PHYSIOLOGICAL MATURITY OF SEEDS AND COLORIMETRY OF FRUITS OF Jatropha curcas L. 1

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1 158 PHYSIOLOGICAL MATURITY OF SEEDS AND COLORIMETRY OF FRUITS OF Jtroph curcs L. 1 João Alexndre Lopes Drnski 2, Artur Sores Pinto Júnior 2, Fábio Steiner 3, Tigo Zoz 4, Ubirjr Contro Mlvsi 5, Mrlene de Mtos Mlvsi 6, Vndeir Frncisco Guimrães 6. ABSTRACT - The objective ws to relte the colortion of fruits of Jtroph curcs L during the mturtion process with morphometric (fruits nd seeds) nd germintive chrcteristics (seeds). Fruits were collected t vrious mturtion stges from mst trees locted in Eldordo/MS, nd six mturtion stges were visully clssified bsed on fruit s epicrp color. Epicrp digitl colorimetry ws used to chrcterize mturtion stges in ddition to morphometric vribles of length nd width of fruits nd seeds, s well their wter content. For determining the physiologicl potentil it ws used germintion test, emergence velocity index nd first count of germintion. Results indicted tht seeds of J. curcs should be hrvested when fruit epicrp shows reding equl or smller thn 82, 70 or 65 nm of red, green or blue scles from digitl color nlyzer. Seeds of fruits with tht colorimetry cn be hrvested in virtue of the mxim cumulultion of dry mss, wter content below 38,5% nd higher physiologicl potentil. Index terms: mturtion stge, germintion, refrctnce, morphometry. MATURIDADE FISIOLÓGICA DE SEMENTES E COLORIMETRIA DE FRUTOS DE Jtroph curcs L. RESUMO - O estudo objetivou relcionr colorção de frutos de Jtroph curcs L. durnte o processo de mturção com s crcterístics morfométrics (frutos e sementes) e germintivs (sementes). Os frutos form coletdos de mtrizes em Eldordo/MS, e visulmente clssificdos em seis estádios de mturção de cordo com colorção do epicrpo. Pr crcterizr mturidde fisiológic utilizou-se colorimetri digitl ssim como s vriáveis morfométrics diâmetro e comprimento, ssim como mss sec e o teor de águ de frutos e sementes. Pr determinção do potencil fisiológico utilizou-se o teste de germinção, o índice de velocidde de emergênci, e o teste d primeir contgem de germinção. Sementes de J. curcs devem ser colhids qundo colorção do epicrpo do fruto for igul ou menor que 82, 70 ou 65 nm ds escls vermelh, verde ou zul de um nlisdor digitl de cor. Sementes de frutos com quel colorimetri podem ser colhids em virtude do máximo cúmulo de mss sec, teor de águ bixo de 38,5%, e mior potencil fisiológico. Termos pr indexção: estádio de mturção, potencil fisiológico, refrtânci, morfometri. 1 Submitted in 17/08/2009 Accepted for publiction on 22/09/ Biologist, Mster in Agronomy, Agriculturl Science Center, UNIOESTE, Rondon, PR. Schdrship from CNPq, e-mil: joodrnski@ yhoo.com.br, rtur_bio@hotmil.com. 3 Agr. MSc. in Agronomy, Agriculturl Science Center, UNIOESTE. Rondon-PR, e-mil: fsteiner_gro@hotmil.com 4 Undergrdute student, Agriculturl Science Center, UNIOESTE. Rondon-PR, e-mil: tigo_zoz@hotmil.com 5 Eng. PhD. Professor Associte, Center for Agriculturl Sciences, UNIOESTE. Rondon-PR, e-mil: mrlenemlvsi@yhoo.com.br; ndeirfg@yhoo.com.br

2 PHYSIOLOGICAL MATURITY OF SEEDS AND COLORIMETRY OF FRUITS IN PHYSIC NUT 159 INTRODUCTION Jtroph curcs L. is perennil nd monoecious species from the Euphorbicee fmily found in severl regions of Brzil (Heller, 1996; Beltrão, 2005). The physic nut is species with gret potentil for biodiesel production due to the high oil content of seeds (Arrud et l., 2004). Disuniform plnttions due to the use of low-qulity seeds limit the cultivtion expnsion in Brzil. Therefore, studies iming to chrcterize the mximum seed qulity re essentil. During the mturtion process, seeds go through physicl, biochemicl nd physiologicl chnges influenced by genetic nd environmentl fctors to rech physiologicl mturity, which offer mximum germintion, vigor nd dry mss (Mrcos Filho, 2005). In the field, hrvesting of forest seeds usully employs physicl spects such s fruit color, size, smell nd spontneous opening s reference for identifying current prctice of hrvesting (Sen nd Grigliio 1998). In erly seed hrvesting, the process of mturtion is interrupted, which my impir seed qulity. The hrvesting of seeds with high levels of vigor nd germintion is of mjor importnce for fst nd efficient production of qulity seedlings (Crneiro, 1995; Crvlho, 2000; Mrcos Filho, 2005). The identifiction of seed qulity relted to the color of fruits is importnt to define the time of hrvest (Crvlho nd Nkgw, 2000). Knowledge of the mturtion process contributes to the estblishment of the idel time to hrvest when seeds hve better physiologicl qulity. Additionlly, in erly seed hrvesting the mturtion process is stopped reducing seed qulity. In physic nut, fructifiction is uneven where growth of lst fruits continues fter ripening of the first ones in the sme inflorescence (Sturnino et l., 2005). Thus, hrvest occurs in repeted cycles during the breeding seson cusing burden on the mnpower. Therefore, the clssifiction of fruits of J. curcs bsed on the fruit color nd not on the flowering phenology (dys fter nthesis) hs economic nd logisticl ppliction to obtin seeds of this species. This study imed to relte the epicrp color of fruits from Jtroph curcs L. t different mturtion stges with morphometric chrcteristics of fruits nd seeds nd with seed germintion chrcteristics indictive of physiologicl mturity of seeds. MATERIAL AND METHODS J. curcs fruits were collected in Jnury 2009 in 07 h of plnttion with popultion density of 1333 plnts per hectre, locted in Eldordo/MS under the geogrphic coordintes of W nd 23º S nd 342 meters of men ltitude. The climte of the region is clssified ccording to Köppen s mesotherml subtropicl humid, with hot summers (verge temperture bove 22 ºC), tendency towrd rinfll concentrtion, winters with frequent frosts (men temperture below 18 C), nd 1,148 mm men nnul precipittion. The soil is clssified s Drk Red Ltosol of sndy texture (Embrp, 2006). Thirty kilogrms of fruit were collected nd visully clssified into six mturity stges ccording to the predominnt color with the use of the Munsell color chrt (Munsell, 1976) s shown in Figure 1. The first stge ws chrcterized by green fruits, stge 2 by trnsition fruits from green to yellow pigment, stge 3 predominntly yellow fruits, stge 4 by fruits in trnsition from yellow to brown pigmenttion, stge 5 by mture fruits, nd stge 6 dry fruits t the beginning of dehiscence. Mesurements in fruits (pericrp + seed) for ech mturity stge involved the nlysis of refrctnce in red, green nd blue spectr of 10 fruits using digitl color nlyzer (ACR-1023 by Instrutherm ); quntifiction of the dry mss of 10 fruits by the oven dry method for 24 h t 105 C with results expressed mg fruit -1 ; the wter content obtined from 10 fruits by the method of oven dry for 24 h t 105 ± 3 C with vlues expressed in percentges on wet bsis; nd morphometric mesurements of length nd dimeter (the lrgest centrl mesure perpendiculr to the length) of 10 fruits. Technologicl prmeters used to chrcterize the physiologicl mturity of seeds comprised morphometric determintions of seed length nd dimeter (referring to the lrgest centrl perpendiculr to the rphel scr) in 10 seeds; wter content obtined from 50 grms of seeds mechniclly grounded by the oven dry method for 24 h t 105 ± 3 C with vlues expressed in percentge on wet bsis (Brsil, 1992), nd the verge dry weight of 10 seeds obtined by the oven dry method for 24 h t 105 C with results expressed mg seed -1. Germintion test ws performed bsed on the germintion test for Ricinus communis recommended by the Rules for Seed Anlysis (Brsil, 1992) with germintion expressed s percentge of norml seedlings obtined 14 dys fter sowing. The test ws conducted in snd substrte, which ws sterilized in n utoclve t 120 C for 15 minutes nd moistened with distilled wter in mount of 60% snd field cpcity (Brsil, 1992), using

3 160 J. A. L. DRANSKI et l. plstic trys (42 x 28 x 6 cm) under constnt temperture of 25 C with 12 hours photoperiod in BOD-type germintor (247 G Fnen ) with 25 seeds per replicte. As vigor tests we lso evluted first count germintion (percentge of norml seedlings 7 dys fter sowing) nd emergence velocity index (EVI) ccording to Mguire (1962). Mturrtion stge Epicrp color Visul Chrcteriztion Munsell chrt* 1 Green Fruit 7,5 GY 6/6 2 Fruit r the beginning of yellow pigmenttion 2,5 GY 5/10 3 Predominntly yellow fruit 5 Y 8/10 4 Fruit t the beginning of brown pigmenttion 5 Y 8/8 5 Mture fruit 7 YR 4/2 6 Dry fruit r the beginning of dehiscence 5YR 3/2 FIGURE 1. Color of physic nut fruits (Jtroph curcs L.) t different mturtion stges. * Munsell color chrts for plnts tissues. Vlues expressed in percentges were trnsformed into squre root rcsine x/100. Dt were checked for normlity of residul distribution nd vrince homogeneity, followed by nlysis of vrince using the sttisticl softwre SISVAR ver. 5.0 (Ferreir, 2000). The experimentl design ws completely rndomized with tretments consisting of six stges of fruit ripening with four replictes. Tretment mens were compred by the Tukey test t 5% probbility. RESULTS AND DISCUSSION The quntifiction of fruit refrctnce with the digitl color nlyzer (Tble 1) showed sttisticl differences (P <0.01) for the different fruit mturtion stges in the three colour scles of the instrument. The redings on the red, blue nd green scles showed higher men vlues when reched stges 2 nd 3, indicting tht fruit s pericrp ws expressed predominntly yellow color, followed by reding reduction towrds stges 5 nd 6, in which no significnt difference existed between mturtion stges in ll scles of the instrument. Digitl quntifiction of colors lines by monochrome scle enbles to distinguish res subject to color vritions. In this context, the gretest vrition of mens in reltion to mturity stges llowed to clssify fruits with greter repetbility. Refrctnce vlues in scles of red nd green of the digitl color nlyzer llowed better differentition in mturtion stges of fruits compred to the refrctnce numericl vlues in the blue scle (Tble 1). Similr behvior ws observed by Effendi et l. (2009) in unripe, ripe nd dried fruits of J. curcs with refrctnce vlues of red, green nd blue scles vi digitized imges. The digitl colorimeter hs dvntges of visul cuity, trining of technicl personnel nd computer mnipultion to seprte mturtion stges of fruits. Sen nd Griglio (1998) nd Germque et l. (2002) reported tht color chnge of fruits is good indictor to ssist in identifying the hrvest stge of severl species.

4 PHYSIOLOGICAL MATURITY OF SEEDS AND COLORIMETRY OF FRUITS IN PHYSIC NUT 161 TABLE 1. Averge refrctnce vlues of fruits of Jtroph curcs L. obtined with digitl color nlyzer. Refrctnce scle Mturtion stge Red green blue nm b b b b b c c c d d c d d MSD CV (%) Averges followed by the sme letter in columns re not sttisticlly different mong themselves t 5% by the Tukey test. 1. Green fruit; 2. Fruit t the beginning of yellow pigmenttion 3. Predominntly yellow fruit, 4. Fruit t the beginning of brown pigmenttion, 5. mture fruit; 6. Dry fruit t the beginning of dehiscence. The criterion of fruit color s n indictor of physiologicl seed qulity ws recommended to hrvest seeds of Ricinus communis when fruits were brown colored (Silv et l., 2009) s well s for severl species s reported by Aquino et l. (2006) in Peltophorum dubium, for Mcherium brsiliense by Guimrães nd Brbos (2007), nd by Aguir et l., (2007) for Ceslpini echint, when fruits of these species were in trnsition from green to brown preceding the spontneous opening. Therefore, the hrvesting of physic nut bsed on fruit color, quntified by digitl instrument will fcilitte the development of pproprite mchinery for fruit selection by color, llowing the obtining of seeds of tht oleginous with mximum physiologicl potentil. Physicl chrcteristics of fruits of J. curcs in the six mturtion stges (Tble 2) indicted tht the mximum length of fruits is reched t stge 2, while the dimeter progressed up to stge 3. Subsequent to those stges, there ws reduction in morphometric vribles. The length nd dimeter rnged from 33.0 mm nd 28.3 mm in fruits clssified s stge 1, to 27.2 mm nd 21.4 mm for fruits clssified s stge 6, respectively. These figures show tht both fruit length nd dimeter were reduced with the mturtion process s function of wter loss. Similr results were reported by Silv (2002) with the fruit size of Cnidosculus phyllcntus which reched mximum vlues 53 dys fter flowering, when fruits were entirely yellow. Sturnino et l. (2005) reported tht the dimeter of J. curcs when ripe rnged between mm, similrly to those mesured in this study. TABLE 2. Length nd dimeter of fruits nd seeds of Jtroph curcs L. depending on the mturtion stges. Fruit Seed Mturtion stge Length Dimeter Length Dimeter mm b bc c b b d c b e d c 8.75 MSD CV (%) Averges followed by the sme letter in columns re not sttisticlly different mong themselves t 5% by the Tukey test. 1. Green fruit; 2. Fruit t the beginning of yellow pigmenttion 3. Predominntly yellow fruit, 4. Fruit t the beginning of brown pigmenttion, 5. mture fruit; 6. Dry fruit t the beginning of dehiscence.

5 162 J. A. L. DRANSKI et l. The dimeter of seeds ws not significntly chnged in mturing stges of fruits (P> 0.05). However, higher vlues for length were reched when fruits showed mostly yellow. Seeds obtined from fruits t stge 1 mesured mm nd 892 mm, while those obtined by fruits t stge 6 mesured mm nd 8.75 mm in length nd dimeter, respectively (Tble 2). Sturnino et l. (2005) reported J. curcs seeds mm long nd 8-13 mm in dimeter which re similr mesures to this test. Studies on seeds morphology re necessry due to the importnce in botnicl identifiction, ecologicl spects, s well s the propgtion of species. Fruit dry weight (Tble 3) did not chnge significntly fter fruit pigmenttion (stge 2). There ws n verge gin of 314 mg fruit -1 from stge 1 to stge 2, where it remined constnt until dehiscence. Therefore, the dry mtter of fruits ws not n effective indictor of physiologicl mturity in seeds of J. curcs. TABLE 3. Mturtion stge Wter content nd dry mtter in fruits nd seeds of Jtroph curcs L. depending on mturtion stges of fruit. Fruit Seed Wter content Dry mss Wter content Dry mss ---- % mg fruit % mg seed b c b b 291 b bc b 299 b c b 294 b d b e c 259 bc MSD CV (%) Averges followed by the sme letter in columns re not sttisticlly different mong themselves t 5% by the Tukey test. 1. Green fruit; 2. Fruit t the beginning of yellow pigmenttion 3. Predominntly yellow fruit, 4. Fruit t the beginning of brown pigmenttion, 5. mture fruit; 6. Dry fruit t the beginning of dehiscence. The seeds of physic nut re chrcterized by three distinct phses concerning dry mss (Tble 3). From the green fruits to those in trnsition to yellow color, there ws n verge biomss gin of 71 mg seed -1 when they weighed 291 mg seed -1, which then remined sttisticlly equl until mturity (stge 5), followed by the third phse with mss reduction to 259 mg seed -1 due to deteriortion. Seed dry weight when combined with other fetures ws good indictor of physiologicl mturity for Torresi cren (Firmino et l., 1996), for Dlbergi nigr (Mrtins nd Silv, 1997), for Cedrel fissilis (Corvello et l., 1999), for Cnidosculus phyllcnthus for Mimos ceslpiniifoli by Alves (2005), for Peltophorum dubium by Aquino et l. (2006), for Mcherium brsiliense by Guimrães nd Brbos (2007), nd for Ceslpini echint by Aguir et l. (2007). However, for Brbos et l. (1992) fruit nd seed dry biomss of Copifer lngsdorffii were not good indictors of physiologicl mturity. The wter content of fruits nd seeds suffered grdul decrese with dvncing mturity (Tble 3). The wter content in seeds decresed from stge 1 to stge 2 whose vlue remined sttisticlly unchnged until the stge 5 of mturtion. The wter content (14.48%) in seeds of J. curcs t stge 6 is the result of less restriction to wter diffusion into the tmosphere s function of dehiscence. Similr behvior ws obtined by Mrtins et l. (2008) studying Jcrnd cuspidifoli. The sme uthors stted tht seeds with mximum germintion nd vigor re those hrvested from ripe fruits with brown color, whose seeds showed grdul dehydrtion process culminting t levels below 24.4% wter content. Firmino et l. (1996), Mrtins nd Silv (1997), Silv (2002), Mrcos Filho (2005) nd Iossi et l. (2007) stted tht wter content when ssocited with other chrcteristics highlights the

6 PHYSIOLOGICAL MATURITY OF SEEDS AND COLORIMETRY OF FRUITS IN PHYSIC NUT 163 physiologicl condition of seeds. Seeds of J. curcs extrcted from fruits t stge 1 did not germinte. From stge 4 chrcterized by the onset of brown pigmenttion, there ws significnt increse in seed germintion from 29% to 66% nd 74% t stges 5 nd 6, respectively (Figure 2). Emergence speed index nd first count germintion reched mximum vlues with seeds collected from fruits clssified s stges 5 nd 6. Seeds hrvesting of J.curcs bsed on fruits color mesured by digitl instrument will fcilitte the development of mchinery suitble for processing seeds of tht plnt. According to Kushik (2003), the mximum seed physiologicl potentil of J. curcs ws chieved in yellow fruits which recorded 89% of germintion with lower results obtined from seeds of green nd dry fruits. For Dgr et l. (2004), the increse in seed germintion ws positively correlted with fresh weight, with higher germintion vlues been chieved in seeds with mss exceeding 300 mg. Germque et l. (2002) studying the idel hrvesting time of purple-ipe (Tbebui impetiginos) seeds reported higher germintion nd vigor of seeds hrvested in the erly dehiscence of fruits when they showed green color with purple spots, nd seeds of green-yellow-brown color with wter content of 69.02% nd 50.98% in fruits nd seeds, respectively. 80 First count (%) b bc bc c Germintion (%) b bc bc c ESI 9 b bc bc c Mturtion stge FIGURE 2. First count of germintion, germintion percentge nd emergence speed index (ESI) of seeds of Jtroph curcs L. t different mturtion stges. Mrechl Cândido Rondon/PR, Mens compred by the Tukey test t 5% probbility. 1. Green fruit; 2. Fruit t the beginning of yellow pigmenttion 3. Predominntly yellow fruit, 4. Fruit t the beginning of brown pigmenttion, 5. Mture fruit; 6. Dry fruit t the beginning of dehiscence.

7 164 J. A. L. DRANSKI et l. CONCLUSIONS Seeds of Jtroph curcs L. with germintion higher thn 60% nd IVE higher thn 15 must be hrvested from ripe fruits with fully brown color of epicrp corresponding to the clssifiction 7 YR 4/2 of the Munsell color chrt, or refrctnce vlues equl to or lower thn 82, 70 or 65 nm in the red, green nd blue scles respectively, obtined with digitl colorimeter. Fruits with epicrp presenting the forementioned colorimetric chrcteristics hve mximum ccumultion of dry biomss, wter content below 38.5%, nd seeds with physiologicl mturity. ACKNOWLEDGEMENTS We express our thnks to the Rurl Biodiesel S/A by the vilbility of plnt mteril, nd the CNPq for finncil support. REFERENCES AGUIAR, F.F.A.; PINTO, M.M.; TAVARES, A.R.; KANASHIRO, S. Mturção de frutos de Ceslpini echint Lm. Pu-Brsil. Revist Árvore, v.31, n.1, p.1-6, ALVES, E.U.; SADER, R.; LUCENA, R.; ALCÂNTARA, B.; ALVES, A.U. Mturção fisiológic de sementes de Sbiá. Revist Brsileir de Sementes, v.27, n.5, p.1-8, AQUINO, N.F. BORTOLINI, M.; CAMPAGNOLO, M.A.; IGNÁCIO, V.L.; KOPPER, A.C.; MALAVASI, M.M. Dormêci de sementes de Peltophorum dubium (Sprengel.) Tubert colhids em diferentes estádios de desenvolvimento. Scienti Agrri Prnensis, v.5, n.2, p.31-37, ARRUDA, F.P.; BELTÃO, N.E.M.; ANDRADE, A.P.; PEREIRA, W.E; SEVERINO, L.S. Cultivo de pinhãomnso (Jtroph curcs L.) como lterntiv pr o semiárido nordestino. Revist Brsileir de Oleginoss e Fibross, v.8, n.1, p , BARBOSA, J.M.; AGUIAR, I.B.; SANTOS, S.R.G. Mturção de sementes de Copifer lngsdorffii Desf. Revist do Instituto Florestl, v.4, n.3, p , BELTRÃO, N.E.M. Agronegócio ds oleginoss no Brsil. Informe Agropecuário, v.26, p.44-78, BRASIL. Ministério d Agricultur e Reform Agrári. Secretári Ncionl de Defes Agropecuári. Deprtmento Ncionl de Defes Vegetl. Coordenção de Lbortório Vegetl. Regrs pr nálise de sementes. Brsíli, DF, p. CARNEIRO, J.G.A. Produção e controle de qulidde de muds florestis. Curitib: UFPR/UENF/FUPEF, p. CARVALHO, P.E.R. Produção de muds de espécies ntivs por sementes e implntção de povomentos. In: GALVÃO, A.P.M. (Org.). Reflorestmento de proprieddes ruris pr fins produtivos e mbientis. Um gui prático pr ções municipis e regionis. Colombo: Embrp Florests, p CARVALHO, N.M.; NAKAGAWA, J. Sementes: ciênci, tecnologi e produção. 2.ed. Jboticbl: FUNEP, p. CORVELLO, W.B.V.; VILLELA, F.A.; NEDEL, J.L.; PESKE, S.T. Mturção fisiológic de sementes de cedro (Cedrel fissilis Vell.). Revist Brsileir de Sementes, v.21, n.2, p.23-27, DAGAR, J. C.; BHAGWAN H.; KUMAR Y. Seed germintion studies of Slvdor persic nd Jtroph curcs. Indin Journl of Forestry, v.27, n.3, p , EFFENDI, Z. RAMLI,; R.; GHANI, J.A.; YAAKOB, Z. Development of Jtroph curcs color grding system for ripeness evlution. Europen Journl of Scientific Reserch, v.30, n.4, p , EMPRESA BRASILEIRA DE PESQUISA AGROPECUÁRIA - EMBRAPA.Centro Ncionl de Pesquis de Solos. Sistem Brsileiro de Clssificção de Solos. 2.ed. Rio de Jneiro: Embrp, p. FERREIRA, D.F. Análise esttístic por meio do SISVAR pr Windows versão 4.0. In: REUNIÃO ANUAL DA REGIÃO BRASILEIRA DA SOCIEDADE INTERNACIONAL DE BIOMETRIA, 45., São Crlos, Anis... São Crlos: UFScr: p FIRMINO, J.L.; SANTOS, D.S.B.; SANTOS FILHO, B.G. Crcterístics físics e fisiológics de sementes de cerejeir (Torresi cren Ducke) qundo s sementes form coletds do chão ou do interior dos frutos. Revist Brsileir de Sementes, v.18, n.1, p.28-32, GERMAQUE, R.C.R.; DAVIDE, A.C.; FARIA, J.M.C. Indicdores de mturidde fisiológic de sementes de ipêroxo (Tbebui impetiginos (Mrt.) Stndl.). Cerne, v.8, n.2, p.84-91, GUIMARÃES, D.M.; BARBOSA, J.M. Colorção

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