JOURNAL OF HORTICULTURE AND POSTHARVEST RESEARCH 2020, VOL. 3(1), 1-10

University Journal homepage: www.jhpr.birjand.ac.ir of Birjand

An optimized protocol for in vitro propagation of and using apical-bud microcuttings

Mariem Lotfi1*, Chokri Bayoudh1, 2, Afifa Majdoub2 and Messaoud Mars1,2

1Research Unit on Agrobiodiversity (UR13AGR05), Department of Horticultural Sciences, Higher Agronomic Institute, IRESA- University of Sousse, 4042 Chott-Mariem, Sousse, Tunisia 2Regional Research Centre on Horticulture and Organic Agriculture (CRRHAB), IRESA-University of Sousse; 4042 Chott-Mariem, Sousse, Tunisia

A R T I C L E I N F O A B S T R A C T

Original article Article history: Received 26 April 2019 Purpose: In Tunisia, cultivars are widely threatened by the Revised 14 June 2019 attack of fire blight disease. Cultivation of tolerant cultivars is an Accepted 22 June 2019 effective control strategy for disease control. For this purpose, a Available online 3 October 2019 reliable protocol was established for micropropagation of local Pyrus communis and Pyrus syriaca L. and for large-scale production Keywords: of high-quality plantlets. Research method: Using apical explants, different media and hormones were tested to establish a acclimatization micropropagation procedure for local Tunisian Pyrus communis apical explants cultivars ‘Arbi’, ʻMaltiʼ, ʻMahdia 6ʼ and ʻMoknine 10ʼ and for Pyrus growth regulators syriaca. Disinfection with 4% HgCl2 treatment for 20 minutes micropropagation showed the highest percentage of survival. Successful initiation of the cultures was achieved on MS basal medium Tunisian pear cultivars supplemented with 0.25 mg L-1 BA. Findings: During the DOI: 10.22077/ jhpr.2019.2420.1055 proliferation stage, optimal shoot multiplication was obtained on P-ISSN: 2588-4883 MS medium with a half concentration of NH4NO3 and KNO3 supplemented with 0.1 mg L-1 IBA and 2 mg L-1 BA, but for maximum E-ISSN: 2588-6169 shoot length the BA concentration needed to be lowered to 1 mg L- 1. A rooting rate of 100% and the highest root length and root

*Corresponding author: number were attained on Cheng medium supplemented with 1.0 Research Unit on Agrobiodiversity -1 mg L IBA. Pear vitroplants were successfully acclimatized on S2 (UR13AGR05), Department of substrate, composed by peat moss. Research limitations: Horticultural Sciences, IRESA-University of Vitroplants acclimatization step needs to be well studied for the Sousse, 4042 Chott-Mariem, Sousse, improvement of the acclimatized vitroplant survival rates by Tunisia. reducing the symptoms of crown rot. Originality/Value: This efficient optimized in vitro protocol will be successfully applied for E-mail: [email protected] large multiplication of high quality of Tunisian Pyrus vitroplants and cultivars. © This article is open access and licensed under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/ which permits unrestricted, use, distribution and reproduction in any medium, or format for any purpose, even commercially provided the work is properly cited.

Chitranshi et al. Micropropagation of Pyrus communis Lotfi et al.

(w/v), myo-inositol (100 mg L-1), thiamine-HCl (1 mg L-1), nicotinic acid (1 mg L-1), Castro, C. D. P. C., Faria, J. & Dantas, INTRODUCTIONT. B. H. (2014). Evaluating the performance of coconut fiber pyridoxine-HCl (1 mg L-1), phloroglucinol (162 mg L-1), and 0.7% (w/v) Difco Bacto-Agar. and wood straw as cushioning materials to reduce injuries of papaya and mango during The pH was adjusted to 5.7 with KOH/HCl and the growth regulators were added before Pear,transportation. belonging Internationalto the genus Journal Pyrus of Advanced, subtribe Packaging Malinae Technology, (corresponding 2(1), 84to-95. the former autoclaving at 121 °C for 20 minutes. Maloideae https://doi.org/10.23953/cloud.ijapt.10), family (Zheng et al., 2014), is one of the oldest temperate crops. It The cultures were kept at 25 ± 1°C under a photoperiod of 16 hours under fluorescent Chandra,is considered D., & as Kumar, a worldwide R. (2012). fruit Qua tree,litative belonging effect of mainlywrapping to andAsian cushioning countries materials and the on Indian guava during storage. HortFlora Research Spectrum, 1(4), 318-322. light (40 µmol m-2 s-1). The explants of nodal segments (≈ 1.5 cm) were cultured in glass Chauhan,subcontinent S. K., (Sharma & Babu, & D.Pramanick, R. (2011). 2012 Use ).of Pearbotanicals: fruits Aare new an excellentprospective source for enhancing of vitamins, fruit tubes (12 cm×2.5 cm) containing 10 ml of M1, M2 and M3. After 3 weeks, the percentage of sugars,quality and over important chemicals phytochemicals in an era of global(Xia climateet al., 2016change.). InAsian Tunisia, Journal local of Environmentallow chilling explants forming shoots was recorded. After 3 weeks, when growth started (Fig. 1- a), the cultivarsScience, of 6 (1)Pyrus, 17- 28communis. L. are cultivated in coastal regions and classical European best-grown explants were shifted to multiplication medium M4, M5 and M6. Subculture was Creelman,cultivars in R. continental A., & Mullet, areas J. ( MarsE. (1995). et al., Jasmonic1994). Pyrus acid syriacadistribution Boiss., in :growing regul spontaneouslyation during done on a fresh medium with the same compositions every 4 weeks. The number of shoots in northdevelopment Tunisia andand responsesconsidered to tobiotic be veryand abioticresistant stress. to drought Proceedings and calcareous of the National soils, Academywas tested of per explant and the shoot length (mm) were measured monthly with a digital caliper at the end as potentialSciences USA,rootstock 92, 4114 for -common4119. https://doi.org/10.1073/pnas.92.10.4114 pear (Brini et al., 2008). However, both wild and local pear of the fourth subculture. Davidson,cultivars haveP. M., not &been Taylor, subjected M. T. to (2007).much researchChemical despite preservatives their interesting and natural characteristics. antimicrobial Maincompounds. threats for Food local Microbiology:pear cultivars Fundamentals and rootstocks and in Frontiers,Tunisia are Washington, urbanization, DC: generalized American Rooting use Societyof introduced for Microbiology cultivars, Pressclimatic, 713 variations,-734. https://doi.org/ and fire 10.1128/9781555815912blight (Rhouma et al., 2013; Gaaliche The rooting experiments were conducted on four rooting media (M7-M10; Table 1) under in DebMandalet al., 2018,) .M., & Mandal, S. (2011). Coconut (Cocos nucifera L.: Arecaceae): in health promotion vitro conditions with micropropagated shoots (approximately 1.5 cm long) obtained from the andTraditional disease prevention. vegetative Asian methods Pacific to Journalpropagate of Tropicalpear plants Medicine, are cutting 4(3), 241and- 247grafting,. https://doi.org/ but they 10.1016/S1995-7645(11)60078-3. fourth subcultures. All media were supplemented with 3% sucrose (w/v), thiamine-HCl (400 do not ensure disease-free plants and have low multiplication rates (Mars et al., 1994). -1 -1 -1 Donsì, F., Annunziata, M., Vincensi, M., & Ferrari, G. (2012). Design of nanoemulsion-based delivery mg L ), inositol (250 mg L ), phloroglucinol (162 mg L ) and 0.6% of (w/v) Difco Bacto- Micropropagationsystems of natural has antimicrobials: proven to be effect an efficient of the emulsifier. way to overcome Journal of these Biotechnology problems, 159from(4), many 342- Agar. After 3 days, shoots were transferred to growth regulator-free medium under standard species350. https://doi.org/and it enables10.1016/j.jbiotec.2011.07.001. rapid multiplication of disease-free plants at a commercial scale -2 -1 growth room conditions for 3 weeks (24±1 °C under 16-h photoperiod with 40 μmol m s Droby,(Bahmani S., etPorat, al., 2009 R.,; AyedCohen, et al.,L.,Weiss, 2018 ). B.,However, Shapiro, the B., pear Philosoph is considered-Hadas, as S.,one &of theMeir, most S. fluorescent light). The percentage of rooted shoots, the number of roots and average root recalcitrant(1999). Suppressing dicotyledonous green species mold decayfor tissue in grapefruit culture manipulations with postharvest (Reed jasmonates et al., 2013 application.; Aygun length per rooted shoot (mm) were measured with a digital caliper and recorded after 14 days. & Dumanoglu,Journal of 2015the) withAmerican low shoot Society multiplication for Horticultural rates, hyperhydricity Science , ,124(2), tissue oxidation,184-188. lackhttps://doi.org/10.21273/JASHS.124.2.184 of consistent adventitious rooting, and loss during acclimatization as the major Acclimatization bottlenecks.Dubey, N., Mishra, Nevertheless, V., & Thakur, micropropagation D. (2018). Plant of basedP. communis antimicrobial OHF formulations. 333, ‘Old Postharvest Home× The roots of the in vitro regenerated pear plants were rinsed with tap water to eliminate FarmingdaleDisinfection 87,’ ‘Hornerof Fruits 51,’ ‘Winterand Vegetables, Nelis’, and 1,OHF Elsevier 51 have beenAcademic reported Pres (Cheng,s, 322 1979-331;. culture medium and immersed in 0.1% fungicide solution (Pelt 500 SC®) for 3 min. The Nachevahttps://doi.org/10.1016/B978 et al., 2009; Reed et al.,-0 -201312-812698) and -P.1.00011 syriaca-X (Shibli et al., 1997). plants were transferred into trays containing one of two substrates: S1, 1/2 perlite and peat or Fallik,Since E., Arhbold,no reports D. D., are Hamil available ton-Kemp, on inT. R.,vitro Clements, micropropagation A. M. , Collins, of R.local W., Tunisian& Barth, M.pear E. (1998). (E)-2-Hexenal can stimulate Botrytis cinerea growth in vitro and on strawberry fruit in S2, peat moss, and kept under a tunnel at 24 ± 2°C, 16-h photoperiod. After 4 weeks, new cultivars,vivo duringthis study storage. was Journal undertaken of the toAmerican develop Society a reliable for Horticulturalin vitro propagation Science, 123,protocol 875-8 for88. leaves emerged and acclimated plants were transferred to a shaded greenhouse at 26/20 °C P. communishttps://doi.org/10.21273/JASHS.123.5.875 cultivars ‘Arbi’, ʻMaltiʼ, ʻMahdia 6ʼ and ʻMoknine 10ʼ and P. syriaca . (day/night), under 70% of relative humidity for hardening. Fenwick, G. R., Heaney, R. K., & Mullin, W. J. (1983). Glucosinolates and their break-down products in food and food plants.MATERIALS Critical Reviews AND in METHODFood ScienceS and Nutrition 18, 123-201. Table 1. Medium composition for Pyrus micropropagation https://doi.org/10.1080/10408398209527361 Medium BA IBA NAA FoodPlant and material Agriculture preparation Organiza tion of the United Nations (FAO), (2011). Global food losses and food Mg L-1 mg L-1 mg L-1 Fourwaste. cultivars United (‘Arbi’, Nations ‘Malti’,Food and ‘MahdiaAgriculture 6’ Organization. and ‘Moknine Rome, 10’) Italy. of http://faostat.fao.orgP. communis and. an Initiation Garcia,accession J. L.of (1982). P. syriaca Storage were of tomatused.oes In (vitroLycopersicon stock cultures esculentum were Mill) established in rice hull from ash apical of different buds M1 MS* 0.25 - - collectedparticle in sizesthe spring and moisture from 20 content-year- old[Philippines]. trees located Thesis. at theIn AgriculturalHigher Agronomic Information Institute Bank forof ** M2 MS 0.25 - - ChottAsia,-Mariem, South Tunisia.-East Asian Apic Regionalal segments Center (3 cmfor- long)Graduate were Study washed and underResearch running in Agriculture tap water M3 MS** 1 - - for 1(AGRIS) hour and s incewere 1985. surface -sterilized with 70% ethanol for 1 min, followed by treatment with Proliferation/Elongation Gyawali, R., & Ibrahim, S. A. (2014). Natural products as antimicrobial agents. Food Control, 46, ** 0.6% Benomyl for 2 min. Then, the explants were dipped in an antioxidant solution (ascorbic M4 MS - - - 412-429. https://doi.org/10.1016/j.foodcont.2014.05.047 ** and citric acid) at 0.2% and 0.1%, respectively, and washed under running tap water to M5 MS 1 0.1 - Hanušová, K., Dobiáš, J., & Klaudisová, K. (2009). Effect of packaging films releasing antimicrobial M6 MS** 2 0.1 - removeagents all on residues.stability of Priorfood products.to culturing, Czech Jounalthe explantsof Food Science were , 27,sterilized 347-349 .by submersion Rooting in commercial https://doi.org/10.17221/958 bleach sodium hy-CJFSpochlorite (NaOCl2; 10, 12, 15 and 20%) (w/v) or mercuric *** M7 Cheng - 1 - chlorideICAR, (2009 (HgCl). Indian2; 1, Council2 and 4%)of Agricultural (w/v) for Research.5, 10, 15, Annual 20 and Report. 30 min, adding a few drops of *** M8 Cheng - - 1 TweenJanjarasskul,-20. Finally, T., & Krochta, explants M. were (2010). rinsed Edible three packaging times with materials. sterile distilledFood Science water and and Technology planted in- * M9 MS - 1 - initiationAnnual medium. Reviews, Explant 1, 415- survival448. https://doi.org/10.1146/annurev.food.080708.100836 was scored 10 days after transfer to the medium. * M10 MS - - 1 Kamble, P. B., & Chavan, J. K. (2005). Effects of post-harvest treatments and storage temperature on * Original MS (Murashige & Skoog, 1962) Cultureshelf initiation-life of custard and apple shoot fruits. proliferation Jounal Food Science Technology, 42 (3), 253-255. ** MS with half the concentration of NH4NO3 and KNO3 ThreeKim, S. media A., & wereRhee, testedM. S. (2016).for culture Highly initiation enhanced (M1, bactericidal M2 and effects M3) ofand medium for proliferation chain fatty acids and *** Cheng medium (Cheng, 1979) elongation(caprylic, (M4, capric, M5, and M6) lauric of acid)the in combined vitro shoots with edible (Table plant 1). essentialThey all oils contained (carvacrol, 3% eugenol, sucrose β-

JOURNAL OF HORTICULTURE AND POSTHARVEST RESEARCH VOL. 3(1) MARCH 2020 JOURNAL OF HORTICULTURE AND POSTHARVEST RESEARCH VOL. 3(1) MARCH 2020

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Micropropagation of Pyrus communis Botanical products for post-harvest management of perishable produce

-1 -1 -1 (w/v),resorcylic myo-in acid,ositol trans (100-cinnamaldehyde, mg L ), thiamine thymol,-HCl and (1vanillin) mg L against), nicotinic Escherichia acid coli (1 O157:mg L H7.), pyridoxineFood Control-HCl (1, 60 mg, 447 L--1454.), phloroglucinol https://doi.org/ 10.1016/j.foodc(162 mg L-1), ont.2015.08.022.and 0.7% (w/v) Difco Bacto-Agar. TheKupwade, pH was S. R., adjusted Manglani, to N.5.7 S., with Wadkar, KOH/HCl S. S., Inamdar,and the F.growth R., Shete, regulators C. C., & were Ghosh, added J. S. before(2017) autoclavingAntimicrobial at 121 °Cactivity for 20 of minutes. Lemon peel extract against the bacteria belonging to the TheGenus cultures Xanthomonas. were kept International at 25 ± 1°CJournal under of aPharmaceutic photoperiodal ofSciences 16 hours Review under and fluorescent Research, light 45(2)(40 ,µmol 251-254 m-.2 https://doi.org/10.22159/ijcpr.2017v9i4.20962s-1). The explants of nodal segments (≈ 1.5 cm) were cultured in glass Lucera,tubes (12 A., cm×2.5 Costa, cm)C., containingConte, A., 10& mlDel of Nobile,M1, M2 M. and A. M3. (2012). After Food 3 weeks, applications the percentage of natural of explantsantimicrobial forming compounds.shoots was Frontiersrecorded. in Microbiology,After 3 weeks 3, ,287 when. growth started (Fig. 1- a), the https://doi.org/10.3389/fmicb.2012.00287. eCollection 2012 bestMalasri,-grown S., Tiapradit,explants P.,were Russell, shifted S., to Poonpurmsiri, multiplication P., mediumTarkarnviroj, M4, C., M5 Pourhashemi, and M6. Subculture A., Moats, was R., done& on Hudson, a fresh B.medium (2015) withInsulation the same effectiveness compositions of rice e veryhull. 4International weeks. The Journal number of of Advance shoots per explantPackaging and T theechnology, shoot length 3(1), 158 (mm)-168. were https://doi.org/ measured 10.23953/cloud.ijapt.20monthly with a digital caliper at the end ofMari, the M.,fourth Leoni, subculture. O., Lori, R., & Cembali, T. (2002) Antifungal vapour-phase activity of allyl isothyocyanates against Penicillium expansum on . Plant Pathology, 51, 231-236. Rooting https://doi.org/10.1046/j.1365 -3059.2002.00667.x TheMcClements, rooting experimentsD. J., & Rao, wereJ. (2011). conducted Food-grade on four nanoemulsions: rooting media formulation, (M7-M10; fabrication, Table 1 )properties, under in vitroperformance, conditions with biological micropropagated fate, and potential shoots toxicity. (approximately Critical Review 1.5 cm Foo long)d Science obtained and Nutrition,from the fourth51(4) subcultures., 285-330. https://doi.org/10.1080/10408398.2011.559558All media were supplemented with 3% sucrose (w/v), thiamine-HCl (400 mgMishra, L-1), B., inositol & Khatkar, (250 mgB. S. L -(2009).1), phloroglucinol Studies on post(162-harvest mg L -1quality) and 0.6%and physiology of (w/v) Difcoof coated Bacto ber- Agar.(Zizyphus After 3 days,mauritiana shoots Lamk) were fruit. transferred Journal to of growth Food Science regulator and-free Technology medium (Mysore),under standard 46(5), 455-458. -2 -1 Mohanty,growth room S., Ramesh, conditions S., & for Muralidharan, 3 weeks (24±1 N. P . °C(2017). under Antimicrobial 16-h photoperiod efficacy with of apple 40 μcidermol vinegarm s fluorescentagainst Enterococcuslight). The percentage faecalis and of Candida rooted albicansshoots,: theAn numberin vitro ostudyf roots. Journal and averageof Advanced root lengthPharmacy per rooted Education shoot (mm) and Research were measured, 7(2), 137 with-141 a. digital caliper and recorded after 14 days. M ustafa, E. , Noah, A. , Beshay, K. , Sultan, L. , Essam, M. & Nouh, O. (2015). Investigating the AcclimatizationEffect of Various Nanomaterials on the Wettability of Sandstone Reservoir. World Journal of The Engineeringroots of the and in Technologyvitro regenerated, 3, 116- 126.pear https://doi.org/ plants were10.4236/wjet.2015.33013. rinsed with tap water to eliminate Neri,culture F., medium Mari, andM., immersedMeniti, A.in 0.1%M., &fungicide Brigati, solutionS. (2006). (Pelt Activity500 SC ®of) fortrans 3 -min.2-hexenal The plantsagainst were transferredPenicillium intoexpansum trays incontaining ‘Conference’ one ofpears. two substrates:Journal of S1,Applied 1/2 perliteMicrobiology and peat 100 or, S2, peat1186 moss,-1193. andhttps://doi.or kept underg/10.1111/j.1365 a tunnel at -242672.2006.02873.x ± 2°C, 16-h photoperiod. After 4 weeks, new Nguyen,leaves emerged V. T. A., and Le, acclimatedT. D., Phan, plants H. N., were & Tran, transferred L. B. (2017). to a shadedAntibacterial greenhouse activity atof 26/20free fatty °C (day/night),acids from under hydrolyzed 70% of virginrelative coconut humidity oil using for hardening.lipase from Candida rugosa. Journal of Lipids, 1- 7. https://doi.org/10.1155/2017/7170 Prasad, K., & Stadelbacher, G. J. (1974). Effect of acetaldehyde vapor on postharvest decay and Table 1. Medium composition for Pyrus micropropagation market quality of fresh strawberries. Phytopathology, 64, 948-951. https://doi.org/10.1094/phyto- 64-948 Medium BA IBA NAA -1 -1 -1 Raorane, G. P. (2003). Studies on growth, flowering, fruitingMg Land somemg aspectL ofmg harvest L handling of kokum (GarciniaInitiation indica choisy). M.Sc. Thesis. Dr. Balasaheb Sawant Konkan Krishi * Vidyapeeth,M1 Dapoli, District. RatnagiriMS. 0.25 - - M2 MS** 0.25 - - Quirós-Sauceda, A. E., Ayala-Zavala, J.F., **Olivas, G. I., & González-Aguilar, G.A. (2014). Edible coatings asM3 encapsulating matrices forMS bioactive compounds:1 a review.- Journal- of Food Science and Technology,Proliferation/Elongation 51(9), 1674- 1685. https://doi.org/10.1007/s13197 -013-1246 -x M4 MS** - - - Ranjan, S., Dasgupta, N., Chakraborty, A. R., Samuel, S. M., Ramalingam, C., Shanker, R., & Kumar, M5 MS** 1 0.1 - A. (2014). Nanoscience and nanotechnologies in food industries: opportunities and research M6 MS** 2 0.1 - trends. Journal of Nanoparticle Research, 16(6), 2464. https://doi.org/10.1007/s11051-014-2464- Rooting 5. M7 Cheng*** - 1 - Saxena, R., Liquido, N. J., & Justo, H. B. (1981). Natural pesticides from the neem tree (Azadirachta M8 Cheng*** - - 1 indica). ProceedingM9 of International MSNeem* Conference- Rattach Engem,1 Germany- . Sembdner, G., M10& Parthier, B. (1993). TheMS biochemistry* and- the physiological- and1 molecular actions of jasmonates. Annual Review of Plant Physiology and Plant Molecular Biology, 44, 569-589. https://doi.org/10.1146/annurev.pp.44.060193.003033 * Original MS (Murashige & Skoog, 1962) ** Shilling, M., Matt, L., MS & with Rubin, half E.the (2013). concentration Antimicr of NHobial4NO effects3 and KNO of virgin3 coconut oil and its medium- chain fatty acids *** Cheng on mediumClostridium (Cheng, difficile 1979) . Journal of Medicinal Food, 16(12), 1079-1085. https://doi.org/ 10.1089/jmf.2012.0303. 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Chitranshi et al. Micropropagation of Pyrus communis Lotfi et al.

Table 4. Effect of culture media on multiplication rate and shoot length in different Pyrus sp. at the end of the StatisticalCastro, C. D. analyses P. C., Faria, J. & Dantas, T. B. H. (2014). Evaluating the performance of coconut fiber fourth subculture. All andexperiments wood straw were as conducted cushioning in materialsa completely to reduce randomized injuries design of papaya with threeand replicatesmango during and Species/cultivar Medium Multiplication rate Shoot length (mm) transportation. International Journal of Advanced Packaging Technology, 2(1), 84-95. c c 20 samples per each experimental unit (n = 60). The data were presented as means ± standard M4 1±0 7.46±0.33 https://doi.org/10.23953/cloud.ijapt.10 b a error (SE). The standard factorial analysis of variance and mean comparisons analysis, using ʻArbiʼ M5 6.6±0.59 23.05±0.42 Chandra, D., & Kumar, R. (2012). Qualitative effect of wrapping and cushioning materials on guava a b Duncan’s test (at P ≤0.05), were done using SPSS (Version 20.0 for windows Inc., Chicago, M6 11.7±0.97 18.76±0.52 fruits during storage. HortFlora Research Spectrum, 1(4), 318-322. c c IL, USA). M4 1±0 7.09±0.36 Chauhan, S. K., & Babu, D. R. (2011). Use of botanicals: A new prospective for enhancing fruit b a ʻMaltiʼ M5 5.85±0.36 21.68±0.47 quality over chemicals in an era of global climate change. Asian Journal of Environmental a b RESULTS Pyrus communis M6 9.05±0.82 18.74±0.59 Science, 6(1), 17-28. c c M4 1±0 7.30±0.48 Creelman, R. A., & Mullet, J. E. (1995). Jasmonic acid distribution in plants: regulation during b a Efficacydevelopment of sterilizing and responses agents to for biotic in vitro and cultureabiotic stress. establishment Proceedings of the National Academy of ʻMahdia 6ʼ M5 6.35±0.48 22.34±0.69 a b NaOClSciences2 and USA, low 92 concentrations, 4114-4119. https://doi.org/10.1073/pnas.92.10.4114 of HgCl2 were efficient to remove bacterial and fungal M6 9.65±0.58 15.48±0.81 c c contaminations,Davidson, P. M., but & later,Taylor, all M.the T.explants (2007). wereChemical lost. HgClpreservatives2 at 4% andfor 20natural min antimicrobialyielded the M4 1±0 9.57±0.51 compounds. Food Microbiology: Fundamentals and Frontiers, Washington, DC: American 6.45±0.51 b 20.04±0.35 a highest sterilization and survival rates for P. communis and P. syriaca explants (Table 2). ʻMoknine 10ʼ M5 Society for Microbiology Press, 713-734. https://doi.org/10.1128/9781555815912 9.35±0.48 a 15.11±0.63 b Extending the exposure time to 30 min was detrimental for the explants, whereas reducing the M6 DebMandal, M., & Mandal, S. (2011). Coconut (Cocos nucifera L.: Arecaceae): in health promotion c c exposure time resulted in increased explant loss due to contamination (Table 3). M4 1±0 10.08±0.26 and disease prevention. Asian Pacific Journal of Tropical Medicine, 4(3), 241-247. https://doi.org/ b a Pyrus syriaca M5 7.25±0.44 22.67±0.33 10.1016/S1995-7645(11)60078-3. a b Effect of medium composition on tissue browning and culture initiation M6 10.4±0.50 12.46±0.83 Donsì,Three F.,different Annunziata, media M., were Vincensi, tested M., to &obtain Ferrari, an G. optimized (2012). Design in vitro of nanoemulsioninitiation of -basedthe sterilized delivery

explants.systems As of shownnatural antimicrobials:in Table 3, the effect medium of the emulsifier.composition Journal had ofa significantBiotechnology impact, 159(4), on 342the- Media effect ** ** 350. https://doi.org/10.1016/j.jbiotec.2011.07.001. Cultivar effect ** ** response of the explants. M1 medium, a full-strength MS with 0.25 mg L-1 BA, was superior Droby, S., Porat, R., Cohen, L.,Weiss, B., Shapiro, B., Philosoph-Hadas, S., & Meir, S. Interaction ‘Cultivars × Media’ ** ** and gave a 100% explant establishment and completely prevented tissue browning in all the The values are compared vertically. Means with a different letter in a row are statistically different (Duncan. P ≤0.01). (1999). Suppressing green mold decay in grapefruit with postharvest jasmonates application. cultivars tested. Decreasing the NH4NO3 and KNO3concentration (M2 and M3) as well as Journal of the American Society for Horticultural Science , 124(2), 184-188. increasing the BA concentration (M3) stimulated necrosis and had a negative impact on Effect of hormone concentrations on shoot proliferation https://doi.org/10.21273/JASHS.124.2.184 Dubey,initiation, N., both Mishra, in the V., P. & communis Thakur, D. cultivars (2018). Plantand in based P. syriaca antimicrobial (Table formulations.3). Although Postharvest the trends After three weeks, the healthy shoots were excised from the initiation media (M1) and in theDisinfection responses wereof comparaFruits bleand for Vegetables,all cultivars, 1,there Elsevier was a significantAcademic interaction Press, between322-331. transferred on to multiplication medium. Shoot proliferation was assessed on three different cultivarhttps://doi.org/10.1016/B978 and medium composition-0- 12(Table-812698 3).- 1.00011-X MS media with half the concentration of NH4NO3 and KNO3 and different concentrations of Fallik, E., Arhbold, D. D., Hamil ton-Kemp, T. R., Clements, A. M. , Collins, R. W., & Barth, M. E. Table 2. Effect of different exposure times to HgCl2 (4%) on explant survival (%) of different Pyrus sp. BA and IBA (Table 4). A significant impact of medium composition and pear variety was (1998). (E)-2-Hexenal can stimulate Botrytis cinerea growth in vitro and on strawberry fruit in Exposure time Species/cultivar noted. Without hormones (M4), none of the genotypes multiplied and the shoot length vivo during storage. Journal of the American5 min Society10 min for Horticultural 15 min 20Science min , 123,30 875 min -888. -1 -1 remained constant after the first subculture (Table 4). When 1 mg L BA and 0.1 mg L IBA https://doi.org/10.21273/JASHS.123.5.875‘Arbi’ 0±0 e 22 ±1.22 c 31±1.30 b 45±1.14 a 6±1.14 d were supplemented to the medium (M5), the number of shoots per subculture increased Fenwick,Pyrus communis G. R., Heaney,‘Malti’ R. K., & Mullin, W.4 ± 1.30J. (1983). d 12±1.14 Glucosinolates c 17±0.83 and b their30±0.83 break a -down1±0.70 products e in food and food‘Mahdia plants. 6’ Critical 10±0.70Reviews d in19±1.14 Food c Science31±1.22 band Nutrition45±1.14 a 18,0±0 123 e -201. considerably and the shoots were longer for all cultivars (Table 4, Fig. 1- b). When the BA d c b a e -1 https://doi.org/10.1080/10408398209527361‘Moknine 10’ 7±1.14 14±0.89 24±0.70 40±1.58 3±0.83 concentration was increased to 2 mg L (M6), the multiplication rate increased with each Pyrus syriaca 34±1.30 d 52±1.22 c 67±1.58 b 85±1.11 a 7±1.14 e subculture for all tested cultivars. However, under these conditions, the shoot length Food and Agriculture Organization of the United Nations (FAO), (2011). Global food losses and food Significancewaste. of United exposure Nations duration Foodeffect and Agriculture** Organization.** Rome,** Italy. http://faostat.fao.org** ** . decreased and leaves turned narrow (Table 4), which made more difficult the further handling Garcia, J. L. (1982). Storage of tomatoes (Lycopersicon esculentum Mill) in rice hull ash of different Significance of interaction ‘CVS×Duration’ ** ** ** ** ** of the shoots multiplication and rooting. particle sizes and moisture content [Philippines]. Thesis. In Agricultural Information Bank for The valuesAsia, are South compared-East horizontally. Asian RegionalMeans with aCenter different for letter Graduate in a row are Study statistically and different Research (Duncan, in PAgriculture ≤0.01). Rooting of in vitro propagated Pyrus plantlets (AGRIS) since 1985. Multiple shoots of high quality were produced on M5 then placed on M4 (without plant Gyawali,Table 3. EffectR., & of Ibrahim, medium S.composition A. (2014). on Natural tissue browningproducts andas antimicrobialPyrus explant agents.establishment Food duringControl, culture 46, growth regulator) for one week to improve shoot elongation and leaf size prior to rooting. As initiation412 -429. https://doi.org/10.1016/j.foodcont.2014.05.047 Tissue browning (%) Healthy explant establishment (%) shown in Table 5, both medium composition and variety significantly affected these Hanušová,Species /cultivar K., Dobiáš, J., & Klaudisová, K. (2009). Effect of packaging films releasing antimicrobial parameters. Overall, Cheng medium with IBA (M7) resulted in the best rooting response for agents on stability of food products.M1 CzechM2 Jounal ofM3 Food ScienceM1 , 27, 347M2- 349. M3 all tested genotypes, whereas, MS medium with NAA (M10) gave the worst results (Table 5). https://doi.org/10.17221/958‘Arbi’ -CJFS0±0 c 5±0.22 b 20±0.41a 100±0 a 90±0.30 b 85±0.36 c ICAR, (2009). Indian‘Malti’ Council of Agricultural0±0 c 5 Research.±0.22 b Annual25±0.44 aReport. 100±0 a 90±0.30 b 80±0.41 c Additionally, the presence of IBA led to the highest root number and length in all cultivars Pyrus communis c b a a b c Janjarasskul, T., & Krochta,‘Mahdia 6’ M. (2010).0±0 Edible10±0.30 packaging 15±0.36 materials. 100±0 Food Science90±0.30 and 85Technology±0.36 - (M7, M9) (Table 5). Although NAA also stimulated rooting to some extent (M8, M10) (Table c b a a b c Annual Reviews,‘Moknine 1, 415- 10’448. https://doi.org/10.1146/annurev.food.080708.1008360±0 15±0.36 25±0.44 100±0 70±0.47 60±0.50 5), the roots were short and fleshy and developed from excessive brown, spongy and friable c b a a b c Kamble,Pyrus syriaca P. B., & Chavan, J. K. (2005).0±0 Effects10±0.30 of post -harvest15±0.36 treatments 100±0 and90 storage±0.30 temperature85±0.36 on callus at the stem base. Medium effect ** ** ** ** ** ** shelf-life of custard apple fruits. Jounal Food Science Technology, 42 (3), 253-255. Kim,interaction S. A., ‘Cultivars & Rhee, × Medium’ M. S. (2016).** Highly enhanced** bactericidal** effects** of medium** chain** fatty acids The values(caprylic, are compared capric, horizontally. and lauric Means acid) with combined a different with letter edible in a row plant are statistically essential differentoils (carvacrol, (Duncan, Peugenol, ≤0.01). β-

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Micropropagation of Pyrus communis Botanical products for post-harvest management of perishable produce

Tableresorcylic 4. Effect of acid, culture trans media-cinnamaldehyde, on multiplication thymol, rate and and shoot vanillin) length againstin different Escherichia Pyrus sp. coliat the O157: end of H7. the fourthFood subculture. Control , 60, 447-454. https://doi.org/10.1016/j.foodcont.2015.08.022. Species/cultivar Medium Multiplication rate Shoot length (mm) Kupwade, S. R., Manglani, N. S., Wadkar, S. S., Inamdar, F. R., Shete,c C. C., & Ghosh, J. S.c (2017) Antimicrobial activity of Lemon peelM 4 extract against1±0 the bacteria belonging7.46±0.33 to the b a Genus Xanthomonas. InternationalʻArbiʼ JournalM 5of Pharmaceutic6.6±0al.59 Sciences Review23. 05±0and. 42Research , a b 45(2), 251-254. https://doi.org/10.22159/ijcpr.2017v9i4.20962M6 11.7±0 .97 18.76±0.52 c c Lucera, A., Costa, C., Conte, A., & Del Nobile,M4 M. A. (2012).1±0 Food applications7.09±0. 36of natural b a ʻMaltiʼ M5 5.85±0.36 21.68±0.47 antimicrobial compounds. Frontiers in Microbiology, 3, 287. a b Pyrus https://doi.org/ communis 10.3389/fmicb.2012.00287. MeCollection6 20129. 05±0.82 18.74±0.59 c c Malasri, S., Tiapradit, P., Russell, S., Poonpurmsiri,M4 P., Tarkarnviroj,1±0 C., Pourhashemi,7.30±0 A.,.48 Moats, R., b a & Hudson, B. (2015) ʻInsulationMahdia 6ʼ effectivenessM5 of rice hull6.35±0. International.48 Journal22.34±0 of.69 Advance a b Packaging Technology, 3(1), 158-168. https://doi.org/M6 10.23953/cloud.ijapt.209.65±0.58 15.48±0.81 c c Mari, M., Leoni, O., Lori, R., & Cembali, MT.4 (2002) Antifungal1±0 vapour-phase9 .activity57±0.51 of allyl isothyocyanates against Penicillium expansum on pears. Plant Pathology b , 51, 231-236. a ʻMoknine 10ʼ M5 6.45±0.51 20.04±0.35 https://doi.org/10.1046/j.1365-3059.2002.00667.x a b M6 9.35±0.48 15.11±0.63 McClements, D. J., & Rao, J. (2011). Food-grade nanoemulsions: formulation,c fabrication, properties,c M4 1±0 10.08±0.26 performance, biological fate, and potential toxicity. Critical Reviewb Food Science and Nutrition,a Pyrus syriaca M5 7.25±0.44 22.67±0.33 51(4), 285-330. https://doi.org/10.1080/10408398.2011.559558 10.4±0.50 a 12.46±0.83 b Mishra, B., & Khatkar, B. S. (2009). Studies Mon6 post-harvest quality and physiology of coated ber

Media( Zizyphuseffect mauritiana Lamk) fruit. Journal of Food Science** and Technology **(Mysore), 46(5), Cultivar455 effect-458 . ** ** Mohanty,Interaction ‘CultivarsS., Ramesh, × Media’ S., & Muralidharan, N. P . (2017). Antimicrobial** efficacy of apple** cider vinegar The valuesagainst are comparedEnterococcus vertically. faecalis Means withand aCandida different letter albicans in a row: Anare statisticallyin vitro differentstudy. Journal(Duncan. Pof ≤0.01). Advanced Pharmacy Education and Research, 7(2), 137-141. EffectMustafa, of E. hormone , Noah, A.concentrations , Beshay, K. , onSultan, shoot L. proliferation , Essam, M. & Nouh, O. (2015). Investigating the AfterEffect three of weeks, Various the Nanomaterials healthy shoots on the were Wettability excised of from Sandstone the initiationReservoir. media World (M1)Journal and of Engineering and Technology, 3, 116-126. https://doi.org/10.4236/wjet.2015.33013. transferred on to multiplication medium. Shoot proliferation was assessed on three different Neri, F., Mari, M., Meniti, A. M., & Brigati, S. (2006). Activity of trans-2-hexenal MS mediaagainst with Penicillium half the expansum concentration in ‘Conference’ of NH4NO 3pears. and KNOJournal3 and of differentApplied Microbiologyconcentrations 100 of, BA and1186 IBA-1193 (.Table https://doi.or 4). A significantg/10.1111/j.1365 impact-2672.2006.02873.x of medium composition and pear variety was Nguyen,noted. Without V. T. A., hormones Le, T. D., (M4),Phan, H.none N., &of Tran,the genotypesL. B. (2017). multiplied Antibacterial and activity the shoot of free length fatty remainedacids fromconstant hydrolyzed after the virgin first coconut subculture oil using (Table lipase 4). fromWhen Candida 1 mg L rugosa-1 BA .and Journal 0.1 mg of Lipids L-1 IBA, 1- were7. supplemented https://doi.org/ 10.1155/2017/7170to the medium (M5), the number of shoots per subculture increased considerablyPrasad, K., & andStadelbacher, the shoots G. were J. (1974). longer Effect for all of cultivars acetaldehyde (Table vapor 4, Fig. on postharvest1- b). When decay the BAand concentrationmarket quality was ofincreased fresh strawberries. to 2 mg Phytopathology,L-1 (M6), the multiplication64, 948-951. https://doi.org/10.1094/phyto rate increased with each- subculture64-948 for all tested cultivars. However, under these conditions, the shoot length decreasedRaorane, G and. P. (leaves2003). turnedStudies narrowon growth, (Table flowering, 4), which fruiting made and more some difficult aspect ofthe harvest further handling handling of of thekokum shoots ( multiplicationGarcinia indica and choisy). rooting. M.Sc. Thesis. Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli, District. Ratnagiri. RootingQuirós-Sauceda, of in vitro A. E.,propagat Ayala-Zavala,ed Pyrus J.F., plantlets Olivas, G. I., & González-Aguilar, G.A. (2014). Edible Multiplecoatings shoots as encapsulatingof high quality matrices were forproduced bioactive on compounds: M5 then aplaced review. onJournal M4 of(without Food Science plant and Technology, 51(9), 1674-1685. https://doi.org/10.1007/s13197-013-1246-x growth regulator) for one week to improve shoot elongation and leaf size prior to rooting. As Ranjan, S., Dasgupta, N., Chakraborty, A. R., Samuel, S. M., Ramalingam, C., Shanker, R., & Kumar, shownA. (2014).in Table Nanoscience 5, both mediumand nanotechnologies composition in and food variety industries: significantly opportunities affected and research these parameters.trends. Journal Overall, of ChengNanoparticle medium Research with IBA, 16(6), (M7) 2464. resulted https://doi.org/10.1007/s11051 in the best rooting response-014-2464 for- all tested5. genotypes, whereas, MS medium with NAA (M10) gave the worst results (Table 5). Saxena,Additionally, R., Liquido, the presence N. J., & ofJusto, IBA H. led B. (1to981). the hiNaturalghest pesticidesroot number from and the lengthneem tree in all(Azadirachta cultivars (M7,indica M9) ).(Table Proceeding 5). Although of International NAA also Neem stimulated Conference rooting Rattach to Engem,some extent Germany (M8,. M10) (Table 5Sembdner,), the roots G., were & Parthier, short andB. (1993). fleshy The and biochemistry developed fromand the excessive physiological brown, and spongymolecular and actions friable of callusjasmonates. at the stem Annual base. Review of Plant Physiology and Plant Molecular Biology, 44, 569-589. https://doi.org/10.1146/annurev.pp.44.060193.003033 Shilling, M., Matt, L., & Rubin, E. (2013). Antimicrobial effects of virgin coconut oil and its medium- chain fatty acids on Clostridium difficile. Journal of Medicinal Food, 16(12), 1079-1085. https://doi.org/ 10.1089/jmf.2012.0303. JOURNAL OF HORTICULTURE AND POSTHARVEST RESEARCH VOL. 3(1) MARCH 2020

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Chitranshi et al. Micropropagation of Pyrus communis Lotfi et al.

Effect of substrates on acclimatization and hardening Castro, C. D. P. C., Faria, J. & Dantas, T. B. H. (2014). Evaluating the performance of coconut fiber In vitro Well-rooted pear plantlets (Fig. 1- c) were transferred to a substrate with peat and and wood straw as cushioning materials to reduce injuries of papaya and mango during perlite (S1) or peat moss (S2) for acclimatization. During the first four weeks, there were 100% transportation. International Journal of Advanced Packaging Technology, 2(1), 84-95. survivals for all accessions on both substrates. Later during the acclimatization process, a https://doi.org/10.23953/cloud.ijapt.10 fungal infestation strongly affected the plants, except for P. communis ‘Arbi’ of which 5% Chandra, D., & Kumar, R. (2012). Qualitative effect of wrapping and cushioning materials on guava fruits during storage. HortFlora Research Spectrum, 1(4), 318-322. survived on S1 and 18% on S2 substrate (Fig. 1- d). On S2 substrate the surviving plants Chauhan, S. K., & Babu, D. R. (2011). Use of botanicals: A new prospective for enhancing fruit achieved a better growth, plant height and branching number, whereas on S1 substrate the quality over chemicals in an era of global climate change. Asian Journal of Environmental highest leaf number was recorded; on S1: plant height: 7.5 cm; branching number: 1.3; leaf Science, 6(1), 17-28. number: 9; on S2: plant height: 5.5 cm; branching number: 1; leaf number: 14; recorded after Creelman, R. A., & Mullet, J. E. (1995). Jasmonic acid distribution in plants: regulation during 10 months. The acclimatized plants were transferred to an insect-proof greenhouse and development and responses to biotic and abiotic stress. Proceedings of the National Academy of subsequently showed good growth and formed thick trunks and new leaves (Fig. 1- e) without Sciences USA, 92, 4114-4119. https://doi.org/10.1073/pnas.92.10.4114 any variation in morphological characteristics. Davidson, P. M., & Taylor, M. T. (2007). Chemical preservatives and natural antimicrobial compounds. Food Microbiology: Fundamentals and Frontiers, Washington, DC: American Society for Microbiology Press, 713-734. https://doi.org/10.1128/9781555815912 DISCUSSION DebMandal, M., & Mandal, S. (2011). Coconut (Cocos nucifera L.: Arecaceae): in health promotion and disease prevention. Asian Pacific Journal of Tropical Medicine, 4(3), 241-247. https://doi.org/ In this study, we established an efficient protocol for tissue culture initiation and propagation 10.1016/S1995-7645(11)60078-3. for local P. communis cultivars and P. syriaca accession. Yeo and Reed (1995) reported that Donsì, F., Annunziata, M., Vincensi, M., & Ferrari, G. (2012). Design of nanoemulsion-based delivery explants from field-grown pear trees are usually difficult to disinfect and better results were systems of natural antimicrobials: effect of the emulsifier. Journal of Biotechnology , 159(4), 342- obtained with explants taken from actively growing plants. Nevertheless, 20-minute 350. https://doi.org/10.1016/j.jbiotec.2011.07.001. incubation with 4% HgCl2 was highly efficient to disinfect the apical explants from old pear Fig.Droby, 1. Overview S., Porat, of the R., complete Cohen, procedure L.,Weiss, for micropropagation B., Shapiro, ofB., pear. Philosoph (a) In vitro-Hadas, establishment S., & ofMeir, explants. S. (b) The(1999). proliferation Suppressing of pear green shoots moldon medium decay containing in grapefruit MS with with half postharvestthe concentration jasmonates of NH4NO application.3 and KNO3 trees. The effectiveness of HgCl2 as a disinfectant is in agreement with previous studies on and 1 mg L-1 BA + 0.1 mg L-1 IBA. (c) In vitro rooting, on Cheng medium with 1 mg L-1 IBA. (d) First-stage other pear cultivars (Bahri-Sahloul et al., 2005), other (Assareh & Sardabi, 2005) and Journal of the American Society for Horticultural Science , 124(2), 184-188. acclimatizationhttps://doi.org/10.21273/JASHS.124.2.184 of pear plantlets after two months. (e) Hardening of acclimatized pear vitroplants under insect- others species (Qin et al., 2017). Further, in accordance with Mihaljevic et al. (2013), NaOCl2 proofDubey, greenhouse N., Mishra, conditions V., & (picture Thakur, taken D. after (2018). 10 months). Plant based antimicrobial formulations. Postharvest proved to be inadequate as a disinfectant in our experiments. In contrast, Shibli et al. (1997) Disinfection of Fruits and Vegetables, 1, Elsevier Academic Press, 322-331. obtained 95% survival with NaOCl2 for P. syriaca, but the explants were harvested from a Tablehttps://doi.org/10.1016/B978 5. Effect of medium composition-0 -on12 -rooting812698 rate-1.00011 number- Xand length in different Pyrus sp. phytotron which might explain the discrepancy with our results. Fallik,Species/cultivar E., Arhbold, D. D., Hamil tonMedium-Kemp, RootingT. R., Clements,rate (%) A.Root M. number, Collins, R. W.,Root &length Barth, (mm) M. E. a a a Optimal culture initiation was accomplished on M1 medium consisting of full-strength (1998). (E)-2-Hexenal can stimulateM7 Botrytis 100±0 cinerea growth 13.71±0.48 in vitro and on29.72±0.94 strawberry fruit in b c MS medium supplemented with 0.25 mg L-1 BA for all tested cultivars, although growth vivo during storage. Journal ofM 8 the American 78.57±0.41 Society for Horticultural8.71±1.11 c Science5.76±0.97, 123, 875-888. ʻArbiʼ a b b medium composition and genotype influenced explant establishment. These findings agree https://doi.org/10.21273/JASHS.123.5.875M9 100±0 10.71±0.48 21.78±0.55 c c d Fenwick, G. R., Heaney, R. K., & Mullin,M10 W. 50±0.50J. (1983). Glucosinolates9.28±1.79 and their break4.06±0.84-down products well with those of Leite et al. (1997) and Karimpour et al. (2013) who studied other European a a a in food and food plants. CriticalM7 Reviews60.71±0.49 in Food Science6.57±0.53 and Nutrition12.73±0.26 18, 123-201. and Iranian pear cultivars. The inclusion of phloroglucinol in all our media likely prevented c c https://doi.org/10.1080/10408398209527361M8 14.25±0.35 b 3.42±0.78 5.16±0.46 excessive phenolic browning, as reported for the in vitro establishment of apple rootstock ʻMaltiʼ a b b Food and Agriculture Organization Mof9 the United46.42±0.50 Nations (FAO),4.24±0.48 (2011). Global food9.98±0.53 losses and food (Sharma et al., 2007). Additionally, as observed by Mamaghani et al. (2010) for roses, b c d M10 10.71±0.31 2.85±0.89 4.28±0.39 Pyruswaste. communis United Nations Food and Agriculture Organization. Rome, Italy. http://faostat.fao.org. lowering the total nitrogen content by halving the NH4NO3 and KNO3 concentration led to a a Garcia, J. L. (1982). Storage of tomatM7 oes (Lycopersicon42.85±0.50 esculentum9.42±0.53 Mill) a in rice hull18.73±0.92 ash of different increased tissue necrosis. ab c c particle sizes and moisture contentM8 [Philippines].21.42±0.41 Thesis. In5±0.81 Agricultural Information5.37±1.02 Bank for ʻMahdia 6ʼ ab b b Concerning propagation, in the absence of cytokinins, no multiplication occurred. Asia, South-East Asian RegionalM9 Center25±0.44 for Graduate Study6.85±0.69 and Research15.27±0.42 in Agriculture b d c Cytokinins are reported to be essential for all pear species proliferation (Karimpour et al., (AGRIS) since 1985. M10 7.14±0.26 3.57±0.78 4.81±0.34 a a 2013; Lotfi et al., 2019). On medium with plant growth regulators, the multiplication rate for Gyawali, R., & Ibrahim, S. A. (2014).M7 Natural39.28±0.49 products as antimicrobial6.85±0.69 a agents. 9.77±0.88Food Control, 46, ab c c the selected pear cultivars increased with each subculture and was the highest on M6 medium 412-429. https://doi.org/10.1016/j.foodcont.2014.05.047M8 21.42±0.41 3.42±0.53 5.91±0.57 ‘Moknine 10ʼ b b b containing MS medium with half the concentration of NH4NO3 and KNO3 supplemented with Hanušová, K., Dobiáš, J., & Klaudisová,M9 K. (2009).10.71±0.31 Effect of packaging4.57±0.78 films releasing7.11±0.47 antimicrobial -1 -1 b d d 2 mg L BA and 0.1 mg L IBA. However, shoot elongation was best on M5 medium, which agents on stability of food products.M10 Czech3.57±0.18 Jounal of Food Science2.71±0.48, 27, 347-3494.36±0.59. a a only has 1 mg L-1 BA. In Bacopa monnieri, the optimal biomass yield with particular https://doi.org/10.17221/958-CJFSM7 50±0.50 7.71±0.75 a 26.82±0.87 b c ICAR, (2009). Indian Council of AgriculturalM8 17.85±0.39Research. Annual Report.5±0.57 b 13.25±0.41 combinations of BA and IBA has been attributed to the synergistic effect of auxin and Pyrus syriaca ab b Janjarasskul, T., & Krochta, M. (2010).M9 Edible28.71±0.46 packaging materials.5.42±0.53 Food Science20.32±0.75 and Technology b - cytokinin on the growth of tissues, cell expansion and cell division (Sakharam et al., 2017). b c d Annual Reviews, 1, 415- 448. https://doi.org/10.1146/annurev.food.080708.100836M10 21.42±0.41 2.57±0.53 6.73±0.54 Overall, our observations are in agreement with those reported for in vitro propagation of Kamble, P. B., & Chavan, J. K. (2005). Effects of post-harvest treatments and storage temperature on other P. communis varieties and pear species (Dimitrova et al., 2016; Lizarraga et al., 2017; Mediashelf effect-life of custard apple fruits. Jounal Food** Science Technology, ** 42 (3) , 253**-255 . Hassan & Zayed, 2018). Kim,Interaction S. A., ‘Cultivars & Rhee, × Media’ M. S. (2016). Highly enhanced** bactericidal ** effects of medium** chain fatty acids The values(caprylic, are compared capric, vertically. and lauric Means acid) with combined a different letterwith in edible a row areplant statistically essential different oils (carvacrol, (Duncan, P≤0.05). eugenol, β-

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Micropropagation of Pyrus communis Botanical products for post-harvest management of perishable produce

Effectresorcylic of substrates acid, trans on -acclimatizationcinnamaldehyde, thymol,and hardening and vanillin) against Escherichia coli O157: H7. In vitroFood Well Control-rooted, 60, pear447- 454.plantlets https://doi.org/ (Fig. 1- 10.1016/j.foodcc) were transferredont.2015.08.022. to a substrate with peat and Kupwade,perlite (S1) S. orR., peat Manglani, moss (S2)N. S., for Wadkar, acclimatization. S. S., Inamdar, During F. R.,the Shete, first fourC. C., weeks, & Ghosh, there J. wereS. (2017) 100% survivalsAntimicrobial for all accessionsactivity ofon Lemonboth substrates. peel extract Later against during thethe bacteriaacclimatization belonging process, to the a fungalGenus infestation Xanthomonas. strongly International affected the Journal plants, of exceptPharmaceutic for P.al communis Sciences Review‘Arbi’ andof which Research 5%, survived45(2) on, 251 S1-254 and. https://doi.org/10.22159/ijcpr.2017v9i4.20962 18% on S2 substrate (Fig. 1- d). On S2 substrate the surviving plants Lucera,achieved A., a betterCosta, growth,C., Conte, plant A., height & Del and Nobile, branching M. A. number (2012)., whereasFood applications on S1 substrate of natural the highestantimicrobial leaf number compounds. was recorded Frontiers; on in S1: Microbiology, plant height: 3, 2877.5. cm; branching number: 1.3; leaf https://doi.org/10.3389/fmicb.2012.00287. eCollection 2012 Malasri,number: S., 9; Tiapradit,on S2: plant P., Russell,height: S.,5.5 Poonpurmsiri, cm; branching P., number:Tarkarnviroj, 1; leaf C., number: Pourhashemi, 14; recorded A., Moats, after R., 10 months.& Hudson, The B. acclimatized (2015) Insulation plants effectiveness were transferred of rice hullto .an International insect-proof Journal greenhouse of Advance and subsequentlyPackaging showed Technology, good 3 growth(1), 158 -and168. formed https://doi.org/ thick trunks10.23953/cloud.ijapt.20 and new leaves ( Fig. 1- e) without Mari,any variation M., Leoni, in morphological O., Lori, R., &characteristics. Cembali, T. (2002) Antifungal vapour-phase activity of allyl isothyocyanates against Penicillium expansum on pears. Plant Pathology, 51, 231-236. https://doi.org/10.1046/j.1365-3059.2002.00667.x DISCUSSION McClements, D. J., & Rao, J. (2011). Food-grade nanoemulsions: formulation, fabrication, properties, performance, biological fate, and potential toxicity. Critical Review Food Science and Nutrition, In this51(4) study,, 285 we-330 established. https://doi.org/10.1080/10408398.2011.559558 an efficient protocol for tissue culture initiation and propagation Mishra,for local B., P. & communis Khatkar, B.cultivars S. (2009). and StudiesP. syriaca on postaccession.-harvest Yeoquality and and Reed physiology (1995) reportedof coated thatber explants(Zizyphus from mauritianafield-grown Lamk) pear fruit.trees Journalare usually of Food difficult Science to disi andnfect Technology and better (Mysore), results 46(5)were, obtained455- 458with. explants taken from actively growing plants. Nevertheless, 20-minute Mohanty,incubation S., with Ramesh, 4% HgClS., & 2Muralidharan, was highly efficientN. P. (2017). to disinfect Antimicrobial the apical efficacy explants of apple from cider old vinegar pear trees.against The effectiveness Enterococcus offaecalis HgCl and2 as Candida a disinfectan albicanst is: inAn agreement in vitro study with. Journalprevious of studies Advanced on otherPharmacy pear cultivars Education (Bahri and-Sahloul Research et, al7(2)., 2005, 137-)141, other. Rosales (Assareh & Sardabi, 2005) and Mothersustafa, species E. , Noah, (Qin A.et al.,, Beshay, 2017). K. Further, , Sultan, in L.accordance , Essam, M. with & MihaljevicNouh, O. (2015). et al. (Investigating2013), NaOCl the2 provedEffect to beof inadequateVarious Nanomaterials as a disinfectant on the inWettability our experiments. of Sandstone In contrast, Reservoir. Shibli World et al.Journal (1997 of) Engineering and Technology, 3, 116-126. https://doi.org/10.4236/wjet.2015.33013. obtained 95% survival with NaOCl2 for P. syriaca, but the explants were harvested from a Neri, F., Mari, M., Meniti, A. M., & Brigati, S. (2006). Activity of trans-2-hexenal phytotronagainst which Penicillium might explainexpansum the in discrepancy ‘Conference’ with pears. our results.Journal of Applied Microbiology 100, Optima1186-1193l culture. https://doi.or initiationg/10.1111/j.1365 was accomplished-2672.2006.02873.x on M1 medium consisting of full-strength -1 Nguyen,MS medium V. T. supplementedA., Le, T. D., Phan,with H.0.25 N., mg & Tran,L BA L. B.for (2017). all tested Antibacterial cultivars, activity although of free growth fatty mediumacids composition from hydrolyzed and virgin genotype coconut influenced oil using lipaseexplant from establishment. Candida rugosa These. Journal findings of Lipids agree, 1- well 7.with https://doi.org/ those of Leite10.1155/2017/7170 et al. (1997) and Karimpour et al. (2013) who studied other European Prasad,and Iranian K., &pear Stadelbacher, cultivars. TheG. J. inclusion (1974). Effect of phloroglucinol of acetaldehyde in allvapor our on media postharvest likely preventeddecay and excessivemarket phenolic quality of browning, fresh strawberries. as reported Phytopathology, for the in 64vitro, 948 establishment-951. https://doi.org/10.1094/phyto of apple rootstock- (Sharma64-948 et al., 2007). Additionally, as observed by Mamaghani et al. (2010) for roses, Raoranelowering, Gthe. P . total(2003 nitrogen). Studies content on growth, by flowering,halving the fruiting NH4 NOand3 someand KNOaspect3 ofconcentration harvest handling led toof increasedkokum tissue (Garcinia necrosis. indica choisy). M.Sc. Thesis. Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli, District. Ratnagiri. Concerning propagation, in the absence of cytokinins, no multiplication occurred. Quirós-Sauceda, A. E., Ayala-Zavala, J.F., Olivas, G. I., & González-Aguilar, G.A. (2014). Edible Cytokininscoatings are as encapsulatingreported to be matrices essential for bioactivefor all pear compounds: species aproliferation review. Journal (Karimpour of Food Science et al., 2013and; Lotfi Technology, et al., 2019 51(9),). On 1674 medium-1685. https://doi.org/10.1007/s13197 with plant growth regulators,-013 the-1246 multiplication-x rate for Ranjan,the selected S., Dasgupta, pear cultivars N., Chakraborty, increased A.with R., each Samuel, subculture S. M., Ramalingam, and was the C., highest Shanker, on R.,M6 & medium Kumar, containingA. (2014). MS mediumNanoscience with and half nanotechnologies the concentration in of food NH 4industries:NO3 and KNOopportunities3 supplemented and research with 2 mgtrends. L-1 BA Journal and 0.1 of mgNanoparticle L-1 IBA. ResearchHowever,, 16 shoot(6), 2464. elongation https://doi.org/10.1007/s11051 was best on M5 medium-014, -w2464hich- only 5.has 1 mg L-1 BA. In Bacopa monnieri, the optimal biomass yield with particular Saxena,combinations R., Liquido, of BA N. J.,and & IBAJusto, has H. B.been (1981). attributed Natural topesticides the synergistic from the neemeffect tree of ( Azadirachtaauxin and cytokininindica on). Proceeding the growth of ofInternational tissues, cell Neem expansion Conference and Rattach cell division Engem, ( GermanySakharam. et al., 2017). Sembdner,Overall, our G., observations& Parthier, B. are(1993). in agreementThe biochemistry with thoseand the reported physiological for in and vitro molecular propagation actions of otherjasmonates. P. communis Annual varieties Review and of pear Plant species Physiology (Dimitrova and Plant et al., Molecular 2016; Lizarraga Biology, et44 ,al., 569 2017-589;. https://doi.org/10.1146/annurev.pp.44.060193.003033 Hassan & Zayed, 2018). Shilling, M., Matt, L., & Rubin, E. (2013). Antimicrobial effects of virgin coconut oil and its medium- chain fatty acids on Clostridium difficile. Journal of Medicinal Food, 16(12), 1079-1085. https://doi.org/ 10.1089/jmf.2012.0303. JOURNAL OF HORTICULTURE AND POSTHARVEST RESEARCH VOL. 3(1) MARCH 2020

JOURNAL OF HORTICULTURE AND POSTHARVEST RESEARCH VOL. 3(1) MARCH 2020 7

Chitranshi et al. Micropropagation of Pyrus communis Lotfi et al.

Bahmani, R., Karami, O., & Gholami, M. (2009). Influence of carbon sources and their concentrations Castro,Optimal C. D. P.rooting C., Faria, of J.the & plantsDantas, micropropagated T. B. H. (2014). Evaluating on M5 medium the performance was achieved of coconut on fibM7er on rooting and hyperhydricity of apple rootstock MM106. World Applied Sciences Journal, 6, mediumand woodconsisting straw ofas Cheng cushioning medium materials with 1 tomg reduce L-1 IBA. injuries Compared of papaya to MS and medium, mango Chengduring 1513-1517. mediumtransportation. has a Internationallower concentration Journal of Advancedof ammonium Packaging and Technology, nitrate ions. 2(1), 84A -95.reduced salt Bahri-Sahloul, R., Mtar, R., Msallem, A., & Ammar, S. (2005). Micropropagation of three "Pyrus" concentration https://doi.org/10.23953/cloud.ijapt.10 has been reported to improve adventitious rooting in diverse plants for rootstocks. Advances in Horticultural Science, 19, 1000-1008. commercialChandra, D., exploitation& Kumar, R., (2012).increasing Qua litativethe root effect number of wrapping and the androot cushioning length (Moncousin, materials on 2012 guava). Brini, W., Mars, M., & Hormaza, I. (2008). Genetic diversity in local Tunisian pears (Pyrus communis fruits during storage. HortFlora Research Spectrum, 1(4), 318-322. L.) studied with SSR markers. Scientia Horticulturae, 115, 337-341. TheChauhan, superior S. K., rooting & Babu, response D. R. of(2011). the selected Use of pearbotanicals: varieties A newwith prospectiveIBA as compared for enhancing to NAA fruit is https://doi.org/10.1016/j.scienta.2007.10.012 in concordancequality over withchemicals the findings in an era of Shibliof global et al.climate (1997 change.), Reed Asian(1995 )Journal and Thakur of Environmental & Kanwar, Cheng, T.V. (1979). Micropropagation of clonal fruit tree rootstocks. Compact Fruit Trees, 12, 127- (2008Science,) for other6(1), 17pears.-28. In contrast, Al-Maarri et al. (1994) obtained the best in vitro rooting -1 137. Creelman,for the pear R. cultivars A., & Mullet, ‘Passe J.Crassane’ E. (1995). and Jasmonic ‘Williams’ acid with distribution NAA at in 0.2 plants: mg L regul, indicatingation during the Dimitrova, N., Nacheva, L., & Berova, M. (2016). Effect of meta-topolin on the shoot multiplication occurrencedevelopment of a andgenotype responses-dependent to biotic response and abiotic. Unfortunately, stress. Proceedings concerning of the Nationalthe acclimatization, Academy of of pear rootstock OHF-333 (Pyrus communis L.). Hortorum Cultus-Acta Scientiarum our Sciencesresults are USA, inconclusive 92, 4114-4119. due https://doi.org/10.1073/pnas.92.10.4114to fungal rots that eradicated most of our plants. Polonorum, 15, 43-53. https://doi.org/10.17660/actahortic.2009.839.23 Davidson, P. M., & Taylor, M. T. (2007). Chemical preservatives and natural antimicrobial Gaaliche, B., Chehimi, S., Dardouri, S., & Hajlaoui, M.R. (2018). Health status of the pear tree compounds. Food Microbiology: FundamentalsCONCLUSIONS and Frontiers, Washington, DC: American following the establishment of Fire blight in Northern Tunisia. International Journal of Fruit Society for Microbiology Press, 713-734. https://doi.org/10.1128/9781555815912 Science, 18, 85-98. https://doi.org/10.1080/15538362.2017.1377670 AnDebMandal efficient, M., tissue & Mandal culture, S. protocol (2011). Coconut was established (Cocos nucifera for the L.: P.Arecaceae): communis in cultivars health promotion ‘Arbi’, Hassan, S.A.M., & Zayed, S.N. (2018). Factor controlling micropropagation of fruit trees: A review. ʻMaltiʼ,and disease ʻMahdia prevention. 6ʼ, and Asian ʻMoknine Pacific 10ʼ Journal and forof Tropical P. syriaca Medicine,. An overview 4(3), 241 -247of the. https://doi.org/ protocol is Science International, 6, 1-10. https://doi.org/10.17311/sciintl.2018.1.10 10.1016/S1995-7645(11)60078-3. given in Figure 1. The recommended procedure consists of 20 minutes explant disinfection Karimpour, S., Davarynejad, G.H., Bagheri, A., & Tehranifar, A. (2013). In vitro establishment and Donsì, F., Annunziata, M., Vincensi, M., & Ferrari, G. (2012). Design of nanoemulsion-based-1 delivery clonal propagation of Sebri pear cultivar. Journal of Agricultural Sciences and Technology, 15, withsystems 4% HgCl of natural2, followed antimicrobials: by transfer effect to MSof the medium emulsifier. supplemented Journal of Bwithiotechnology 0.25 mg, 159L (4), BA 342 for- 1209-1217. initiation350. https://doi.org/ and to MS10.1016/j.jbiotec.2011.07.001. basal medium with half the concentration of NH4NO3 and KNO3 -1 -1 Leite, G.B., Finardi, N.L., & Fortes, G.R.L. (1997). The effect of BAP and NAA concentration on supplementedDroby, S., Porat, with R.,2 mg Cohen, L BA L.,Weiss, and 0.1 mgB., LShapiro, IBA for B., optimal Philosoph shoot-Hadas, multiplication; S., & Meir, rooting S. multiplication “in vitro” of pears cv. Bartlett and clone OH × F 97. Ciencia-e-Agrotecnologia, 21, of the(1999). propagated Suppressing shoots green should mold be decay executed in grapefruit on Cheng with medium postharvest with 1 jasmonatesmg L-1 IBA. application. Pending 436-441. the Journaloptimization of ofthe the acclimatizationAmerican Society and hardeningfor Horticultural process and theScience molecular , 124(2), validation 184-188. of Lizarraga, A., Fraga, M., Ascasibar, J., & Gonzalez, M.L. (2017). In Vitro propagation and recovery of the https://doi.org/10.21273/JASHS.124.2.184genetic stability of the regenerated plants , this protocol can be applied for a large-scale eight apple and two pear cultivars held in a germplasm bank. American Journal of Plant Sciences, Dubey,production N., Mishra,of good V.,quality & Thakur, and healthy D. (2018). plants Plant to be based used antimicrobialfor the establishment formulations. of a Postharvestsuccessful 8, 2238-2254. https://doi.org/10.4236/ajps.2017.89150 commercialDisinfection production of Fruitsof local andTunisian Vegetables, pear cultivars. 1, Elsevier Academic Press, 322-331. Lotfi, M., Mars, M., & Stefaan, W. (2019). Optimizing pear micropropagation and rooting with light https://doi.org/10.1016/B978-0-12-812698-1.00011-X emitting diodes and trans-cinnamic acid. Plant Growth Regulation. 1-8. Fallik, E., Arhbold, D. D., Hamil ton-Kemp, T. R., Clements, A. M. , Collins, R. W., & Barth, M. E. ACKNOWLEDGMENTS https://doi.org/10.1007/s10725-019-00498-y (1998). (E)-2-Hexenal can stimulate Botrytis cinerea growth in vitro and on strawberry fruit in Mamaghani, A.B., Ghorbanli, M., Assareh, M.H., & Ghamari, Z.A. (2010). In vitro propagation of vivo during storage. Journal of the American Society for Horticultural Science, 123, 875-888. three Damask Roses accessions. Iranian Journal of Plant Physiology, 1, 85-94. Authorshttps://doi.org/10.21273/JASHS.123.5.875 thank Danny Vereecke for critical reviewing of the manuscript. Mars, M., Carraut, A., Marrakchi, M., Gouiaa, M., & Gaaliche F. (1994). Ressources génétiques Fenwick, G. R., Heaney, R. K., & Mullin, W. J. (1983). Glucosinolates and their break-down products fruitières en Tunisie (poirier, oranger, figuier, grenadier). Plant Genetic Ressources Newsletter, in food and food plants. CriticalCONFLICT Reviews OF in INTEREST Food Science and Nutrition 18, 123-201. 100, 14-17. https://doi.org/10.1080/10408398209527361 Mihaljevic, I., Dugalic, K., Tomas,V., Viljevac, M., Pranjic, A., Cmelik, Z., Puskar,B., & Jurkovic, Z. FoodThe authors and Agriculture have no Organizaconflict tionof interest of the United to report. Nations (FAO), (2011). Global food losses and food (2013). 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Micropropagation of Pyrus communis Botanical products for post-harvest management of perishable produce

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Original Article Droby,https://doi.org/10.15835/nbha36278 S., Porat, R., Cohen, L.,Weiss, B., Shapiro, B., Philosoph-Hadas, S., & Meir, S. Xia,(1999). L., Xuejiao, Suppressing L., Wang, green T., mold& Gao, decay W. (2016).in grapefruit Nutritional with compositionpostharvest ofjasmonates pear cultivars application. (Pyrus Article history: Journalspp.). In Nutritionalof the CompositionAmerican Societyof Fruit Cultivarsfor Horticultural (Preedy V.R. Science & Simmonds , 124(2), M.S.J184. -ed.).188. Received 19 July 2019 Purpose: The aim of this study was to investigate the effects of https://doi.org/10.21273/JASHS.124.2.184Elsevier, USA, 573-608. https://doi.org/10.1016/b978 -0-12-408117-8.00024-6 Revised 29 August 2019 nano-chitosan and chitosan coating on physico-chemical properties Yeo,Dubey, D.Y., N., &Mishra, Reed, B.M.V., & (1995). Thakur, Micropropagati D. (2018). Planton of based three antimicrobialPyrus rootstocks. formulations. HortScience, Postharvest 30, 620- Accepted 30 August 2019 of strawberries during storage. Research methods: Fresh 623.Disinfection https://doi.org/10.21273/hortsci.30.3.620 of Fruits and Vegetables, 1, Elsevier Academic Press, 322-331. Available online 3 October 2019 strawberries were coated with different concentrations of chitosan Zheng,https://doi.org/10.1016/B978 X., Cai, D., Potter, D., Postman,-0-12-812698 J., Liu,-1.00011 J., & Teng,-X Y. (2014). Phylogeny and evolutionary (1%, 1.5%. 2%) or nano-chitosan (0.2%, 0.4%, and 0.8%) and stored Fallik,histories E., Arhbold, of Pyrus D. L.D., revealed Hamil ton by- Kemp,phylogenetic T. R., Clements,trees and networksA. M. , Collins, based onR. W.,data & from Barth, multiple M. E. Keywords: in 20C for 21 days. Findings: Coating strawberry with 0.2% and 0.4% nano-chitosan preserved the overall quality index of the fruit up to DNA(1998). sequences. (E)-2-Hexenal Molecular can Phylogeneticsstimulate Botrytis and Evolutioncinerea growth, 80, 54 in-65. vitro and on strawberry fruit in anthocyanin content 21 days. The treatments reduced weight loss, retained firmness, https://doi.org/10.1016/j.ympev.2014.07.009vivo during storage. Journal of the American Society for Horticultural Science, 123, 875-888. edible coating titratable acidity and L-ascorbic acid, significantly retarded https://doi.org/10.21273/JASHS.123.5.875 post-harvest losses malondialdehyde production and inhibited polyphenol oxidase Fenwick, G. R., Heaney, R. K., & Mullin, W. J. (1983). Glucosinolates and their break-down products post-harvest quality activity of the stored fruit. The 0.2% nano-chitosan treatment in food and food plants. Critical Reviews in Food Science and Nutrition 18, 123-201. reserved total soluble solid and total anthocyanin content better total phenolic content https://doi.org/10.1080/10408398209527361 than the 0.4% nano-chitosan. Although 2% chitosan coating showed Food and Agriculture Organization of the United Nations (FAO), (2011). Global food losses and food the positive effects, the overall quality index of the coated fruit was DOI: 10.22077/ jhpr.2019.2698.1082 waste. United Nations Food and Agriculture Organization. Rome, Italy. http://faostat.fao.org. reduced below the acceptable level after 18 days, shorter as P-ISSN: 2588-4883 Garcia, J. L. (1982). Storage of tomatoes (Lycopersicon esculentum Mill) in rice hull ash of different compared to the others coated with the lower concentrations of E-ISSN: 2588-6169 nano-chitosan. Research limitations: Nano-chitosan, showing to be particle sizes and moisture content [Philippines]. Thesis. In Agricultural Information Bank for the effective coating material in this study, is not popular traded in Asia, South-East Asian Regional Center for Graduate Study and Research in Agriculture

*Corresponding author: the industry. Originality/Value: The combination of 0.2 % nano- Food Technology Department, (AGRIS) since 1985. chitosan coating and storing fresh strawberry at 2°C preserved the Biotechnology School, International Gyawali, R., & Ibrahim, S. A. (2014). Natural products as antimicrobial agents. Food Control, 46, quality of fruits up to 21 days. The much lower concentrations of University, Vietnam National University nano-chitosan showed higher positive effects as compared to the 412-429. https://doi.org/10.1016/j.foodcont.2014.05.047 HCMC . higher concentrations of chitosan. This would help to reduce the Hanušová, K., Dobiáš, J., & Klaudisová, K. (2009). Effect of packaging films releasing antimicrobial cost of postharvest handlings for the strawberry industry. agents on stability of food products. Czech Jounal of Food Science, 27, 347-349. E-mail: [email protected] https://doi.org/10.17221/958-CJFS

© This article is open access and licensed under the ICAR, (2009). Indian Council of Agricultural Research. Annual Report. terms of the Creative Commons Attribution License Janjarasskul, T., & Krochta, M. (2010). Edible packaging materials. Food Science and Technology- http://creativecommons.org/licenses/by/4.0/ which Annual Reviews, 1, 415- 448. https://doi.org/10.1146/annurev.food.080708.100836 permits unrestricted, use, distribution and reproduction in any medium, or format for any Kamble, P. B., & Chavan, J. K. (2005). Effects of post-harvest treatments and storage temperature on purpose, even commercially provided the work is shelf-life of custard apple fruits. Jounal Food Science Technology, 42 (3), 253-255. properly cited. Kim, S. A., & Rhee, M. S. (2016). Highly enhanced bactericidal effects of medium chain fatty acids (caprylic, capric, and lauric acid) combined with edible plant essential oils (carvacrol, eugenol, β-

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