Studies on In vitro Surface Sterilisation and Antioxidants on Pomegranate (Punica granatum L.) cv. Bhagwa
Keerthana S. G.
Department of Fruit Science, College of Horticulture, Bagalkot, University of Horticultural Sciences, Bagalkot, Karnataka, India.
Raghavendra Gunnaiah *
Department of Biotechnology and Crop Improvement, College of Horticulture, Bengaluru, University of Horticultural Sciences, Bagalkot, Karnataka, India.
S. N. Patil
Department of Fruit Science, College of Horticulture, Bagalkot, University of Horticultural Sciences, Bagalkot, Karnataka, India.
Sateesh Pattepur
Department of Fruit Science, College of Horticulture, Bagalkot, University of Horticultural Sciences, Bagalkot, Karnataka, India.
D. P. Prakasha
Department of Fruit Science, College of Horticulture, Sirsi, University of Horticultural Sciences, Bagalkot, Karnataka, India.
Anand Nanjappanavar
Department of Fruit Science, College of Horticulture, Bagalkot, University of Horticultural Sciences, Bagalkot, Karnataka, India.
Raghavendra K. Mesta
Regional Office, UHSB Campus, COE FPO, Bengaluru, Karnataka, India.
*Author to whom correspondence should be addressed.
Abstract
Pomegranate, regarded as the “Fruit of Paradise”, is one of the important fruit crops of tropical and subtropical regions. Pomegranate fruits are delicious and possess significant nutritional and medicinal benefits. Due to the increase in demand, the proliferation of pomegranate through tissue culture is essential to get high-quality planting material. However, microbial contamination in plant tissue culture is one of the bottlenecks for establishing aseptic cultures. Although multiple explant sterilisation methods have been developed for pomegranate, explant sterilisation methods can indeed vary from region to region due to the local environment and the mother plant. In the present study, 1-2 drops of tween 20 (20 min) + 500 mg/L carbendazim (30 min) + 500 mg/L streptomycin sulphate (10 min) + 100 mg/L citric acid (30 min) + 100 mg/L tebuconazole (folicur) (30 min) + 0.1% HgCl2 (1 min) + dipping explant after cutting the edges in sterile water (30 min) + dipping in 500 mg/L PVP (5 min) was found to be optimum in the prevention of microbial contamination with minimal cell death. Nodal segments-initiated callus at 54 days after culture, showing a callus induction percentage of 55.55% and a calli weight of 80 mg/explant.
Keywords: Bhagwa, fungicide, bactericide, antioxidants, callus
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References
Levin GM. Pomegranate, first edition, third millennium publishing, East Libra Drive Tempe, AZ; 2006.
Anonymous, Pomegranates- Raw Food. U. S. Department of Agriculture; 2018 a. Available:https://fdc.nal.usda.gov
Jurenka, J. Therapeutic applications of pomegranate (Punica granatum L.). Altern. med. rev., 2008;13(2):128-144.
Ghavipour M, Sotoudeh G, Tavakoli E, Mowla K, Hasanzadeh J and Mazloom Z. Pomegranate extract alleviates disease activity and some blood biomarkers of inflammation and oxidative stress in Rheumatoid Arthritis patients. Eur. J. Clin. Nutr. 2017;71(1):92-96.
Neurath AR, Strick N, Li YY and Debnath AK. Punica granatum (Pomegranate) juice provides an HIV-1 entry inhibitor and candidate topical microbicide. BMC Infect. Dis. 2004;4: 41.
Westwood MN. Temperate Zone Pomology. W. H. Freeman and Company, San Francisco; 1978.
Kumar R, Meena R, Sharma BD and Saroj PL. Production Technology of Pomegranate in Arid Region, 1st edition, ICAR-Central Institute for Arid Horticulture. 2018; 2.
Anonymous. Solapur Pomegranate.
Available:https://www.nhb.gov.in. 2020-21.
Anonymous. Project report on export promotion of pomegranate from India, UNDER GOI-UNCTAD DFID Project on Strategies and Preparedness for Trade and Globalization in India, Agricultural Finance Corporation, New Delhi; 2007.
Vasane SR and Patil JL. Multiplying Benefits with Acclimatization Practices (Gacp) In Tissue Culture Propagated Mrudula Pomegranate. In V International Symposium on Acclimatization and Establishment of Micropropagated Plants oct. 2011;988:77-84.
Tayade SA, Joshi PS, Raut HS and Shete MB. Effect of time and air layer per shoot on rooting and survival of air layers in pomegranate cv. Bhagwa. Int. j. med. aromat. Plants. 2017;3(1):20-24.
Chater JM, Merhaut DJ, Preece JE and Blythe EK. Rooting and vegetative growth of hardwood cuttings of 12 pomegranate (Punica granatum L.) cultivars. Sci. Hortic. 2017;221:68-72.
Kahramanoglu I and Umar K. Effects of cutting length, time and growing media on the sprouting of dormant semi-hardwood cuttings of pomegranate cv. Wonderful. African. J. Agric. Res. 2018;13(45): 2587-2590.
Chandra R., Babu KD, Jadhav VT, Jaime A and Silva TD. Origin, history and domestication of pomegranate. Fruit, Veg. Cereal Sci. Biotech. 2010;1-6.
Chauhan RD, and Kanwar K. Biotechnological advances in pomegranate (Punica granatum L.). In Vitro Cell. Dev. Biol. Plant. 2012;48:579-594.
Satheesh M, Sridharan T, Standardization of Punica granatum explant and callus induction through micropropagation-indirect organogenesis. Int. J. Renew. Energy Technol. 2014;3:19-27.
Choudhary R., Jakhar ML, Kumar R, Jat H, Shekhawat K, Kumawat S and Choudhary RN. In vitro callus induction from different explants in Pomegranate (Punica granatum L.). J. pharmacogn. phytochem. 2018;7(4):2656-2659.
Murkute A, Patil S, Patil BN and Kumari M. Micropropagation in pomegranate, callus induction and differentiation. South Indian Hort. 2002;50(1-3):49-55.
Deepika R and Kanwar K. In vitro regeneration of Punica granatum L. plants from different juvenile explants. J Fruit Ornamental Plant Res. 2010;18(1):5- 22.
Bonyanpour A and Khosh-Khui M. Callus induction and plant regeneration in Punica granatum L. ‘nana’ from leaf explants. J. Cent. Eur. Agric. 2013;14(3):75-83.
Guranna P, Hoolageri HC. Studies on establishment of aseptic culture in pomegranate cv. Bhagwa. Annu.res.rev.biol. 2017;21(5): 1-7.
Verma V, Zinta G and Kanwar K. Optimization of efficient direct organogenesis protocol for Punica granatum L. cv. Kandhari Kabuli from mature leaf explants. In vitro Cell. Dev. Biol., 2021;57: 48-59.
Song J, Han C, Zhang S, Wang Y, Liang Y, Dai Q, Huoand Z, Xu K. Hormetic Effects of Carbendazim on Mycelial Growth and Aggressiveness of Magnaporthe oryzae. J. Fungus., 2022;8 (10):1008.
Patil VM, Dhande GA, Thigale DM and Rajput JC. Micropropagation of pomegranate (Punica granatum L.) ‘Bhagava’ cultivar from nodal explant. African. J. Biotechnol. 2011;10(79):18130-18136.
Guranna P, Hosamani I, Sathyanarayana R, Hegde R and Hipparagi K. Micropropagation in pomegranate (Punica granatum L.) cv. ‘Bhagwa’ through indirect organogenesis and assessment of genetic fidelity by RAPD marker. Biotechnol. J. Int. 2018;20(3):1-8.
Kumari G, Kanwar K, Soni M and Sharma D. In vitro selection of cell lines in Punica granatum L. (Daru) against bacterial blight. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 2015;87:59.
Murkute AA., Patil S and Singh SK. In vitro regeneration in pomegranate cv. Ganesh from mature trees. Indian J Hortic. 2004; 61(3):206-208.
Kumar K, Arora PK, Brar JS, Bhatia D and Kumar A. Influence of explant collection period, antibrowning strategy and growth regulators composition on in vitro propagation of Bhagwa pomegranate. Indian J Hortic. 2019;76(2): 273-278.
Lakshmi PC, CN Reddy and B Srinivas. Effects of Bavistin and Cefotaxime on in vitro Contaminant free Shoot Regeneration of Ruellia tuberosa L. Plant Tissue Cult Biotechnol. 2021;31(1):1-12.
Anonymous. Fungicide Folicur; 2018 Available:https://www.cropscience.bayer.in.
Hillebrands L, Lamshoeft M, Lagojda A, Stork A and Kayser O. Evaluation of Callus Cultures to Elucidate the Metabolism of Tebuconazole, Flurtamone, Fenhexamid, and Metalaxyl-M in Brassica napus L., Glycine max (L.) Merr., Zea mays L., and Triticum aestivum L. J. Agric. Food Chem. 2020;68(48):14123-14134.
Pandey S, Prakash S, Prasad Y, Kumar A, Vaishali PC and Singh G. Studies on the effects of different surface sterilization agents under in vitro culture of Pepino (Solanum muricatum Ait.) cv. Valentia. J. Pharma. Innov. 2021;10:371-377.
Gerolino EF, Chierrito TPC, Filho AS, Souto ER, Goncalves RAC and Oliveira AJBD. Evaluation of limonoid production in suspension cell culture of Citrus sinensis. Rev. bras. farmacogn. 2015;25: 455-461.
Chongloi L, Gunnaiah R, Hipparagi K, Guranna P, Prakasha DP, Chittapur R, Kamble A and Vishweshwar S. Economic Analysis of Micropropagation of Dragon Fruit (Hylocereus undatus (Haw.) Britton and Rose). Int. j. plant soil sci., 2022;34(22):1267-1275.
Schatz A, Bugie E, Waksman SA, Hanssen AD, Patel R and Osmon DR. The classic: streptomycin, a substance exhibiting antibiotic activity against Gram-positive and Gram-negative bacteria. Clin. Orthop. Relat. Res. 2005;437:3-6.
Das MP, Rebecca LJ, Sharmila S and Chatterjee S. Study on the effect of mercury (II) chloride as disinfectant on mixed culture. J Chem Pharm Res. 2012;4: 4975-4978.
Pal D, Kumar M, Kumar A, Sengar RS, Yadav MK, Pal A, Singh KP and Pandey V. Standardization of surface sterilization for in vitro cloning of pomegranate (Punica granatum L.) cv. Bhagwa. Biol Forum. 2022;14(4):653-656.
Zamir R, Shah ST, Ali N, Khattak GSS and Muhammad T. Studies on in vitro surface sterilization and antioxidants on guava shoot tips and nodal explants. Pakistan. J. Biotechnol. 2004;1(2):12-16.
Concepcion O, Napoles L, Perez AT, Hernandez M, Peralta N and Trujillo R. Efecto de tres antioxidantes en el cultivo in vitro de apices de guayaba (Psidium guajava L.). Cult. Trop. 2005;26: 33-39.
Chavan SS, Ranade SS, Deore AC, Deshpande RS and Dhonukshe BL. Cloning Alphonso mango through vegetative explants. Annals of Plant Physiology. 2000;14(2):178-181.
De Assis FA, Rodrigues FA, Pasqual M, De Assis GA, Luz JMQ, Janoni F, Costa I, Costa BNS and Soares JDR. Antioxidants in the control of microorganism contamination and phenol oxidation in Eugenia pyriformis. J. Biosci. 2018;34(1); 49-58.
Cai X, Wei H, Liu C, Ren X, Thi LT and Jeong BR. Synergistic effect of NaCl pretreatment and PVP on browning suppression and callus induction from petal explants of Paeonia lactiflora Pall. ‘Festival Maxima’. Plant J. 2020;9(3):346.
Kona P, Kumar MH, Reddy KHP, Reddy DM, Reddy NPE, Latha P and Balaji MS. Effect of 2,4-D and EMS on in vitro regeneration in sugarcane cultivar, Co86032. Int. J. Curr. Microbiol. App. Sci. 2019;8:1228-36.
Karale AR, Kadam JH, Kamble AB, Kaledhonkar DP and Patil MT. Micropropagation of Emblica officinalis Gaertn. through callus culture. In Recent Trends in Horiculture Biotechnology, Raghunath and Keshvachandran (Eds) New Indian Publishing Agency, New Delhi (India). 2007:239-244.
Sunitibala H, Damayanti M and Sharma GJ. In vitro propagation and rhizome formation in Curcuma longa Linn. Cytobios. 2001;71-82.
Savita R, Shasthree T and Mallaiah B. High frequency of plantlet regeneration and multiple shoot induction from leaf and stem explant of C. jambhiri (L). Asian. J. Exp. Biol. Sci. 2010;1(3):575-579.
Azim F, Rahmna MM, Prodham SH, Sikdar SU, Zobayer N and Ashrafuzzaman M. Development of efficient callus initiation of Malta (Citrus sinensis) through tissue culture. Int. J. Agril. Res. Innov. & Tech. 2011;1(1&2):64-68.