APPLICATION OF RESPONSE SURFACE METHODOLOGY TO PREDICT THE OPTIMAL CONDITIONS FOR REDUCING BROWNING IN THE IN VITRO CULTURE OF MUSA (AA GROUP) ‘KLUAI KHAI’ PLANTLETS
DOI:
https://doi.org/10.14456/lsej.2026.1Keywords:
Lethal browning, Musa (AA group) ‘Kluai Khai’, In vitro culture, response surface methodologyAbstract
One of the major problems of Musa (AA group) ‘Kluai Khai’ tissue culture is the browning from the oxidation of phenolic compounds. This problem can be resolved by adding antioxidants or phenolic–adsorbing agents to the culture medium. A response surface methodology (RSM) approach was employed in combination with a central composite design (CCD) to model the effects of three factors: ascorbic acid (AA), polyvinylpyrrolidone (PVP), and activated charcoal (AC), each at three concentration levels. A total of 17 experimental treatments were conducted for 12 weeks. The levels of phenolic compounds, tannins, and flavonoids in the culture medium were analyzed, and the four treatments yielding the lowest concentrations were selected for further validation. The optimized conditions were subsequently tested for 12 weeks. The results indicated that Murashige and Skoog (1962) medium supplemented with 0.03 g/L AA, 3.325 g/L PVP, and 1.48 g/L AC effectively reduced browning in in vitro cultures of Musa (AA group) ‘Kluai Khai’.
References
Ahmad I, Hussain T, Ashraf I, Nafees M, Maryam RM, Iqbal M. Lethal effects of secondary metabolites on plant tissue culture. American–Eurasian Journal of Agricultural & Environmental Sciences 2013;13(4):539–547.
Amente G, Chimdessa E. Control of browning in plant tissue culture: A review. Journal of Scientific Agriculture 2021;5:67–71.
Babu P. An efficient protocol for in vitro regeneration of banana var. Nanjangudu rasabale (Musa spp. AAB). International Journal of Current Microbiology and Applied Sciences 2019;8(6):3392–3402.
Halee A, Supavititpatana P, Ruttarattanamongkol K, Jittrepotch N, Rojsuntornkitti K, Kongbangkerd T. Effects of solvent types and citric acid concentrations on the extraction of antioxidants from the black rice bran of Oryza sativa L. cv. Hom Nin. Journal of Microbiology, Biotechnology and Food Sciences 2018;8(2):765–769.
Kaewbon P. Inhibition of browning agents in bisected protocorm-derived callus of pigeon orchid (Dendrobium crumenatum Swartz). Ph.D. Thesis. Prince of Songkla University; 2015.
Ko WH, Su CC, Chen CL, Chao CP. Control of lethal browning of tissue culture plantlets of Cavendish banana cv. Formosana with ascorbic acid. Plant Cell, Tissue and Organ Culture 2009;96:137–141.
Kongphan P, Theanhom AA, Jaruwattanaphan T. Size and cutting pattern of explant for micropropagation of Musa (AA Group) ‘Kluai Khai Kasetsart 2’. Thai Science and Technology Journal 2020;28(12):2164–2172.
Luecha P, Trunjaruen A, Suraporn S, Yaowachai W, Maneerattanarungroj P, Kunpratum N, Taratima W. et al. Optimization of phytochemical content and DPPH scavenging activity from pokeweed (Phytolacca americana L.) callus using Response Surface Models. International Journal of Agriculture and Biosciences 2024;13(3):356–366.
Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum 1962;15:473–497.
Ndakidemi CF, Mneney E, Ndakidemi PA. Effects of ascorbic acid in controlling lethal browning in in vitro culture of Brahylaena huillensis using nodal segments. American Journal of Plant Sciences 2014;5:187–191.
Ngomuo M, Mneney E, Ndakidemi P. Control of lethal browning by using ascorbic acid on shoot tip cultures of a local Musa spp. (banana) cv. Mzuzu in Tanzania. African Journal of Biotechnology 2014;13(16):1721–1725.
Pradubsri W, Chomtee B, Thongteeraparp A. A study of small response surface designs for the full second order model and a set of reduced models in a spherical region. Thai Science and Technology Journal 2015;23(3):362–376.
Silayoi, B. Banana. (2nd ed.). Bangkok: Kasetsart University; 1995.
Somjaipeng S, Krasang K, Sopa P, Noychaipruk C. The application of response surface methodology to predict the radial growth rate of aflatoxigenic Aspergillus section Flavi: the effect of climate change factors. Khon Kaen Agriculture Journal 2017;4(1):1140–1416.
Thomas TD. The role of activated charcoal in plant tissue culture. Biotechnology Advances 2008;26(6):618–631.
Vuylsteke DR. Shoot-tip culture for the propagation conservation, and distribution of Musa germplasm. Nigeria: International Institute of Tropical Agriculture; 1998.
Wattanapan N, Srichuay W, Klinnawee L, Meesawat U. Effects of anti-browning agents on protocorm-like body induction using thin cell layers of endangered lady’s slipper orchid: Paphiopedilum callosum var. sublaeve. Princess of Naradhiwas University Journal 2019;11(1):107–116.
Wongsa T, Inthima P, Kongbangkerd A. Effect of cytokinins and explant preparation on in vitro shoot multiplication in Musa (AA group) ‘Khai Kamphaeng Phet’. YRU Journal of Science and Technology 2021a;6(2):171–180.
Wongsa T, Jacksri, N, Kongbangkerd A. Effect of culture media and sucrose concentrations on in vitro young shoots storage of Musa (AA group) ‘Khai Kamphaeng Phet’. Udon Thani Rajabhat University Journal of Sciences and Technology 2021b;9(2):139–151.
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