KUBOTA, T. M. K.

MILORI, D. M. B. P.

VILAS-BOAS, P. R.

Repository Open Access to Scientific Information from Embrapa

POTENCIAL DA ESPECTROSCOPIA DE FLUORESCÊNCIA INDUZIDA POR LASER PARA SEPARAR VARIEDADES DE LARANJA GENETICAMENTE PARECIDAS T.M.K. Kubota1, A.B. Magalhães2, M. Cristofani-Yaly3, P.R. Vilas-Boas2, D.M.B.P. Milori2(1) Universidade de São Paulo, Avenida Trabalhador São Carlense, 400, 13560-970, São Carlos, SP, thiagomassaiti.k.k@gmail.com(2) Embrapa Instrumentação, XV de Novembro, 1452, 13560-970, São Carlos, SP, habibe.aida@gmail.com, paulino.villas-boas@embrapa.br, debora.milori@embrapa.br (3) Centro APTA Citros Sylvio Moreira, IAC, Rodovia Anhanguera Km 158, 13490-970, Cordeirópolis, SP, mariangela@centrodecitricultura.brResumo: As laranjas doces (Citrus sinensis L. Osbeck) concentram as espécies mais importante do grupo GRV FLWURV RQGH D SURGXomRPXQGLDO HP DWLQJLX PLO WRQHODGDV(ODV VmR FODVVL¿FDGDV FRPR ODUDQMDVpigmentadas, comum, de baixa acidez e de umbigo. Entre esses grupos existem grandes diferenças fenotípicas, como a forma do paio, cor, tamanho do fruto e época de maturação do fruto. Apesar de todas estas diferenças fenotípicas, existe uma grande semelhança entre os genótipos destas espécies. Nota-se uma pequena variabilidade PROHFXODUGR'1$RTXHGL¿FXOWDDVXDFODVVL¿FDomR9DULRVPDUFDGRUHVGH'1$WHPVLGRXWLOL]DGRSDUDWHQWDUdiferenciar estas variedades, porém com pouco sucesso. Pensando nisso a Embrapa Instrumentação junto com o ,QWLWXWR$JURQ{PLFRGH&DPSLQDV,$&YHPGHVHQYROYHQGRXPDQRYDWHFQLFDSDUDUHDOL]DUDLGHQWL¿FDomRHQWUHDVYDULHGDGHVGHODUDQMDVJHQpWLFDPHQWHSDUHFLGDV(VWDWHFQLFDWHPFRPREDVHDHVSHFWURVFRSLDGHÀXRUHVFHQFLDinduzida por laser e algumas ferramentas estatisticas. Os resultados são obtidos de forma rápida, sem preparo de amostras e com baixo custo. Neste trabalho foi utilizado um conjunto de folhas de tres variedades de laranjas genéticamente parecidas: Laranja Pera Abril, Laranja Pera Bianchi e Laranja Pera Roberto Gullo. Estas folhas IRUDPOLPSDVPHGLGDVFRPRHTXLSDPHQWRGHÀXRUHVFHQFLDLQGX]LGDSRUODVHU3KRWRQ&LWUXVHFRPRDX[LOLRGRVRIWZDUH:HNDIRUDPIHLWRVRVWUDWDPHQWRVHVWDWLVWLFRFRPRVHVSHFWURVGHÀXRUHVFHQFLDGDVIROKDV&RPLVVRDpartir de uma validação cruzada, foi possível separar corretamente as três classes de Laranja Pera com um acerto de 88 %.Palavras-chaveHVSHFWURVFRSLDÀXRUHVFrQFLD3KRWRQ&LWUXVODUDQMDVGRFHPOTENTIAL OF THE LASER INDUCED FLUORESCENCE SPECTROSCOPY IN SEPARATING CITRUS VARIETIES VERY CLOSE GENETICALLY Abstract: The sweet oranges (Citrus sinensis l. Osbeck) concentrate the most important species of the citrus JURXS,QWKHVZHHWRUDQJHZRUOGSURGXFWLRQKDVUHDFKHGWKRXVDQGWRQV7KH\DUHFODVVL¿HGDV3LJPHQWHGcommon, low acidity and navel oranges. Major differences between these groups are the phenotypic size, shape of the paio, its color, size of the fruit and the fruit maturation season. Despite all these phenotypic differences, there is a large similarity between the genotypes of these species. It is possible to observe a small molecular YDULDELOLW\RIWKH'1$ZKLFKGLI¿FXOWWKHLUFODVVL¿FDWLRQ6HYHUDO'1$PDUNHUVKDYHEHHQXVHGWRGLIIHUHQWLDWHthese varieties, but they were not successful. Thinking about this, the Embrapa Agrocultural, together with the Instituto Agronômico de Campinas (IAC), has developed a new technique to perform the differentiation between JHQHWLFDOO\VLPLODURUDQJHYDULHWLHV7KLVWHFKQLTXHLVEDVHGRQODVHULQGXFHGÀXRUHVFHQFHVSHFWURVFRS\/,)6DQGVRPHVWDWLVWLFVWRROV7KH/,)6WHFKQLTXHFDQSURYLGHDIDVWDQGORZFRVWDQDO\VLVZLWKRXWVDPSOLQJSUHSDUDWLRQcan be applied to a large number os samplesandprovide a quality database. In this work, it was used a set of leaves of three different orange varieties genetically simlar: “Laranja Pera Abril”, “Laranja Pera Bianchi” and “Laranja 3HUD5REHUWR*XOOR´7KHVHOHDYHVZHUHFOHDQHGDQGWKHQPHDVXUHGZLWKWKH/,)6HTXLSPHQWDOVRQDPHG3KRWRQ&LWUXV:LWKWKHDLGRIWKH:HNDVRIWZDUHVWDWLVWLFDOWUHDWPHQWVZHUHPDGHIURPWKHÀXRUHVFHQFHVSHFWUDRIWKHleavesand it was possible to separate the three classes correctly, with a success rateof 88%.KeywordsVSHFWURVFRS\ÀXRUHVFHQFH3KRWRQ&LWUXVVZHHWRUDQJHV1. IntroduçãoNo setor agropecuário a produção de citros tem grande destaque econômico. São produzidos aproxima-damente 122 milhões de toneladas de frutas, que correspondem a 17 bilhões de dólares em vendas da fruta fresca AT1-14.indd   49 18/12/14   11:27T.M.K. Kubota, A.B. Magalhães, M. Cristofani-Yaly, P.R. Vilas-Boas, D.M.B.P. Milori50HVXFRVFRQFHQWUDGRVQRPXQGR)$22%UDVLOpRPDLRUSURGXWRUHH[SRUWDGRUGHODUDQMDGRSODQHWDUHV-SRQViYHOSRUPLOK}HVGHWRQHODGDVDQXDLV2SDtVWDPEpPpRPDLRUH[SRUWDGRUGHVXFRFRQFHQWUDGR2VHWRUpUHVSRQViYHOSRUJHUDUPDLVGHPLOHPSUHJRVGLYLGLGRVHPPDLVGHPXQLFtSLRV1(9(6$VODUDQMDVGRFHVCitrus sinensis /2VEHFNFRQFHQWUDPDVHVSpFLHVPDLVLPSRUWDQWHGRJUXSRGRVFL-WURVFXMDSURGXomRPXQGLDOHPFKHJRXDPLOWRQHODGDV)$2(ODVVmRFODVVL¿FDGDVFRPRODUDQMDVSLJPHQWDGDVFRPXPGHEDL[DDFLGH]HGHXPELJR(QWUHHVVHVJUXSRVH[LVWHPXPDJUDQGHGLIHUHQoDIHQRWtSLFDVFRPRRWDPDQKRDIRUPDGRSDLRFRUWDPDQKRGRIUXWRHpSRFDGHPDWXUDomRGRIUXWR$SHVDUGH WRGDVHVWDVGLIHUHQoDV IHQRWtSLFDVH[LVWHXPDJUDQGHVHPHOKDQoDHQWUHRVJHQyWLSRVGHVWDVHVSpFLHV1RWDVHXPDSHTXHQDYDULDELOLGDGHPROHFXODUGR'1$RTXHGL¿FXOWDDVXDGLIHUHQFLDomRHFODVVL¿FD-omR(VWDSHTXHQDGLIHUHQoDQDYDULDELOLGDGHPROHFXODUGR'1$pFDXVDGDSRUXPDHVWUHLWDEDVHJHQpWLFDFRPXPGHVWHJUXSRHDVFDUDFWHUtVWLFDVPRUIROyJLFDVGLIHUHQWHVpGHYLGDDPXWDo}HVHPDQWLGDSRUSURSDJDomRYHJHWDWLYD+(55(529DULRVPDUFDGRUHVGH'1$WHPVLGRXWLOL]DGRSDUDWHQWDUGLIHUHQFLDUHVWDVYDULHGDGHVSRUpPFRPSRXFRVXFHVVR3HQVDQGRHPGHVHQYROYHUXPPpWRGRUiSLGRGHEDL[RFXVWRTXHQmRSURGX]DUHVLGXRVTXHSRVVDPDJUH-GLURPHLRDELHQWHD(PEUDSD,QVWUXPHQWDomRMXQWRFRPR,QWLWXWR$JURQ{PLFRGH&DPSLQDV,$&YHPGHVHQ-volvendo XPDQRYDWHFQLFDSDUDUHDOL]DUDLGHQWL¿FDomRHQWUHDVYDULHGDGHVGHODUDQMDVJHQpWLFDPHQWHSDUHFLGDV(VWDWHFQLFDWHPFRPREDVHDHVSHFWURVFRSLDGHÀXRUHVFHQFLDLQGX]LGDSRUODVHU/,)6HDOJXPDVIHUUDPHQWDVHVWDWLVWLFDV0,/25,2. 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Resultados e Discussão2VHVSHFWURVFDUDFWHUtVWLFRVGHFDGDYDULHGDGHVREWLGRVSHOR3KRWRQ&LWUXVHVWiPRVWUDGRQD)LJXUDD)D]HQGRXPDDQDOLVHDSHQDVYLVXDOSRGHPRVYHUTXHQDIDL[DHQWUHQPDQPRVHVSHFWURVGDYDULHGDGH3HUD$EULOVmRPDLVLQWHQVRVTXHRVGD3HUD5REHUWR*XOOR-iQDIDL[DHQWUHQPDQPDVLWXDomRVHLQYHUWH(SDUDRFDVRGDYDULHGDGH3HUD%LDQFKLRVHVSHFWURVSRVVXHPXPDLQWHQVLGDGHLQWHUPHGLDULDHFRPLVVRVHFRQ-IXQGHPFRPDVRXWUDVGXDVFODVVHV(VWDGLIHUHQoDQRVHVSHFWURVSRGHVHUH[SOLFDGDSHODGLIHUHQWHFRQFHQWUDomRGDFORUR¿ODHRXWURVÀXRURIRURVFDUDFWHUtVWLFRSDUDFDGDYDULHGDGH$JRUDDQDOLVDQGRR3&$PRVWUDGRQD)LJXUDESRGHVHQRWDUTXHRVSRQWRVUHODFLRQDGRjVDPRVWUDVGDYDULHGDGH3HUD5REHUWR*XOORWHQGHPDVHDFXPXODUDRODGRHVTXHUGRGR]HURGD3&HRVSRQWRVUHODFLRQDGRVj3HUD$EULOHVWmRVHDJUXSDQGRQRODGRGLUHLWRGR]HUR(QRFDVRGD3HUD%LDQFKLHODVHVWmRVHDJUXSDQGRQRFHQWURSUy[LPRDR]HUR)LJXUDD(VSHFWURVREWLGRVFRPR3KRWRQ&LWUXVE$QDOLVHGH3&$GDVWUrVYDULHGDGHVGHODUDQMD3HUD(VWHWLSRGHDQDOLVHGHUHFRQKHFLPHQWRGHSDGU}HVHQWUHDVYDULHGDGHVpH[WUHPDPHQWHLPSRUWDQWHSRLVPRVWUDRSRWHQFLDOGHVHFULDUXPFODVVL¿FDGRU(FULDGRHVWHFODVVL¿FDGRUSRGHVHLGHQWL¿FDUHFODVVL¿FDUSODQWDVGHVFRQKHFLGDVGHVWDVWUrVYDULHGDGHVGHODUDQMD3HUD3DUDPRQWDUXPFODVVL¿FDGRUIRLXWLOL]DGRRVRIWZDUH:HNDRQGHIRLXVDGDDIHUUDPHQWDHVWiWLFD3/65SDUDID]HUXPDYDOLGDomRFUX]DGDFRPRVGDGRVREWLGRVGR3KRWRQ&LWUXV2UHVXOWDGRGHVWDYDOLGDomRHVWDPRV-WUDGDQDPDWUL]FRQIXVmRGD)LJXUD&ODVVL¿FDGRFRPR9DULHGDGH5HDOAbril %LDQFKL *XOORAbril   %LDQFKL   *XOOR   )LJXUD0DWUL]FRQIXVmRGRVUHVXOWDGRVGDYDOLGDomRFUX]DGD1DPDWUL]FRQIXVmRDVOLQKDVPRVWUDPDVWUrVYDULHGDGHVTXHHVWmRVHQGRFODVVL¿FDGDTXHIRUDPFKD-PDGDVGH9DULHGDGH5HDO-iDVFROXQDVUHSUHVHQWDPFRPRHVWDVYDULHGDGHVIRUDPFODVVL¿FDGDVSHORFODVVL¿FDGRU&RPLVVRDGLDJRQDOSULQFLSDOUHSUHVHQWDRDFHUWRGRFODVVL¿FDGRUQDYDOLGDomRFUX]DGD$QDOLVDQGRHVWDGLDJRQDOSRGHVHREVHUYDUTXHRFODVVL¿FDGRUDFHUWRXGDVFODVVL¿FDo}HV4. Conclusões3RGHPRVFRQFOXLUDQDOLVDQGRRVHVSHFWURVFDUDFWHUtVWLFRVGHFDGDYDULHGDGHTXHHOHVSRVVXHPXPDGLIH-UHQoDQDVVXDVLQWHQVLGDGHV(DQDOLVDQGRRWHVWHGH3&$SRGHPRVYHUTXHDVYDULHGDGHVWHQGHPDVHDJUXSDUHPXPDGDGDUHJLmRGDV3&VPDVSRGHQGRRFRUUHUXPDSHTXHQDFRQIXVmRHQWUH%LDQFKLH$EULOH%LDQFKLH5REHUWR*XOOR0DVPHVPRDVVLPSRGHPRVYHUTXHRVHVSHFWURVWHPRSRWHQFLDOSDUDVHUHPGLIHUHQFLDGRV2UHVXOWDGRGDYDOLGDomRFUX]DGDREWHYHXPDFHUWREDVWDQWHDOWRXPDYH]TXHHVWDVYDULHGDGHVVmRJH-QHWLFDPHQWHSDUHFLGDVPDVPHVPRDVVLPQRVVDWpFQLFDIRLFDSD]GHGLIHUHQFLDUHFODVVL¿FDUDVYDULHGDGHVRTXHPRVWUDRJUDQGHSRWHQFLDOGDQRVVDWpFQLFD siagro-2014.indb   51 16/12/14   18:27T.M.K. Kubota, A.B. Magalhães, M. Cristofani-Yaly, P.R. Vilas-Boas, D.M.B.P. Milori52Agradecimentos*RVWDULDGHDJUDGHFHUWRGRVRVDXWRUHVSHODFRODERUDomRQRERPUHVXOWDGRGRWUDEDOKR&RQVHOKR1DFLR-QDOGH'HVHQYROYLPHQWR&LHQWt¿FRH7HFQROyJLFR&13TH)XQGDomRGH$PSDURjSHVTXLVDGR(VWDGRGH6mR3DXOR)DSHVSSHORDSRLR¿QDQFHLURHD(PEUDSD,QVWUXPHQWDomRReferências)22'$1'$*5,&8/785(25*$1,=$7,212)7+(81,7('1$7,216)$2 Produção total de citros em 2012'LVSRQtYHOHPKWWSZZZIDRRUJ!$FHVVRHP VHW1(9(60)HWDO2UHWUDWRGDFLWULFXOWXUDEUDVLOHLUD'LVSRQtYHOHPKWWSZZZFLWUXVEUFRPEUGRZQORDGELEOLRWHFD$SUHVHQWDFDR0DUFRV)DYDHYHQWRYDORUSGI!$FHVVRHPVHW+(55(525$6Ë160-&$5%21(//($1$9$552/*HQHWLFGLYHUVLW\ LQ WKHRUDQJHVXEIDPLO\$X-UDQWLRLGHDH ,QWUDVSHFLHV HQG LQWUDJHQXVJHQHWLF YDULDELOLW\7KHRUHWLFDO DQG$SSOLHG*HQHWLFV Y S 0,/25,'pERUD0DUFRQGHV%DVWRV3HUHLUD5$<1$8'0LFKHO%2$63DXOLQR5LEHLUR9LOODV9(1Æ1&,2$QGUp/HRQDUGR0281,(56WpSKDQH%$66$1(=,5HQDWR%HR]]RH5('215RODQG,GHQWL¿FDWLRQRIFL-WUXVYDULHWLHVXVLQJODVHULQGXFHG)OXRUHVFHQFH6SHFWURVFRS\/,)6&RPSXWHUDQG(OHFWURQLFVLQ$JULFXOWXUHYSsiagro-2014.indb   52 16/12/14   18:27

Potencial da espectroscopia de fluorescência induzida por laser para separar variedades de laranja geneticamente parecidas.

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