Effect of CaCl 2 and Exogenous Putrescine on Post-harvest Life and Quality of Peach (Prunus persica (L.) Batsch) Fruit, cv. J. H.

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1 Fruit, Vegetble nd Cerel Siene nd Biotehnology 211 Globl Siene Books Effet of CCl 2 nd Exogenous Putresine on Post-hrvest Life nd Qulity of Peh (Prunus persi (L.) Btsh) Fruit, v. J. H. Hle Mhbub Alizde-Dshqpu 1 Mhmood Esn-Ashri 1* Jfr Hjiloo 2 Mohmmd Asghrpur 3 1 Deprtment of Hortiulturl Sienes, Fulty of Agriulture, Bu-Ali Sin University, Hmedn, Irn 2 Deprtment of Hortiulturl Sienes, Fulty of Agriulture, Tbriz University, Est Azrbyjn, Irn 3 Deprtment of Hortiulturl Sienes, Fulty of Agriulture, Urmi University, West Azrbyjn, Irn Corresponding uthor: * m.esnshri@bsu..ir ABSTRACT The effet of, 6 nd 8 mm lium hloride (CCl 2 ) nd.5, 1 nd 2 mm putresine (Put) on post-hrvest life nd qulity of peh (Prunus persi L. v. J. H. Hle ) fruit ws studied. This experiment ws rried out by immersing fruits in solutions t 25 C for 5 min, then trnsferring them into storge t 1-2 C nd 75% reltive humidity, together with untreted fruits (dry) s well s fruits immersed in distilled wter only. Ethylene prodution, soluble solids ontent (SSC), titrtble idity (TA), ph, flesh firmness, weight loss nd sorbi id (AA) of fruits were mesured t regulr intervls (5 dys for old storge nd 2 dys for shelf life) throughout the experiment. Qulity of fruits ws lso tested through tste pnel t the end of experiment. During the whole storge time in whih 5 mesurements were performed, 6 mm CCl 2 ws the best tretment for keeping fruit qulity in terms of flesh firmness, SSC, AA, TA, ph nd weight loss. The highest sore in terms of fruit qulity, s given by pnelists, orresponded to 6 mm CCl 2 nd 2 mm Put. Among the Put solutions, 2 mm ould mintin the qulity of fruits preventing ethylene prodution t the highest rte ompred to other tretments. The results of mirobiologil test showed tht 8 mm CCl 2 ws the best tretment to ontrol yest nd mold popultions signifintly (P <.5). Put solutions were better thn CCl 2 tretments in terms of preventing ethylene prodution by fruits. Lower qulity fruit ws observed when fruit ws immersed in distilled wter only. Keywords: ethylene, fruit qulity, shelf life, storge time, yest nd mold popultion Abbrevitions: AA, sorbi id; CFU, olony forming unit; FW, fruit weight; PA, polymine; Put, putresine; ROS, retive oxygen speies; SE, stndrd error; SSC, soluble solid ontent; TA, titrtble idity INTRODUCTION Peh is limteri fruit hving short storge-life. Post-hrvest dey is the mjor ftor limiting the extension of storge nd/or shelf-life of fruits. Rpid ripening lso shortens the shelf-life of ommodities nd represents serious onstrin for effiient hndling nd trnsporttion (Bonghi et l. 1999). Considering the vrying response of different produts to post-hrvest tretments, development of suitble tehniques beomes neessry to hieve optimum qulity for ny produt. In reent yers, the use of physiologilly tive ompounds hs ttrted food industries nd onsumers (Alzmor et l. 25). Clium is one of the importnt elements ffeting qulity nd post-hrvest life of mny fruits. Clium defiieny in some fruits uses ertin diseses suh s bitter pit, ork spot, wter ore nd senesent brekdown. Fruits with lium defiieny fll erlier thn the expeted time of ripening nd nnot be kept in store longer ompred to those with enough lium (Vlero nd Serrno 21). Clium redution in the ell middle lmell is the mjor ftor of ell wll softening in pple during post-hrvest storge (Stow 1999). Severl other roles hve been reported for lium inluding tissue firmness preservtion in pple (Chrdonnet et l. 23), peh (Prussi et l. 25), strwberry (Sftner et l. 23; Verdini et l. 28) nd fresh-ut lettue (Mrtin-Din et l. 26); fruit ripening nd senseene retrdtion (Pihioni et l. 1996; Lester nd Grusk 1999); redution of ethylene prodution (Ben-Aries et l. 1982); CO 2 nd respirtion rte redution (Singh et l. 1993; Lun-Guzmán 1999); flesh browning ontrol (Hewjulige et l. 23; Sftner et l. 23; Mngnris et l. 27) nd molds dey prevention (Smilnik nd Sorenson 21; Lr et l. 2; Verdini et l. 28). Internl brekdown nd flesh browning in peh fruits hve lso been observed in old storge due to lium defiieny (Hewjulige et l. 23). Polymines (PAs) re biologil tive ompounds with low moleulr weight nd liphti nitrogen groups ourring in ll living things inluding nimls nd plnts (Vlero et l. 22). Severl studies hve shown tht PA umultion ours under bioti stresses inluding drought, slinity, extreme temperture, hypoxi, UV-B irrdition, hevy metls, mehnil wounding nd herbiide tretments (Gropp nd Benvides 28). At the ellulr ph vlues, PAs behve s utions. In order word, these ompounds hve positive moleulr hrge, nd this mkes them to ret with nioni mromoleules suh s DNA, RNA, Phospholipids nd ertin proteins (Heby nd Persson 199). A strong inorportion of PAs with the ell wll poly-nioni strutures suh s petins nd their exhnge with peti polyshrides ontributing to petin signl modultion in pthogenesis nd differentition hve been reported (D Orzi nd Bgni 1987). Mny benefiil roles of PAs in post-hrvest physiology of fruits nd vegetbles ould lso be found in litertures whih inlude senesene retrdtion nd deresing ethylene prodution (Sftner nd Bldi 199; Kkkr nd Ri 1993; Bregoli et l. 22; Torrigini et l. 2); tissue firmness nd membrne sfety preservtion (Krmer et l. 1991; Wng et l. 1993; Pon- Reeived: 18 June, 21. Aepted: 3 My, 211. Originl Reserh Pper

2 Fruit, Vegetble nd Cerel Siene nd Biotehnology 5 (Speil Issue 2), Globl Siene Books pp et l. 1993; Serrno et l. 23); old dmge redution (Shen et l. 2; Mrténez-Téllez et l. 22; Mirdehghn et l. 26) nd mehnil dmge resistne indution (Vlero et l. 1998; Mrtínez-Romero et l. 2; Perez-Viente et l. 22). PAs should not be tken only s protetive moleules but rther like double-fed moleules tht likely serve s mjor re for further reserh efforts (Hussin et l. 211). The im of this reserh ws to study the effet of different CCl 2 nd putresine (Put) onentrtions on posthrvest life nd ertin qulity hrteristis of peh fruit, ultivr J. H. Hle during storge period. J. H. Hle is one of the most desirble peh ultivrs to onsumers in Irn with inresed prodution rte. MATERIALS AND METHODS Plnt mteril Peh (Prunus persi (L.) Btsh) fruits, v. J. H. Hle were hrvested in lte August from privte orhrd (Moghn Industry nd Culture Co., Azerbyjn, Irn). About 13-1 g fruits with uniform size were seleted t ommeril mturity stge when 1/2 to 2/3 fruit surfe hd red olor. Fruits were then pked in polyethylene bgs, put into 1 kg plsti bskets nd trnsferred to supplementry urriulum lbortories of Tbriz University. All dmged nd physiologilly injured fruits were finlly eliminted nd divided into 2 lots of 3-35 fruits. Tretments Experiment ws onduted in omplete rndomized design, inluding eight tretments (.5, 1 nd 2 mm Put (Merk, Germny);, 6 nd 8 mm CCl 2 (Merk), distilled wter for the ontrol nd dry tretment (without wter, Put or CCl 2 ) nd three replitions in eh tretment. Fruits were immersed in Put nd CCl 2 solutions s well s distilled wter ll ontining 2 g l -1 Tween-8 (Sigm Chemil Co., USA) surftnt t 2-25 C for 5 min. Fruits were then dried in room temperture for 2 hrs nd trnsferred into old storge with 1-2 C nd 75% reltive humidity. Qulity prmeters were mesured weekly (5 dys old storge + 2 dys shelf life t 25 C) during 35-dy storge period. Weight loss In order to determine fruit weight loss during the storge time, fruits from ny lot were seprted t the beginning of storge nd weighed using digitl blne (Srtorius, GM612, Germny). Fruit weight loss ws then mesured weekly (5 dys old storge + 2-dys shelf-life) through the following eqution: Fruit weight loss (%) = 1 (pre-storge FW - fter-storge FW) pre-storge FW Fruit firmness Flesh firmness determintion ws rried out using Penetrometer (Model FDK32; Wgner, Miln, Itly) to mesure the fore required for 11 mm-probe to penetrte the fruit tissue fter removing the peel nd expressed s Newton. ph, soluble solids ontent nd titrtble idity The ph of fruit juie (blended 3 fruits) ws mesured using PHS-25C ph-meter (Shnghi, Chin). SSC ws determined using portble refrtometer (ATAGO N1, Tokyo, Jpn) t 25 C nd expressed s perentge (%). TA ws determined by titrtion to ph 8.2 with.1 N NOH nd expressed s g mli id per 1 g fruit weight. Asorbi id AA ws determined by 2,6-dihlorophenolindophenol (Merk) titrtion method (AOAC 198) nd expressed s mg sorbi id per 1 g fruit weight. Tste pnel In order to ompre the qulity of treted fruits with the ontrol nd dry tretment, tste pnel exmintion ws rried out by 1 dults, ged 2-36 yers inluding 5 mles nd 5 femles. Fruit pperne, firmness, juiiness, tste nd sweetness were evluted by the pnelists t the end of storge period bsed on rnking of 1 to 5, where 5 = superb; = very nie; 3 = nie; 2 = verge; 1 = bd. Mirobiologil nlysis One fruit from eh lot ws onsidered for mirobiologil nlysis t the beginning s well s the end of the experiment. To do this, 1 g fruit ws smpled by sliing from peel to deep flesh in vrious prts nd homogenized in 9 ml peptone wter using sterile mortr under septi ondition. 1 ml of the homogenized solution ws pled on Potto Dextrose medium (ph = 3.5) nd trnsferred into inubtor t 3 C for 3 dys. The number of fungi nd yests olonies formed on the medium ws then ounted nd only 3-3 olony forming units (CFU) were tken into ount nd expressed s Log CFU g -1 FW -1. Ethylene prodution Ethylene prodution ws mesured by pling one fruit in 1 l glss jr tightly filled in with rubber p for 1 h. One ml of the holder tmosphere ws withdrwn using gs syringe, nd the ethylene ws quntified by gs hromtogrph (Shimdzu, C-RA, Jpn) pprtus. Results were expressed in nl of ethylene relesed per grm of fruit flesh per hour (nl g -1 h -1 ). Sttistil nlysis The dt ws nlyzed using SAS sttistil softwre nd mens were ompred by Dunn s multiple rnge test (P <.5). RESULTS AND DISCUSSION ph The ph of fruit juie ws generlly inresed during storge nd the vrious tretments signifintly redued the extent of this inrese (Fig. 1). At the end of storge the highest nd the lowest ph vlues were belonged to distilled wter nd 6 mm lium hloride respetively. Inrese in ph vlue my be due to the brekup of ids with respirtion during storge whih hs been reported in peh v. J. H. Hle (To rul nd Arsln 2); Tokhm-Sefid, lol vriety of priot (Zokee-Khosroshhi nd Esn-Ashri 27; Liu et l. 29) nd green sprgus (Bhowmik et l. 22). Fruit firmness Fruit firmness ontinuously deresed during storge nd vrious tretments signifintly redued fruit softening (Fig. 2). The highest nd the lowest derese in fruit firmness were belonged to distilled wter nd 6 mm lium hloride respetively t the end of storge. However, Put tretments ould lso mintin fruit firmness during storge whih is in greement with the results of Zokee-Khosroshhi nd Esn-Ashri (27) nd Liu et l. (29) in priot (Prunus rmeni L.) nd Zokee-Khosroshhi et l. (27) in strwberry (Frgri nnss Duh.). Mintining flesh firmness by lium nd PAs hs lredy been reported in different fruits inluding pples (Krmer et l. 1991; Wng et l. 1993; Chrdonnet et l. 23), Pehes (Bregoli et l. 22; Prussi et l. 25) nd strwberries (Ponpp et l. 1993; Sftner et l. 23; Shfiee et l. 21). The mehnism involved seems to be the result of their bounds with petin ompounds leding to physilly stbilized ell wll whih is detetble immeditely fter tretment (Vn Buren 1979; Heby nd Persson 199; Grí et l. 1996). Signifint effet of polymines nd CCl 2 tretments on mintining fruit firmness hs shown tht 1

3 Effet of CCl 2 nd putresine on post-hrvest life of peh. Alizde-Dshqpu et l. ph these ompounds hve similr bilities in terms of using senesene dely in fruit softening in bounding ell wll or ell membrne (Ponpp et l. 1993). Weight loss C mm C 6 mm C 8 mm Put.5 mm (+2) Fig. 1 The effet of vrious tretments on ph of fruit juie during storge (Cold storge + Shelf life). Dt re the men ± S.E. Firmness (N) C mm C 6 mm C 8 mm Put.5 mm (+2) Fig. 2 The effet of vrious tretments on fruit firmness during storge (Cold storge + Shelf life). Dt re the men ± S.E. Weigh Loss (%) C mm C 6 mm C 8 mm Put.5 mm Fig. 3 The effet of vrious tretments on fruit weight loss during storge (Cold storge + Shelf life). Dt re the men ± S.E. Considerble weight loss ourred during storge (Fig. 3) but, vrious tretments showed signifintly less weight loss thn the ontrol. Fruits treted with nd 6 mm CCl 2 showed lowest weight loss thn the other tretments. However, no signifint differene in weight loss ws found between the nd 6 mm CCl 2 tretments. The mximum weight loss observed in the ontrol ws probbly beuse of loosing piles from the fruit surfe during immersion in wter my ffet peh storge life nd the mehnism by whih lium prevents fruit weight loss is Asobi id (mg/1g Fruit) due to the inhibition of respirtion. Singh et l. (1993) reported tht pre-hrvest lium (nitrte nd hloride) tretment in mngoes (Mngifer indi L.) delyed fruit ripening nd redued respirtion rte resulting in fruit qulity improvement during storge. Immersion of fresh-ut ntloupe (Cuumis melo L., vr. retiultus) hunks in CCl 2 hs lso deresed CO 2 prodution nd respirtion rte (Lun-Guzmán et l. 1999). The effets of Put nd CCl 2 tretments on the prevention of fruit weight loss in this study re in greement with the results of Vlero et l. (1998), Mrtínez-Romero et l. (22), Serrno et l. (23), Zokee-Khosroshhi nd Esn-Ashri (27), Shfiee et l. (21) nd Chen et l. (211). Asorbi id (AA) Continuous redution in sorbi id ontent of treted nd untreted fruits during storge is shown in Fig.. Vrious tretments signifintly ffeted the mount of AA; however, the lowest extent of redution ws observed in 6 mm CCl 2 tretment while the highest one belonged to the ontrol. Reduing trend in AA ontent in fruits ws deresed with inresing Put onentrtion, most likely beuse Put tretments prohibit fruit ethylene prodution followed by metbolism retrdtion. Clium-treted ppy (Cri ppy L.) fruit led to mintin higher AA onentrtion during storge ompred to non-treted ones (Mhmud et l. 28). Delining AA ontent during storge hs lso been reported by To rul nd Arsln (2) in peh nd Tulio Jr. et l. (22) in Corhorus olitorius whih re in greement with the results of this study. Retive oxygen speies (ROS) prodution is ommon plnt response to both bioti nd bioti stresses (Miller et l. 28). ROS is hrterized by the umultion of toxi moleules 2, H 2 O 2 nd OH in tissues whih re pble of using dmge to plnt ell, membrnes nd mromoleules (Apel nd Kirt 2), due to inrese in oxidtive metbolism during ripening, espeilly in limteri fruits, ROS re inresed dmging ell membrnes. Plnts therefore need to prevent ROS dmges to their ells membrnes through developing enzymti nd/or non-enzymti nti-oxidnt gents suh s sorbte peroxidse, vitmin C nd vitmin E (Spinrdi 25). Anti-oxidnt gents ret themselves with retive oxygen speies nd prevent their oxidtive dmges to the ell membrnes. Titrtble idity nd soluble solids ontent C mm C 6 mm C 8 mm Put.5 mm (+2) Fig. The effet of vrious tretments on sorbi id ontent of fruits during storge (Cold storge + Shelf life). Dt re the men ± S.E. TA nd SSC were deresed in ll treted nd untreted fruits during storge (Figs. 5, 6), but the differene between tretments ws signifint in this respet. TA nd SSC were highest in 6 mm CCl 2 tretment t ll mesuring times, while their lowest mounts belonged to the ontrol. These 2

4 Fruit, Vegetble nd Cerel Siene nd Biotehnology 5 (Speil Issue 2), Globl Siene Books TA (g mli id /1g Fruit) results re in greement with the findings of Hung et l. (25) in lithi (Lithi hinensis Sonn), Mngnris et l. (27) in peh (Prunus persi (L.) Btsh), Zokee Khosroshhi nd Esn-Ashri (27) nd Liu et l (29) in priot (Prunus rmeni L.) nd Chen et l. (211) in strwberry (Frgri nnss Duh.) Deresing totl SSC seems to be the result of rbohydrtes nd petins brekdown, prtil protein hydrolysis nd deomposition of glyosides into subunits during respirtion (Lr et l. 2; Mhmud et l. 21). Mirobiology test C mm 6 mm C 8 mm Put.5 mm (+2) Fig. 5 The effet of vrious tretments on titrble idity of fruits during storge (Cold storge + Shelf life). Dt re the men ± S.E. Soluble Solids (%) C mm C 6 mm C 8 mm Put.5 mm (+2) Fig. 6 The effet of vrious tretments on soluble solids ontent of fruits during storge (Cold storge + Shelf life). Dt re the men ± S.E. Yest nd Mold (Log CFU/g) prestorge e All treted nd untreted fruits showed onsiderble inrese in yest nd mold popultion during the storge ompred to the hrvesting time (Fig. 7), but there ws signifint differene between the tretments in terms of yests nd fungi CFUs ounted t the end of storge. The highest inrese in yest nd mold popultions ws observed in dry fruits showing signifint differene with ll tretments. No signifint differene ws found between the ontrol,.5 nd 1 mm Put s well s 6 nd 8 mm CCl 2. The best result in this respet belonged to 6 nd 8 mm CCl 2. These findings re in greement with the results of Mngnris et l. (25) who pplied lium pre-hrvest spry on peh nd found derese in brown rot nd lso Sftner et l. (23) who reported redution in yest nd mold popultion in honeydew (Cuumis melo L., v. Inodorus Group) hunks using lium tretments. It is ssumed tht lium ompounds strongly preserve the ell wll whih ts s powerful brrier ginst pthogens ttks nd prevents miroorgnisms to penetrte into the ells nd tissues nd to inrese their popultion. Suh ell wll preservtion effet of lium hs been reported by Smilnik nd Sorenson (21); Lr et l. (2), Hernández-Munoz et l. (26) nd Chen et l. (211). Cmpnell et l. (22) hve lso shown tht lium slts not only derese the growth of Phytophthor niotine in vitro but lso redue the severity of pthogen inoultion in vivo. Tste pnel b C mm d d d C 6 mm C 8 mm Tretments Put.5 mm b b b Fig. 7 Effets of vrious tretments on yest nd mold popultion during storge. Sores given by the pnelists to some fruit qulity hrteristis were signifintly different. Untreted (dry) nd ontrol fruits lost their qulity 28 dys fter storge when they trnsferred from old storge to the shelves. They lso beme very soft nd sweet gining different tste (like priots) when kept in the old storge (1-2 C) for 35 dys. Untreted (dry) nd ontrol fruits did therefore not obtin ny sore by the pnelists nd omitted from pnel tste (Fig. 8). In summry, pnelists gve the highest sores to 6 mm CCl 2, wheres mm CCl 2 nd 2 mm Put obtined the seond ple. No signifint differene observed between 8 mm CCl 2 nd 1 mm Put tretments. Sore d Fruit spet Firmness Juiiness Tste Sweetness d d d b b b b b b b b b b b d d b b b C mm C 6 mm C 8 mm Put.5 mm Tretments Fig. 8 Sores given by the pnelists to some fruit qulity hrteristis t the end of storge. 3

5 Effet of CCl 2 nd putresine on post-hrvest life of peh. Alizde-Dshqpu et l. Ethylene (nl/g.h) Ethylene prodution Ethylene prodution rte in treted nd untreted fruits inresed during storge (Fig. 9). The effets of vrious tretments on fruit ethylene prodution were signifintly different t mesuring times. Untreted (dry) nd ontrol fruits produed the highest rte of ethylene, while the best result ws obtined from 2 mm Put tretment in terms of reduing ethylene prodution. The solutions of 1 mm Put nd 8 mm CCl 2 were remined the next ple. Through reduing ethylene prodution PAs prevent ethylene-relted effets suh s hlorophyll loss, senesene, membrne deteriortion nd RNAse nd Protese tivities in plnts (Evns nd Mlemberg 1989). Post-hrvest pplition of PAs on number of fruits like vodoes (Perse merin L.), pers (Pyrus ommunis L.) nd tomtoes (Lyopersiom esulentum Miller.) hs shown high perentge of reduing ethylene prodution through inhibition of ACC synthse (Sftner nd Bldi 199; Kkkr nd Ri 1993). In some other fruits like priots (Prunus rmeni L.), pehes (Prunus persi L.), netrines (Prunus persi L.), strwberries (Frgri nnss Duh.) nd severl vrieties of plums (Prunus domesti L.) deresing ethylene prodution by PAs hs lso reported (Vlero et l. 22; Bregoli et l. 22; Torrigini et l. 2; Zokee- Khosroshhi nd Esn-Ashri 27; Zokee-Khosroshhi et l. 27). The results of this study re supported by the bove findings but in ontrst, re not in greement with the repot of Krmer et l. (1991) regrding ethylene prodution in pples. There is n inverse reltion between ethylene prodution rte nd the onentrtion of lium in pple fruit tissues (Fust 1989). Ben-Aries et l. (1982) reported tht lium nd spermine redue ell membrne fluidity through the inhibition of ethylene biosynthesis in pple (Mlus domesti Borkh.) diss, nd this hppens more with lium if the temperture is below 12 C, wheres spermine n prevent ethylene prodution more t the tempertures bove 12 C. Lun-Guzmán et l. 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