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An Optimized Protocol for in Vitro Propagation of Pyrus Communis and Pyrus Syriaca Using Apical-Bud Microcuttings

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An Optimized Protocol for in Vitro Propagation of Pyrus Communis and Pyrus Syriaca Using Apical-Bud Microcuttings 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 Pyrus communis and Pyrus syriaca 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, pear 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 plant 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 Rosaceae (Zheng et al., 2014), is one of the oldest temperate fruit 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 fruits 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 cultivars of Pyrus communis L. are cultivated in coastal regions and classical European Science, 6(1), 17-28. 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 plants: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.
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