COFFEE SEEDLING PRODUCTION FROM STORED SEEDS Ros, S.D.V.F. d, Crvlho,.M., McDonld, M.., Von Pinho, E.R.V., Silv,.P. nd Veig,.D. (2011), Seed Sci. & Technol., 39, 151-164 The effect of storge conditions on coffee seed nd seedling qulity S.D.V.F. D ROS 1,.M. CRVLHO 2, M.. McDONLD 3, E.R.V. VON PINHO 2,.P. SILV 2 ND.D. VEIG 2 1 Emrp, PqE, v. W3 Norte (finl), Ed. Sede, CEP 70770-901, rsíli, DF, rzil (E-mil: sttelveig@dg.ufl.r) 2 Universidde Federl de Lvrs, CEP 37200-000, Lvrs, MG, rzil 3 Seed iology Progrm, Deprtment of Horticulture nd Crop Science, Ohio Stte University, 2021 Coffey Rd, Columus, OH 43210, US (ccepted Octoer 2010) Summry Otining commercilly useful coffee seedlings is hindered y slow, uneven germintion nd low tolernce to desicction s well s reduced coffee seed longevity. Coffee seeds hve een considered reclcitrnt, orthodox nd even intermedite with vrying results. Current recommendtions suggest tht coffee seeds cn e sfely stored etween 10-11% f.w.. t 15 C. However, fter drying nd storge, coffee seeds lose vigour, nd seeds stored fter drying cnnot e used for producing seedlings. Coffee seedling producers usully sow seeds immeditely or lightly dry them fter hrvest for short storge period, if necessry. It is highly desirle tht seeds re stored sfely to optimize coffee seedling production t the pproprite time nd seson with idel climtic conditions for plnting in the field. The ojective of this study ws to determine the qulity of coffee seedlings produced from seeds stored with high, medium nd low moisture levels under hermetic conditions t 10 nd 20 C. Seed nd seedling qulity were ssessed efore nd fter nine months storge. Only the germintion of seeds hrvested t the cherry stge, evluted efore nd fter the storge t 10 C, ws not ffected y moisture content, ut these seeds lost vigour nd did not produce suitle seedlings for plnting when stored for nine months. Seedlings produced in the nursery from seeds with 47 nd 12% moisture content performed the sme s those from greenish-yellow seeds, ut they produced lef re tht ws five times smller, stem height three times shorter, nd hd 1.7 times fewer pirs of true leves thn coffee seedlings produced from fresh seeds. Storge t 20 C ws not suitle for coffee seeds, especilly those t 18% moisture content whose qulity declined drsticlly. These results suggest tht coffee seeds do not tolerte desicction nd tht their storge ehviour clssifiction should e reviewed. Introduction Coffee is significnt griculturl product of economic nd socil importnce for rzil nd is importnt in world griusiness. Coffee trees re propgted using seedlings derived from seeds. However slow nd uneven germintion nd poor storge potentil slows coffee seedling development t the time nd seson when idel climtic conditions exist for plnting in the field in the principl rzilin coffee producing regions. Therefore, it is highly desirle tht coffee seeds e stored sfely to optimize coffee seedling 151
S.D.V.F. D ROS,.M. CRVLHO, M.. McDONLD, E.R.V. VON PINHO,.P. SILV ND.D. VEIG production. The intermedite storge ctegory hs een proposed for coffee seeds sed on the fct tht coffee seeds survived storge for out 10 months t 15 C t 10/11% (f.w..) moisture content nd decresed germintion ws otined with progressive reductions in seed moisture content nd storge temperture (Ellis et l., 1990; 1991). However, the idel moisture content for coffee seed conservtion is still eing studied. In comprehensive review of literture from the 1920s to the present, more thn 60 ppers hve provided contrdictory informtion regrding storing wet or dry coffee seeds. The results re inconsistent y showing tht idel storge moisture contents rnged from wet (greter thn 40% f.w..) to dry (10 to 12% f.w..), or even oth conditions. Studies conducted in the 1980s nd 1990s hve clssified Coffe ric L. seeds s reclcitrnt (King nd Roerts, 1979), orthodox (Roerts et l., 1984) nd intermedite (Ellis et l., 1990). More recent studies hve lso shown inconsistent clssifictions (Guimrães et l., 2002; Dussert et l., 2006; rghini nd Fzuoli, 2007; Veig et l., 2007; rújo et l., 2008). For commercil coffee seedling production, rzilin producers usully do not dry the seeds to 10-11% moisture content. Insted, they immeditely sow the seeds or only lightly dry them fter hrvest to enle short storge period. It is recommended tht coffee seeds e hrvested t the cherry mturtion stge. However, it hs een reported tht coffee seeds cquire mximum germintion cpcity when the fruits re etween the yellow-green nd cherry stges (Ellis et l., 1991; Guimrães et l., 2002). Severl uthors (Finch-Svge, 1996; Frrnt et l., 1988; Pmmenter nd erjk, 1999) hve suggested tht completely mture seeds of species sensitive to desicction my not possess mximum vigour ecuse the germintion process my strt t the end of the mturtion process. The ojective of this study ws to evlute the effects of seed moisture content nd storge temperture on coffee seed nd seedling qulity nd to determine the seeding production potentil of seeds stored t high, intermedite nd low moisture content. Mteril nd methods Seed hrvesting nd processing Coffe ric seeds, cultivr Rui, were hrvested from fruits t the cherry nd greenish-yellow mturtion stges in n experimentl field t the Federl University of Lvrs, MG, rzil, in the 2006/2007 growing seson. Fruits were pulped in hnd pulp seprtor nd the mucilge removed y nturl fermenttion in wter t 30 C for 24 h, wshed in running wter, nd left on n sorent pper towel to remove excess surfce wter efore drying. Seeds were dried in smll-scle sttionry dryer (Nvrtil nd urris, 1982) with 12.5m 3 min -1 ton -1 ir flow t 30 C (i.e. slow drying; 0.15%. h -1 ) to otin three levels of moisture (47, 18 nd 12% [f.w..]). sed on previous studies, these moisture contents were optimum for determining the effects of high, intermedite nd low moisture contents on seed nd seedling qulity. Seed moisture content ws determined efore storge on seeds without prchments (i.e fter removl of the endocrp, the remnnt of the fruit) in chmer t 105 C ± 3 C ccording to the Rules for Seed 152
COFFEE SEEDLING PRODUCTION FROM STORED SEEDS nlysis (rzil, 1992). Seeds were stored with their prchment in wterproof pckges t 10 C nd 20 C for nine months to test the potentil of the seeds to produce coffee seedlings in fvorle climtic seson for plnting in the field since six months re necessry for coffee tree formtion. Seed qulity evlution Seed physiologicl qulity ws ssessed efore nd fter the storge period when the stndrd germintion test ws used for determining the percentge rdicle protrusion, stndrd germintion (norml seedlings), nd seedlings with unfolded cotyledonry leves. The stndrd germintion test ws crried out with four susmples of 50 seeds ech without the outer skin, spred on moist germintion test pper with quntity of wter equivlent to two nd hlf times the weight of the dry sustrte, t 30 C with fluorescent white light t 750lux. Stndrd germintion percentge ws ssessed 30 dys fter sowing following the Rules for Seed nlysis (rzil, 1992). Physiologicl germintion ws ssessed t 15 dys fter the strt of the germintion test when seeds with t lest 2.0 mm root protrusion were counted nd the results expressed in percentge. Unfolded cotyledonry leves (jgur er stge) were ssessed 45 dys fter the strt of the germintion test nd the results expressed in percentge. Seedling production fter the seed storge period, coffee seedling production ws tested in nursery under mient environmentl conditions. Seeds were sown with ttched prchments s in commercil seedling production in 11 22 cm perforted polyethylene gs with stndrd sustrte (CFSMG, 1999) consisting of soil, mnure, simple super phosphte, nd potssium chloride t rtio of 70, 30, 0.5 nd 0.1%, respectively. fter purging the sustrte with methyl romide (Guimrães nd Mendes, 1998) nd prepring the gs, two seeds were sown directly per g nd covered with 2 cm sustrte. Ech experimentl plot consisted of 20 seedlings tht were wtered dily nd pproprite crop tretments nd weeding were crried out whenever necessry. Seedling qulity evlution The qulity of coffee seedlings produced from the stored seed ws ssessed in the nursery y the following: emergence percentge (finl stnd), emergence speed index, nd seedling developmentl prmeters (numer of pirs of true leves, stem dimeter, seedling height, lef re, nd the root nd cnopy dry mtter). The totl numer of emerged seedlings t 120 dys fter sowing in ech experimentl plot of 20 coffee seedlings t the jgur er stge ws expressed s percentge. The speed of emergence index ws clculted using dt from the dily count of emerged seedlings ccording to the formul y Edmond nd Drpl (1958). Coffee seedling developmentl prmeters were determined for six coffee seedlings in ech experimentl plot nd the following mesurements tken 180 dys fter sowing. The pirs of true leves were clculted nd the men pir of true leves determined. Stem dimeters were mesured in the plnt culm region using pchymeter nd the results expressed in millimeters per plnt. Seedling height ws otined y mesuring the region etween the culm nd the point of the terminl sprouts 153
S.D.V.F. D ROS,.M. CRVLHO, M.. McDONLD, E.R.V. VON PINHO,.P. SILV ND.D. VEIG of the coffee seedlings on the orthotropic rnch. Seedling height mens were clculted nd the results expressed in centimeters per seedling. Length nd width of one lef of ech pir of true leves were mesured nd the lef re of ech pir of leves otined multiplying the width length 0.667 2 (pir of leves) proposed y rros et l. (1973). Coffee seedling lef re ws otined y the sum of the re of the pirs of leves of ech coffee seedling. verge lef re ws otined from the lef res vlues of the coffee seedlings in the experimentl plot nd the results expressed in cm 2. Root nd cnopy dry weight determintions for the coffee seedlings were otined y removing the coffee seedlings from the gs, wshing the root system in running wter nd the roots were seprted from the cnopy y cutting the stem t the culm; the weights of the root system nd cnopy were otined fter drying in forced ir chmer t 60 C to constnt weight nd the men results expressed in g per coffee seedling. Sttisticl design complete rndomized lock design ws used to ssess lortory seed qulity with four replictions nd rndomized lock design ws used with three replictions to ssess nursery coffee seedling qulity. two (mturtion stges) three (seed moisture content) two (storge temperture) fctoril scheme ws used for totl of 12 tretments. The men results of the ssessments were compred y the Scott nd Knott test t 5% proility y trnsforming the percentge dt using [(rc sen X100-1/2 ) + 0.5]. The nlyses were crried out using the Sisvr progrm (Ferreir, 2000). Results Seed qulity evlution results fter hrvest nd drying, the mturtion stge hd significnt effect on the stndrd germintion; no significnt effects were oserved for physiologicl germintion, moisture content or interction mong these fctors. The effect of mturtion stge on the percentge of stndrd germintion ws significnt for the 18 nd 47% moisture contents (figure 1), ut not for the 12% moisture content. fter nine month s storge, however, the interction etween mturtion stge, seed moisture content, nd storge temperture ws significnt for ll seed physiologicl qulity ssessment vriles. With respect to fruit mturtion stge (figure 2), significnt differences were detected in ll ssessments, except for the physiologicl germintion (rdicle protrusion) vlues of 47% moisture content nd open cotyledonry leves t 18 nd 12% moisture content. Seeds t the greenish-yellow mturtion stge hd poorer physiologicl performnces thn those t the cherry mturtion stge nd the gretest reductions in seed qulity were oserved in seeds stored t 18% moisture content t 20 C. Considering storge temperture, seeds of the sme mturity level stored t 12% moisture content hd the sme physiologicl performnce (tle 1). The physiologicl performnce of seeds stored t 18% moisture content decresed when stored t 20 C t oth mturtion stges. mong seeds stored t 47% moisture content, there were no differences in greenish-yellow seed qulity following storge t the two tempertures, ut 154
COFFEE SEEDLING PRODUCTION FROM STORED SEEDS PG PG PG SG SG SG 100 90 80 70 60 50 40 30 20 10 0 12 18 47 Wter Content (%) Figure 1. Comprisons etween cherry ( ) nd greenish-yellow ( ) mturity stges within ech moisture contents. Seed physiologicl qulity ws ssessed y physiologicl germintion ( ) nd stndrd germintion ( ), fter drying of the seeds to different moisture contents (12, 18 nd 47%). The results re from one-wy NOV s. Lower cses re used for compring physiologicl germintion (PG) vlues nd upper ones for stndrd germintion (SG). verges with equl cse letters re not significntly different t P = 0.05 ccording to the Scott Knott test. Emrp/UFL/OSU, 2008. (%) seeds t the cherry mturtion stge hd lower stndrd germintion nd percentge open cotyledonry leves when stored t 20 C. Coffee seed performnce ws ffected very little y moisture content when seeds were stored t 10 C, regrdless of the mturtion stge (tle 1). However, t 20 C, oth ripe (cherry) nd non-ripe (greenish-yellow) seeds showed mrked reductions in physiologicl qulity when stored t 18% moisture content. Tle 1. ssessment results of the Coffe ric L. seeds otined from cherry fruits nd greenish-yellow fruits dried to different moisture contents, nd stored t different tempertures during nine months. The results re from three-wy NOV s. verges showing the sme letters were not significntly different t P = 0.05 ccording to the Scott Knott test. UFL, Lvrs, MG, 2007. Emrp / UFL / OSU, 2008. Moisture content Cherry Greenish-Yellow 10 C 20 C 10 C 20 C Physiologicl germintion 12 93 90 81 81 18 92 36 77 6 C 47 88 91 86 91 Stndrd germintion 12 86 90 81 76 18 90 40 C 77 11 C 47 91 58 77 87 Open Cotyledonry Leves 12 69 60 72 63 18 67 5 C 72 2 47 81 48 67 72 Cpitl letters re used for comprisons within columns nd lower cses letters within lines. 155
S.D.V.F. D ROS,.M. CRVLHO, M.. McDONLD, E.R.V. VON PINHO,.P. SILV ND.D. VEIG Rdicle Protrusion (%) 120 100 80 60 40 20 0 12 18 47 Wter Content (%) Viility (%) 120 100 80 60 40 20 0 12 18 47 Wter Content (%) (%) Leves Cotyledonry Open 100 80 60 40 20 0-20 12 18 47 Wter Content (%) Figure 2. Comprisons etween cherry (, ) nd greenish-yellow (, ) mturity stges within ech moisture contents (12, 18 nd 47%) nd storge temperture (10 nd 20 C). Seed physiologicl qulity ws ssessed fter nine months of storge, nd the results re from three-wy NOVs. Lower cses re used for compring within 10 C nd upper ones within 20 C. verges with equl cse letters re not significntly different t P = 0.05 ccording to the Scott Knott test. Emrp/UFL/OSU, 2008. Generlly, when seeds were dried to 12% moisture content, the mturtion stge nd storge temperture hd more influence on physiologicl qulity. However, when seeds were stored t 47% moisture content t 20 C, reductions in seed qulity were oserved only for the cherry mturtion stge. 156
COFFEE SEEDLING PRODUCTION FROM STORED SEEDS Seedling qulity ssessment Seeds hrvested t the cherry nd greenish-yellow mturtion stges nd dried to different moisture contents followed y storge t 10 C nd 20 C were tested for their cpcity to produce coffee seedlings tht were cple of propgting coffee plnts. Generlly, seed moisture content followed y storge temperture, were the most importnt fctors tht influenced coffee seedling qulity wheres the fruit mturtion stge lest influenced qulity. There ws triple interction of fctors tht influenced finl stnd nd speed of emergence index of seedlings produced in the nursery, nd lso n interction etween seed mturity nd storge temperture for the other vriles in the ssessment of the qulity of coffee seedlings. The est performnce s ssessed y the smllest emergence speed index vlues clculted ccording to the formul y Edmond nd Drpl (1958) were from coffee seedlings derived from greenish-yellow seed t ll the storge moisture contents nd tempertures (figure 3). Forty dys fter sowing, coffee seedlings derived from seeds t Finl Emergency (%) 100 90 80 70 60 50 40 30 20 10 0 10 o C 12 18 47 Wter Content (%) Emergence speed index 80 70 60 50 40 30 20 10 0 12 18 47 Wter Content (%) Figure. 3 Comprisons etween cherry (, ) nd greenish-yellow (, ) mturity stges within ech moisture contents (12, 18 nd 47%) nd storge temperture (10 nd 20 C). Seedlings were produced in the nursery from seeds stored for nine months. The results of seedling physiologicl qulity re from three-wy NOVs. Lower cses re used for compring within 10 C nd upper ones within 20 C. verges with equl cse letters re not significntly different t P = 0.05 ccording to the Scott Knott test. Emrp/UFL/OSU, 2008. 157
S.D.V.F. D ROS,.M. CRVLHO, M.. McDONLD, E.R.V. VON PINHO,.P. SILV ND.D. VEIG the greenish-yellow mturtion stge nd stored t 47% moisture content hd produced open cotyledonry leves (dt not shown) tht normlly occurs t 90 dys fter sowing. lthough these seedlings hd higher emergence speed indices t the end of plnting period, coffee seedlings derived from greenish-yellow seeds hd finl emergence percentges similr to those derived from cherry stge seeds (figure 3). There ws significnt difference only in coffee seedlings derived from greenish-yellow seeds stored t 18% moisture content t 20 C tht resulted in the poorest performnce. Coffee seedlings produced from cherry or greenish-yellow fruit seeds stored t 18% moisture content t 20 C produced the poorest qulity seedlings nd this finding ws consistent with the seed qulity ssessments in the lortory. There ws significnt interction etween seed moisture content nd storge temperture for ll other vriles for coffee seedling qulity ssessment, ut no significnt effect ws oserved for fruit mturtion stge. Seed moisture content nd storge temperture hd less impct on coffee seedling production from seed stored t 18% moisture content t 20 C (tle 2). In contrst, coffee seedlings produced from seeds hrvested t the greenish-yellow or cherry stge stored t 12 or 47% moisture content did not hve differences in seed qulity following storge t either 10 C or 20 C. Poorer physiologicl qulity ws oserved in most of the tretments in coffee seedlings stored t 18% moisture content. Seeds stored t 18% moisture content t 20 C for oth mturtion stges consistently gve poor results. These results re further supported t 85 dys, when the finl stnd hd lredy een estlished nd most seedlings lredy hd unfolded cotyledonry leves (dt not presented). The est seedling performnce ws from seedlings derived from seeds t the cherry mturtion stge t 47% moisture content nd stored t 10 C. Discussion Slow nd uneven germintion of coffee seeds gretly hinders otining high qulity coffee seedlings to form coffee plnttion. Furthermore, in the min rzilin coffee producing regions, tempertures re in the 15 to 19 C rnge during the seed germintion nd seedling emergence period cusing complete seedling emergence to tke up to 120 dys (Guimrães nd Mendes, 1998). The idel temperture for coffee seed germintion is 30 C (rzil, 2009). However, this temperture results in coffee seedling dmge tht is lredy slow process, tking t lest six months to e concluded. Thus, in ddition to germintion cpcity, it is highly desirle tht coffee seeds hve high vigour index tht leds to etter performnce during germintion nd coffee seedling formtion in the nursery, especilly under dverse conditions, so tht more vigorous seedlings re otined. Under these conditions, vigorous coffee seedlings hve fster estlishment in productive plnttion nd fster return for the producer in invested cpitl. The results presented here show tht coffee seeds cn germinte when they re dried to 12% moisture content, n intermedite moisture level mong the levels tolerted y reclcitrnt nd orthodox seeds. fter nine months storge they still hve the ility to germinte. lthough the fruit mturtion stge nd seed moisture content fter drying 158
COFFEE SEEDLING PRODUCTION FROM STORED SEEDS Tle 2 ssessment results of the Coffe ric L. seedlings otined from fruits dried to different moisture contents, nd stored t different tempertures during nine months. The results re from two-wy NOVs. verges showing the sme letters were not significntly different t P = 0.05 ccording to the Scott Knott test. Dt re the verge ±SE from six replictions. UFL, Lvrs, MG, 2007. Emrp / UFL / OSU, 2008. Storge Wter content (%) Temperture P ( C) 12 18 47 Pirs of true leves 10 C 2.6 2.5 2.9 0.5752 20 C 2.8 0.8 2.8 <0.0000 Stem dimeter 10 C 1.57 1.59 1.50 0.6220 20 C 1.59 0.70 1.67 <0.0000 Seedling height 10 C 5.40 5.35 5.21 0.9329 20 C 5.40 2.10 5.24 <0.0000 Root dry mtter 10 C 0.98 0.88 1.00 0.6885 20 C 1.10 0.18 0.98 <0.0000 Cnopy dry mtter 10 C 2.23 2.10 2.13 0.9688 20 C 2.46 0.64 2.32 0.0027 Lefy re 10 C 44.11 45.58 44.37 0.9889 20 C 51.55 12.36 49.69 0.0010 Lower cses re used for comprisons within lines nd cpitl letters within columns. hd significnt effects on seed qulity, seeds sumitted to the different tretments hd physiologicl germintion (rdicle protrusion) nd stndrd germintion (norml seedlings) vlues similr to ech other nd greter thn 80%. Seeds hrvested t the cherry mturtion stge hd sttisticlly equl germintion percentge t ll moisture contents, indicting tht coffee seeds tolerte drying to 12% moisture content, supporting the results reported y others (Ellis et l., 1990 nd 1991; Hong nd Ellis, 1992 nd 1995; Dussert et l., 1999; Eir et l., 1999; Dussert et l., 2006). However, it is importnt to note tht in ll the former studies to define coffee seed response following drying nd storge, seeds were tested without the prchment lyer under lortory conditions. These results filed to determine the potentil of dry stored seeds for producing seedlings under nursery conditions s done in this study. In ddition to slow nd uneven coffee seed germintion chrcteristics, in commercil prctice, coffee seeds re plnted with the prchment ecuse there is no sfe method for its removl without dversely ffecting seed qulity. Tht is the reson why coffee 159
S.D.V.F. D ROS,.M. CRVLHO, M.. McDONLD, E.R.V. VON PINHO,.P. SILV ND.D. VEIG seeds were plnted in this study with the prchment ttched to evlute their potentil for producing seedlings in nursery. The outer skin is mechnicl rrier to seed germintion tht mkes coffee seedling development even more difficult (Válio, 1976; Ren nd Mestri, 1986; Guimrães, 1995; Ros et l. 2002 or 2005). Ellis et l. (1990) lso indicted tht not ll plnted seeds re le to emerge from the soil surfce nd grow, nd some hve just their rdicles emerged ut remin under the soil. lthough seeds germinted fter drying nd fter nine months storge, they hd low vigour index vlues tht reduced the cpcity to form coffee seedlings. Coffee seeds with high vigour nd storge cpcity for t lest nine or ten months re highly desirle ecuse the germintion nd emergence processes cn strt under more fvorle temperture conditions nd successful coffee seedlings cn e otined in climte seson more suitle for plnting from Septemer to Novemer. Otining coffee seedlings t the strt of Octoer, s long s they hve commerciliztion stndrds, with three to seven pirs of true leves, increses the likelihood of successful coffee seedling production ecuse the climtic conditions re more fvorle to plnt the coffee seedling in the field during this period. Unfortuntely, storing coffee seeds t intermedite moisture content (18%) reduced physiologicl qulity, especilly when the seeds were stored t 20 C. Respirtion dmge from free rdicls cn occur t n intermedite moisture content level (etween -15 nd -5 MP), ut the injury repir process is not ctivted. Seeds stored or sumitted to slow drying so they remin t n intermedite moisture content level for long period of time deteriorte in similr fshion to the physiologicl mechnisms ssocited with the ccelerted ging test (Wlters et l., 2001; Leprince et l., 2000). Dussert et l. (2006) showed tht, lthough slow drying did not directly influence coffee seed viility, it induced oxidtive injury nd deteriortion processes in the seed ecuse the seeds remined t n intermedite moisture content level. ll the tretments, except for the seeds with 18% moisture content stored t 20 C, produced coffee seedlings with sttisticlly equl developmentl prmeters. Coffee seedlings produced in the present study, including those derived from seeds processed nd stored ccording to the recommendtions y Hong nd Ellis (1992, 1995), i.e., 10-11% (f.w..) moisture content nd 10-15 C storge temperture, did not produce coffee seedlings suitle for plnting. lthough these coffee seedlings were produced during the summer under higher tempertures, they hd chrcteristics very different from seedlings produced during the winter, the usul seson for commercil coffee seedling production. Coffee seedlings produced in this study hd, on verge, lef re five times smller, stem height three times shorter, nd 1.7 times fewer pirs of true leves thn those produced in the winter (Ros et l., 2007). These results show tht even though coffee seeds germinted fter drying, their vigour decresed during storge. These results confirm tht the idel pproch to processing nd storing coffee seeds in the wet or dry condition still remins to e determined. Desicction-tolernt seeds whose longevity is enhnced y reduced moisture content, temperture, nd decresed reltive humidity hve desicction tolernce mechnisms such s lte emryonic undnt protein synthesis nd ccumultion (le or le like) (Wlters et l., 1997; Wolkers et l., 2001) nd sugrs such s sucrose nd some oligoscchrides 160
COFFEE SEEDLING PRODUCTION FROM STORED SEEDS (Oendorf, 1997; uitink et l., 2000; Oliver et l., 2001; uitink et l., 2002; uitink nd Leprince, 2004) involved in the formtion of queous glsses, s well s the presence of enzymtic nd non-enzymtic nti-oxidnt systems (erjk, 2006). These mechnisms should e completely expressed nd ctivted in the loctions where they re required to protect ginst free rdicl ttck (erjk, 2006). Sugr ccumultion (Chrillnge et l., 2000; Guimrães et l., 2002) nd cytoplsmtic glss formtion (dt eing prepred for puliction) do not pper to exist in coffee seeds following drying nd storge. These dt indicte tht desicction tolernce mechnisms in coffee my not ssocited with wter removl s occurs in desicction-tolernt seeds (ewley nd lck, 1994; Leprince et l., 1993). It hs een suggested tht the reduction in orthodox seed moisture content during mturtion drying elicits switch from developmentl to germintive mode s shown in chnges in genome ctivity resulting in the synthesis of germintion-specific enzymes (Evns et l., 1975; Dsgupt nd ewley, 1982; rmstrong et l., 1982; Dsgupt et l., 1982; Kermode nd ewley, 1985; Misr et l., 1985; Kermode et l., 1989; ewley et l., 1989; Oishi nd ewley, 1990; urris et l., 1997). Seeds of other species, which develop within fleshy fruit nd frequently do not dry out prior to dispersl, cn germinte when removed from the fruit without ny requirement for drying. Thus,removl of the seeds from the constrints of the fruit is sufficient for germintion (ewley nd lck, 1994). In this study, coffee seeds hrvested in the greenish-yellow mturtion stge, germinted s they were removed from the fruits without ny drying tretment indicting tht drying is not requirement for germintion. Recent reserch concerning the effects of post-hrvesting processing of coffee fruits (Selmr et l., 2004; Selmr et l., 2006; ytof et l., 2007) show tht in the course of processing, fter the fruit flesh is removed, the seeds hve the germintion process initited on the sis of the expression of the germintion-specific isocitrte lyse nd n increse in the undnce of β-tuulin, mker for cell division or elongtion. Other studies on coffee seed performnce during drying nd storge nd in wet nd dry conditions hve not shown n interction mong physiologicl nd pthologicl effects on seed qulity. ecuse coffee seeds germinte slowly nd hve low storge potentil, it is importnt to consider their stte of helth, ecuse interctions mong the physiologicl nd pthologicl prmeters cn led to erroneous interprettions nd contrdictory results. It is known tht this interction cn influence the ssessments of seed performnce sensitive to desicction during storge, especilly when they re stored under wet conditions (Mycock nd erjk, 1995). The influence of this interction on the ssessment of seed qulity ws oserved in this study. Coffee seeds derived from fruits t the greenish-yellow stge fter nine months storge performed etter thn those extrcted t the cherry stge. Figure 3 shows enhnced speed of emergence indices nd finl stnd percentge for greenish-yellow seeds compred to cherry seeds. In other ssessments of coffee seedling performnce, differences were not oserved etween the greenishyellow nd cherry seeds, nd only the effects of moisture content nd storge temperture previled. This etter performnce of seedlings produced from greenish-yellow seeds is lso oserved for seedlings from seeds stored t 47% moisture content t 20 C. The pthologicl qulity of the seeds ws not determined, ut greenish-yellow seeds were 161
S.D.V.F. D ROS,.M. CRVLHO, M.. McDONLD, E.R.V. VON PINHO,.P. SILV ND.D. VEIG visily less infected y fungi thn seeds t the cherry stge nd this my explin their enhnced performnce when stored t 20 C due to the etter pthologicl helth qulity. Thus, ccording to the dt presented nd discussed here, dry stored coffee seeds do not produce suitle seedlings for plnting nd the sttement they re tolernt to desicction to 10-11% moisture content should e reviewed. References rmstrong, C., lck, M., Chpmn, J.M., Normn, H.E. nd ngold, R. (1982). The induction of sensivity to gierellin in leuronic tissue of developing whet grins. I. The effect of dehydrtion. Plnt, 154, 573-577. rros, R.S., Mestri, M. nd rg Filho, L.J. (1973). Determinção d áre de folhs de cfé (Coffe ric L., cv. ouron mrelo). Revist Ceres, 20, 44-53. erjk, P. (2006). Unifying perspectives of some mechnisms sic to desicction tolernce cross life forms. Seed Science Reserch, 16, 1-15. ewley, J.D. nd lck, M. (1994). Seeds: physiology of development nd germintion. New York: London Plenum Press, 367 p. ewley, J.D., Kermode,.R. nd Misr, S. (1989). Desicction nd miniml drying tretments of seeds of cstor en nd Phseolus vulgris which terminte development nd promote germintion cuse chnges in protein nd messenger RN syntheses. nnls of otny, 63, 3-17. rsil (1992). Ministério d gricultur e Reform grári. Secretri Ncionl de Defes gropecuári. Regrs pr nálise de Sementes. rsíli, DF. 365 p. uitink, J. nd Leprince, O. (2004). Glss formtion in plnt nyhdroiotes: survivl in the dry stte. Cryoiology, 48, 215-228. uitink, J., Hemming, M.. nd Hoekstr, F.. (2000). Is there role for oligoscchrides in seed longevity? n ssessment of intrcellulr glss stility. Plnt Physiology, 122, 1217-1224. uitink, J., Hoekstr, F.. nd Leprince, O. (2002). iochemistry nd iophysics of tolernce systems. pp. 293-318 in lck, M.; Pritchrd, H.W. (Eds) Desicction nd survivl in plnts: Drying without dying. Wllingford, CI Pulishing. rghini, M.T. nd Fzuoli, L.C. (2007). rmzenmento de sementes de cfé. O gronômico, 59, 44-45. urris, J.S., Peterson, J.M., Perdomo,.J. nd Feng, D.S. (1997). Morphologicl nd physiologicl chnges ssocited with desicction in mize emryos. In: Ellis, R.H.; lck, M.; Murdoch,.J. nd Hong, T.D. sic nd pplied spects of seed iology. Dordrecht: Kluwer cdemic Pulishers 1997. 103-111. ytof, G., Knopp, S., Krmer, D., reitenstein,., ergervoet, J.H., Groot, S.P.C. nd Selmr, D. (2007). Trnsient occurrence of seed germintion processes during coffee post-tretment. nnls of otny, 100, 61-66. CFSMG-Comissão de Fertilidde do Solo do Estdo de Mins Geris (1999). Recomendções pr o uso de corretivos e fertilizntes em Mins Geris, 5ª proximção. Lvrs, 1999. 359p. Chrillnge, N., Dussert, S., Engelmnn, F., Douldeu, S. nd Hmon, S. (2000). Desicction tolernce in reltion to solule sugr contents in seeds of ten coffee (Coffe L.) species. Seed Science Reserch, 10, 393-396. Dsgupt, J. nd ewley, J.D. (1982). Desicction of xes of Phseolus vulgsis during development cuses switch from developmentl pttern of protein synthesis to germintion pttern. Plnt Physiology, 70, 1224-1227. Dsgupt, J., ewley, J.D. nd Yeung, E.C. (1982). Desicction-tolernt nd desicction-intolernt stges during the development nd germintion of Phseolus vulgris seeds. Journl of Experimentl otny, 33, 1045-1057. Dussert, S., Chrillnge, N., Engelmnn, F. nd Hmon, S. (1999). Quntittive estimtion of seed desicction sensitivity using quntl response model: ppliction to nine species of the genus Coffe L. Seed Science Reserch, 9, 135-144. 162
COFFEE SEEDLING PRODUCTION FROM STORED SEEDS Dussert, S., Dvey, M.W., Lffrgue,., Douleu, S., Swennen, R. nd Etienne, H. (2006). Oxidtive stress, phospholipids loss nd lipid hydrolysis during drying nd storge of intermedite seeds. Physiologi Plntrum, 127, 192-204. Edmond, J.. nd Drpl, W.S. (1958). The effects of temperture, snd nd cerone on germintion of okr seed. Proceedings of mericn Society for Horticulturl Science, 71, 428-434, June. Eir, M.T.S., Wlters, C., Clds, L.S., Fzuoli, L.C., Smpio, J.. nd Dis, M.C.L.L. (1999). Tolernce of coffe spp. seeds to desicction nd low temperture. Revist rsileir de Fisiologi Vegetl., 11, 97-105. Ellis, R.H., Hong, T.D. nd Roerts, E.H. (1990). n intermedite ctegory of seed storge ehvior? I. Coffee. Journl of Experimentl otny, 41, 167-1174, Sept. Ellis, R.H., Hong, T.D. nd Roerts, E.H. (1991). n intermedite ctegory of seed storge ehviour? II. Effects if provennce, immturity nd imiition on desicction tolernce in coffee. Journl of Experimentl otny, 42, 653-657, My. Evns, M., lck, M. nd Chpmn, J.M. (1975). Induction of hormone sensivity y dehydrtion is one positive role for drying in cerel seed. Nture, 258, 144-145. Frrnt, J.M., Pmmenter, N.W. nd erjk, P. (1988). Reclcitrnce current ssessment. Seed Science nd Technology, 16, 155-166. Ferreir, D.F. (2000). Sistem de nálise esttístic- SISVR. Lvrs: DCE, UFL. Finch-Svge, W.E. (1996). The role of developmentl studies in reserch on reclcitrnt nd intermedite seeds. In: Workshop on Improved Methods for Hndling nd Storge of Intermedite/Reclcitrnt Tropicl Forest Tree Seeds, 1995, Humleek, Denmrk. Proceedings... Humleek, Denmrk, 1996. p.83-97. Guimrães, R.J. (1995). Formção de muds de cfeeiro: (Coffe rid L.): Efeitos de reguldores de crescimento e remoção do pergminho n germinção de sementes e do uso de N e K em coertur, no desenvolvimento de muds. 133p. Tese (Doutordo em Fitotecni) Universidde Federl de Lvrs. Lvrs. Guimrães, R.J. nd Mendes,.N.G. (1998). Produção de muds de cfeeiro. In: Mendes, NG, Guimrães, RG (eds). Cfeicultur empresril - produtividde e qulidde. Lvrs: UFL/FEPE, p.1-60. Guimrães, R.M., Vieir, M.G.G.C., Frg,.D., Von Pinho, E.V.R. nd Ferrz, V.P. (2002). Tolerânci à dessecção em sementes de cfeeiro (Coffe ric L.). Cienci e grotecnologi, 26, 128-139. Hong, T.D. nd Ellis, R.H. (1992). Optimum ir-dry seed storge environments for ric coffee. Seed Science nd Technology, 20, 547-560. Hong, T.D. nd Ellis, R.H. (1995). Interespecific vrition in seed storge ehviour within two gener Coffe nd Citrus. Seed Science nd Technology, 23, 165-181. Hong, T.D. nd Ellis, R.H. (2002). Optimum moisture sttus for the exceptionl survivl of seeds of ric coffee (Coffe ric L.) in medium-term storge t -20 C. Seed Science Technology, 30, 131-136. Kermode,.R. nd ewley, J.D. (1985). The role of mturtion drying in the trnsition from seed development to germintion. I. cquisition of desicction-tolernce nd germinility during development of Ricinus communis L. seeds. Journl of Experimentl otny, 36, 1906-1915. Kermode,.R., Oishi, M.Y. nd ewley, J.D. (1989). Regultory roles for desicction nd scisic cid in seed development: comprison of the evidence from whole seeds nd isolted emryos. In: Stnwood, P. C. nd Mcdonld, M.. (eds.). Seed Moisture. Mdison: CSS, 1989. p.23-50. (Specil Puliction, 14). King, M.W. nd Roerts, E.H. (1979). The storge of reclcitrnt seeds: chievements nd possile pproches. Interntionl ord for Plnt Genetic Resources, Rome. Leprince O., Hrren, F.J.M., uitink J., lerd, M. nd Hoekstr, F.. (2000). Metolic dysfunction nd unted respirtion of germinting rdicles. Plnt Physiology, 112, 597-608. Leprince, O., Hendry, G..F. nd McKersie,.D. (1993). The mechnims of desecction tolernce in developing seeds. Seed Science Reserch, 3, 231-246. Misr, S., Kermode,. nd ewley, J.D. (1985). Moleculr form nd function of the plnt genome. New York: Plenum, 1985. (NTO dvnced Studies Institute Proceedings). Mturtion drying s the switch tht termintes seed development nd promotes germintion, p.113-128. Mycock, D.J. nd erjk P. (1995). The implictions of Seed-ssocited Mycoflor During Storge. In: Kigel, J. nd Glili, G., ed. Seed Development nd Germintion. oc Rton, NY: CRC Press, 747-766. Nvrtil, R.J. nd urris, J.S. (1982). Smll-scle dryer designer. gronomy Journl, 74, 159-161. Oendorf, R.L. (1997). Oligoscchrides nd glctosyl cyclitols in seed desicction tolernce. Seed Science Reserch, 7, 63-74. 163
S.D.V.F. D ROS,.M. CRVLHO, M.. McDONLD, E.R.V. VON PINHO,.P. SILV ND.D. VEIG Oishi, M.Y. nd ewley, J.D. (1990). Distinction etween the responses of developing mize kernels to fluridone nd desicction in reltion to germinility, α-mylse ctivity, nd scisic cid content. Plnt Physiology, 94, 592-598. Oliver,.E., Leprince, O., Wolkers, W.F., Hinch, D.K., Heyer,.G. nd Crowe, J.H. (2001). Non-discchridesed mechnisms of protection during drying. Cryoiology, 44, 151-167. Pmmenter, N.W. nd erjk, P.. (1999). Review of reclcitrnt seed physiology in reltion to dessiction tolernce mechnisms. Seed Science Reserch, 9, 13-37. Ren,.. nd Mestri, M. (1986). Fisiologi do Cfeeiro. In: Simpósio sore ftores que fetm produtividde do cfeeiro, Poços de Clds, 1986. nis... Pircic: POTFÓS, 13-85. Roerts, E.H. (1973). Predicting the storge life of seeds. Seed Science nd Technology, 1, 499-514. Roerts, E.H., King, M.W. nd Ellis, R.H. (1984). Reclcitrnt seeds: their recognition nd storge. In: Crop genetic resources: conservtion nd evlution. Eds. J.H.W. Holden nd J.T. Willims. George llen nd Unwim, London, pp.38-52. Ros, S.D.V.F., rndão Junior, D.S., Von Pinho, E.V.R., Veig,.D. nd Silv, L.H.C. (2005). Effects of different drying rtes on the physiologicl qulity of Coffe cnephor Pierre seeds. rzilin Journl Plnt Physiology, 17, 199-205. Ros, S.D.V.F., Melo, L.Q., Veig,.D., Oliveir, S., Souz, C..S. nd guir, V.. (2007). Formção de muds de Coffe ric L., cv Rui, utilizndo sementes e frutos em diferentes estádios de desenvolvimento. Ciênci e grotecnologi, 31, 349-356. Selmr, D., ytof, G., Knopp, S.E., rdury,., Wilkens, J. nd ecker, R. (2004) iochemicl insights into coffee processing: qulity nd nture of green coffee re interconnected with n ctive seed metolism. In: 20 th Interntionl Conference in Coffee Science. nglore 2004, SIC, Indi cd-room. Selmr, D. ytof, G., Knopp, S.-E. nd reitenstein,. (2006). Germintion of coffee seeds nd its significnce for coffee qulity. Plnt iology, 8, 260-264. Vlio, I.F.M. (1976). Germintion of coffee seeds (Coffe ric L., cv. Mundo Novo). Journl of Experimentl otny, 27, 983-991. Veig,.D., Guimrães, R.M., Ros, S.D.V.F. d, Von Pinho, E.V. de R, Silv, L.H. de C. e, Veig,.D. (2007). rmzenilidde de sementes de cfeeiro colhids em diferentes estádios de mturção e sumetids diferentes métodos de secgem. Revist rsileir de Sementes, 29, 83-91. Wlters, C., Ried, J.L. nd Wlker-Simmons, M.K. (1997). Het-solule proteins extrcted from whet emryos hve tightly ound sugrs nd hydrtion properties. Seed Science Reserch, 7, 125-134. Wlters, W.F., McCredy, S., rndt, W.F., Lindsey, G. nd Hoekstr, F.. (2001). Isoltion nd chrcteriztion of D-7 LE protein from pollen tht stilizes glsses in vitro. iochimic et iophysic ct, 1544. Wolkers, W.F., McCredy, S., rndt, W.F., Lindsey, G.C. nd Hoekstr, F.. (2001). Isoltion nd chrcteriztion of D-7 LE protein from pollen tht stilizes glsses in vitro. iochimic et iophysic ct, 1544, 196-206. 164