Table Of Content,
Rodriguésia 61(3):505-518.2010
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Leaf anatomy and micromorphology of six
Posoquería Aublet species (Rubiaceae)
AnatomiaemicromorfologiafoliardeseisespéciesdePosoqueriaAublet(Rubiaceael
RosanidoCarmode OliveiraArruda Dona MariaSaiterGomes2
Aline CarvalhodeAzevedo'3 MichelleLima Magalhães ' & Mario Gomes4
,
Abstract
ThepresentstudydealswiththeleafanatomyandleafsurfaceofPosoqueríaaaitifoliaMart.,P. latifoliaMart.,P.
longiflora Aublet,P. macropusMait,P.palustrís(Rudge) Roem. and Posoqueriasp„ collectcd in ffagments of
Atlanticrainforest,RiodeJaneiro,Brazil.Theepicuticularwaxmayoccurintheformoffilaments,granulesorcrusts.
Theleavesarecoveredbyathickcuticularlayerthatmaybesmoothorstriatcd.Paracyticstomata,andnon-glandular
trichomesarelimitedtotheabaxialsurface;thelattcrarenumerousinP.palustrís,andrareinP. longifloraandP.
latifolia. Leaveshaveadorsiventralstructure,withonlyonelayerofpalisadeparenchymaandvaricdamountsof
spongyparenchyma.IdioblasLscontainingcrystalliferoussandwcreobserved,andweremoreabundantinP.latifolia.
Theleafbladevascularsystemisformedbycollateralbundlcswithaparenchymatoussheath,associatedwithfibers.
Thevascularsystemofthepetioleandtheleafbladefomtsanarch.SomeofÜtcanatomicalfcaturesobscrvcdcanbe
uscdtodistinguishthespeciesstudied.Anatomicalleafcharactcrscouldbeusedintherccognitionofsixspeciesof
Posoqueriastudied,suchasanticlinalwallofepidemia!cells,waxdeposition,trichomesandshapeoftheleafntargin.
Kcy words: leafanatomy, scanningclcctron microscopy,taxonomy, Cinchonoidcae.
Resumo
S3oapresentadasinfomtaçòcssobreaanatomiacasuperfíciefoliardePosoqtvriaaaitifoliaMart.,P.latifoliaMart.,P.
longifloraAublet,P.macropusMart.P.palustrís(Rudge)RoemePosoqueriasp..espéciesocorrentesanfragmentosda
FlorestaPluvialAtlântica,RiodeJaneiro,Brasil.Aceraepicuticularpodeocotrernaformadefilamentos,grânulosou
crostas Asfolhassãorecobertasporespessacamadaarticular,quepodeserlisaouestriada.Nafaceabaxialacutícula
apresenta-seestriadaemP. latifoliaePpalustrís.Osestômatosestãorestritosàfaceabaxialc sãodotipoparacítico.
TricomastectoresocorremapenasnestafacesendonumerososcmP.palustrís rarosemP.longifloraeP. latifolia.As
folhastêmestruturadorsiventral,comapenastunacamadadeparènquimapaliçádicocvariadonúmeronoparênquima
esponjoso Nomesofiloforamobservadosidioblastoscontendoareiacristalífera,maisabundantescmP.latifolia.Na
lâmina foliaro sistema vascularé formado por feixesdo tipocolateral, envolvidosporbainhaparenquimâtica, e
apresentamfibrasassociadas.Nopcciolocnanervuraprincipalosistemavascularapresenta-secmarco.Características
anatômicasfoliarespodemserusadasnadistinçãodasseisespéciesdePosoqueriaestudadas,taiscomo:aparede
anticlinaldascélulasepidérmicas,deposiçãodecera.tricomaseaformadamargemdafolha.
Palavra-chave: anatomia foliar, microscopia eletrônica de varredura, taxonomia. Cinchonoidcae.
recently divided into two subfamilies, lhe
Introduction &
Cinchonoideae and the Rubioidcac (Robbrecht
Rubiaceae (Gentianales) is one of the four
Manen2006).
iargestfamiliesofangiosperms,withrepresentatives Belonging to Cinchonoidcae subfamily,
hawainbddietlisynacdnliudsdtelrisifebaufbtoeordumtsw.o1rT0l.hd7ew0i0fdaesm,pielpcyrieeissse,mnotwinhnoigpchhdyilvweeetrrisece aPnodsoiqsuceortínapAousbeldeotfisapfporuonxdiimnattheelyNco14trsoppeicciersegtihoant
1UniversidadeFederaldoEstadodoRiodejaneiro(UNIRJO).InstitutodeBioeiíncias.Dep.o,Boliiniea.Av.Pasteur458,22290-540.RiodeJaneiro.RJ.Autorapara
correspondência:rosaniarrudaáTgmail.com
:UniversidadcFederalRuraldoRiodeJaneiro(UFRJU).InstitutodeBiologia.DeptO.Botânica.Rod.465.km7,25890-000.Scropídica.RJ.
IniciavàoCientífica.
4ExtractaMoléculasNaturaisS.A.,Av.CarlosChagasFilho791.CidadeUniversitária,21941-904,RiodeJaneiro.RJ.
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aredifficulttoberecognizedinvegetativeStateand (Bidegain & Michael 2003). In this site, the plants
presentgreatmoiphologicalsimilarity(Macias 1988). are submitted to seasonal flooding in the rainy
ToBrazil,sixspeciesaredescribed(Macias 1988); season, when the water table rises considerably.
mostofthemareendangered(Leonhardtetal.2008). Posoqueria acutifolia was collected at the Parque
Some speciesofthis group are used as omaments, Estadual da Serra da Tiririca, Niterói, RJ
because of their decorative leaves and perfumed (22°48’S,42°57’W),afragmentoftheAtlanticRain
flowers. Most species of Posoqueria display the Forest.Inthisregiontheannualaveragetemperature
characteristic pollen catapult mechanism, which is is24 C,withannualrainfaliof1300mmconcentred
thetrademarkfeatureofthePosoquerieae(Delprete in the summer. Herborized leaves ofP. macropus
2009). According to Ariza et al. (2007), indolic (Sucre7678)andPosoqueriasp.(Martinelli8804),
alkaloids,iridoids,andacetatesofhexanileandhexile, obtained from the Instituto de Pesquisas Jardim
among other compounds, have been identified as Botânico do Riode Janeiro (RB), were rehydrated
perfume sources in Posoqueria latifolia (Rudge) accordingtoGomes(2002).
Roem.,popularlyknownas“açucena-do-mato”.For The fresh leaves were fixed in F.A.A.
P. acutifolia Mart., laboratory tests have identified (formalin, acetic acid, ethanol 70%) for48h and
analgesic and anti-inflammatory activities of the preserved in 70% ethanol (Johansen 1940). For
methanolic extracts, which may be related to this light microscopy, sections of the leafblade and
speciesuseinpopularmedicine(Souzaetal.2007). Petioleweredehydratedingradedethanol-butanol
Among the organs that can be useful to series, embedded in paraffin, sectioned
taxonomic propositions, the leaf is the most transversely and longitudinally in rotatory
examined onebecause ofits internai architecture, microtomeat athicknessof 12 mm. The sections
which is a classic source ofuseful information in were stained with 1% safranin - 1% astra blue in
the systematics, specially the epidermis and the tartaricacid(Luqueetal. 1996).
cuticle(Stuessy 1990;Juddetal. 2009). Theepidermis wasstudied usingsectionsof
MantoAvbanoiut&mViiecirroam(o19r9p3h/o1l9o9g7)yatnhdeKosctusdiiseestaolf. tpheerolxeiadfeb(lFardaenkdliisnso1c9i4a5t)edThieneapceitdiecrmaicsidw/ahsysdtraoigneend
(2004) stand out and are source of important with 1% safranin, alcian blue, or 0.5% fuchsin
informationabouttheleafsurfaceofdiversespecies (modificationsinKraus&Arduin 1997).Inorderto
ofthisfamily.Descriptiveanatômica]studiesofthe remove wax and to confirm its presence, sections
vegetative or reproductive parts also have been ofherborizedleaveswereboiledinchlorofonmthree
&c(aMrGarnoitemTodehvsioasun1ti9s9etw6tui.adtlyhD.ed1oet9Tsh9oce5nrr;iibGB&erosaMmzaetirhlsieiaaetlntehaalr2f.u0b0a2i80no)0ai.0td;osmApyrercauindedas ewtlieemrceetsrcforonirtoimcniaeclr-mpoiosniucntotepd,yroi(nedSaEthhMort)opulaganhtael.aysFgiorsra,dtehldeeassfcearpinienesicneogsf
micromorphology of six Brazilian Posoqueria ethyl alcohols. Leaf fragments were mounted on
rtMRsaoeporecetcoim.igd,e.nesiPn:&.ztipPiSna.fclgyhuautsclthfutreet.isia,seftPoMus.larpirleeatocs.niM,geatisarh.fntal.dot,rPaomPs.Aaouqyblault.eib,rfeioPla.iuamsspa.(ecRfaruuioldmpgiuenis)gn JcfsotrSuaoMbtm-se5dhFc3ewoori1rab0tttahehsrdegcioahuwlnimidnstitwbhnoeagcfsghoeoerlalmeeldcisbtcoeraaomilnnnoditgucesnoomtbbtsiss,eceerdfrrorvovseenecedodhppaireinnon.dtphMseaeaertcJSestEErtiiuoOManb.Llss
of leaf blade were treated with the following
Materials and Methods reagents: starch grains were localized by Lugol
staining(Johansen 1940), lipidiccompoundswere
Adult leaves of Posoqueria latifolia and P. recognized by Sudan Red 7B in glicerol/ethanol
longiflora were collected from plants found at the (Kraus & Arduin 1997), pectin substances with
ParqueNacionaldaFlorestadaTijuca(22°57’Sand 0.02% ruthenium red (Kraus & Arduin 1997);
43°18’W),inRiodeJaneiro,(RJ).Inthisregion,the primary walls were identified with zinc iodine
annualavaragetemperaturevariesbetween 19.3°Cin chloride(Jensen 1962)andthesecondarywallswith
thewinteranmdm25.5°Cinthesummer,andtherainfallis acidphloroglucinforlignifiedcells(Foster 1949);
about2000 annually(Vieira 1994).Posoqueria phenolic compounds were confirmed with ferric
palustriswascollectedinRiodasOstras(RJ,22°31S'- chloride(Johansen 1940).Testswithhydrochloric
41°55'W)wheretheannualaveragetemperatureis acid were used to detect calcium oxalate crystals
22°Candtheannualrainfallbetween1500and2000mm (Jensen 1962).
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Leafanatomyandmicromorphology ofPosoqueriaAublet (Rubiaceae)
cuticular-wax strata (Fig 5 a-d). On the abaxial
Results
surface, above the stomatal pore, the cuticle layer
Inmostofthestudiedspccies,theepidemiis, mayformconspicuousledges(Fig.5c).Thestomata
inadaxialsurfaceiscomposedofordinaryepidcrmal arelocatedon thesame leveiorslightlybelow the
cells,withsinuousanticlinalwalls(Fig. 1 a-f).and other epidermal cell and the subsidiary cells can
thick, notably in Posoqueria macropus(Fig. 1 d). form projections in the rcgion ofthe substomatal
Overvascularbundles,suchasintheabaxialsurface chamber(Fig.5c).Thetrichomeshavethickwalls
ofPosoqueria sp. the anticlinal walls can present (Fig. 5 b), and in Posoqueriapalustris, the basal
themselvesstraightorcurved(Fig. 1 f).Theabaxial regionoftheseptatetrichomc ispitted(Fig. 5d).
surface is composed by ordinary epidermal cells, All the species studied show dorsiventral
stomataandtrichomes(Fig. 2a-f)- Theepidermal mesophyll, with one layerofcells in the palisade
anticlinalcellswallsaresinuousinallspecies(Fig. parenchymaandabout 10to 14layersinthespongy
2a-e),withtheexceptionofPosoqueriaspinwhich parenchyma(Fig.6a-e).Thepalisadecellsarewidcr
theyarestraight. P. acutifoliahasglabrousleaves andshorterinPosoqueriaacutifoliaandP.latifolia
(Fig.2a),inP.macropusnumerous,non-glandular, (Fig.6a-b)andlongerandnarrowerinthercmaining
unicellulartrichomes,withthicklignifiedwallswere species(Fig.6c-e).Idioblastscontainingtinypolyhedral
observed (Fig. 2 d). In P. palustris there are crystalsofcalciumoxalate(crystallifcroussand)are
pluricellularuniseriatedtrichomes,long,nonlignified observedontheentiremesophyllinallspeciesstudied
(Fig. 2e). Posoqueria sp is markedby presenceof (Fig.6c,0-TheyaremoreabundantinP. latifolia
numerouspapillae(Fig. 2f). Rare shortunicellular and P. palustris. These cells occur in a greater
non-glandulartrichomesareobservedontheabaxial frequcncybeneaththepalisadeparenchyma,ornear
surface ofthe leafofP. latifolia and P. longiflora, theepidermisoftheabaxial surface,isolatedorin
restricted to midvein; in P. palustris, numerous groups, especially in P. latifolia and P. palustris
pluricellulartrichomesareobserved. (Fig. 6 c-f). The histochemical tests showed that
Thestomatabelongtotheparacytictypeand the crystallifcrous cells also accumulate phenolic
arearrangedrandomly(Fig2).Thesubsidiarycells compounds.
may bc of different sizes; some stomata may be The vascular system of the leaf blade is
flankedinparallelbymorethanonesubsidiarycell. formedbybundlesofthecollateral type,ofvaried
The six species investigated present some large calibers (Fig. 6 a, d). The vascular bundles are
stomata, around which the epidermal cells are surrounded by a parenchymatous sheath that may
organized radially, and the cuticle layer forms a contain chloroplasts, crystalliferous sand, and
radiate pattem (Fig. 2 b). In P.macropus and P. phenolic compounds. Internally to this sheath
palustris the stomata are partly covered by lignified fibers occur in variable proportions. In
numeroustrichomes(Fig.2d-e). Posoqueriaacutifolia,P. latifoliaandP.macropus
Theepicuticularwaxcanoccurwithoutdefincd thesclcrenchymaoccursinalargerproportionthan
omamentation,intheformoffilaments,granulesand thevasculartissues (Fig. 6a,d).
incrusts(Figs.3a-h,4a-g).Thegranulescanbcof Thecross scction showedthattheoutline of
variedsizesandarepresentinPosoqueriaacutifolia leaf margin is truncatc-revolutc in Posoqueria
(Fig. 4a), P. longiflora (Fig. 3 d,4d), P. macropus longiflora and slightly revolute in the othcrs
(Fig.4e), P. palustris(Fig.4 0 andPosoqueriasp species(Fig.7a-d).Intheepidermis,characteristics
(tFriigc.h4omge),si(nFcigl.ud4ien)g.oTvheerctrhuesststoomcactuar(iFnigP..4mad,crg)opaunsd osibmsielravretdo,thnoosteicoifntghetrheasttothfethceutleiacfulbalradelacyaenrbies
(Fig. 3 0.P.palustris(Fig. 3g) andare thick in P. stronglythickcned, formingconspiciouscuticular
oltahfteiPfa.odlaliaaxti(iFafilogl.siu3arb(f)Fa;ictgeh.eo4ffiacll)al.mseTpnethsceioeccscutu(irFciilgne.ai3bsaaxs-imha)loosautnrhdfaoocnne thflhaaevnnegieans.ptahTpeihrleelsoctsoeofmaptmhpoeenalreaeanfpcibedl.eardIemnataelnrndcaelilnllysP,a.trlheoensgmmiaafrllgloiernar
the abaxial surfaceofP. longiflora (Fig.4d), and isfilledwithparenchymatouscellswiththickcned
striatedonthe abaxial surfaceoftheotherspecies walls; among them idioblasts withcrystals occur.
(Fig.4b,e-g),speciallyinP. latifolia(Fig.4c)and Lignified fibers were observed associated to the
inPosoqueriasp(Fig.4g). vascular system.
Inall speciesinvestigatedthe leafepidcrmis In transversal scction the outline oí petiole
cisovoenree-dlabyyreadlfayoerrmeofdcbuytictlheicakn-dwawlellcld-dceevlelsloapnedd c8aan-cb)e.sOunlcatthee,ppeltaino-lceoannvdexaolrocngonocfatvhce-cmoindvvecixn(,Fiogn.
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cF.iPg.urleon1gi-flLoeraaf\edp.iPd.ermmaicsroopfuPso-soeq.uPe.rpiaaluAsutbrlies;tfi.nPf.rolnotnaglivjiloerwa,,aidnaxtihealmsiudrvfeaicne-reag.iPo.na(c{u)t.ifolia;b P latifolia
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—
co , Anhletinfrontalview,ab*axia*lswurifnaBce,-hca.,Ph.eacutifolia; b. P°.rlsa“tijol’ia;
'
trichomes (d, e) andpapillae (0-
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Figure3-ScanningelectronmicroscopyofleafbladeofPosoqueriaAublet-a.P. acutifolia;b-c.P. latifolia;
d-e.P. longijlora; f.P. macropus; g. P.palustris; h. Posoqueria sp. Adaxial surface showingwax in granules
(d), in crusts(f) and inthickcrusts (b).
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Figure4-ScanningelectronmicroscopyofleafbladeofPosoqueriaAublet-a P. acutífolia;b.P.acutifoliaboiled;
teriPchloamtiefsoliinaP4.dmPaclroongpiufslo(rea);aen.dP.P.mapcalruosptursis;(f.0P.anpdalpuasptriilsl;aeg.inPoPsoosqouqeureinaasps.p.Ab(ag)x..aTlhesusrufrafeaceschcoawn.nbgesstmoomoat.ha(a-go)r,
striated(b-c,e-g). Epicu.icularwax(arrow)in filamcnts(c)andgranules(e).
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Figure5-CrosssectionsofPosoqueriaAubletleaf,detailsofepidermis-a.P. latifolia withathickcuticularlayer
(arrow); b.P.longiflora,trichomeinmidveinregion;c.P. longiflora,stomatawithdevelopedrimsandsubsidirycells
inthesubstomatalchamber;d.P.palustris,lignifiedtrichomewithpittedwall(arrow).
both surfaces of the leaf, the epidermal cells are palisade parenchymaare shorterand widerthan the
organizedinrowsandhavethicker,nearlystraight remainingpartsoftheleafblade.Inthecorticalportion
walls. Stomata can be found in this region ofthe belowtheabaxialsurfacethereisaparenchymawhich
leafblade.Inthecorticalregionofthepetiolethere shows progressively more-voluminous cells with
are two to five layers of angular collenchyma thinnerwallstowardsthevascularsystem.
beneaththeepidermis.Afundamentalparenchyma In this study the most relevant leaf
containing idioblasts with crystals, phenols, or characteristics tothe distinguishing ofthe species
somesclereidsarelocatedintemally(Fig.8d,h).In arerelatedintheTable 1
all species, the vascular system is arranged in an
arch(Fig.8a-c,e),withthexylemcellsarrangedin Discussion
rows andthephloem in the externai position, like Fromalltheanatômica]featuresexamined.the
thecollateraltype.Additionalbundlesareobserved
ones that showed ahigherpotencial ofutilization
attheextremitiesofthevascularsystem.
to identify the studied species were: cuticularand
In the region of the midvein, beneath the wax sculpturing, aspect ofthe anticlinal walls of
epidermis,thecorticalregionisoccupiedbythreeto epidermal cells and presence and types of
five layers ofcollenchyma that varies between the trichomes. The variation on the thickness em
angularandlamellartypes,andundemeaththereisa sinuosityfromtheanticlinalwallsisquotedfotthe
palisadeparenchymaconnectingthetwopartsofthe RubiaceaebyMetcalfe&Chalk(1950)andGomes
leafblade(Fig. 8f-g). Inthisportion,thecellsofthe etal. (2000). Harberlandt (1928)believesthat the
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Figure7 CrosssectionsofPosoqueriaAubletshowingvariationoutlineoftheleafmargin-a.P. acutifolia-.
b.P. latifolia\c.P. longiflora\ d. P.palustris.
Table 1 DistmctivecharacterestotheidentificationofsixPosoqueriaAublet(Rubiaceae)species.
SCpheacriaecsters/ Posoqueria Posoqueria Posoqueria Posoqueria Posoqueria Posoqueria
acutifolia latifolia longiflora macropus palustris sp.
Epidermis,
adaxial surface
Ouwtallilne of anticlinal sinuous sinuous sinuous sinuous sinuous straight
Cuticular surface smooth smooth smooth smooth smooth striated
Epicuticular wax smooth crusts granules crusts crusts smooth
Epidermis,
abaxial surface
Ouatnltiincleinoafl wall sinuous sinuous sinuous sinuous straight straight
to curves
Cuticular surface striated striated smooth striated striated striated
Epicuticular wax granules filaments granules granules granules granules
Fofortmriacnhdomdeisstribution absent unicellular, long, long, long, papillae,
restrict to unicellular, unicelular, pluricelular, intercoastal
midvein restrict to intercoastal intercoastal region
region midvein region region region
Ouitnlitnreanosfvetshael lseeacftimoanrgin revolute revolute trreuvnoclauttee/ revolute revolute revolute
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