com);andtheBotanicalGardenofJagiellonianUn iversityinKraków,Poland.TheGenlisearepensBenj.andG.p ygmaeaA.St.-

Hil.MaterialwascollectedbyBJPandVFOMfrompopulatio nsthatarelocatedinthesouthernregionintherockyfieldsofthe cerradoofMinasGeraisStateintheSerradaCanastra(south- easternBrazil),permitno.SISBI0#26938.Flowersofthesespec ieswerefixedinthefieldcondition.

Floralstructureandhistochemistry

Thedistributionofthesecretoryglandulartrichomeswasde- terminedbyexaminingwholeflowersusinganOlympusSZX1 6stereoscopicmicroscope(equippedwithanOlympusDP72ca meraandthecellSensStandard1.4program).Floralpartsthatbo reglandulartrichomes,namelythepalateof

G.hispidula,wereexaminedusinglightmicroscopy(LM),scan ningelectronmicroscopy(SEM)andtransmissionelec- tronmicroscopy(TEM)asfollows.First,theepidermisoftheflor alpalatewasexaminedduringanthesisandpiecesofthefloraltis sueswereexcisedandfixedin2.5%glutaraldehydeand2.5%fo rmaldehydeina 0.05M cacodylatebuffer(Sigma)

(pH7.2)overnight(materialfromcultivatedplants)orforsevera ldays(materialfromplantscollectedinthefield

condition),washedthreetimesina0.1Msodiumcacodylatebuff erandpost-fixedina1%osmiumtetroxidesolutionfor

1.5hatroomtemperature.Thiswasfollowedbydehydrationusi ngagradedethanolseriesandinfiltrationandembeddingusinga nepoxyembeddingmediumkit(Fluka).Followingpolymeri sationat60°C,sectionswerecutat70nmforTEMusingaLeicau ltracutUCTultramicrotome,stainedwithuranylacetateandlea dcitrate(Reynolds1963)andexaminedusingaHitachiH500tra nsmissionelectronmicroscopeatanacceleratingvoltageof75k V.

Semi-thinsections(0.9–1.0μmthick)werepreparedfor LMandstainedforgeneralhistologyusingaqueousmethy- leneblue/azureII(MB/AII)for1–

2min(HumphreyandPittman1974)andexaminedwithanOly mpusBX60lightmicroscope.Theperiodicacid-

Schiff(PAS)reactionwasalsousedtorevealthepresenceofinso lublepolysaccharides,andSudanBlackBwasusedtodetectthe presenceoflipids(Jensen1962).Stainingfortotalproteinswasp erformedusingCoomassiebrilliantblueR250orPonceau2R(F isher1968;Ruzin1999).

NikonEclipseNi-

UandOlympusBX60microscopeswereusedforthegeneralph otographyandmicrometry/photomi-crography,respectively.

ForSEM,therepresentativefloralpartsofsixGenliseasp ecieswerefixed(asabove)andlaterdehydratedandsub- jectedtocritical-

pointdryingusingliquidCO2.Theywerethensputter- coatedwithgoldandexaminedatanacceleratingvoltageof20k VusingaHitachiS-

4700scanningelectronmicroscope(Hitachi,Tokyo,Japan),w hichishousedattheInstituteofGeologicalSciences,Jagiello nianUniversityinKraków.

Result s

G.hispidula(Figs.1,2and3)

Thepalatewasprominentwithclearlyvisibletrichomesonthea daxialsurface(Fig.1a–d).Althoughthepalateepider- misformedpapillae(Fig.1d,e),theywerenotontheentiresurfa ceofthepalate.Therewerecapitate,glandulartri-

chomesbetweenthepapillae(Fig.1e–g)

(lengthabout=70μm)consistingofaunicellularbasalcell,ave ryshort,pedestalcell(=barriercell)

(lengthabout=11μm)andaheadcomprisingtwoglandularcell s(diameteroftheheadabout=40μm)

(Fig.1g,h).Thelateralwallofthebasalcellwaspartlyembedde dintheepidermis,butmostofthiscellprotrudedtoformalongst alk,whichhadathickcuticle.However,insomeofthetrichome s,thebasalcellhadamoretrapezoidshape(Fig.1h).Thebasalc ellwashighlyvacuolated,andthevacuolewastranslucentorco

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ntainedpinkanthocyanins.Thecytoplasminthebasalcellfor medathinlayerthatcontainedtheusualorganelles.

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Fig.2U ltrastructureofthepalatetr ichomesofGenliseahispidulafro manimmatureflower.aLongitudi nalsectionshowingtheheadcells(

Hc),pedestalcell(Pc),basalcell(B c),thickenedanticlinalwallofstal kcell(star);bar=2μm.bUltrastruct ureofthepedestal(Pc)andheadcell (Hc):nucleus(N),microbody(Mi) ,dictyosome(d),plasmodesmata(

encircled);bar=0.75μm.c,dUltra structureoftheheadcells:plastids(

P),vacuole(V),mito-

chondria(m),cellwallsbetweenth eterminalcells(cw),cuticle(c);bar

=0.75and1μm

Thepedestalcellhada thickradialwallinwhichcutinisat ionoccurredthatledtoitstransformationintoCasparianstrip -likestructures(Fig.2a).Thesecuticular

depositsofthepedestalcellwerecontinuouswithawell- developedcuticleoftheglandularcells(Fig.2a).Simpleplasm odesmataoccurredinthetransversewallsbetween

Fig.3StructureofthepalatetrichomesofGenliseahispidula.a Longitud inalsectionshowingtheheadcellsoftrichomefromanimmatureflower:n ucleus(N),vacuole(V);bar=2μm.bUltrastructureofthepedestal(Pc)andh eadcells(Hc)ofthetrichomefromamature

flower:nucleus(N);bar=1.7μm.C.Ultrastructureoftheheadcellofthetrich omefromamatureflower:cuticle(c),subcuticularspace(asterisk),vacuol e(V);bar=1.7μm.d,eThestrongauto-fluorescenceofthecuti-

clesoftheheadcellsunderUV;florescencemicroscopy,bars=25μm

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thebasalcellandthepedestalcell(Fig.2a).Therewerebranche dplasmodesmatainthetransversewallsbetweenthepedestalc ellandtheheadcells.

Thereweretwoglandularheadcells.Thecytoplasmofthehea dcellswasmostlyconcentratedtowardsitsbaseandradialwalls(

Figs.2aand3a,b).Aprominentnucleuswaslocalisedhere.Ther ewerenumerousmitochondria,microbodiesandsmalldictyos omesinthecytoplasm(Fig.2b–

d).Therewerenumerousplastidsthatcontainedsmallplastoglo buliandla-

mellaewithosmiophilicinclusions.Althoughsmalllipiddropl etswerevisibleinthecytoplasmoftheheadcells,theywerenotfr equent(notshown).Themostconspicuouscharac-

teroftheheadcellwasareticulatevacuolarsystem(Figs.2a–

dand3a–c).Analysisofultra-

thinsectionssuggestedthatitwasdividedintovarioussmallbuti nterconnectedparts(e.g.Fig.3d).Vacuolisationincreasedduri ngflowerdevelopment(immatureflowerFig.2a;matureflower Fig.3b,c).Thecu-

ticleoftheheadcellwasthick(meanthickness=0.225μm;Figs.

2cand3c)andcontainedmicrochannels.Thesubcuticulars paceswereformedbytheseparationofthecu-

ticlefromthecellwall(Fig.3c).Thefoldedcuticlewasalsoclearl yvisibleinSEM(Fig.1f),aswellwasclearlyvisibleinthelivingtr ichomes(Fig.1h).Thecuticlesoftheheadcellsemittedastronga uto-

fluorescenceunderUV(Fig.3d,e),andthismightreflectdiffere ntialimpregnationofcellwallswithcutinandothermaterialsbe tweenglandularcellsandothercelltypes.Cytochemicaltestsdi dnotshowthatthecellheadproducedinsolublepolysaccharides orproteinproducts(Sup.Material1).Treatmentforlipidsstaine dtheplastidsandcuti-

cleofheadcellsandlateralwallofpedestalcell(notshown).

Otherspecies(Fig.4a–f)

Inthepalatesofalloftheotherspecies(G.subglabra,

G.roraimensis,G.africana,G.repens(Fig.4a)andGpygmaea(Fi g.4d))thatwereexamined,theadaxialepidermalsurfaceconsist edofconicalpapillae(Fig.4b,c,e).Therewerecapitate,glandulartr ichomesbetweenthepapillae(Fig.4c,e,f).Eachtrichomeconsist edofabasalcell,apedestalcellandamulti-

celledhead.ComparedtoG.hispidula,theotherspeciesthatwere examinedhadamoreroundedheadshape(Fig.4c,e,f).

Discussion

Fleischmann(2012a)interpretedtheglandulartrichomesonpa latesurfaceofG.subglabraas nectar glands.However, invivononectarwasobservedonthesetrichomesinthis speciesoronthepalatetrichomesoftheotherspeciesthatwere examinedinthisstudy.Also,PASreactionresultsdidnotreveal thatthesehairsproducepolysaccharidesecretion.Wealsodidn

otfindanyprooftoconfirmFleischmann’ssug-

gestion.Thesetrichomesdidnotproducepolysaccharidesor

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proteinproducts.Themicro-

morphologyoftheGenliseaviolaceapalatewasanalysedrec ently(Arangurenetal.2018).However,thisspeciesfromthes ubgenusTayloriadidnothavecapitatetrichomesonitspalat e.FuturestudiesofotherspeciesofthesubgenusTayloriasho uldrevealwhetherthereisanystructuralpatternofpalatemo rphologyinthissubgenus.Weshouldmentionthathere,weex aminedinde-

tailsonlyG.hispidulatrichomes(TEMdata);thus,otherspe- ciesshouldbestudiedincaseofultrastructureinthefuture.Thi scouldincreasevalueofthecomparativeanalysis.

Stalkedglandulartrichomeswithabicellularhead,whicha resimilartothepalatetrichomesofG.hispidulaand

G.subglabra,weredescribedfromthepalateofUtriculariaco rnigeraandU.nelumbifolia(Płachnoetal.2017b).Howeve r,thesetrichomesdidnothaveafoldedcuticleofthehead cells,whichwasobservedintheGenliseatrichomes.Inourprev iousstudies,wedescribedtheultrastructureoftheshort- stalkedorsessileglandulartrichomesfromanareaoftheosmoph ores(unguentarius)inUtriculariadunlopii(Płachnoetal.201 6),U.bremiiandU.minor(Płachnoetal.2017a).Insomecharac ters,thesetrichomesaresimilartothepalatetri-

chomesofGenlisea.Thismightsuggestthattheyhaveasim- ilarfunction.Forexample,plastidsthathavelipidglobulesofth eheadcellsofG.hispidulaweresimilartothosethatwerefoundi nU. dunlopiiandU. bremii.Moreover,inthetri-

chomesofthesethreespecies,TEMobservationsrevealed thatthecuticlebecamedistendedandseparatedfromthecellw allsoftheheadcellsandformedasubcuticularspace.Aleuco plastthatcontainedlamellaewithosmiophilicinclusionswasd escribedfromthesecretorycellofthetrichomesthatproduce phenolsandterpenoidsinTussilagofarfaraandotherAsterace aespecies(Muravniketal.2016,andliteraturetherein).Mura vniketal.(2016)suggestedthattheblackde-

positioninthelamellaecorrespondtoterpeneprecursors.

IntheheadcellsofGenlisea,thetrichomeswerenumerous microbodies.Itisbelievedthatmicrobodiesactivelypartici- pateintheproductionofsecondarymetabolitessuchasterpe- noidsandphenolicsubstances(e.g.Vassilyev2000;Muravni ketal.2016).Thelipidbodiesinthecytoplasmand/orplastidsw ithlipidglobuleswerefrequentlyrecordedinosmophoretiss ues(e.g.PridgeonandStern1983;Sternetal.1987;Vogel199 0;Ascensaoetal.2005;Płachnoetal.2010;Antońetal.2012).

AlthoughthetissueoftheUtriculariadunlopiiungue ntariuswasrichinstarchgrains,whichwassimilartothestarch thatwasrecordedintheparenchymacellsofthe

U.bremiipalate,thisisincontrasttotheG.hispidulapalate,inwhi chwedidnotobservedanystarch.Intheglandulartri- chomesfromtheareaoftheosmophoresfromU.dunlopii(Pł achnoetal.2016)and thepalateofU.bremii(Płachnoetal.2 017a),boththebarriercellandheadcellsweretransfercells(th eoccurrenceofcellwallingrowths).Inthe

G.hispidulapalatetrichomes,wedidnotfindanytransfer

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Fig.4M orphologyofGenliseaflo wers.aFloralmorphologyofGenl isearepensinthesouthernregiono fMinasGeraisStateintheSerrada Canastra.bMorphologyofthelow erlipofa

G.repensflower;bar=1mm.cAsect ionoftheG.repenspalatewithaglan dulartrichomeandpapillae;bar=20 μm.dFloralmorphologyofGenlise a

pygmaeainthesouthernregionof MinasGeraisStateintheSerradaC anastra.eAsectionofthe G.pygmaeapalatewitha glandulartrichomeandpapillae;

bar=50μm.fGlandulartrichome frompalateof

G.africana;bar=20μm

cells.However,cellwallingrowthswererecordedinthecellsoft hetrichomesthatoccurinGenliseatraps(Heslop-

Harrison1976;Płachnoetal.2007).

Themostunusualcharacteroftheheadcellsofthepalatetri chomesinG.hispidulawasthespecificvacuolisation.Alth oughavacuolewithsomecytoplasmicbridgeswasobservedin theheadcellsofUtriculariadunlopiitrichomes(seeFig.3ainP łachnoetal.2016).Observationsofultra-

thinsectionssuggestedthatthevac-

uoleinGenliseawasextremelydividedintovarioussmallbutin terconnectedparts(e.g.Fig.3c).However,theanal-

ysisofseriesofTEMsectionsandpreparationof3Dreconstru ctionhavetobedonetoproofwhetherthesesmallvacuolesarer eallyinterconnected.Suchareticulatevacuolemaybea charac terofcelldevelopment,e.g.inthecellsofthebarleyrootmeriste m(Guilliermond1941)

oritmaybea symptomofspecialcellactivity.LazzaroandTho mson(1996)describeda complicatedvacuolar-

tubularsysteminthecellsofCicerarietinumtrichomes.Thesea uthorsproposedthatthevacuolar-

tubularsysteminthesetrichomesfunctionstorapidlydelivers olutefromthebaseofthetrichometothesecretoryheadcells.A specificprocesscalledaggregationwasdescribedincar- nivorousplantglands(Juniperetal.1989;Peroutkaetal.2008,a ndliteraturetherein).Afterthechemicalandme-

chanicalstimulationoftheglands,alargecentralvacuolewascl eavedintovarioussmallbutinterconnectedpartsbythecytopla smintheglandcells.Althoughtherearevar-

ioushypothesesaboutthefunctionofthisprocess,thesechange sprovideevidenceofa highlevelofactivityofthesecells.Thus, wealsobelievethattheoccurrenceofacomplexvacuolarsyste mintheheadcellsofGenlisea

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trichomesisconnectedwitha highlevelofmetabolicactivity(

productionandsecretion).

Conclusion

Basedonourmorphologicalandultrastructuralobservations,w esuggestth atthepalateinGenliseafromthesubgenusGenlis eamayfunctionasanareaofosmophoresintheflowerthatprovid eanolfactorystimulusforpollinators.

AcknowledgementsWededicateourpapertoProf.Dr.StefanPorembski(

UniversityofRostock),whohasbeeninvolvedinstudyingLentibular iaceaeformanyyears.ThestayofBJPinBrazilwasfinancedbyUniversidad eEstadualPaulista.Someofequipmentthatwasusedinthisstudywasfunde dbyNationalScienceCentre,Poland.contractgrantnumberDEC- 2013/09/B/NZ8/03308.Theauthorswouldliketoexpresstheirsincereand warmthankstoDr.MiroslavStudnička(directorofLiberecBotanicalG arden,CzechRepublic),Dr.VlastikRybka(PragueBotanicalGarden,Cze chRep.),andDr.LubomirAdamecandKamilPasekforprovidingsomeof theplantmaterialforthestudy.WealsothankhorticulturistLucynaKurlet oforherconscientiouscareoftheliv-

ingcollectionofcarnivorousplantsthatarelocatedintheBotani calGardenofJagiellonianUniversityinKraków.

Compliancew ithe thicalstandards

Conflictof interestTheauthorsdeclaret hattheyh aven o conflictofinterest.

OpenAccessThisarticleisdistributedunderthetermsoftheCreativeCo m mo nsAt tribu t ion4. 0In t ernat iona lLicen se(ht tp://creativecommo ns.org/licenses/by/4.0/),whichpermitsunrestricteduse,distribution,and reproductioninanymedium,providedyougiveappropriatecredittotheori ginalauthor(s)andthesource,providealinktotheCreativeCommonslicens e,andindicateifchangesweremade.

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