MORPHOLOGICAL AND PHYSIOLOGICAL SCREENING OF ETHYL METHANESULFONATE-TREATED SUGARCANE CALLI AND PLANTLETS FOR In vitro MANNITOL-INDUCED OSMOTIC STRESS TOLERANCE
PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY,
Page 107-120
Abstract
Sugarcane production in South Africa is limited by drought stress and its impact is expected to increase due to climate change. However, limited research is conducted to develop cultivars that are suitable for cultivation under water-stressed conditions. This study aimed to select, through morphological and physiological traits, the ethyl methanesulfonate treatment(s) that can produce calli and plantlets that are resistant to mannitol-induced osmotic stress. Sugarcane calli were exposed to ethyl methanesulfonate for 0.5, 1, 2, and 3 hours. To determine optimal selection lethal doses, calli were cultured on media containing 0, 150, 225 and 300 mM mannitol for further eight weeks under dark and light conditions. Incubation periods of half an hour and one hour induced genomic mutations without inhibiting callus growth and plant regeneration abilities. EMS-treated calli retained whiter, compact and with yellowish friable texture when compared with the control after the two-weeks recovery period post exposure to osmotic stress. Callus that was exposed for one hour was able to recover and regenerate plantlets at 225 and 407 mM mannitol stress. However, the two-hour incubation period and above resulted in stunted and albino plantlets. The LD50 and LD90 for selection were calculated as 225 and 407 mM mannitol. The EMS mutagenesis and in vitro selection for osmotic stress using mannitol can be used successfully to select sugarcane plantlets with better morphological and physiological responses to water stress in a very short time.
Keywords:
- Ethyl methanesulfonate mutagenesis
- mannitol osmotic stress
- sugarcane
How to Cite
DLAMINI, P. J., SCHULTZ-VILJOEN, T., & NTULI, N. R. (2020). MORPHOLOGICAL AND PHYSIOLOGICAL SCREENING OF ETHYL METHANESULFONATE-TREATED SUGARCANE CALLI AND PLANTLETS FOR In vitro MANNITOL-INDUCED OSMOTIC STRESS TOLERANCE. PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY, 21(37-38), 107-120. Retrieved from https://www.ikppress.org/index.php/PCBMB/article/view/5420
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Ahuja I, de Vos RC, Bones AM, Hall RD. Plant molecular stress responses face climate change. Trends in Plant Science. 2010;15:664-674.
Ghaheri M, Kahrizi D, Bahrami G. Effect of mannitol on some morphological characteristics of In vitro Stevia rebaudiana Bertoni. Biharean Biologist. 2017;11:94-97.
Snyman S, Mhlanga P, Watt M. Rapid screening of sugarcane plantlets for In vitro mannitol-induced stress. Sugar Tech. 2016; 18:437-440.
Masoabi M, Lloyd J, Kossmann J, van der Vyver C. Ethyl methanesulfonate mutagenesis and In vitro polyethylene glycol selection for drought tolerance in sugarcane (Saccharum spp.). Sugar Tech. 2018;20:50-59.
Lutts S, Almansouri M, Kinet JM. Salinity and water stress have contrasting effects on the relationship between growth and cell viability during and after stress exposure in durum wheat callus. Plant Science. 2004; 167:9-18.
Beck CB. Structure and development of the cell wall. In "An introduction to plant structure and development: Plant anatomy for the twenty-first century". Cambridge Univerisity press, Cambridge, UK. 2012; 58-82.
Koiwa H. Pathways and genetic determinants for cell wall-based osmotic stress tolerance in the Arabidopsis thaliana root system. In "Genes for Plant Abiotic Stress" (Jenks MA, Wood AJ, eds.). Wiley-Blackwell. 2010;35-53.
Zingaretti SM, Rodrigues FA, Graça JP da, Pereira L de M, Lourenço MV. Sugarcane responses at water deficit conditions. In "Water Stress". In Tech. 2012;255-276.
Snyman SJ, Meyer GM, Koch AC, Banasiak M, Watt MP. Applications of In vitro culture systems for commercial sugarcane production and improvement. In vitro Cellular & Developmental Biology-Plant. 2011;47:234-249.
Mahlanza T, Rutherford RS, Snyman SJ, Watt MP. In vitro generation of somaclonal variant plants of sugarcane for tolerance to Fusarium sacchari. Plant Cell Reports. 2013;32:249-262.
Koch AC, Ramgareeb S, Rutherford RS, Snyman SJ, Watt MP. An In vitro mutagenesis protocol for the production of sugarcane tolerant to the herbicide imazapyr. In Vitro Cellular & Developmental Biology-Plant. 2012;48: 417-427.
Koetle MJ, Evans DL, Singh V, Snyman SJ, Rutherford RS, Watt MP. Agronomic evaluation and molecular characterisation of the acetolactate synthase gene in imazapyr tolerant sugarcane (Saccharum hybrid) genotypes. Plant Cell Reports. 2018;37:1201-1213.
Purnamaningsih R, Hutami S. Increasing Al-Tolerance of sugarcane using ethyl methanesulfonate and In vitro selection in the low pH media. Hayati Journal of Biosciences. 2016;23:1-6.
Sega GA. A review of the genetic effects of ethyl methanesulfonate. Mutation Research/Reviews in Genetic Toxicology. 1984;134:113-142.
Hofmann NE, Raja R, Nelson RL, Korban SS. Mutagenesis of embryogenic cultures of soybean and detecting polymorphisms using RAPD markers. Biologia Plantarum. 2004;48:173-177.
Grivet L, Arruda P. Sugarcane genomics: Depicting the complex genome of an important tropical crop. Current Opinion in Plant Biology. 2002;5:122-127.
Hassan NS, Shaaban LD, Hashem E, Seleem EE. In vitro selection for water stress tolerant callus line of Helianthus Annus L. cv. Myak. International Journal of Agriculture and Biology. 2004;6:13-18.
Zhou M. Conventional sugarcane breeding in South Africa: Progress and future prospects. American Journal of Plant Sciences. 2013;4:189-196.
Snyman SJ. Sugarcane transformation. In "Transgenic Crops of the World: Essential Protocols" (Curtis IS, ed.). Springer Netherlands, Dordrecht. 2004;103-114.
Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum. 1962;15:473-497.
Shekhawat GS, Verma K, Jana S, Singh KL, Teotia P, Prasad A. In vitro biochemical evaluation of cadmium tolerance mechanism in callus and seedlings of Brassica juncea. Protoplasma. 2009;239: 31-38.
Errabii T, Gandonou CB, Essalmani H, Abrini J, Idaomar M, Senhaji NS. Effects of NaCl and mannitol induced stress on sugarcane (Saccharum spp.) callus cultures. Acta Physiologiae Plantarum. 2007;29:95–102.
Taylor PWJ, Ko HL, Adkins SW, Rathus C, Birch RG. Establishment of embryogenic callus and high protoplast yielding suspension cultures of sugarcane (Saccharum spp. hybrids). Plant Cell, Tissue and Organ Culture. 1992;28:69-78.
Gocke E, Bürgin H, Müller L, Pfister T. Literature review on the genotoxicity, reproductive toxicity, and carcinogenicity of ethyl methanesulfonate. Toxology Letters. 2009;190:254–265.
Kenganal M, Hanchinal R, Nadaf H. Ethyl methanesulfonate (EMS) induced mutation and selection for salt tolerance in sugarcane In vitro. Indian Journal of Plant Physiology. 2008;13:405-410.
Khalil F, Naiyan X, Tayyab M, Pinghua C. Screening of EMS-induced drought-tolerant sugarcane mutants employing physiological, molecular and enzymatic approaches. Agronomy. 2018;8:11-13.
Svetleva L. Effect of ethyl methanesulfonate (EMS) and N-nitrose-N-ethyl Urea (ENU) on callus growth of common bean. Journal of Central European Agriculture. 2005;6:59-64.
Silué S, Diarrasouba N, Fofana IJ, Muhovski Y, Toussaint A, Mergeai G, Jacquemin JM, Baudoin JP. Description of Phaseolus vulgaris L. aborting embryos from ethyl methanesulfonate (EMS) mutagenized plants. Biotechnology, Agronomy, Society and Environment. 2013;17:563-571.
Novak F, Afza R, Duren MV, Omar M. Mutation induction by gamma irradiation of in vitro cultured shoot-tips of banana and plantain (Musa cvs). Tropical Agriculture. 1990;67:21-28.
Yadav P, Meena H, Meena P, Kumar A, Gupta R, Jambhulkar S, Rani R, Singh D. Determination of LD50 of ethyl methanesulfonate (EMS) for induction of mutations in rapeseed-mustard. Journal of Oilseed Brassica. 2016;1:77-82.
Rai MK, Kalia RK, Singh R, Gangola MP, Dhawan A. Developing stress tolerant plants through In vitro selection—an overview of the recent progress. Environmental and Experimental Botany. 2011;71:89-98.
Daub ME. Tissue culture and the selection of resistance to pathogens. Annual Review of Phytopathology. 1986;24:159-186.
Butterfield M, D’hont A, Berding N. The sugarcane genome: A synthesis of current understanding, and lessons for breeding and biotechnology. Proceedings of the South African Sugar Technologists Association. 2001;75:1-5.
Sengwayo S, Zhou M, Labuschagne M. Trends and magnitudes of genotype × environment interaction variance components for yield, quality and agronomic traits among coastal short cycle sugarcane breeding populations. South African Journal of Plant and Soil. 2017; 35:41-50.
Begum M, Islam M, Miah M, Hossain M, Islam N. Production of somaclone in vitro for drought stress tolerant plantlet selection in sugarcane (Saccharum officinarum L.). The Agriculturists. 2011;9:18-28.
Ferreira THS, Tsunada MS, Bassi D, Araújo P, Mattiello L, Guidelli GV, Righetto GL, Gonçalves VR, Lakshmanan P, Menossi M. Sugarcane water stress tolerance mechanisms and its implications on developing biotechnology solutions. Frontiers in Plant Science. 2017;8:1-18.
Ahuja I, de Vos RC, Bones AM, Hall RD. Plant molecular stress responses face climate change. Trends in Plant Science. 2010;15:664-674.
Ghaheri M, Kahrizi D, Bahrami G. Effect of mannitol on some morphological characteristics of In vitro Stevia rebaudiana Bertoni. Biharean Biologist. 2017;11:94-97.
Snyman S, Mhlanga P, Watt M. Rapid screening of sugarcane plantlets for In vitro mannitol-induced stress. Sugar Tech. 2016; 18:437-440.
Masoabi M, Lloyd J, Kossmann J, van der Vyver C. Ethyl methanesulfonate mutagenesis and In vitro polyethylene glycol selection for drought tolerance in sugarcane (Saccharum spp.). Sugar Tech. 2018;20:50-59.
Lutts S, Almansouri M, Kinet JM. Salinity and water stress have contrasting effects on the relationship between growth and cell viability during and after stress exposure in durum wheat callus. Plant Science. 2004; 167:9-18.
Beck CB. Structure and development of the cell wall. In "An introduction to plant structure and development: Plant anatomy for the twenty-first century". Cambridge Univerisity press, Cambridge, UK. 2012; 58-82.
Koiwa H. Pathways and genetic determinants for cell wall-based osmotic stress tolerance in the Arabidopsis thaliana root system. In "Genes for Plant Abiotic Stress" (Jenks MA, Wood AJ, eds.). Wiley-Blackwell. 2010;35-53.
Zingaretti SM, Rodrigues FA, Graça JP da, Pereira L de M, Lourenço MV. Sugarcane responses at water deficit conditions. In "Water Stress". In Tech. 2012;255-276.
Snyman SJ, Meyer GM, Koch AC, Banasiak M, Watt MP. Applications of In vitro culture systems for commercial sugarcane production and improvement. In vitro Cellular & Developmental Biology-Plant. 2011;47:234-249.
Mahlanza T, Rutherford RS, Snyman SJ, Watt MP. In vitro generation of somaclonal variant plants of sugarcane for tolerance to Fusarium sacchari. Plant Cell Reports. 2013;32:249-262.
Koch AC, Ramgareeb S, Rutherford RS, Snyman SJ, Watt MP. An In vitro mutagenesis protocol for the production of sugarcane tolerant to the herbicide imazapyr. In Vitro Cellular & Developmental Biology-Plant. 2012;48: 417-427.
Koetle MJ, Evans DL, Singh V, Snyman SJ, Rutherford RS, Watt MP. Agronomic evaluation and molecular characterisation of the acetolactate synthase gene in imazapyr tolerant sugarcane (Saccharum hybrid) genotypes. Plant Cell Reports. 2018;37:1201-1213.
Purnamaningsih R, Hutami S. Increasing Al-Tolerance of sugarcane using ethyl methanesulfonate and In vitro selection in the low pH media. Hayati Journal of Biosciences. 2016;23:1-6.
Sega GA. A review of the genetic effects of ethyl methanesulfonate. Mutation Research/Reviews in Genetic Toxicology. 1984;134:113-142.
Hofmann NE, Raja R, Nelson RL, Korban SS. Mutagenesis of embryogenic cultures of soybean and detecting polymorphisms using RAPD markers. Biologia Plantarum. 2004;48:173-177.
Grivet L, Arruda P. Sugarcane genomics: Depicting the complex genome of an important tropical crop. Current Opinion in Plant Biology. 2002;5:122-127.
Hassan NS, Shaaban LD, Hashem E, Seleem EE. In vitro selection for water stress tolerant callus line of Helianthus Annus L. cv. Myak. International Journal of Agriculture and Biology. 2004;6:13-18.
Zhou M. Conventional sugarcane breeding in South Africa: Progress and future prospects. American Journal of Plant Sciences. 2013;4:189-196.
Snyman SJ. Sugarcane transformation. In "Transgenic Crops of the World: Essential Protocols" (Curtis IS, ed.). Springer Netherlands, Dordrecht. 2004;103-114.
Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum. 1962;15:473-497.
Shekhawat GS, Verma K, Jana S, Singh KL, Teotia P, Prasad A. In vitro biochemical evaluation of cadmium tolerance mechanism in callus and seedlings of Brassica juncea. Protoplasma. 2009;239: 31-38.
Errabii T, Gandonou CB, Essalmani H, Abrini J, Idaomar M, Senhaji NS. Effects of NaCl and mannitol induced stress on sugarcane (Saccharum spp.) callus cultures. Acta Physiologiae Plantarum. 2007;29:95–102.
Taylor PWJ, Ko HL, Adkins SW, Rathus C, Birch RG. Establishment of embryogenic callus and high protoplast yielding suspension cultures of sugarcane (Saccharum spp. hybrids). Plant Cell, Tissue and Organ Culture. 1992;28:69-78.
Gocke E, Bürgin H, Müller L, Pfister T. Literature review on the genotoxicity, reproductive toxicity, and carcinogenicity of ethyl methanesulfonate. Toxology Letters. 2009;190:254–265.
Kenganal M, Hanchinal R, Nadaf H. Ethyl methanesulfonate (EMS) induced mutation and selection for salt tolerance in sugarcane In vitro. Indian Journal of Plant Physiology. 2008;13:405-410.
Khalil F, Naiyan X, Tayyab M, Pinghua C. Screening of EMS-induced drought-tolerant sugarcane mutants employing physiological, molecular and enzymatic approaches. Agronomy. 2018;8:11-13.
Svetleva L. Effect of ethyl methanesulfonate (EMS) and N-nitrose-N-ethyl Urea (ENU) on callus growth of common bean. Journal of Central European Agriculture. 2005;6:59-64.
Silué S, Diarrasouba N, Fofana IJ, Muhovski Y, Toussaint A, Mergeai G, Jacquemin JM, Baudoin JP. Description of Phaseolus vulgaris L. aborting embryos from ethyl methanesulfonate (EMS) mutagenized plants. Biotechnology, Agronomy, Society and Environment. 2013;17:563-571.
Novak F, Afza R, Duren MV, Omar M. Mutation induction by gamma irradiation of in vitro cultured shoot-tips of banana and plantain (Musa cvs). Tropical Agriculture. 1990;67:21-28.
Yadav P, Meena H, Meena P, Kumar A, Gupta R, Jambhulkar S, Rani R, Singh D. Determination of LD50 of ethyl methanesulfonate (EMS) for induction of mutations in rapeseed-mustard. Journal of Oilseed Brassica. 2016;1:77-82.
Rai MK, Kalia RK, Singh R, Gangola MP, Dhawan A. Developing stress tolerant plants through In vitro selection—an overview of the recent progress. Environmental and Experimental Botany. 2011;71:89-98.
Daub ME. Tissue culture and the selection of resistance to pathogens. Annual Review of Phytopathology. 1986;24:159-186.
Butterfield M, D’hont A, Berding N. The sugarcane genome: A synthesis of current understanding, and lessons for breeding and biotechnology. Proceedings of the South African Sugar Technologists Association. 2001;75:1-5.
Sengwayo S, Zhou M, Labuschagne M. Trends and magnitudes of genotype × environment interaction variance components for yield, quality and agronomic traits among coastal short cycle sugarcane breeding populations. South African Journal of Plant and Soil. 2017; 35:41-50.
Begum M, Islam M, Miah M, Hossain M, Islam N. Production of somaclone in vitro for drought stress tolerant plantlet selection in sugarcane (Saccharum officinarum L.). The Agriculturists. 2011;9:18-28.
Ferreira THS, Tsunada MS, Bassi D, Araújo P, Mattiello L, Guidelli GV, Righetto GL, Gonçalves VR, Lakshmanan P, Menossi M. Sugarcane water stress tolerance mechanisms and its implications on developing biotechnology solutions. Frontiers in Plant Science. 2017;8:1-18.
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