The Use of Alcohol and Sodium Hypochlorite as Single Sterilizing Agents for Oil Palm Explant Sterilization
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Published:
Apr 20, 2021
Keywords:
alcohol, explants, oil palm, sodium hypochlorite, sterilization
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Abstract
Oil palm micropropagation through tissue culture is initiated with explants sterilization. Sterilization is a crucial stage because it determines the production rate of sterile plant cultures. Concentration and exposure time of sterilizing agents must be determined empirically to gain effective method which produces explants with low mortality. This research aimed to obtain effective protocols for unopened-leaves sterilization of oil palm using single sterilizing agents. Alcohol and sodium hypochlorite (NaOCl) at certain concentrations and duration of exposure were used as sterilization treatments. Treatments of alcohol did not show any significant differences on contamination, browning, and growth response rate based on analysis of variance (ANOVA) as well as sodium hypochlorite. The best results were shown by 70% alcohol for 5 minutes and 10% NaOCl for ten minutes. These treatments were sufficiently effective in reducing contamination with the lowest percentage of browning explants and high growth response rate. Application with higher concentration of alcohol (80% and 90%) caused death of explants, while higher concentration of sodium hypochlorite increased browning explants. The type of contamination found in culture were bacteria and fungi. Domination of bacteria was found in alcohol treatment while fungi contamination dominated in NaOCl treatment.
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Pratiwi, D. R., Wening, S., Nazri, E., & Yenni, Y. (2021). The Use of Alcohol and Sodium Hypochlorite as Single Sterilizing Agents for Oil Palm Explant Sterilization . Jurnal Penelitian Kelapa Sawit, 29(1), 1-10. https://doi.org/10.22302/iopri.jur.jpks.v29i1.120
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This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
- Indonesian Journal of Oil Palm Research can be accessed freely by anyone (open access) to introduce more journals to the public.
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References
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Caliskan, O., J. Radusiene, K.E. Temizel, Z. Staunis, C. Cirak, D. Kurt, and M.S. Odabas. 2017. The effects of salt and drought stress on phenolic accumulation in greenhouse-grown Hypericum pruinatum. Italian Journal of Agronomy. 12(918): 271-275.
Cobrado, J.S., and A.M. Fernandez. 2016. Common fungi contamination affecting tissue-cultured Abaca (Musa textiles Nee) during initial stage of micropropagation. Asian Research Journal of Agriculture. 1(2):1–7.
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Omamor, I.B., A.O. Asemota, C.R. Eke, and E.I. Eziashi. 2007. Fungal contamination of the oil palm tissue culture in Nigerian institute for oil palm research (NIFOR). African Journal of Agricultural Research. 2(10): 534-537.
Onwubiko N.C., C.S. Nkogho, C.P. Anyanwu, and G.C. Onyeishi. 2013. Effect of different concentration of sterilant and exposure time on sweet potato (Ipomoea batatas Lam) explants. International journal of current microbiology and applied sciences 2(8): 14-20.
Orlikowska, T., K. Nowak, and B. Reed. 2017. Bacteria in the plant tissue culture environment. Plant Cell, Tissue and Organ Culture 128(3): 487–508.
Oyebanji, O.B., O. Nweke, O. Odebunmi, N.B. Galadima, M.S. Idris, U.N. Nnodi, A.S. Afolabi, and G.H. Ogbadu. 2009. Simple, effective and economical explant-surface sterilization protocol for cowpea, rice and sorghum seeds. African Journal of Biotechnology 8(20): 5395-5399.
Pádua, M.S., R.S. Santos, C.R.G. Labory, V.C. Stein, E.G. Mendonça, E. Alves, and L.V. Paiva. 2018. Histodiffentiation of oil palm somatic embryo development at low auxin concentration. Protoplasma. 255(1): 285-295.
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Pierik, R.L.M. 1987. In Vitro Culture of Higher Plants. Dordrecht : Martinus Nijhoff.
Ray, S.S., and N. Ali. 2016. Biotic contamination and possible ways of sterilization review with reference to bamboo micropropagation. Brazilian Archives of Biology and Technology. 59: 1-10.
RStudio Team. 2018. RStudio: Integrated Development for R. RStudio, Inc., Boston, MA URL http://www.rstudio.com/.
Saad, A.I.M., and A.M. Elshahed. 2012. Plant Tissue Culture Media. Di dalam: Recent Advances in Plant in vitro Culture. Leva, A., and L.M.R. Rinaldi, editor. Available from: https://www.intechopen.com/books/recent-advances-in-plant-in-vitro-culture/plant-tissue-culture-media.
Sun, R.Z., G. Cheng, Q. Li, Y.N. He, Y. Wang, Y.B. Lan, S.Y. Li, Y.R. Zhu, W.F. Song, X. Zhang, X.D. Cui, W. Chen, and J. Wang. 2017. Light-induced variation in phenolic compounds in cabernet sauvignon grapes (Vitis vinifera L.) involves extensive transcriptome reprogramming of biosynthetic enzymes, transcription factors, and phytohormonal regulators. Frontiers in Plant Science. 8(547): 1-18.
Yoo, Jin-Hong. 2018. Review of Disinfection and Sterilization – Back to the Basics. Infection and Chemotherapy. 50(2): 101-109.
Zamir, R., and Abdur-Rab. 2014. Effect of exposure time and incubation period of various sterilants and antioxidants on the in vitro morphogenis of guava explants. IOSR Journal of Agriculture and Veterinary Science 7(5): 81-86.
Abdi, Gholamreza, Hassan Salehi, and Morteza Khosh-Khui. 2008. “Nano Silver: A Novel Nanomaterial for Removal of Bacterial Contaminants in Valerian (Valeriana Officinalis L.) Tissue Culture.” Acta Physiologiae Plantarum 30 (5): 709–14.
Ankur V., B. Meena, and N.S.K. Harsh. 2014. Identification and bioassay of fungal contaminants observed during in vitro propagation of Saracaasoca (Roxb.) De Wilde. Biotechnology International. 7(2): 35-42.
Benzoni T, and Hatcher JD. 2019. Bleach Toxicity. [Diakses 3 September 2019]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing. Available from: https://www.ncbi.nlm.nih.gov/books/NBK441921/.
Brandl, M.T. 2008. Plant lesions promote the rapid multiplication of Escherichia coli O157:H7 on postharvest lettuce. Applied and environmental microbiology 74(17): 5285-5289.
Caliskan, O., J. Radusiene, K.E. Temizel, Z. Staunis, C. Cirak, D. Kurt, and M.S. Odabas. 2017. The effects of salt and drought stress on phenolic accumulation in greenhouse-grown Hypericum pruinatum. Italian Journal of Agronomy. 12(918): 271-275.
Cobrado, J.S., and A.M. Fernandez. 2016. Common fungi contamination affecting tissue-cultured Abaca (Musa textiles Nee) during initial stage of micropropagation. Asian Research Journal of Agriculture. 1(2):1–7.
Eziashi, E.I., O. Asemota, C.O. Okwuagwu, C.R. Eke, N.I. Chidi, and E.A. Oruade-Dimaro. 2014. Screening sterilizing agents and antibiotics for the elimination of bacterial contaminants Journal of Applied Oral Science from oil palm explants for plant tissu e culture. European Journal of Experimental Biology. 4(4): 111-115.
Fang, J.Y., and Y.R. Hsu. 2012. Molecular identification and antibiotic control of endophytic bacterial contaminants from micropropagated Aglaonema cultures. Plant Cell, Tissue and Organ Culture, 110(1): 53–62.
Ibrahim, M.H., H.Z.E. Jaafar, A. Rahmat, and Z.A. Rahman. 2011. The relationship between phenolics and flavonoids production with total non structural carbohydrate and photosynthetic rate in Labisia pumila benth. under high co2 and nitrogen fertilization. Molecules 16: 162-174.
Junairiah, N.I., Zuraidassanaaz, F.N. Izdihar, and Y.S.W. Manuhara. Callus induction of leaf explant Piper betle L. Var Nigra with combination of plant growth regulators indole-3-acetic acid (IAA), benzylamino purin (BAP) and kinetin. AIP Conference Proceedings. 1888: 020028-1–020028-7.
Kane, M.E., P. Kauth, and T. Johnson. 2011. Culture indexing for bacterial and fungal contaminants. Di Dalam: Plant Tissue Culture, Development, and Biotechnology. Editor : Trigiani, R.N., and D.J. Gray. Boca Raton (US): CRC Press.
Kasnak, C. 2020. Effects of anti-browning treatments on the polyphenol oxidase and antioxidant activity of fresh-cut potatoes by using response surface methodology. Potato Research 1-14.
Klayraung, S., P. Niamsup, P. Poonnoy, and N. Topoonyanont. 2017. Diversity and control of bacterial contamination of plants propagated in temporary immersion bioreactor system. Acta Horiticulturae 1155: 439–446.
Laukkanen, H., H. Häggman, S. Kontunen-Soppela, and A.Hohtola. 1999. Tissue browning of in vitro cultures of Scots pine: Role of peroxidase and polyphenol oxidase. Physiologica Plantarum 106: 337-343.
McDonnell, G., and A.D. Russell. 1999. Antiseptics and disinfectants: activity, action, and resistance. Clinical Microbiology Reviews 12(1): 147-179.
McDonnell G. General mechanism of action. In: McDonnel GE, eds. Antisepsis, Disinfection, and Sterilization. 2nd ed. Washington DC: ASM Press; 2017;255-69.
Modeste, K.K., M.T. Eliane, K. Daouda, S.A. Brahima, K. Tchoa, K.E. Kouablan, and K. Mongomake. 2017. Effect of antioxidants on the callus induction and the development of somatic embryogenesis of cocoa [Theobroma cacao (L.)]. Australian Journal of Crop Science 11(1): 25-31.
Nadha, H.K., R. Salwan, R.C. Kasana, M. Anand, and A. Sood. 2012. Identification and elimination of bacterial contamination during in vitro propagation of Guadua angustifolia Kunth. Pharmacognosy Magazine. 8(30): 93–97.
Nakagarwara, S.,T. Goto, M. Nara, Y. Ozawa, K. Hotta, and Y. Arata. 1998. Spectroscopic characterisation and the pH dependence of bacterial activity of the aqueous chlorine solution. Analytical Sciences. 14: 691-698.
Odutayo, O.I., N.A. Amusa, O.O. Okutade, and Y.R. Ogunsanwo. 2007. Sources of microbial contamination in tissue culture laboratories in southwestern Nigeria. African Journal of Biotechnology. 2: 67-72.
Omamor, I.B., A.O. Asemota, C.R. Eke, and E.I. Eziashi. 2007. Fungal contamination of the oil palm tissue culture in Nigerian institute for oil palm research (NIFOR). African Journal of Agricultural Research. 2(10): 534-537.
Onwubiko N.C., C.S. Nkogho, C.P. Anyanwu, and G.C. Onyeishi. 2013. Effect of different concentration of sterilant and exposure time on sweet potato (Ipomoea batatas Lam) explants. International journal of current microbiology and applied sciences 2(8): 14-20.
Orlikowska, T., K. Nowak, and B. Reed. 2017. Bacteria in the plant tissue culture environment. Plant Cell, Tissue and Organ Culture 128(3): 487–508.
Oyebanji, O.B., O. Nweke, O. Odebunmi, N.B. Galadima, M.S. Idris, U.N. Nnodi, A.S. Afolabi, and G.H. Ogbadu. 2009. Simple, effective and economical explant-surface sterilization protocol for cowpea, rice and sorghum seeds. African Journal of Biotechnology 8(20): 5395-5399.
Pádua, M.S., R.S. Santos, C.R.G. Labory, V.C. Stein, E.G. Mendonça, E. Alves, and L.V. Paiva. 2018. Histodiffentiation of oil palm somatic embryo development at low auxin concentration. Protoplasma. 255(1): 285-295.
Pais, A.K., A.P. da Silva, J.C. de Souza, S.L. Teixeira, J.M. Ribeiro, A.R. Peixotol, and C.D. da Paz. 2016. Sodium hypochlorite sterilization of culture medium in micropropagation of Gerbera hybrida cv. Essandre. African Journal of Biotechnology 15(36): 1995-1998.
Pierik, R.L.M. 1987. In Vitro Culture of Higher Plants. Dordrecht : Martinus Nijhoff.
Ray, S.S., and N. Ali. 2016. Biotic contamination and possible ways of sterilization review with reference to bamboo micropropagation. Brazilian Archives of Biology and Technology. 59: 1-10.
RStudio Team. 2018. RStudio: Integrated Development for R. RStudio, Inc., Boston, MA URL http://www.rstudio.com/.
Saad, A.I.M., and A.M. Elshahed. 2012. Plant Tissue Culture Media. Di dalam: Recent Advances in Plant in vitro Culture. Leva, A., and L.M.R. Rinaldi, editor. Available from: https://www.intechopen.com/books/recent-advances-in-plant-in-vitro-culture/plant-tissue-culture-media.
Sun, R.Z., G. Cheng, Q. Li, Y.N. He, Y. Wang, Y.B. Lan, S.Y. Li, Y.R. Zhu, W.F. Song, X. Zhang, X.D. Cui, W. Chen, and J. Wang. 2017. Light-induced variation in phenolic compounds in cabernet sauvignon grapes (Vitis vinifera L.) involves extensive transcriptome reprogramming of biosynthetic enzymes, transcription factors, and phytohormonal regulators. Frontiers in Plant Science. 8(547): 1-18.
Yoo, Jin-Hong. 2018. Review of Disinfection and Sterilization – Back to the Basics. Infection and Chemotherapy. 50(2): 101-109.
Zamir, R., and Abdur-Rab. 2014. Effect of exposure time and incubation period of various sterilants and antioxidants on the in vitro morphogenis of guava explants. IOSR Journal of Agriculture and Veterinary Science 7(5): 81-86.