Bioactive Compounds and Antioxidant Activity of Tomato High Lycopene Content Advanced Breeding Lines

Transcription

1 The Afrin Journl of Plnt Siene nd Biotehnology 9 Glol Siene Books Biotive Compounds nd Antioxidnt Ativity of Tomto High Lyopene Content Advned Breeding Lines Ridh Ilhy 1,2 Chfik Hdider 1* Imen Tlili 1,2 1 Lortory of Biotehnology nd Plnt Physiology, Ntionl Agriulturl Reserh Institute of Tunisi. Hedi Krry street, 249 Arin, Tunis, Tunisi 2 Deprtment of Biology, Fulty of Sienes, Bizerte, Tunisi Corresponding uthor: * hdider.hfik@ires.grinet.tn ABSTRACT In Tunisi, tomto (Lyopersion esulentum Mill.) is the min vegetle grown nd onsumed ll yer round nd is therefore of strtegi importne. Tomto fruits re eoming then n importnt soure of nturl ntioxidnts primrily lyopene, phenolis nd vitmin C, whih re involved in inhiiting retive oxygen speies responsile for mny ner nd rdiovsulr diseses. Breeding for high nutritionl tomto vlue is eoming n inresingly importnt im. In this ontext, tomto high lyopene ontent dvned reeding lines (HLT-F51 nd HLT-F52) were evluted for their totl rotenoid, lyopene, totl phenolis, flvonoids, sori id, dehydrosori id s well s their hydrophili nd lipophili ntioxidnt tivities, ompred to the vriety Rio Grnde ommonly grown in Tunisi. All tested high lyopene ontent vrieties showed generlly stisfying gronomi hrteristis. The totl rotenoid, lyopene, totl phenolis, flvonoids, dehydrosori id, totl vitmin C nd hydrophili nd lipophili ntioxidnt tivities in tomto fruit vried signifintly etween the studied vrieties. Compred to the ontrol, the seleted line HLT-F51 showed 2.65-, 2.62 nd fold higher totl rotenoid, lyopene nd flvonoids, respetively. Also, HLT-F51 showed 2.9 nd 2.24-fold higher hydrophili nd lipophili ntioxidnt tivities respetively. HLT-F52 exhiited prtiulrly higher dehydrosori id nd totl vitmin C ontents ompred to the ontrol vriety Rio Grnde. These results emphsize the promising use of suh dvned reeding lines for helthy qulity produts. Keywords: rotenoids, hydrophili nd lipophili ntioxidnt tivities, Lyopersion esulentum Mill, phenolis, tomto fruit qulity, vitmin C INTRODUCTION In Tunisi, tomto (Lyopersion esulentum Mill.) is the min vegetle grown nd onsumed nd is therefore of strtegi importne. In 7, the tomto rop mounted to 78, t, inluding 558, t for proessing. Moreover, Tunisi is the min tomto-proessing ountry in Afri nd mong the world s leders in terms of per pit nnul onsumption, with up to 54 kg on fresh weight sis (Hdider et l. 7). During the pst dedes, mny tomto vrieties were developed with the im of inresing yield potentil, disese tolerne nd extending shelf life without enough ttention to their level of helth-promoting ompounds (Doris et l. 8). Nowdys, the demnd for high nutritionl qulity food is inresing euse of the inresing onsumer wreness of the reltionship etween foods nd helth nd the ommeril opportunities offered y suh produts. The regulr onsumption of fresh tomto or tomto produts hs een inversely orrelted to the development of widespred humn diseses when tken dily in dequte mounts (Ro nd Agrwl 1998). This protetive effet hs een minly ttriuted to the rotenoid onstituents of the fruit prtiulrly lyopene nd -rotene (Sies nd Sthl 1998). These ompounds my ply n importnt role in inhiiting retive oxygen speies responsile for mny importnt diseses (Clinton 1998). Besides rotenoids, other ntioxidnts present in tomtoes, suh s phenoli ompounds nd sori id, lso ply n importnt role in preventing diseses (Rords et l. 1999; Krky et l. 1). Therefore, mximizing the level of these iotive ompounds in tomto fruit vrieties is eoming one of the gols of mjor importne for tomto reeders (Atnssov et l. 7; Doris et l. 8). In order to stisfy the demnd of growers, proessors nd onsumers for high nutritive qulity food, lrge numer of new tomto vrieties with inresed lyopene ontent (high lyopene ontent tomto) hve een developed y onventionl plnt reeding tehniques. However, it hs een reported tht the seletion of plnts whih nturlly over express genes improving lyopene ontent my led to the formtion of plnt with mny undesirle hrteristis suh s redued level of other ntioxidnts suh s -rotene on ehlf of lyopene (Sks nd Frnis 1), low fruit size nd redued produtivity thn in urrently ville vrieties (Atnssov et l. 7). In this ontext, there is need to investigte the ehviour of these high lyopene ontent tomto vrieties under vrious environmentl growing onditions. In ft, Dums et l. (3) nd Atnssov et l (7) reported tht lyopene ontent in tomto fruit vry not only in reltion to genotype, ut lso depends on the environmentl ftors nd griulturl tehniques used. Some reserhers hve studied new high lyopene ontent tomto vrieties in order to evlute their qulity hrteristis. However, few studies hve investigted their ntioxidnt tivity (Lenui et l. 6). In Tunisi, lthough informtion on the urrently grown tomto vrieties is widely ville (Hdider et l. 7), little is known out high lyopene ontent tomto vrieties. In n erly study, we evluted rnge of high lyopene ontent tomto vrieties nd onluded tht despite the high lyopene ontent of some seleted vrieties, they were hrterized y low fruit size (Ilhy nd Hdider 7), hrteristi generlly not ppreited y the Tunisin fresh mrket onsumers (Hdider et l. 7). In ddition, some vrieties showed oth rdil nd onentri rking whih leves Reeived: 31 July, 9. Aepted: 11 Novemer, 9. Originl Reserh Pper

2 The Afrin Journl of Plnt Siene nd Biotehnology 3 (Speil Issue 1), Glol Siene Books the ffeted tomtoes unmrketle nd quikly deteriorting. Therefore, nd sed on the fts mentioned ove the im of this study ws to evlute two dvned tomto reeding lines for yield nd fruit qulity hrteristis ompred to the vriety Rio Grnde ommonly grown in Tunisi. MATERIALS AND METHODS Field experiment The field experiment ws rried out t Mnnou support reserh sttion in northern Tunisi during the 8 growing seson (Mrh July). Three tomto vrieties were used in this experiment. Two dvned tomto high lyopene ontent lines (F5 genertion), with the ssigned nmes HLT-F51 nd HLT-F52, seleted y the Ntionl Agriulturl Reserh Institute of Tunisi nd the open pollinted vriety Rio Grnde (Petoseed, Stioy, CA, USA) ommonly grown in Tunisi whih ws used s ontrol. The HLT-F51 nd HLT-F52 lines were seleted y single seed desendnt from essions otined from Isreli origin on the sis of lrger fruit size nd tolerne to rking. The high-lyopene tomto essions derive from spontneous mutnt hrterized y deeply pigmented fruits due to their exggerted light responsiveness with respet to wild-type plnts (Mustilli et l. 1999; Atnssov et l. 7). Sowing ws rried out in Mrh 8 in plug-seedling trys. Trnsplnting in doule rows took ple on April 8 t density of out 33, plnts h 1. Sping within rows nd etween doule rows ws.4 nd 1.5 m, respetively. Tomto vrieties were grown in three replited plots. Irrigtion ws pplied using drip method with 4 L h 1 drippers pled t.4 m intervls long the irrigtion line. Drip irrigtion my run for h t time, fter 1 2 dy intervl, depending on potentil evpotrnspirtion for reserh sttion limte dt nd rop oeffiient. The prodution method ws in ordne with the proedures utilized y the Mnnou support reserh sttion nd high-yielding Tunisin onventionl frmers. It inluded fertiliztion with syntheti hemil fertilizers (186 kg N h 1, kg P 2 O 5 h 1, 44 kg K 2 O h 1 nd 83.2 kg MgO h 1 ). Chemil fertilizer solution ws dded to wter irrigtion y pump injetion one week. The methods lso inluded weed ontrol with syntheti hemil heriides nd plnt pthogen ontrol with syntheti hemil pestiides. Phoslone (35 g L 1 ) nd triforine (19 g L 1 ) were utilized to redue phids nd powdery mildew, respetively. Proprgite (57 g L 1 ) nd sulfur (8 g L 1 ) were used to redue/prevent mites. Mnozee (8 g L 1 ) ws pplied to prevent mildew. All these pestiides were pplied one yle. All vrieties were grown simultneously in the sme field nd sujeted to identil ulturl prties nd, of ourse, environmentl onditions in order to minimize the influene of pre- nd post hrvest ftors on genotype-relted vriility of field-grown tomtoes. Fruit smpling Tomto fruits were hnd hrvested one in July 8 nd totl yield ws reorded. A smple of pproximtely 2 kg of mrketle ripe tomtoes ws olleted from eh vriety, weighed nd delivered to the lortory the sme dy. The tomtoes were ut into smll piees nd sequentilly homogenized in mixer. Prt of eh smple ws immeditely used for some nlyses ( Brix, ph nd titrtle idity) nd the remining prt ws frozen t 2 C nd used for lyopene, totl phenolis, flvonoids, sori id, dehydrosori id s well s the hydrophili nd lipophili ntioxidnt tivities determintions within 1 week. The resons for doing this were the signifint volume of work involved nd the limited shelf life of tomtoes under refrigertion. Chemils 2,2'-Azinois(3-ethylenzothizoline-6-sulphoni id) dimmonium slt (ABTS), sori id, rutin nd 6-hydroxy-2,5,7,8- tetrmethylhromn-2-roxyli id (trolox) were otined from Sigm-Aldrih, Chemil Co., Miln. Other regents were of nlytil grde. Determintion of gronomi hrteristis Solule solid ontent ws determined in tomto juie using digitl refrtometer (Atgo PR-1, NSG Preision Cells, In., Frmingdle, NY, USA) nd expressed in Brix. ph ws mesured using n eletroni ph meter (WTW, Miroproessor ph Meter, PH 539 Weilheim, Germny). Titrtle idity ws estimted y titrtion t ph 8.1 with.1 mol L 1 sodium hydroxide solution nd expressed s % itri id (Ntionl Cnners Assoition 1968). Determintion of totl rotenoid nd lyopene ontent Totl rotenoid nd lyopene were extrted with hexne/ethnol/etone (2: 1: 1, v/v/v) ontining utylted hydroxytoluene (BHT) nd nlysed using spetrophotometer (Bekmn DU 65, Bekmn Coulter, In., CA, USA) t 45 nd 53 nm, respetively s desried y Fish et l. (2) nd Lee (1). Totl rotenoids were expressed s mg of -rotene equivlents kg 1 fresh weight (FW) (mg -rotene eq kg 1 FW). A molr extintion oeffiient = L mol -1 m -1 ws used for lyopene ontent determintion nd results were expressed in mg kg 1 FW. Determintion of totl phenoli ontent Totl phenoli ontent ws determined ording to the Folin Ciolteu olorimetri method s modified y Eerhrdt et l. () nd Singleton et l. (1999). Eh smple (.5 g) ws extrted with 1 ml of methnol for 24 h, fter whih 125 L of this extrt ws diluted 1: 5 (v/v) with distilled wter. Then 125 L of the diluted extrt ws mixed with 5 L of distilled wter in test tue, 125 L of Folin Ciolteu regent ws dded nd the mixture ws llowed to stnd for 3 min. Therefter, 1.25 ml of 7 g L 1 sodium ronte solution ws dded nd the finl volume ws mde up to 3 ml with distilled wter. Eh smple ws llowed to stnd for 9 min t room temperture efore mesurement t 76 nm ginst lnk in spetrophotometer (Behmn DU 65). The liner reding of the stndrd urve ws from to 3 g glli id ml 1. Results were expressed in mg glli id equivlent (GAE) kg 1 FW. Determintion of flvonoid ontent The flvonoid ontent ws determined s desried y Ji et l. (1999) on triplite liquots of the homogenous juie (.3 g). Fifty miroliter liquots of the methnoli extrt were used for flvonoids determintion. Smples were diluted with distilled wter to finl volume of.5 ml, nd 3 μl of 5% NNo 2 ws dded. After 5 min, 6 μl of 1% AlCl 3 ws dded nd finlly μl of 1 M NOH ws dded fter 6 min. The sorne ws red t 51 nm in spetrophotometer (Behmn DU 65) nd flvonoid ontent ws expressed in milligrms of rutin equivlents (RE) kg 1 FW. Determintion of sori id (AsA) nd dehydrosori id (DHA) AsA nd DHA ontents were determined s repeorted y Kmpfenkel et l. (1995) on triplite smples of the homogente juie (.1 g). AsA nd DHA were extrted y using 6% metphosphori id nd deteted t 525 nm in spetrophotometer (Bekmn DU 65) nd expressed in mg kg -1 FW. Trolox equivlent ntioxidnt pity (TEAC) ssy The mesurement of the hydrophili nd lipophili ntioxidnt tivity ws performed using the trolox equivlent ntioxidnt pity (TEAC) ssy. The ntioxidnt tvity ws mesured using the ABTS deolortion method (Miller nd Rie-Evns 1997). Hydrophili nd lipophili ntioxidnts were extrted from.3 g homogenous juie (three replites) with 5% methnol or 5% etone respetively t 4 C under onstnt shking (3 rpm) for 12 h. Smples were entrifuged t 1, g for 7 min nd the 2

3 Antioxidnt ompounds nd tivity in tomto reeding lines. Ilhy et l. different superntnts were reovered nd used for ntioxidnt tivity mesurements. The ntioxidnt tivities were mesured t 734 nm in spetrophotometer (Bekmn DU 65). Two different lirtion urves were onstruted using freshly prepred trolox solutions for HAA nd LAA determintions. Vlues were otined from three replites s M Trolox 1 g -1 FW. Sttistil nlysis Effets of vriety on yield, physiohemil nd nutritionl properties of tomto lines were ssessed y nlysis of vrine (ANOVA). When signifint differene ws deteted, mens were ompred using the lest signifint differene (LSD) test (P <.5). All sttistil omprisons were performed using SAS Version 6.1 softwre (SAS Institute, Cry, NC, USA). Correltions were done using Person s orreltion oeffiient, p <.5. RESULTS AND DISCUSSION Agronomi hrteristis Tomto plnts grown under the experimentl onditions were vigorous with exellent folige over. The high lyopene ontent tomto reeding lines were hrterized y drk folige nd drk green immture fruit without morphologil errtions. The most importnt gronomi hrteristis of the studied tomto vrieties re reported in Tle 1. The results showed tht differenes in tomto totl yield were signifint etween the studied tomto vrieties (P<.1). The vriety HLT-F52 rehed omprle yield to the ontrol Rio Grnde. The solule solids ontent ws signifintly different etween the studied tomto vrieties (P<.5). HLT-F51 hs omprle vlue to Rio Grnde. With regrds to verge fruit weight, ph nd titrtle idity, the dvned reeding lines showed lso omprle vlues to the ontrol vriety Rio Grnde. Generlly, the tomto high lyopene ontent dvned reeding lines showed eptle gronomi hrteristis nd yield. Totl rotenoid nd lyopene ontent The totl rotenoid nd lyopene ontents of the different tomto vrieties re presented in Fig. 1. The results showed signifint differenes mong tomto vrieties in totl rotenoid nd lyopene (P<.1). Totl rotenoid vlues vried from mg kg 1 FW in the stndrd Rio Grnde to mg kg 1 FW in HLT-F51. HLT-F51 nd HLT-F52 were determined to e the rihest vrieties in rotenoids with ontent nd 1.89-fold higher, respetively ompred to the stndrd Rio Grnde. Our vlues fll within the rnge reported y Rffo et l. (2) Cookson et l. (3) nd reently Lenui et l. (6) vrying from 14 to 299 mg kg 1 FW. Conerning lyopene, vlues rnged from 97.1 mg kg 1 FW in Rio Grnde to mg kg 1 FW in HLT-F51. As for totl rotenoids, HLT-F51 nd HLT-F52 were found to e lso the rihest soure of lyopene with ontent nd 1.94-fold higher respetively ompred to Rio Grnde. Our vlues re omprle with those reported for field-grown tomtoes y Aushit et l. (), Gomez et l. (1), nd Tkeok et l. (1), rnging from 52 to 236 mg kg 1 FW, nd onfirm tht field-grown tomtoes generlly present higher levels of lyopene with respet of greenhouse-grown tomtoes, in whih it rnges etween 1 nd 18 mg kg 1 FW (Leonrdi et l. ; Rffo et l. 2). These results re lso in line nd onfirm those of Lenui et l. (6) who reported tht high-pigment tomtoes re hrterized y the highest lyopene ontent ompred to the other studied tomto vrieties nd ttined more thn mg kg 1 FW. The differenes etween highlyopene nd ordinry tomto vrieties hve een ttriuted to different growing onditions nd ultivrs (Dums et l. 3). From the moleulr point of view, the onsiderle lyopene umultion in high-lyopene tomto vrieties n e due to the redued yling rte of this moleule to Tle 1 Agronomi hrteristis of the studied tomto vrieties Cultivrs Yield (t/h) ph Averge fruit weight (g) Solule solids ontent ( Brix) synthesize rotenes. Thus, these high-lyopene fruits seem exeptionl for humn nutrition. Consuming one serving of HLT-F52 or HLT-F51 vrieties (1 g) provides to 11.7% of the reommended dily intke of lyopene ompred to only 38.8% for Rio Grnde (Ro nd Agrwl 1998). Totl phenoli nd flvonoid ontent Titrtle idity (%) HLT-F HLT-F Rio Grnde Signifine ** ns ** ns ns Signifition: ** Proility level of 1%; ns: not signifint. Vlues in the sme olumn followed y the sme letters do not differ signifintly (LSD test, P<.5). Totl rotenoid Totl rotenoid HLT-F51 HLT-F52 Rio Grnde The totl phenoli nd flvonoid ontents of the studied tomto vrieties re presented in Fig. 2. The results showed signifint differenes mong tomto vrieties in totl phenolis (P<.5) nd flvonoids (P<.1). Totl phenoli vlues rnged from 171. mg GAE kg 1 FW in Rio Grnde to mg GAE kg 1 FW in HLT-F51. HLT-F52 vriety exhiited similr phenoli level to Rio Grnde. Our vlues re lose to those of Toor nd Svge (5) who reported tht the pulp totl phenoli ontent men vlue (oth hydrophili s well s lipophili) of three tomto vrieties ws 15 mg GAE kg 1 FW during the investigtion of ntioxidnt omponent in different tomto frtions. Even so, Lenui et l. (6) reported higher pulp totl phenoli vlues in high-pigment tomto vrieties. Although geneti ontrol is the primry ftor in determining the mount of phenols in fruits nd vegetles, vritions ould lso depend on ripening stges t the moment of hrvesting, environnementl ftors (minly light nd temperture) (Mheix et l. 199; Dums et l. 3) nd nlytil methodology. Moreover, the often ontrditory results ould e ttriuted to different pttern of hnges in different lsses of phenoli during the ripening of tomto fruits s ws reported y But nd Spulding (1997) nd Rffo et l. (2). Regrding flvonoids, vlues rnged from mg Lyopene Fig. 1 Totl rotenoid (mg -rotene eq kg -1 FW) nd lyopene (mg kg -1 FW) ontent of the studied tomto vrieties. Vlues re men ± stndrd error. Vlues for eh vriety with the sme letters re not signifintly different (LSD test, P<.5). Totl phenolis 6 4 Totl phenolis Flvonoids HLT-F51 HLT-F52 Rio Grnde Fig. 2 Totl phenolis (mg GAE kg -1 FW) nd flvonoids (mg RE kg -1 FW) ontent of the studied tomto vrieties. Vlues re men ± stndrd error. Vlues for eh vriety with the sme letters re not signifintly different (LSD test, P<.5). Lyopene Flvonoids 3

4 The Afrin Journl of Plnt Siene nd Biotehnology 3 (Speil Issue 1), Glol Siene Books RE kg 1 FW in Rio Grnde to mg RE kg 1 FW in HLT-F51 nd were in line with the reent results of Lenui et l. (6) for high-pigment tomto vrieties rnging from 168 to 47 mg RE kg 1 FW. In our nlysis, HLT-F51 nd HLT-F52 vrieties otined respetively more thn nd 1.58-fold higher flvonoids ompred to Rio Grnde. Tomtoes hve een onsidered to e reltively rih soure of flvonoids, with n verge of 5 mg kg 1 FW (Stewrt et l. ). In the present study, the stndrd vriety Rio Grnde otined more thn 2.65-fold the verge tomto fruit flvonoid ontent reported y Stewrt et l. (). Interestingly, HLT-F51 nd HLT-F52 otined in verge respetively nd 4.2-fold the reported flvonoid verge ontent. Vritions n e sried to the high lyopene trits. In ft, it hs een reported tht in red ripe tomto fruits, nturlly ourring muttions tht inrese rotenoid ontent, inluding lyopene, re lso hrterized y drmti inrese in plstid iogenesis nd in the prodution of other ompounds suh s vitmin C nd flvonoids (Mohizuki nd Kmimur 1984). Moreover, it hs een reported tht field-grown tomto fruits, whih reeive higher mount of light nd UV rdition ontin higher mount of flvonoids (queretin nd kempferol) in omprison to geenhouse-grown tomto. In our experiment, one serving of HLT-F51 or HLT-F52 fruit provided respetively 189 nd 84% of the reomended dily intke of flvonoids ompred to 53% for Rio Grnde whih sertin the importne of suh tomto vrieties s dietry flvonoids suppliers (Hertog et l. 1992). Although phenoli ontents in tomtoes re only moderte ompred with those in other vegetles suh s onion, their high onsumption in the Tunisin diet (Hdider et l. 7) due to their yer-round vilility nd high utility in ulinry preprtions, mkes them good soure of phenols, s reported for Indin nd Amerin diets (Vinson et l. 1998; George et l. 4) Our results onfirmed tht vriety signifintly ffets totl phenoli ontents in tomto, s reported y other uthors (Aushit et l. ; George et l. 4). Vitmin C ontent The totl vitmin C, AsA, nd DHA ontents of the different tomto vrieties re presented in Fig. 3. Exept for AsA, the results showed signifint differenes mong tomto vrieties in DHA nd totl vitmin C (P<.1). DHA, the oxidized form, ounted for etween 4 nd 68% of the totl vitmin C in line with the reent result of Lenui et l. (6) for high-pigment tomto vrieties rnging from to 85%. It is well known tht the mount of DHA is strongly influened y experimentl proedure (De Gr 3). HLT-F52 vriety exhiited the highest mount of DHA with mg kg 1 FW. The mount of DHA in HLT-F51 ws omprle to Rio Grnde. In our experiment, the mounts of AsA nd DHA were similr to those reported y Lenui et l. (6), George et l. (4), Lipty et l. (1986) nd Soui et l. (1994). Regrding totl vitmin C, vlues rnged from mg kg 1 FW in Rio Grnde to mg kg 1 FW in HLT-F52. HLT-F51 otined similr totl vitmin C vlue to Rio Grnde. The gretest vrition in totl vitmin C ws 1.7-fold higher oserved in HLT-F52 vriety. Although the sori id ontent for new seleted tomto vrieties hs een only srely reported, our result fll within the rnge reported reently y Lenui et l. (6) ttining 3 mg kg 1 FW in different high-pigment tomto vrieties. However, higher vlues, depending on sesons, nd rnging from 31 to 71 mg kg 1 FW were reported y Rffo et l. (6) for the herry tomto ultivr Nomi grown under greenhouse ondditions. Generlly, it is widely reognized tht field-grown tomto hve higher AsA levels (until 258 mg kg 1 FW) when ompred to those produed under shde (155 mg kg 1 FW). Differenes were lso oserved in the redox stte of the system AsA/DHA of the studied tomto vrieties. For the reson tht mny environmentl onditions hnge the redox stte of this system, it hs een hypothesized tht it ould funtion s sensor Vitmin C Asori id Dehydrosori id Totl vitmin C modulting ellulr metolism nd hormone sensitivity in response to exogenous ftors (De Gr 3). This suggests tht different tomto vrieties n pereive the surrounding environment in different wy, ording to their genotype. These vrieties seem therefore exeptionl for humn onsumption. In ft, onsuming one serving of either HLT-F52 or HLT-F51 vrieties provides from 43 to 64% of the reomended dily intke of vitmin C ompred to only 37% for the stndrd vriety Rio Grnde (Soietà Itlin di Nutrizione Umn 1998). Hydrophili nd lipophili ntioxidnt tivities TEAC ssy The hydrophili (HAA) nd liophili (LAA) ntioxidnt tivities of the different tomto vrieties re presented in Fig. 4. The results showed signifint differenes mong tomto vrieties in hydrophili nd lipophili ntioxidnt tivities (P<.1). HAA vlues rnged from μm Trolox 1 g 1 FW in Rio Grnde to μm Trolox 1 g 1 FW in HLT-F51. Those findings fill in the rnge reported y mny uthors. In ft, Rffo et l. (6) reported reently tht the HAA of the herry tomto vriety Nomi rnged etween 191 nd 42 μm Trolox 1 g 1 FW depending on sesons. Cno et l. (3) reported lso tht the hydrophili ntioxidnt tivity ttined 218 μm Trolox 1 g 1 FW for the greenhouse grown tomto vriety Mrmnde-Curenteno. In our experiment, totl ntioxidnt tivity (the sum of oth hydrophili nd lipophili ntioxidnt tivities) vried from μm Trolox 1 g 1 FW to μm Trolox 1 g 1 FW. The hydrophili ntioxidnt ivity ontriuted from 42-49% of the totl ntioxidnt tivity. The HAA vlues in mture fruits of HLT-F52 nd HLT- F51 were 1.28 nd 2.9-fold the HAA of the ontrol vriety Rio Grnde. The hydrophili ntioxidnt tivity hs een ttriuted to the presene of phenoli ompounds, suh s ffei nd hlorogeni id in the methnoli frtion (Gomez et l. 1). After onsidering dt from ll tomto vrieties, no signifint orreltion (R 2 = -.76; P>.5) etween TEAC nd totl vitmin C ontent ws otined (Tle 2). The lk of orreltion ould e due to the ontent of phenolis, whih my ount for most of the TEAC vlues. In ft, there ws good liner orreltion (R 2 =.823; P<.1) etween flvonoids nd TEAC. The totl hydrophili ntioxidnt tivity ws ertinly orrelted HLT-F51 HLT-F52 Rio Grnde Fig. 3 Asori id, dehydrosori id nd totl vitmin C ontent (mg kg -1 FW) of the studied tomto vrieties. Vlues re men ± stndrd error. Vlues for eh vriety with the sme letters re not signifintly different (LSD test, P<.5). Antioxidnt tivity Hydrophili ntioxidnt tivity Lipophili ntioxidnt tivity HLT-F51 HLT-F52 Rio Grnde Fig. 4 Hydrophili nd lipophili ntioxidnt tivities (μm Trolox eq 1 g -1 FW) of the studied tomto vrieties. Vlues re men ± stndrd error. Vlues for eh vriety with the sme letters re not signifintly different (LSD test, P<.5). 4

5 Antioxidnt ompounds nd tivity in tomto reeding lines. Ilhy et l. Tle 2 Correltion oefiient nd relted signifine etween ntioxidnt ompounds nd ntioxidnt tivity Compounds TEAC ssy Corr oeff P Hydrophili frtion Asori id.469 ns Dehydrosori id.19 ns Totl vitmin C -.76 ns Totl phenolis.823 <.1 Flvonoids.977 <.1 Lipophili frtion Totl rotenoids.956 <.1 Lyopene.954 <.1 ns, no signifint orreltion with the level of ll of the min ntioxidnts (vitmin C, totl flvonoids nd phenoli ompounds), rther thn eing the mere sum of their ontent. Furthermore, it is likely tht it depends upon synergisti effets mong ll hydrophili ntioxidnts nd their intertion with other onstituents of the frtion (Diplok et l. 1998; Lenui et l. 6). The test retion used for ntioxidnt tivity mesurement might e influened y other ompounds involved in omplex ntioxidnt system of tomto fruits suh s glutthione nd enzymti omponents (Jiménez et l. 2). Regrding LAA, vlues rnged etween μm Trolox 1 g 1 FW in Rio Grnde to μm Trolox 1 g 1 FW in HLT-F51. The lipophili ntioxidnt tivity vlues were higher thn those otined y Rffo et l. (6) nd Cno et l. (3) rnging etween 26 nd 88 μm Trolox 1 g 1 FW for different tomto vrieties grown under greenhouse onditions. In our experiment, the lipophili ntioxidnt tivity represented 51-58% of the totl ntioxidnt tivity. The higher lipophili ntioxidnt tivity n e minly ttriuted to the lrge mount of lyopene deteted prtiulrly in high lyopene ontent tomto vrieties. In ft, the lipophili ntioxidnt tivity vlues in mture fruits of HLT-F52 nd HLT-F51 were 1.66 nd fold higher ompred to the stndrd vriety Rio Grnde fruits. The lipophili ntioxidnt tivity hs een minly ttriuted to the presene of rotenoids prtiulrly lyopene (Mrtìnez-Vlverde et l. 2; Rffo et l. 2; Cno et l. 3; George et l. 4). 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