Research Article Ruta chalepensis L. Essential Oil: A New Antisprouting Agent for Potatoes Bioconservation

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1 Hindwi Journl of Chemistry Volume 218, Artile ID , 6 pges Reserh Artile Rut hlepensis L. Essentil Oil: A New Antisprouting Agent for Pottoes Bioonservtion Olf Lengliz, 1,2 Jmel Mejri, 1 Mnef Aderr, 1 Rhid Khlif, 3 nd Mondher Mejri 4 1 Lortory of Mterils, Moleules nd Applitions, IPEST, Rod Sidi Bou Sid, BP 51, L Mrs, Tunisi 2 University of Crthge, Ntionl Institute of Applied Siene nd Tehnology, BP 676, 18 Tunis Cedex, Tunisi 3 Tehnil Center of Potto nd Artihoke in Mnou, Street Jedid Km17, Essidi, Mnou, Tunisi 4 Higher Institute of Biotehnology of Bej, University of Jendou, Avenue Hi Bourgui 9, BP 382, Bej, Tunisi Correspondene should e ddressed to Olf Lengliz; olflenglizridene@gmil.om Reeived 24 Otoer 217; Aepted 14 Deemer 217; Pulished 8 Jnury 218 Ademi Editor: Mri Ro Copyright 218 Olf Lengliz et l. This is n open ess rtile distriuted under the Cretive Commons Attriution Liense, whih permits unrestrited use, distriution, nd reprodution in ny medium, provided the originl work is properly ited. The min onern of this work ws to find new iologil ntisprouting produt for pottoes during storge. Rut hlepensis L. essentil oil (RCEO) ws for the first time used s potentil sprouting inhiitor for pottoes. To onfirm RCEO ntisprouting effet, evolution of sprout length nd weight loss of potto tuers ws determined during storge period of six weeks under three RCEO different tretments (2%, 4%, nd 6%). Sprout finl weight ws lso omputed. Then, sreening of RCEO hemil omposition ws performed. An outstnding ntisprouting effet of 6% emulsions onentrtions ws oserved s ompred to 2% nd 4% emulsions onentrtions. Finl sprout weight (SW) of untreted smples ws 4.66%, while 6% treted smples sored.98%. These results were endorsed y high Person orreltion oeffiients (>.9). Indeed, inresing tretment onentrtion enhned the sprout growth inhiition. Identifition of RCEO mjor omponents showed tht 2-undenone represented 87.18% of totl identified omponents suggesting tht this omponent is the possile tive gent of RCEO ginst pottoes sprouting. 1. Introdution Potto (Solnum tuerosum L.) rnks s the world s fourth most importnt food rop, fter mize, whet, nd rie [1]. Being the world s numer one nongrin food ommodity, gret onsidertion should e ttriuted to its onservtion. Potto tuers re usully stored for severl months t low temperture prior to plnting to dely sprouting [2]. However,whendormnyisrokenndsproutingistivted[3], importnt eonomi dmge is reorded. Therefore, effetive sprout ontrol is mjor omponent of mnging stored potto qulity [4]. By fr, the mostused hemil sprout inhiitor worldwide is hlorprophm (isopropyl-(n-3-hlorophenyl) rmte (CIPC), CAS # ) [5]. However, it n negtively impt the environment nd humn helth [6]. Therefore, sfety onsidertions hve prompted the EU to introdue oth restritions on pplition s well s mximum residue levels (mss of CIPC per mss of potto.63 gkg 1 nd.1 gkg 1,resp.)[7]. Ethylene is lso n effetive potto sprout inhiitor, ut it often drkens fry olour [8]. Nowdys, it is very importnt to use nturl produts ompounds suh s essentil oils s well s the pure ompound derived from essentil oils [9]. Bioonservtion ould e good lterntive for pottoes qulity preservtion during storge without further environmentl dmge used y the use of grohemils. Indeed, severl essentil oils hve een identified with sprouting inhiition potentil on pottoes: the rwy (Crum rvi L.) essentil oil [1, 11], the orinder (Corindrum stivum L.) essentil oil [12], nd the mint (Menth spit L.) essentil oil [6]. However, until now no previous studies were foused on the RCEO sprouting potentil inhiition on pottoes. As mtter of ft, Rut hlepensis L.iswildplntwidespredinthe Mediterrnen region [13] known minly for its ortifient, nthelminti, ntispsmodi, phytotoxi, ntimiroil, nd ntioxidnt tivities [14 19]. Yet, no dt ws found on pottoes ioonservtion using Rut hlepensis L. essentil oil.

2 2 Journl of Chemistry Thus, the min purpose of the present work ws to identify RCEO s novel ntisprouting gent for pottoes to sustitute the existing grohemil produts for regenertion inhiition during storge. In ft, severl sprout inhiition ssys were performed to optimize RCEO emulsions onentrtions to provide pioneering pottoes ioonservtor. Then, RCEO ntisprouting omponents were identified using gs hromtogrphy oupled to mss spetrometry. 2. Mteril nd Methods 2.1. Plnt Mteril Tuer Mteril. Tuer mteril used in this study ws Spunt potto vriety. This ultivr represented the urrent Tunisin pottoes mrket (widely ultivted). Smples were olleted during the summer of 215 (June) nd then stored in the Tehnil Center of Potto nd Artihoke in Mnou, Tunisi Rut hlepensis Mteril. Aeril prts of Rut hlepensis were olleted from El Fhs region, loted t the north of Tunisi, during My nd June 215 t flowering stge Rut hlepensis Essentil Oil. RCEO ws extrted from the eril prts y hydrodistilltion for 12 min using levenger pprtus type. Then, the otined essentil oil ws stored t 4 C s previously desried y Mejri et l. [2] Sprout Inhiition Assys. RCEO emulsions (2%, 4%, nd 6%) were prepred in sterilized wter nd Empiol ESB 7 s surftnt (2.5%). A ontrol tretment ws pplied y prepring n emulsion of sterilized wter nd Empiol ESB 7 (2.5%) without essentil oil. Three oxes ontining 1 pottoes were rrnged for eh tretment. Smples were immerged for 1 min in the emulsion for eh onentrtion seprtely. Inhiition of potto sprouting ssy ws rried out during 6 weeks. Potto smples were kept in the drkness t 25 C to simulte trditionl storge onditions Tuer Storge Qulity Prmeters Sprout Length. Sprout length ws mesured weekly for eh smple of eh tretment Weight Loss. Sprouted potto smples were weighted eh week for eh tretment. Moreover, sprouts were seprted from the potto tuers nd weighted seprtely only for the lst week. Perentge of totl weight loss (TWL), perentge of physiologil weight loss (PhWL), nd perentge of sprouts weight (SW) were omputed using TWL = 1 (initil weight weight t week 6 with out sprouts) (initil weight) Sprouting (%) Control 25 C Treted 2% Storge durtion (weeks) Treted 4% Treted 6% Figure 1: Effet of RCEO on potto sprouts length during storge: evolution of perentge of sprouting during storge t different tretments. (initil weight weight t week X) PhWL = 1 (initil weight) (sprouts weight) SW = 1 (initil weight). (1) PhWL illustrtes the weight loss due to rething nd sweting. SW evlutes the loss due to sprouting. TWL is the sum of PhWL nd SW Determintion of Rut hlepensis Essentil Oil Biotive Components. RCEO omposition ws determined using gs hromtogrphy oupled to mss spetrometry (GC-MS) using mss spetrometer HP 5975 C (Agilent Tehnologies) with eletron impt ioniztion (7 ev). A pillry olumn HP-5 ms ws used (3 m 25 um oted with 5% phenyl methyl silione, 95% dimethyl polysiloxne, nd the film thikness.25 um). The oven temperture ws progrmmed from 6 to 22 Ctrteof4 Cmin 1,ndthetempering temperture ws 23 C. The rrier gs ws helium with flow rte of.8 mlmin 1 nd split rtio of 5 : 1. Snning time nd rnge mss were 1 s nd 5 55 m/z (moleulr mss/hrge), respetively. RCEO omponents identifition ws performed y ompring their retention indies (RI) with orresponding dt of ommeril ville stndrds Dt Anlysis. Stndrd one-wy nlysis of vrine (ANOVA) ws onduted to ssess the effet of RCEO emulsions on potto sprout inhiition nd weight loss using Dunn test. Person s orreltion test ws performed etween prmeters. All sttistil tests were performed using the softwre SPSS version 13. (SPSS, Chigo, USA). 3. Result nd Disussions 3.1. Effet of Rut hlepensis L. Essentil Oil on Potto Sprouts Length during Storge. Sprouting experiments were onduted during storge period of six weeks using three different tretments (onentrtions of emulsions). Figure 1 presents sprouting perentge evolution during pottoes storge. An overll ssessment of sprouting evolution urves showed

3 Journl of Chemistry 3 Sprout length verge (mm) d Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Control Treted 2% Storge durtion (weeks) Treted 4% Treted 6% Figure 2: Effet of RCEO on potto sprouts length during storge. (,,)Dt(fortheeffetofRCEOonsproutlengthforehweek) with different letters re signifintly different (p.1). (%) Control Treted 2% Treted 4% Treted 6% TWL PhWL Tretments Figure 3: Effet of RCEO t different onentrtions on weight loss of potto fter 6 weeks of storge. TWL: totl weight loss; PhWL: physiologil weight loss; nd SW: sprouts weight. (,, ) Dt hving different letters re signifintly different (p.1) ording to Dunn test results. SW signifint differenes etween the urves shpes suggesting signifint differenes etween the studied smples intertion mehnism with the different tretments. Regrding treted smples, sprouting perentge inresed smoothly to reh their mximum fter four weeks of storge. In the se of smples treted y 6% emulsions, n pprent inhiitor effet on sprouting ws oserved sine week 3. In ft, notle differene ws shown etween tretment t 6% nd the other tretments. However, less differene ws deteted etween tretments t 2 nd 4%. In order to investigte the impt of RCEO on sprouting, pottoes sprout lengths were sttistilly heked for eh week under eh tretment (Figure 2). ANOVA results reveled high signifint differenes etween sprout length evolutions under the different tretments. In ft, different tretments of RCEO hd signifint effet (p.5) on sprout inhiition. Untreted potto tuers egn to sprout sine the first week of storge, wheres the treted smples sprouting were irrelevnt t this point. Remrkle differene in sprout length ws found fter two weeks of storge etween the three different tretments, wheres tretment 6% ws more effiient in inhiiting potto tuers sprouting. This result suggested tht RCEO ould e onsidered s possile lterntive for sprouting inhiition. The sprout ontinued to grow fter five weeks of storge under treted nd untreted onditions, wheres Dunn s test showed no signifint differene etween tretments 2% nd 4%. After six weeks, sprout finl length rehed n verge of mm, mm, nd 12.2 mm under 2%, 4%, nd 6% tretments, respetively. As expeted, the ontrol smple reorded the highest finl sprout length (59.23 mm). Furthermore, Dunn s test results onfirmed tht no relevnt differene ws deteted etween sprout length of potto tuers treted y 2% nd 4% inditing the outstnding ntisprouting effet of emulsions t 6%. Indeed, y inresing the tretment onentrtion, the sprout growth ws proven to e deresing under the effet of essentil oil. These findings ould e ttriuted to diret effet of RCEO on ell division induing sprout growth inhiition. Romgni et l. findings estlished tht essentil oils nd their omponents inhiit ell division in the root tips growth nd interfere with DNA synthesis in growth meristem [21, 22]. However, Tworkoski et l. works indited tht essentil oils inhiit plnt growth y disrupting memrne integrity [23, 24]. Furthermore, other reserhes suggested tht essentil oils indue oxidtive stress nd inhiit root growth [24 27]. In ft, essentil oil improves lipid peroxidtion nd umultion of hydrogen peroxide nd inreses the eletrolyte lekge in root tissues [24, 27]. Moreover, Btish work showed tht plnts treted with essentil oil of Eulyptus showed signifint degrees of ion lekge inditing loss of memrne permeility leding to serious dmge to the plnt [28]. Furthermore, severl studies [29 31] reveled tht essentil oils hd diret effet on the omposition of hlorophyll nd ell respirtion Effet of Rut hlepensis Essentil Oil on Potto Weight Loss during Storge. Effet of RCEO tretments on potto weight loss ws determined y weighing the tuers during storge period s presented in Figure 3. Weight loss strted effetively fter two weeks of storge. In ft ording to Fernie nd Willmitzer, strh degrdtion is n importnt event relted to the indution of sprouting [32]. Sprouting tuer needs to otin energy from the mother tuer, whih ould use weight loss. As mtter of ft, signifint differene etween tretments ws notiele strting from week 3 s onfirmed y ANOVA results. More prtiulrly, Dunn s test reveled tht until four weeks of storge, there ws no signifint differene etween untreted nd 2% treted smples (PhW). Remrkly, smples treted with4%nd6%emulsionsretedsimilrlyuntilweek4. However, only fter five weeks of storge, high signifint differene ws shown etween 4% nd 6% tretments. In ft, 4% tretment effet on tuers weight loss deresed, while it persisted for the tretment 6% (Tle 1). These findings suggested tht the effet of RCEO on physiologil weight loss strted from the 4% onentrtion. Moreover, Figure 4 onfirmed the gret impt of 6% RCEO tretment

4 4 Journl of Chemistry Tle 1: Evolution of the verge of physiologil weight loss during storge. Tretment Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Control % % % Dt re expressed s men from three replites. Mens of the sme olumn followed y different letters differ signifintly (p <.1). Tle 2: Chemil omposition of Rut hlepensis L.essentil oil. Control smple Emulsion 6% Figure 4: Effet of Rut hlepensis essentil oil on potto weight loss during storge: pottoes during the week 6 of storge: t the right, potto treted with emulsion 6%; t the left, ontrol smple potto. on sprouting. Indeed, we reorded totl weight loss of 11.88%fortheontrolsmple,whileitwsonly6.2%forthe tretment 6%, onsistently with sprout weight ssessment. In ft, untreted tuers finl sprout weight ws 4.66% of the initil weight, wheres for the tretment 6%, it ws.98%. Furthermore, results showed tht untreted smples sored physiologil weight loss of 7.22% of the initil tuer weight, whileforthetretment6%,itws5.4%(figure3). Thus, it ould e onluded tht sprouting inhiition proly delys tuer weight loss, s previously reported y Teper-Bmnolker et l. [6]. In ft, Person orreltion oeffiient results onfirmed high signifint orreltion etween totl weight loss nd physiologil weight loss (.98), totl weight loss nd sprout finl weight (.99), nd physiologil weight loss nd sprout finl weight (.94). These orreltions ould sustin the hypothesis inditing tht RCEO ffeted the ell respirtion, whih ould e the leding mehnism of the sprouting inhiition Rut hlepensis Essentil Oil Biotive Components Identifition. Ative ntisprouting gents of RCEO were nlyzed y GC-MS. Tle 2 presented the identified omponents in RCEO. Results showed tht RCEO omposition ws rih in etoni omponents (95.17%), while 2.15% remined unidentified. The identified omponents were otne, 4- isopropenyl-1-methyl-1-ylohexene, 2-nonnone, nonnl, 1-methyl-5,6-divinyl-1-ylohexene, 2-denone, denl, 2- undenone, 2-dodenone, 2-tridenone, 1-heptdeene, pentosne, nd 1,2-enzenediroxyli id. Moreover, 2- undenone (87.18%) ws the min omponent of RCEO, onsistently with previous findings [2, 33 35] (68.95%, 47.7%, 69.23%, nd 77.18%, resp.). Chemil omposition of KI Rt Proportion [%] Otne Nonnone Nonnl Methyl-5,6-divinyl-1-ylohexene Denone Denl Undenone Dodenone Tridenone Heptdeene Pentosne ,2-Benzenediroxyli id Unidentified omponents 2.1 KI: Kovt index; Rt: retention time. the essentil oil n e ffeted y limti, sesonl, rinfll nd geogrphi onditions, distilltion tehnique, nd hrvest period [36]. RCEO ws pplied using three doses to enle sreening of tretments impt on sprouting level. Tretments reorded signifint impt s ntisprouting gent even t low doses. Therefore, the identifition of RCEO tive omponents ould e onsidered s n importnt disovery. 2-Undenone presented 87.18% of totl identified omponents. So, we ould ssume tht this omponent ould e the possile tive gent of RCEO. This ntisprouting gent hd different tion ompred to hemil produts. In ft, it is more orretly lled sprout suppressnts euse they physilly dmge developing sprouts with high onentrtion of the produt in the surrounding hedspe in the potto storge [4]. On the other hnd, other pottoes ntisprouting omponents in essentil oils hve een lredy identified in previous works. For instne, Brdow nd Connik determined the effet of nine methyl ketones from Amrnthus plmeri S. Wts. (AMAPA) residues on seed germintion of onion, rrot, AMAPA, nd tomto [37]. In ddition, R-rvone ws the tive sprouting inhiitor of mint essentil oil during pottoes storge s previously reported y Teper-Bmnolker et l. [6]. 4. Conlusion The potentil ntisprouting effet of RCEO on pottoes during storge ws investigted. Sprout length nd weight loss

5 Journl of Chemistry 5 of potto tuers urves s funtion of storge durtion (6 weeks) reveled high signifint effet of the different tretmentssonfirmedyanovaresults.anoveltehnique ws suessfully implemented to pply RCEO on pottoes with different emulsions onentrtions. Tuers treted with 6% RCEO emulsions showed distinguished sprouting inhiition s ompred to the other tretments, onsistently with finl sprouts mesurements. Therefore, sprouting of pottoes during storge ws ssoited with weight loss nd sprout length s onfirmed y high Person orreltion oeffiients (>.9). RCEO omposition sreening reveled tht 2-undenone ws the mjor omponent of RCEO (87.18%) of the totlidentifiedomponentswhihouldlikelyethepossile ntisprouting gent of RCEO. These findings suggested RCEO s pioneering lterntive for pottoes ioonservtion without hrmful drwks on environment. Further reserh is expeted to onfirm the 2-undenone hypothesis s the RCEO tive omponent. Conflits of Interest The uthors delre tht there re no onflits of interest regrding the pulition of this pper. Aknowledgments The uthors thnk the Tehnil Center of Potto nd Artihoke for prtiipting in the reliztion of this work. Referenes [1] FAOSTAT, Food nd Agriulture Orgniztion of the United Ntions, Sttistis Division. y regions/e, 213. [2] P. Delple, Y. Brostux, M.-L. Fuonnier, nd P. D. Jrdin, Potto (Solnum tuerosum L.) tuer physiologil ge index is vlid referene frme in posthrvest geing studies, Posthrvest Biology nd Tehnology,vol.5,no.1,pp.13 16,28. [3] W.K.Colemn, Dormnyreleseinpottotuers:Areview, Amerin Journl of Potto Reserh, vol. 64, no. 2, pp , [4] M. J. Frzier, N. Olsen, nd G. Kleinkopf, Orgni nd lterntive methods for potto sprout ontrol in storge, CIS 112, in UniversityofIdho,CollegeofAgriulturendLifeSienes, Idho, USA, 24. 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