Combination of the Auxins NAA, IBA, And IAA With GA3 Improves the Commercial Seed-Tuber Production of Potato (Solanum Tuberosum L.) Under in Vitro Conditions

Research Article Combination of the Auxins NAA, IBA, and IAA with GA 3 Improves the Commercial Seed-Tuber Production of Potato (Solanum tuberosum L.) under In Vitro Conditions Ahmet Metin Kumlay Department of Field Crops, Faculty of Agriculture, Igdir University, 76000 Igdir, Turkey Correspondence should be addressed to Ahmet Metin Kumlay; Received 18 April 2014; Accepted 19 May 2014; Published 17 June 2014 Academic Editor: Khalid Mahmood Khawar Copyright © 2014 Ahmet Metin
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  Research Article Combination of the Auxins NAA, IBA, and IAA with GA  3 Improves the Commercial Seed-Tuber Production of Potato( Solanum tuberosum  L.) under  In Vitro  Conditions  Ahmet Metin Kumlay  Department of Field Crops, Faculty of Agriculture, Igdir University, 󰀷󰀶󰀰󰀰󰀰 Igdir, urkey  Correspondence should be addressed to Ahmet Metin Kumlay; akumlay@hotmail.comReceived 󰀱󰀸 April 󰀲󰀰󰀱󰀴; Accepted 󰀱󰀹 May 󰀲󰀰󰀱󰀴; Published 󰀱󰀷 June 󰀲󰀰󰀱󰀴Academic Editor: Khalid Mahmood KhawarCopyright © 󰀲󰀰󰀱󰀴 Ahmet Metin Kumlay. Tis is an open access article distributed under the Creative Commons AttributionLicense, which permits unrestricted use, distribution, and reproduction in any medium, provided the srcinal work is properly cited.Testudycomparedtheeffectso  1.0× MSmediumcontainingvariousconcentrationso   -naphthaleneaceticacid(NAA),indole-󰀳-acetic acid (IAA), and indole-󰀳-butyric acid (IBA), alone or in combination with gibberellic acid (GA 3 ) in micropropagationo three potato ( Solanum tuberosum  L.) cultivars Pasinler, Granola, and Caspar using binodal stem cuttings. Te results testi󿬁edimproved regeneration on  1.0 ×  MS medium containing variants o NAA, IAA, and IBA plus GA 3  on all cultivars. Te minimumdays to shoot induction on three cultivars ranged 󰀴.󰀲󰀵–󰀵d on 1.0× MS medium containing 󰀰.󰀲󰀵mg L −1 GA 3  +1 mg L −1 NAA. Telongest shoots (󰀱󰀱.󰀸cm), maximum number o nodes (󰀱󰀳.󰀵󰀰), and maximum number o leaves (󰀱󰀱.󰀰󰀰) were recorded on cv. Casparon  1.0 ×  MS medium containing 󰀱mg L −1 NAA  + 0.25 mg L −1 GA 3 . Te minimum time to root induction (󰀱󰀲.󰀲󰀵d) was noted oncv. Pasinler on the same medium. All o the regenerated shoots could be easily rooted. Te results showed that the combined effecto various concentrations o NAA, IAA, and IBA plus GA 3  was more pronounced compared to the auxins used alone. Te resultso this research are o signi󿬁cant importance or potato breeders. 1. Introduction Te potato ( Solanum tuberosum  L.) is an economically important plant used as ood in many countries o the worldandalsoveryimportantplantortheurkisheconomygrownover a large area. Te potato is the 󿬁rst major ood crop inwhich biotechnology has been successully implemented orseed production. Previous studies show that micropropaga-tion o potatoes depends on the biological value o cultivars,explant type (lea, node, shoot tip, etc.), type o culturemedium, season, temperature, photoperiod, and a balancedcombinationoplantgrowthregulators(PGRs)intheculturemedia [󰀱–󰀳]. Axillary buds, nodal tissues, stem explants, roots, leaves, tubers, shoots or stems, and cell suspensioncultures have been utilized to micropropagate potatoes [󰀱, 󰀴]. Te successul  in vitro  multiplication o potatoes dependson the presence o a suitable combination o auxins withgibberellic acid (GA 3 ) in the propagation medium [󰀴–󰀷]. Roest and Bokelmann [󰀸] suggested that a lower concen-tration o auxin with GA 3  (󰀰.󰀲󰀵mgL −1 ) had a positiveimpactontheshootandrootdevelopmentopotatoplantletsgrown  in vitro . Ghaffoor et al. [󰀹] have also suggested thatthe longest shoots were noted on   -naphthaleneacetic acid(NAA), the maximum number o nodes on indole-󰀳-butyricacid (IBA), and the maximum number o leaves on indole-󰀳-aceticacid-(IAA-)containingmedia.Zhangetal.[󰀱󰀰]suggestthat increased shoot length was promoted among potatoexplants with increasing concentrations o IAA; however, thestimulating effect o IAA was enhanced by the addition o GA 3 . Badoni and Chauhan [󰀴] showed that the integrationo 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀰.󰀰󰀱mgL −1 NAA had a positive effecton morphological plantlet characters o cv. Kuri Himalini.Dancietal.[󰀱󰀱]obtainedthebestresultsrom󰀱.󰀰 × MSmediacontaining 󰀱mgL −1 IAA, 󰀱mgL −1 IBA, and 󰀰.󰀳mgL −1 GA 3 rom Amelia, Cristian, Nicoleta, and Roclas potato cultivars.Since each individual hormone has its own unique effecton regeneration, it is vital to determine the combined effectso these on the regeneration o shoots and roots. Tereore,thepresentstudyaimedtodeterminethebestcombinationo  Hindawi Publishing CorporationBioMed Research InternationalVolume 2014, Article ID 439259, 7 pages  󰀲 BioMed Research InternationalNAA,IBA,andIAA,usedsinglyorincombinationwithGA 3 ,or successul  in vitro  commercial seed-tuber production o potato cvs. Pasinler, Granola, and Caspar using meristem-derived stem node cultures. 2. Materials and Methods 󰀲.󰀱. Te Concentrations of PGRs and Media Preparation.  󰀱.󰀰 ×  MS medium (󰀱󰀹󰀶󰀲) containing 󰀰.󰀲󰀵mgL −1 GA 3 , 󰀱mgL −1 NAA, 󰀱mgL −1 IAA, and 󰀱mgL −1 IBA with and without󰀰.󰀲󰀵mgL −1 GA 3  complemented with 󰀳% (w/v) sucrose and󰀰.󰀸% (w/v) agar was used. Te pH was calibrated to 󰀵.󰀶–󰀵.󰀸 by 󰀱N HCl or 󰀱N NaOH afer adding all mediumcomponents except the agar. GA 3 , IAA, and IBA are thermo-labile;thereore,theywere󿬁lter-sterilizedbypassingthrough󰀰.󰀲 󽠵 m Millipore 󿬁lters (Schleicher & Schuell, FP 󰀳󰀰/󰀰.󰀲 CA-S; 󰀰.󰀲 󽠵 m; 󰀷bar max) inside a laminar 󿬂ow cabin beoreadding to each o culture medium afer autoclaving at 󰀴󰀵 ∘ C.Teculturesweresterilizedbyautoclavingat󰀱󰀲󰀰 ∘ Cor󰀲󰀰minand 󰀱󰀰󰀴kPa pressure. 󰀲.󰀲. Plant Material and Micropropagation of Explants.  Bin-odal explants rom three potato cultivars, namely, Pasinler(locally improved and registered mid-early maturing culti- var),Granola(mid-late-maturingcultivar),andCaspar(late-maturing cultivar), were used in the present study. Binodalcuttings were aseptically cultured on 󰀱.󰀰  ×  MS mediumcontaining󰀱󰀰binodalexplantsperreplicationand󰀴󰀰binodalexplantspertreatment.Eachreplicationconsistedooneglass jar and all experiments were replicated our times. Cultureswere incubated at a temperature o   24 ± 2 ∘ C under 󰀲,󰀰󰀰󰀰 luxlight intensity with 󰀱󰀶h o day light photoperiod or 󰀶 weeks. 󰀲.󰀳. Statistical Analysis.  Te data were recorded or meanregeneration percentage (%), days to shoot and root induc-tion, shoot and root length (cm), and the number o nodes,leaves, and roots per shoot. A completely randomized design(CRD) was used to evaluate the three cultivars, eight plantgrowth regulator combinations, and our replications. Datawere subjected to analysis o variance and the means wereseparated by Duncan’s multiple range test. Results on allparameters are the means and standard errors ( ± SE) romour replications. 3. Results and Discussion Te effect o variants o auxins plus GA 3  on the binodalexplants regeneration rate, shoot and root length, days toshoot and root induction, number o nodes, leaves and rootswere signi󿬁cantly different ( 􍠵 < 0.01 ). Te interactionbetween treatmentsandcultivarpotentialityorthelengtho shoots and roots, the number o nodes and leaves, and rootinduction days also showed signi󿬁cant variations ( 􍠵 < 0.01 ).Te results o each treatment and cultivar interaction arepresented below under their individual subheadings. 󰀳.󰀱. Regeneration Rate.  PGRs combinations in 󰀱.󰀰  ×  MSmedium affected the regeneration o binodal explants vari-ably( 􍠵 < 0.01 ).Centpercentregeneration(󰀱󰀰󰀰%)othethreecultivars was observed on agar-solidi󿬁ed 󰀱.󰀰  ×  MS mediumcontaining each o 󰀱mgL −1 NAA, IAA, and IBA with󰀰.󰀲󰀵mgL −1 GA 3 . However, 󰀱.󰀰  ×  MS medium containingNAA, IAA, IBA, and 󰀰.󰀲󰀵mgL −1 GA 3  singly was inhibitory and produced a reduced regeneration rate (Figure 󰀱). Teseresults are in agreement with a study by Webb et al. [󰀱󰀲]in which they suggested that the sequential application o hormones(IAAandNAA),inthepresenceoGA 3 ,enhancedshootregenerationromtheleadiscsoexplantsosixpotatocultivars. Miller et al. [󰀱󰀳] using the slow-growing Desiree,Record, Foxton, and Golden Wonder cultivars ound that acombination o 󰀱mgL −1 GA 3  and 󰀰.󰀱mgL −1 NAA effectively increased the number o nodes which could be culturedthereafer. Te results o this study are also similar to thoseoBadoniandChauhan[󰀴],whichsuggestedthat󰀰.󰀰󰀱mgL −1 NAA with 󰀰.󰀲󰀵mgL −1 GA 3  was the best substrate or shootregeneration in the potato cultivar “Kuri Himalini.” 󰀳.󰀲. Shoot Length.  Variants o NAA, IAA, and IBA plus GA 3 affectedshootlengthvariably,showingsigni󿬁cantdifferencesamong them ( 􍠵 < 0.01 ). Te longest shoots were notedon cv. Caspar (󰀱󰀱.󰀸cm) using 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀱mgL −1 NAA ollowed by Granola (󰀱󰀰.󰀴󰀳cm) on the same medium,and Caspar (󰀹.󰀴󰀳cm) using 󰀱.󰀰  ×  MS medium containing󰀰.󰀲󰀵mgL −1 GA 3 + 󰀱mg L −1 IAA. Te minimum shoot lengthwasrecordedoncv.Caspar(󰀲.󰀱󰀸cm)using󰀱.󰀰 × MSmediumcontaining 󰀰.󰀲󰀵mgL −1 GA 3  (Figure 󰀱). Te results in linewith 󿬁ndings o Webb et al. [󰀱󰀲] testi󿬁ed that IAA and NAA,incombinationwithGA 3 ,enhancedshootelongation.Zhanget al. [󰀱󰀰] suggested that the GA 3  and IAA positively affectedthe shoot length in cv. Zihubai. Farhatullah and Sayeed [󰀱󰀴]obtained󰀹.󰀱cmlongshootson󰀰.󰀲󰀴󰀸mgL −1 GA 3 .BadoniandChauhan [󰀴, 󰀶] ound that the combination o 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀰.󰀰󰀱mgL −1 NAA concentrations increased shootlength.Incontrast,O.M.DanciandM.Danci[󰀲]andHoque[󰀷] observed the best shoot regeneration on IAA, and Shiblietal.[󰀱󰀵]andAl-alebetal.[󰀱󰀶]obtainedtheirbestresultson MS medium containing IBA. 󰀳.󰀳. Days to Shoot Induction.  Te effect o all PGR applica-tionsonshootinductiondayswassigni󿬁cantlydifferent( 􍠵 <0.01 ). Te minimum days to shoot induction were noted oncv. Granola (󰀴.󰀲󰀵d), ollowed by cv. Pasinler (󰀴.󰀷󰀵d) and cv.Caspar (󰀵.󰀰󰀰d) on 󰀱.󰀰  ×  MS medium containing 󰀰.󰀲󰀵mgL −1 GA 3  +󰀱mgL −1 NAA.However,late-maturingcv.Casparhadthe most delayed shoot induction period o 󰀱󰀹.󰀲󰀵d, ollowedby󰀱󰀷.󰀵󰀰oncv.Granolaand󰀱󰀵.󰀷󰀵doncv.Pasinleron󰀱.󰀰 × MSmedium containing 󰀰.󰀲󰀵mgL −1 GA 3  (Figure 󰀲). Te resultsshow the experimental treatments were effective to reducetime to shoot induction compared to Yasmin et al. [󰀱󰀷], whonoted the minimum days to shoot induction on cvs. Desiree(󰀴.󰀳d) and Patrones (󰀵.󰀱d) using 󰀰.󰀵mgL −1 GA 3  + 󰀱mgL −1 pantothenic acid. Although the minimum number o days toshoot induction was determined on 󰀱.󰀰  ×  MS medium con-taining󰀰.󰀲󰀵mgL −1 GA 3  +󰀱mgL −1 NAAinthepresentstudy,Hoque [󰀷] observed that IAA-containing media acceleratetime to regenerate with the least time or shoot regeneration.  BioMed Research International 󰀳 020406080100T1 T2 T3 T4 T5 T6 T7PasinlerGranolaCaspar    R  e  g  e  n  e  r  a   t   i  o  n  r  a   t  e    (   %    ) Treatments (a) T1 T2 T3 T4 T5 T6 T7PasinlerGranolaCasparTreatments 024681012    S    h  o  o   t    l  e  n  g   t    h    (  c  m    ) (b) F󰁩󰁧󰁵󰁲󰁥 󰀱: Effects o 󰀱.󰀰  ×  MS medium containing 󰀱mgL −1 o NAA, IAA, and IBA, with and without 󰀰.󰀲󰀵mgL −1 GA 3 , on the regenerationrate and shoot length o cvs. Pasinler, Granola, and Caspar. Means o different values rom our replications ( 󝠵 = 4 ) are statistically differentusing Duncan’s multiple range test at the 󰀰.󰀰󰀱 level o signi󿬁cance. (󰀱: 󰀰.󰀲󰀵mgL −1 GA 3 , 󰀲: 󰀱mgL −1 NAA, 󰀳: 󰀱mgL −1 IAA, 󰀴: 󰀱mgL −1 IBA, 󰀵: 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀱mgL −1 NAA, 󰀶: 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀱mgL −1 IAA, 󰀷: 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀱mgL −1 IBA.) 05101520    D  a  y  s   t  o  s    h  o  o   t   i  n    d  u  c   t   i  o  n T1 T2 T3 T4 T5 T6 T7PasinlerGranolaCasparTreatments (a) 010203040    D  a  y  s   t  o  r  o  o   t   i  n    d  u  c   t   i  o  n T1 T2 T3 T4 T5 T6 T7PasinlerGranolaCasparTreatments (b) F󰁩󰁧󰁵󰁲󰁥 󰀲: Effects o 󰀱.󰀰 × MS medium containing 󰀱mg L −1 o NAA, IAA, and IBA, with and without 󰀰.󰀲󰀵mg L −1 GA 3 , on the number o daysto shoot and root induction o cvs. Pasinler, Granola, and Caspar. Means o different values rom our replications ( 󝠵 = 4 ) are statistically different using Duncan’s multiple range test at the 󰀰.󰀰󰀱 level o signi󿬁cance. (󰀱: 󰀰.󰀲󰀵mg L −1 GA 3 , 󰀲: 󰀱mg L −1 NAA, 󰀳: 󰀱mg L −1 IAA, 󰀴:󰀱mg L −1 IBA, 󰀵: 󰀰.󰀲󰀵mg L −1 GA 3  + 󰀱mg L −1 NAA, 󰀶: 󰀰.󰀲󰀵mg L −1 GA 3  + 󰀱mg L −1 IAA, and 󰀷: 󰀰.󰀲󰀵mg L −1 GA 3  + 󰀱mg L −1 IBA.) 󰀳.󰀴. Days to Root Induction.  Statistically signi󿬁cant effectswererecordedorallPGRapplicationsonthenumberodaysto root induction ( 􍠵 < 0.01 ). Te minimum number o daysrequired or root induction was noted on 󰀱.󰀰  ×  MS mediumcontaining 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀱mgL −1 NAA (󰀱󰀲.󰀲󰀵d or cv.Pasinler) and 󰀱󰀲.󰀵󰀰d each or cv. Granola and cv. Caspar(Figure 󰀲). Te treatment shown was signi󿬁cantly effectivein decreasing the days to root induction compared to theprevious reports, where Yasmin et al. [󰀱󰀷] observed the leastnumber o 󰀱󰀶.󰀷d to root induction in cv. Desiree and 󰀲󰀵.󰀹din cv. Patrones potato meristems by using 󰀰.󰀵mgL −1 GA 3  +󰀱mgL −1 pantothenic acid and control, respectively. 󰀳.󰀵. Number of Nodes.  Plant growth regulators affected thenumberonodesperbinodalexplantsigni󿬁cantly( 􍠵 < 0.01 ).Te maximum number o nodes was obtained on cv. Casparon 󰀱.󰀰  ×  MS medium containing 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀱mgL −1 NAA (󰀱󰀳.󰀵󰀰) and 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀱mgL −1 IAA (󰀱󰀲.󰀷󰀵).Te minimum nodes on each cultivar were obtained on 󰀱.󰀰 × MSmediumcontainingonly󰀰.󰀲󰀵mgL −1 GA 3  (Figure 󰀳).Te  󰀴 BioMed Research International 03691215    N  u  m    b  e  r  o    f  n  o    d  e  s T1 T2 T3 T4 T5 T6 T7PasinlerGranolaCasparTreatments (a) 036912    N  u  m    b  e  r  o    f    l  e  a  v  e  s T1 T2 T3 T4 T5 T6 T7PasinlerGranolaCasparTreatments (b) F󰁩󰁧󰁵󰁲󰁥 󰀳: Effects o 󰀱.󰀰  ×  MS medium containing 󰀱mg L −1 o NAA, IAA, and IBA, with and without 󰀰.󰀲󰀵mgL −1 GA 3 , on the number o nodes andleaves ocvs.Pasinler, Granola,and Caspar.Means odifferent values romourreplications ( 󝠵 = 4 )are statisticallydifferentusingDuncan’s multiplerange testat the󰀰.󰀰󰀱 level osigni󿬁cance. (󰀱:󰀰.󰀲󰀵mg L −1 GA 3 ,󰀲: 󰀱mgL −1 NAA, 󰀳:󰀱mgL −1 IAA, 󰀴:󰀱mg L −1 IBA,󰀵:󰀰.󰀲󰀵mg L −1 GA 3  + 󰀱mg L −1 NAA, 󰀶: 󰀰.󰀲󰀵mg L −1 GA 3  + 󰀱mg L −1 IAA, and 󰀷: 󰀰.󰀲󰀵mg L −1 GA 3  + 󰀱mg L −1 IBA.) resultsareimprovementoverpreviousreportsbyBadoniandChauhan [󰀴], Badoni and Chauhan [󰀶], Miller et al. [󰀱󰀳], Hassan et al. [󰀱󰀸], and Zaman et al. [󰀱󰀹]. Miller et al. [󰀱󰀳] noted that the combination o 󰀱mgL −1 GA 3  + 󰀰.󰀱mgL −1 NAA was effective in increasing the number o nodes (󰀷.󰀶).Zaman et al. [󰀱󰀹] reported that a higher concentration o auxinsresultedinahighernumberonodes(󰀷.󰀳).BadoniandChauhan[󰀴,󰀶]oundthatthe󰀰.󰀲󰀵mgL −1 GA 3  +󰀰.󰀰󰀱mgL −1 NAA concentrations increased the number o nodes (both󰀹.󰀴 nodes). Ghaffoor et al. [󰀹] obtained a higher number o nodes (󰀹.󰀷) on MS medium containing IBA. Te results o Shiblietal.[󰀱󰀵]showedthatthetotalnumberonodesrangedrom󰀱󰀰.󰀲(at󰀲.󰀰mgL −1 IBA+󰀱mgL −1 GA 3 )to󰀳.󰀵nodes/testtube (at 󰀲.󰀰mgL −1 IAA + 󰀱mgL −1 GA 3 ). In another study,Armin et al. [󰀲󰀰] also speculated that the application o NAAcompletely inhibited the growth o single nodes o   in vitro grown potato plantlets. 󰀳.󰀶. Number of Leaves.  Te number o leaves per binodalexplant showed signi󿬁cant variation ( 􍠵 < 0.01 ) afer treat-ment with 󰀱.󰀰  ×  MS medium containing 󰀱mgL −1 NAA,󰀱mgL −1 IAA,and󰀱mgL −1 IBA,withandwithout󰀰.󰀲󰀵mgL −1 GA 3 .Temaximumnumberoleavesperbinodalexplantoncv. Caspar (󰀱󰀱.󰀰󰀰) and on cv. Granola (󰀱󰀰.󰀰󰀰) was obtainedon 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀱mgL −1 NAA and on 󰀰.󰀲󰀵mgL −1 GA 3  +󰀱mgL −1 IAA,respectively,whichwasollowedclosely by 󰀶.󰀷󰀵 leaves per explant on cv. Granola using 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀱mgL −1 NAA. Te minimum leaves per explant(󰀱.󰀷󰀵) on each o the three cultivars were noted on 󰀱.󰀰  × MS medium containing 󰀰.󰀲󰀵mgL −1 GA 3  (Figure 󰀳). Teresults have edge over previous results with more numbero leaves per explant. Farhatullah and Sayeed [󰀱󰀴] reportedthe maximum number o leaves (󰀷.󰀳) on 󰀰.󰀲󰀴󰀸mgL −1 GA 3 .Similarly, Zaman et al. [󰀱󰀹] regenerated maximum numbero leaves (󰀸.󰀹) using 󰀰.󰀵mgL −1 NAA. Ghaffoor et al. [󰀹]regeneratedthemaximumnumbero󰀶.󰀱󰀴󰀳leavesperexplanton 󰀰.󰀲󰀵mgL −1 IBA containing 󰀱.󰀰  ×  MS medium. 󰀳.󰀷. Number of Roots.  Tere was a signi󿬁cant ( 􍠵 < 0.01 )effect o all PGR applications on the number o roots. Temaximum number o roots was observed on cv. Caspar(󰀲󰀷.󰀰󰀰), ollowed by cv. Pasinler (󰀲󰀵.󰀵󰀰) and cv. Granola(󰀲󰀵.󰀵󰀰) on medium containing 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀱mgL −1 IBA. Te minimum roots (󰀱.󰀰) grew on medium contain-ing 󰀰.󰀲󰀵mgL −1 GA 3  (Figure 󰀴). Te results presented hereare improvement over the previous results. Sanavy andMoeini [󰀲󰀱] suggested that the application o 󰀱.󰀵mgL −1 NAAdecreased the number o roots rom 󰀵 to 󰀳. In another study,Al-aleb et al. [󰀱󰀶] recorded the highest the number o 󰀱󰀰.󰀴󰀰roots per explant rom cv. Spunta on media containing IBA.Zamanetal.[󰀱󰀹]reported󰀲󰀳.󰀷rootson󰀱mgL −1 IBAcontain-ing medium. Shibli et al. [󰀱󰀵] obtained the best results (󰀱󰀶.󰀲roots) rom IBA-containing media. Hoque et al. [󰀷] obtainedtheirmaximumnumberoroots(󰀱󰀷.󰀴)using󰀰.󰀲󰀵mgL −1 IAA.Dhital et al. [󰀳] observed that 󰀱.󰀰mgL −1 NAA gave rise toa greater number o roots (󰀹.󰀵) than 󰀱.󰀰mgL −1 IAA (󰀴.󰀰).Uddin [󰀵] determined the highest number o roots (󰀴.󰀴) on󰀰.󰀵mgL −1 IBA-containing medium. 󰀳.󰀸. Root Length.  Te PGR combinations affected the rootlength signi󿬁cantly ( 􍠵 < 0.01 ). Te longest roots were notedon cv. Pasinler (󰀹.󰀹󰀸cm) on 󰀰.󰀲󰀵mgL −1 GA 3  + 󰀱mgL −1 NAA, ollowed by cv. Granola (󰀸.󰀶󰀳cm) and cv. Caspar(󰀷.󰀷󰀸cm) on 󰀱.󰀰  ×  MS medium containing 󰀰.󰀲󰀵mgL −1 GA 3
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