Rev. bras. farmacogn. vol.27 número2

RevistaBrasileiradeFarmacognosia27(2017)158–161

w ww.e l s e v i e r . c o m / l o c a t e / b j p

Original

Article

Leaf

venation

pattern

to

recognize

austral

South

American

medicinal

species

of

“cow’s

hoof”

(

Bauhinia

L.,

Fabaceae)

Renée

H.

Fortunato

_,

_Beatriz

_G.

_Varela

_,

_María

_A.

_Castro

_,

_María

_J.

_Nores

a_{InstitutodeRecursosBiológicos(InstitutoNacionaldeTecnologíaAgropecuaria–ConsejoNacionaldeInvestigacionesCientíficasyTécnicas)andFacultaddeAgronomíayCiencias}

Agroalimentarias,UniversidaddeMorón,BuenosAires,Argentina

b_{FacultaddeFarmaciayBioquímica,UniversidadNacionaldeBuenosAires,BuenosAires,Argentina} c_{FacultaddeCienciasExactasyNaturales,UniversidadNacionaldeBuenosAires,BuenosAires,Argentina}

d_{InstitutoMultidisciplinariodeBiologíaVegetal(UniversidadNacionaldeCórdoba–ConsejoNacionaldeInvestigacionesCientíficasyTécnicas)andFacultaddeCienciasExactas,}

FísicasyNaturales,UniversidadNacionaldeCórdoba,Córdoba,Argentina

a

r

t

i

c

l

e

i

n

f

o

Articlehistory:

Received6September2016 Accepted28October2016 Availableonline8December2016

Keywords:

Areolation

Botanicalidentification Freelyendingveinlets Leafvenationpattern Qualitycontrol

Schnella

a

b

s

t

r

a

c

t

TheleavesextractsofsomespeciesofBauhiniaL.s.l.areconsumedtotreatdiabetes,inflammation, painsandseveraldisordersintraditionalmedicineinaustralSouthAmerica.Despiteitswideuseand commercialization,saleisnotcontrolled,andbotanicalqualityofsamplesisnotalwaysadequatebecause ofplantmisidentificationandadulteration.Here,wecharacterizedleafveinpatterninnineteentaxato contributetotherecognitionandcommercialqualitycontrolofplantmaterialcommerciallyavailable. Theveincharactersintercostaltertiaryandquinternaryveinfabric,areoledevelopmentandshape,free endingveinletbranchingandmarginalultimatevenationallowedtodistinguishthemainmedicinal speciesintheregion.

©2016SociedadeBrasileiradeFarmacognosia.PublishedbyElsevierEditoraLtda.Thisisanopen accessarticleundertheCCBY-NC-NDlicense(http://creativecommons.org/licenses/by-nc-nd/4.0/).

Introduction

TheleavesextractsofsomespeciesofBauhiniaL.,Fabaceae,so calledcow’shoof,“pata-de–vaca”,cow’spawororchidtrees,are consumedtotreatdiabetes,inflammation,painsandseveral dis-ordersintraditionalmedicineinaustralSouthAmerica(Oliveira and Saito, 1989; Bortoluzzi et al., 2006; Barboza et al., 2009; Pizzioloetal.,2011;Fortunato,2012).With300speciesworldwide,

Bauhinias.l.isrepresentedbyfifteenspeciesinArgentina,Paraguay, UruguayandSantaCatarina,ParanáandRioGrandedoSulstates ofBrazil(Fortunato,1986,1996;Fortunatoetal.,2008;Vazetal., 2010).

Bauhiniabauhinioides(Mart.)J.F.Macbr.,B.microstachya(Raddi) J.F.Macbr.,B.ungulataL.,B.holophylla(Bong.)Steud.,B.forficata

Link, B.cheilantha (Bong.)Steud. and B. rufa (Bong.) Steud.are themainspeciesforwhichpharmacologicalactivitieshavebeen reported.Hypoglycemic,antioxidant,analgesic,antiinflammatory andlarvicidalactivitiesalongwithantitumorpotentialityletthem auspicioustargetsfornewbioactivesubstancesresearchingand phytotherapy(SilvaandCechinelFilho,2002;Sousaetal.,2004;

∗ Correspondingauthor.

E-mail:jnores@imbiv.unc.edu.ar(M.J.Nores).

Gadottietal.,2005;Lunaetal.,2005;Nakahataetal.,2006;Cechinel Filho,2009;Paulaetal.,2014;Rozzaetal.,2015;Martins-Olivera etal.,2016).

Thesetaxaareextensivelyemployedandsoldinnative Amer-ican,ruralandurbancommunitiesintheregionandsurrounding areas(Bortoluzzietal.,2006;Barbozaetal.,2009;Hurrelletal., 2011;IbarrolaandDegendeArrúa,2011).Despiteitswideuseand commercialization,saleisnotcontrolled,andbotanicalqualityof samplesisnotalwaysadequatebecauseofplantmisidentification andadulteration.Commercialsamples–crudeherbsor industrial-izedherbalmedicines–includeentireorbrokendriedleaves,stems andoftenpods.Ingeneral,plantsareharvestedfromtheirnatural habitats,beingpopularlyrecognizedbytheirbilobedorbifoliolate leaves.Speciesaredifficulttoidentify,especiallyfromvegetative stagesorinfragmentedmaterial,andsamplelabelingandtrading aremostlyaccomplishedbyusingcommonnames,genericname orincorrectnames(Meloetal.,2004).Theseoftenresultin adul-terated,falsifiedorsubstitutesamples,whichwouldinterferewith theeffectivenessandeventhesafetyoftheproduct(Ferreresetal., 2012).Leaf anatomicalfeatures areusefulforidentificationand adequatesamplebotanicalquality,inBauhinias.l.,venationpattern hasbeenutilizedintaxonomy,phylogeneticanalysesandevenin palaeobotany(Vaz,1979;Fortunato,1986;Zhang,1994;Linetal., 2015).

http://dx.doi.org/10.1016/j.bjp.2016.10.007

R.H.Fortunatoetal./RevistaBrasileiradeFarmacognosia27(2017)158–161 159

Fig.1.LeafvenationinsouthernSouthAmericanBauhinia.(AandB)Generalveinpattern.(C–G,I–P,RandS)Areolation.(HandQ)Loopedmarginalultimatevenation.(A)B. bauhinioides.(B)B.hagenbeckii.(C)Bauhiniaaffinis.(D)B.angulosa.(E)B.argentinensis.(F)B.bauhinioides.(GandH)B.campestris.(I)B.cheilantha.(J)B.forficatassp.pruinosa. (K)B.hagenbeckii.(L)B.holophylla.(M)B.microstachya.(N)B.mollisvar.mollis.(O)B.mollisvar.notophila.(PandQ)B.rufa.(R)B.ungulatavar.ungulata.(S)B.uruguayensis. Arrowhead:basalveins.Arrow:agroaphicveins.Scalebar:AandB:3mm;C–G,I–P,RandS:300␮m;H,Q:1mm.

Inthis work,wecontributetotherecognitionand commer-cialqualitycontrolofaustralSouthAmericanmedicinalBauhinia

throughastudyofleafveinpattern.

Materialsandmethods

Weanalyzed19australSouthAmericantaxa(Appendix1).The studyofleafvenationpatternwascarriedoutfollowingEllisetal. (2009).For this,freshorherbariumleaveswerecleared accord-ingtoDizeo deStrittmatter (1973)and stainedinsafranin/80% ethanol.SamplesweremountedinDPX(AldrichChemical Com-pany,Gillingham,UK)andexaminedusingZeissStereoMicroscope aZeissAxiolablightmicroscope(CarlZeiss,Germany)witha trinoc-ularphototubeandacolordigitalimagingcamera.Somevenation datawereextractedfrompreviouswork(Fortunato,1986,1996).

Resultsanddiscussion

Alltaxa analyzedpresent basal actinodromous primary vein framework, withnonaked basal veins,3–5(7) basal veins per lobeorleafletandagrophicveinssimple (Fig.1A andB). Previ-ously,someofthese taxahave beendescribed asacrodromous (Fortunato, 1986, 1996) or campylodromous (Vaz, 1979), and recentlybothAmericanBauhinias.s.andSchnellaareconsidered actinodromous or acrodromous by Lin et al. (2015).Main vein characters(Fig.1,Table1)provideacriteriontoidentificationof medicinalspecies anddetectionoffalsificationsor adulteration.

Theareoledevelopmentallowsdifferentiatingthreemaingroups, someofthem,butnotall,taxonomicallyrelated.

Thefirstgroupischaracterizedbyagoodareoledevelopment that is foundin some medicinalmembers of sect.Pauletia ser. Cansenia,inthelianaBauhiniaangulosa(=S.angulosa)andinthe spinytreeB.uruguayensis(Fig.1D,L,P–S).ThemedicinalB. holo-phyllaandB.rufabothdisplayquadrangularandpentagonal,rarely triangularareoles,withabsentandunbranchedfreelyending vein-lets (FEV),the firstone canbe furtherrecognizedsince FEVin generalcrosstheareolaandmarginalultimatevenationand quin-ternaryveinfabricvenationareabsent(Fig.1L).B.rufa,instead, presentsloopedmarginalultimatevenationandquinternaryvein fabricpresent(Fig.1PandQ).ThemedicinalB.ungulatahasabsent, unbranchedandonebranchedFEVandareolashapemore vari-able(Fig.1R).Somesimilarfeatureshavebeendescribedinothers taxafromser.Cansenia,andZhang(1994)nameditasthe Canse-niavenationtype.B.angulosaiseasilydistinguishedbecauseofthe verysmallareoleslackingFEVandmarginalultimateveinabsent (Fig.1D).B.uruguayensisistheonlytaxonthatpresentspaxillate areoledevelopmentwithoutFEV,orjustveryfew FEV(Fig.1S), thesefeaturesaresimilartotheElayunatypedescribedbyZhang (1994).

160 R.H.Fortunatoetal./RevistaBrasileiradeFarmacognosia27(2017)158–161

Table1

LeafvenationpatterninaustralSouthAmericanBauhinia.FEV,freelyendingveinlets;P,pentagonal;PE,percurrent;Q,quadrangular;R,reticulate;T,triangular.

Species Intercostal tertiaryvein fabric Quinternary veinfabric Areole development

Freeendingveinlet (FEV)branching

Areoleshape Marginal ultimate venation

Fig.1

B.affinis PE Present Moderate Unbranchedandoneor twobranched;rarely absent

variable Absent C

B.amambayensis Ratapex,PEat base

Present Moderate Unbranchedandwith oneormorebranches; sometimesabsent

variable Absent

B.angulosa(=S. angulosa)

Ratapex,PEat base

Absent Good Absent;rarely

unbranched

QandP,rarelyT Absent D

B.argentinensis Ratapex,PEat base

Present Moderate Mostlywithoneor morebranches; sometimesunbranched

Variable Absent E

B.bauhinioides R Absent,topoor distinct

Moderateto good

Mostlyunbranched andonebranched; rarelytwobranchedor absent

Variable Absent A,F

B.campestris PEtoR Absent Moderate, rarelygood

Mostlywithoneor morebranches; sometimes unbranched;rarely absent

Variable Looped GandH

B.cheilantha PE Absent Goodto moderate

Absent,unbranched andonebranched; rarelytwobranched.

Variable Absent I

B.forficata PE,notablyat base

Present Moderate Mostlywithoneor morebranches; sometimesunbranched

Variable Absent J

B.hagenbeckii Ratapex,PEat base

Present Moderate Mostlywithoneand morebranches;rarely unbranched

Variable Absent B,K

B.holophylla PE Absent Good Absentand unbranched(FEVs mostlycrossing areoles);rarelyone branched

QandP,rarelyT Absent L

B.microstachya(=S. microstachya)

Ratapex,PEat base

Absent,topoor distinct

Moderate Mostlyunbranched andonebranched; sometimeswithmore branches

Variable Loopedand absent

M

B.mollisvar.mollis PEtoR Present Moderate Mostlyunbranched andonebranched; sometimeswithmore branchesorabsent

Variable Absent N

B.mollisvar.

notophila

PE Poordistinct Moderate Mostlyunbranched

andwithoneormore branches;rarelyabsent

Variable Absent O

B.rufa PE Present Good Absentand

unbranched

QandP,rarelyT Looped PandQ

B.ungulatavar.

cuiabensis

PEtoRatapex Absent Good Absent,unbranched andonebranched

VariabletoQ,P,T Loopedand absent

B.ungulatavar.

ungulata

PEtoRatapex Absent,topoor distinct

Good Absent,unbranched andonebranched

VariabletoQ,P,T Looped R

B.uruguayensis Ratapex,PEat base

Present Good/paxillate Absent;rarely unbranchedandone branched

QandP,rarelyT Loopedand inpart absent

S

group. The medicinal B. forficata (both subspecies), B. argenti-nensis and B. hagenbeckii show almost the same pattern, with mostlybranchingFEV(oneormorebranches)andlessfrequently, unbranchedFEV(Fig.1J,E,K).B.forficatacanbedistinguishedby thepercurrenttertiaryveinfabric,whiletheothersarepercurrent atthebaseand reticulateat apex.B. affinis(Fig.1C), B. amam-bayensis(notshown)andB.mollis(Fig.1N,O)haveingeneralboth unbranchedandbranchedFEV,indifferentfrequency.The medici-nallianaB.microstachya(=S.microstachya)possessesalsomoderate areoledevelopment,mostlyunbranchedandonebranchedFEV,but isrecognizedbecauseofthepresenceofbothloopedandabsent marginalultimatevenation(Fig.1M),thisreportissimilartothe onebyFortunato(1986)butdiffersfromprevious(Zhang,1994).

Finally, the medicinal B. cheilantha and B. campestris (sect.

Pauletiaser.Cansenia)presentareoledevelopmentintermediate between good and moderate. B. cheilantha alternates between absent,unbranchedandonebranchedFEVandultimatevenation absentwhileB.campestriscanbedistinguishedbecauseFEVare in generalbranchedandultimatevenation is looped(Fig.1I,G andH).ThemedicinalB.bauhinioides(sect.Pauletiaser.Perlebia) alsopossessareoledevelopmentmoderatetogoodbutareolesare mostlyunbranchedandonebranched(Fig.1F).

Interestingly,allspeciespresentprismaticcrystalsofcalcium oxalateassociatedtovascularbundlesandoftendrusesin meso-phyllcells,morenotableinB.affinis,B.argentinensis,B.mollisand

R.H.Fortunatoetal./RevistaBrasileiradeFarmacognosia27(2017)158–161 161

Hereweprovideahelpfultoolfortherecognitionofthemain regionalmedicinalplantsthroughtheanalysisoftheleafvein pat-ternbasedontheintercostaltertiaryandquinternaryveinfabric, areoledevelopmentandshape,freeendingveinletbranchingand marginalultimatevenationcharacters.

Authors’contributions

RHF collected and identified the plants, performed the lab-oratory work and data analysis and participated in the results discussionandthemanuscriptfinalwriting.MACparticipatedin thelaboratoryworkanddataanalysis.BGVhelped performthe analysiswithconstructivediscussions.MJNdraftedthemanuscript.

Conflictsofinterest

Theauthorsdeclarenoconflictsofinterest.

Acknowledgements

We thank Cristo S. for technical assistance. CONICET (PIP 112-201101-00250), INTA (PNHFA-1106094) and SECyT-UNC (32720160200161CB)areacknowledgedforfinancialsupport.

AppendixA. Supplementarydata

Supplementarydataassociatedwiththisarticlecanbefound,in theonlineversion,atdoi:10.1016/j.bjp.2016.10.007.

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