Table Of ContentRodriguésia61(3):569-574.2010
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Nota Científica / Short Communication:
The formation of the stigmatic surface in
Passiflora elegans (Passifloraceae) 1
Aformaçãodasuperfícieestigmátícaem Passifloraelegans (Passifloraceae)
AdrianoSilvério2 &JorgeErnestodeAraújo Maríath
Abstract
The stigma surface is acomplex multicellularstructure where the developmentofthe pollen tube begins. This
developmentisnecessaryforsucessinfertilizationanddependsonrecognitíonprocessesthatinvolvetheanatomyof
thestigma.Passifloraisaneconomicallyimportantgenusbecauseofitsediblefhiits.Manyauüiorshavedescribedthe
stigmaofPassiflorabutnothingisknownabouttheontogenesisofthisstructure.Thisworkaimedtodescribethe
formationofthestigmaticsurfaceofPassifloraelegans.Resultsshowedthat,inbud,thestigmaticsurfaceofthisspecies
piserfilcaltinwailthwaslmlsa.llDcuerlilsn.gTihtsedceevllesloinpmtehnetstuhbedesrtmiaglmalasyuerrfahcaevbeelcaorgmeesvaucnueolveesnadnudetthoetnhuecleeluosn,ganteiaorntooftcheellesxtmertnhaei
subdermallayer.Elongationresultsinanincreaseofexternaisecretorysurfaceareaofthestigmas,andprobablyplays
animportantroleinpollenrecognitíon.Thepolysaccharidecontentfoundintheinnerwallsofthesestmcturesmight
beinvolvedinthesignalprocessforpollentubegrowthduringitsearlydevelopmentThemorphologicalevidence
presentedhereshowsthat asthestigmaofPassiflora is formedbydermalandsubdermalcells, itshouldnotbe
characterizedascolletersorpapillaeand,therefore,itisdefinedhereasstigmaemergences.
Key-words: anatomy,stigmadevelopment,stigmaemergence,pollination.
Resumo
Asuperfícieestigmátícaéumaestruturamulticelularcomplexa,ondeotubopolínicoiniciaoseudesenvolvimento,
necessáriaparaafecundação.Estedesenvolvimentodependedecondiçõesfavoráveisqueenvolvemaanatomiado
estigmaduranteoprocessodereconhecimento.Passifloraéumgêneroeconomicamenteimportantedevidoaosseus
frutoscomestíveis.OestigmadePassifloratemsidodescritoporváriosautores,masoseuprocessodeformaçaoé
desconhecido EssetrabalhotemporobjetivodescreveroprocessodeformaçãodasuperfícieestigmátícadePassiflora
elegans Osresultados demonstram que durante a fase debotãojovem, a superfície estigmátíca é compostapor
pequenascélulaseapresentasuperfícieplana.Ascélulasdacamadasubdep.déimicaapresentamgrandesvacuolose
nirúrcelgeuolàrpdreóvxiidmooaodaalpoanrgeadmeenpteoridcelicnéalluleaxstedranca.amDaudraanstuebdoepsieduérdmeiscean.vEoslsvaismmeondtiof,icaasçuõpeesrrfíecsiuelteasmteigmmáutmícaactroémsac-ismco
dasuperfíciesecretoraexternadoestigma,eprovavelmentedesempenhamumimportantepapelnoreconhecimento
dopólen.Osconteúdospolissacaridicosencontradosnasuperfícieinternadessasestruturaspodemestarenvolvidos
comosprocessosdesinalizaçãodotubopolinicoduranteseudesenvolvimentoinicial.Asevidenciasmorfológicas
observadasnessetrabalhodemonstramqueasestruturaspresentesnasuperfíciedoestigmadePassiflorasãoconstituídas
porcélulasdeorigemdérmicaesubdérmica,enãodevemsercaracterizadascomocolétcresoupapilas,sendoassim,
caracterizadasnessetrabalhocomoemergênciasestigmáticas.
Palavras-chave: anatomia,desenvolvimentodoestifma, emergênciaestigmátíca,polinização.
ihnacvoimnpSgapteiacbiiselpsiotoryfospPyhassysttieifmclo(arRanêdLg.ogaaremetecathloa.rpah1c9yt9te9ir,ciz2es0de0lb0f;y- Spsotulialgsemsna,utniaacseistnuarolft.ah2ce0er0a(3nR)g,êigaoonspdeettrhamels.r,2eoc0co0cg0nu)ir.tsTímohaneirneslatycitgoimnoantooheff
2UnivcreidadcFederaldoRioGrandedoSul,Deplo.Botânica,Lab.AnatomiaVegetal.Av.Ben ç
!CNPqProductivityinResearchScholarshiprecipienL
*[email protected]
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Passifloraceae has been described in previous 1%Os04 washedin0.1 Msodiumphosphatebuffer
studies. Puri (1947)describedthefloweranatomy at pH 7.2 (Weber 1992), dehydrated in acetone,
ofthisgenus,andconsideredthestigmatobelarge, criticai pointdried (Gersterberger& Leins 1978),
with massive structures; however, no furthcr sputter-coatedwithgoldusingaBalzersSCD050.
anatomicalcommentswereprovided.Raju(1956) andexaminedusingaJeol6060SEM.
classified these structures as projections that It was found, during the initial stages of
facilitate pollengrain retention during pollination development, thatthe apicalportion ofthe stigma
etvuebnetssd,uarinndgatlhseoirapsaasssaitgeeftoorwtahredsgrtohewttrhaonsfmpiotltlienng hasaslightlysinuoussurface(Fig. 1a),thedermal
layerhascellswithanevidentnucleusandportions
twiasssuec.laIsnsiofnieedstausdyd,rtyhewistthigumnaicoefllPualsasrifplaoprialcleaaee ofcondensedchromatin(Fig. 1b),andthesubdermal
& layerhas cells with large vacuoles and respective
(Heslop-Harrison Shivanna 1977),andinanother nucleusdisplacedneartheexternaipericlinalwalls
study Passiflorci racemosa Brot. and other
(Fig. lb).
Passiflora species and genera in the family were Subsequenttotheinitialdevelopmentalphase,
reported to have multicellular papillae (Bemhard
theexternaisurfaceofthestigmabeginstoformmultiple
1999).TheclassificationusedbyBemhard(1999) dome-shaped projections, as a result ofanticlinal
wasalsousedbySouzaetal.(2006)forP. edulisf. divisionsfollowedbyanticlinalandradialelongation
fplaapviillcaatrepastDreugcteunreers.hTahdesceellasutwhiotrhslnaortgeedvathcautoltehse Tofhtehheediegrhmtaolfaenadchsoemmeerogfetnhceesuobndtehremsatligcemlaltsi(cFsiug.rf1acc)e.
andthinwalls.Howeverthesecharacterizationsof
continuestoincrease,whiletheexpandingsubdermal
the pappilate stigmatic surface ofPassiflora were
cellsdividetransversallygivingrisetoprojectionsthat
basedonlyonthefinalstagesofstigmadevelopment. have anapicaland abasal cell (Fig. ld). The apical
Twhheircehisisstipllrnoobacbolnyseinnsduuscoendthbeyortihgeinloafckthoisfsstrpuecctiufriec, cwehlilschretmhaeininitnerdniariecpterciocnltiancatlwwiatlhleapniddetrhmealprcoelxlism,ailn
ontogenetic studies. The goal of this work is to portionoftheanticlinalwallaccumulatecompounds
analyzetheontogeneticprocessofthesestructures ofpecticnature(Fig. 1d).Attheendofdevelopment,
atthestigmaticsurfaceofPassifloraelegansMast., a specialized structure is formed on the stigma,
anendemicspeciesofSouthernBrazil. comprisedofcellsfromthedermalandsubdermallayer
Stigmasof50floralbuds,measuring0.3to2cm, (Fig. 1e).Oncethesestructureshaveformedthestigma
and 20 buds in pre-anthesis were collected from surface, it appears papillate, but in cross-section it
plantsfoundontheCampusdoVale,atRioGrande can be seen that each projection has a multicellular
doSulFederalUniversity.Avoucherspecimenwas organization around a central axis formed by the
depositedintheICNHerbarium(1CN52108). subdermalcell(Fig. 1f).Scanningelectronmicroscopy
and 2T.h5e%magtleurtiaarlawladsehfyidxeedsionlaut2io%n,foirnmaald0e.h1ydMe rsteivgemaalteidcnsuurmfearcoeu(sFimgu.lltgi)c.ellularprojectionson the
sodiumphosphatebuffer,at7.2pH(Roland&Vian Duringpollination,thepollengerminatesonthis
1991).Forthebright-fieldmicMroscopyanalysis,the pappilatesurface(Fig.2a)andthe pollentubepath
material was washed in 0.1 sodium phosphate followsthecentralregionofthestructure, which is
buffer, at7.2pH, dehydrated in an ethanol series, rich in pectic compounds that have accumulated
a&ndSmeimdbe1d9d8e3)d.iSnehcytdiroonxsybetehtywlemeenta2carynlda4tep(mGetrhriictks aBleoynogntdhetahneticsltiingamlaa,ndthpeerpiocllilneanlwtaulbless(gFirgo.w2b-icn)t.o
weremadeusingaZeissMikronrotationmicrotome parenchyma(Fig.2c-e)andthetransmittingtissue
andstai&nedwith0.05%ToluidineBlueO,at4.4pH thathavecellswithsimilarChemicalproperties.
(Feder 0’Brien 1968).Histochemicaltestswere The surface of the stigma is crucial during
performedusingfreshmaterial,incombinationwith pollination,becausepollenrecognitiondependson
Ruthenium red to detect pectic acids (Johansen the lipids stored in the stigmatic cells and on the
T1h9e4s0)eatensdtsSwuedraenoIbIIsetrovteedstwfiotrhlaipLiedsic(aSaDsMsR19-5H1)C. gsulryfcaocpero(tTeiilntsonseectrealt.ed19f8r4o)m. tAfhteemrohnytdoratthieono,uttehre
LmiecircoascDopFeC,a5n0d0t.heFiomragtehsewesrceanonbtianigneedluescitnrgona paonldlgenrogwrasionvgeerrtmhienasttiegsmaa.ndDuthreinpgoltlheinsttiumbee.esmpeercigfeisc
microscopyanalysis,thematerialwaspost-fixedin enzymes loosen the cell wall of the papillae
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The stigma surface oí Passiflora elegans
Figure1—DevelopmentofthePassifloraelegansstigma—a.youngstigmaunderscanmngelectronmicroscopy,b.longitudinal
sectionoftheyoungstigmawiththecellsofthesubdeimallayerwithlargevacuoles(*);c.longitudinalsectionofthestigma
showingepidermalcellspushedbycellsfromthesubdermallayer(*);d.cross-sectionalofthedermalcellwithpectinswalls
(white asterisk) and division ofsubdeimal cell (*); e. longitudinal section ofthe stigma emergence in the final phase of
development;f.cross-sectionofthestigmaemergenceregioninthefinalphaseofdevelopment,showingthecellofthesubdermal
layerpositionedinthecentralregion;g.electromicrographyofthestigmasurfaceshowingthestigmaemergences.
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Figure 2- Stigmaand style ofthe PassiJIora elegansflower-a. stigma surfacewith a pollen tube on the stigma
emergence.Detailunderscanningelectronmicroscopy;b.histochemicaltestforthepresenceofpectins-c longitudinal
section ofthe stigma surface and the way ofthe pollen tube penetration through the stigma emergences and
parenchymatoustissue; d. longitudinal sectionofthe stigmaandstyle showingthe transmittmgtissue attheapical
centerofthestyle(Tt);e.stigmaandstyleunderscanningelectronmicroscopy.
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The stigma surtaee of Passiflora elegans
p(rMeipcahreilnig2i0t01fo)r.tChaelpceinuemtriastpiornoboafbltyheapmoellsesnentguebre stigmaTthiceprmoojrepchtoiolnosgirceavelalcehdariancttehirsiswtoircskdoof ntohte
duringthisprocess,inducingenzymesecretionand agree with the previous descriptions of the
lheconsequentiallooseningofthecellwall(Elleman literature.thatusedthetermpapillatodefinethese
&Dickinson 1986;Hiscocketal.2002). “projections of epidermal cells.” In addition,
In this study, a large amount of pectin was previous studies did not classify these structures
observedonthedermalandsubdermalcellwallsof as colleters. For this reason, we conclude that
thestigma,whichcoincidesatthecellularleveiwith “stigmaemergence”isabettertermtoclassifythe
thepollentubepathduringitsgermination.Pectins structures found on the stigmas of Passiflora, as
probably stimulate the pollen tube growth of P. they are formed from the dermal and subdermal
elegans, and calcium is made available to this layers and they do notsecrete mucilagc.
structurealong itscourse. Calcium (Ca3*) is akey Bemhard(1999)consideredthecharacteristic
element inthis process, regulatingelongation and stigma ofPassifloraceae to be largely distributed
orientationofthepollentubeduringitsdevelopment among the genera of this family, but rare in the
otherfamilies ofangiosperms. From a taxonomic
(Malhóetal.2006).
Pectins are synthesized in dictyosomes, in a perspective, the stigmatic surface also appears to
methyl-sterified form. The mcthyl-sterification of be an important trait that could be used to help
carboxylicgroupspreventsCa3+binding,makingthe describing Passifloraceae. Additional studies on
cellwalllessrigid.Asmethyl-sterificationincreases, othertaxainthefamilyareneededtoconfirmthis.
thefluidityofthepectingelalsoincreases,allowing
thecelltoexpandwhiletheintegrityofitsstructure References
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