Carotenoids in roots indicated the level of stress induced by mannitol and sodium azide treatment during the early stages of maize germination
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Chemical mutagens, such as sodium azide, have attracted the interest of plant breeders. Azide creates DNA point mutations and affects plant growth and development, disturbs metabolic activity and inhibits protein and DNA replication, whereas mannitol is used to simulate drought stresses in tissue culture. To identify biochemical markers for stress tolerance, maize seeds were germinated under mannitol and sodium azide induced stress in controlled conditions for 7 days. Then levels of chlorophyll, carotenoids, phenolics and aldehydes produced were subsequently determined. Germination percentage was not affected by either mannitol or sodium azide and was always above 85%. However, total fresh weight decreased by 50% with the application of 153.4 mM mannitol and 0.26 mM azide in combination. This treatment significantly reduced plantlet growth from 0.94 g in the control to 0.53 g in the treated materials. Root weight reduced by 68.1%, cotyledons by 14.3%, stems by 65.0% and leaves by 70.0% in treated samples. The level of carotenoids in roots was the clearest biochemical indicator of stress produced by the mannitol and sodium azide treatment. Carotenoids increased from 0.01 µg g− 1 fresh weight in the control to 9.03 µg g− 1 fresh weight in the treated materials. A large-scale seed treatment with mannitol and sodium azide was carried out. 2296 seeds were placed in magenta containers with 153.4 mM mannitol and 0.26 mM NaN3. At 7 days of germination, the heaviest seedlings (450) (450/2296 = 20%) were transferred to soil environment. Forty-two plants (42/450 = 9.3%) were off-type phenotypes at 45 days. Genetic variants may have been obtained following the novel procedure described here which combines chronic treatment with sodium azide and selection pressure with mannitol to simulate drought conditions.
KeywordsPlant breeding Induced mutagenesis Drought tolerance
Overall coefficient of variation
Analysis of variance
Reactive oxygen species
This research was supported by the Bioplant Center (University of Ciego de Avila, Cuba), the Thünen Institute of Biodiversity (Germany), The University of Sydney (Australia), and the National Institute for Agricultural Sciences (Cuba). Authors are grateful to Mrs. Julia Martínez for her excellent technical assistance.
Compliance with ethical standards
Conflict of interest
Authors do not have any conflict of interests.
Human and animal rights
This research did not involve experiments with human or animal participants.
Informed consent was obtained from all individual participants included in the study. Additional informed consent was obtained from all individual participants for whom identifying information is included in this article.
- Dubey S, Bist R, Misra S (2017) Sodium azide induced mutagenesis in wheat plant. World J Pharm Pharm Sci 6:294–304Google Scholar
- FAO/IAEA (2017) Joint FAO/IAEA Programme: Nuclear Techniques in Food and Agriculture (2017). http://www-naweb.iaea.org/nafa/pbg/mutation-breeding.html; Accessed Jan 2018
- Gomathi R, Rakkiyapan P (2011) Comparative lipid peroxidation, leaf membrane thermostability, and antioxidant system in four sugarcane genotypes differing in salt tolerance. Int J Plant Physiol Biochem 3:67–74Google Scholar
- Gómez D, Hernández L, Valle B, Martínez J, Cid M, Escalona M, Hernández M, Beemster GTS, Tebbe CC, Yabor L, Lorenzo JC (2017) Temporary immersion bioreactors (TIB) provide a versatile, cost-effective and reproducible in vitro analysis of the response of pineapple shoots to salinity and drought. Acta Physiol Plant. https://doi.org/10.1007/s11738-017-2576-5 CrossRefGoogle Scholar
- Gurr S, McPherson J, Bowles D (1992) Lignin and associated phenolic acids in cell walls. In: Wilkinson DL (ed) Molecular plant pathology. Oxford Press, Oxford, pp 51–56Google Scholar
- Hairuddin R, Yamin M, Hama S (2017) Application methods for conventional and modern development of onion (Allium cepa L.) in abiotic stresses. In: Junaid R (ed) Proceeding international conference on natural and social science, ICONSS. Palopo Cokroaminoto University, Makassar, pp B8-51–B8-57Google Scholar
- Hussain S, Khan WM, Khan MS, Akhtar N, Umar N, Ali S, Ahmed S, Shah SS (2017) Mutagenic effect of sodium azide (NaN3) on M2 generation of Brassica napus L. (variety Dunkled) Pure. Appl Biol 6:226–236Google Scholar
- ISTA (2005) International Rules for Seed Testing International Seed Testing Association, Bassersdorf, SwitzerlandGoogle Scholar
- Kothekar VS (2011) Effects of sodium azide on yield parameters of chickpea (Cicer arietinum L.). J Phytol 3:39–42Google Scholar
- Mendiondo GM, Gibbs DJ, Szurman-Zubrzycka M, Korn A, Marquez J, Szarejko I, Maluszynski M, King J, Axcell B, Smart K, Corbineau F, Holdsworth MJ (2016) Enhanced waterlogging tolerance in barley by manipulation of expression of the N-end rule pathway E3 ligase. Plant Biotechnol J 14:40–50CrossRefPubMedGoogle Scholar
- Mensah JK, Obadoni B (2007) Effects of sodium azide on yield parameters of groundnut (Arachis hypogaea L.). African J Biotech 6:668–671Google Scholar
- Olawuyi OJ, Okoli SO (2017) Genetic variability on tolerance of maize (Zea mays L.) genotypes induced with sodium azide mutagen. Mol Plant Breed 8:27–37Google Scholar
- Olawuyi OJ, Bello OB, Abioye AO (2016) Mutagenic effects of ultraviolet radiation on growth and agronomic characters in maize cultivars. Mol Plant Breed 7:1–10Google Scholar
- Salim K, Fahad A, Firoz A (2009) Sodium azide: a chemical mutagen for enhancement of agronomic traits of crop plants. Environ Int J Sci Tech 4:1–21Google Scholar
- Salvi S, Druka A, Milner SG, Gruszka D (2014) Induced genetic variation, TILLING and NGS-based cloning. In: Kumlehn J, Stein N (eds) Biotechnological approaches to barley improvement, biotechnology in agriculture and forestry 69. Springer, BerlinGoogle Scholar
- Szarejko I, Maluszynski M (1999) High frequency of mutations after mutagenic treatment of barley seeds with NaN3 and MNH with application of inter-incubation germination period. Mutat Breed Newslett 44:28–30Google Scholar
- Szarejko I, Szurman-Zubrzycka M, Nawrot M, Marzec M, Gruszka D, Kurowska M, Chmielewska B, Zbieszczyk J, Jelonek J, Maluszynski M (2017a) Creation of a TILLING population in barley after chemical mutagenesis with sodium azide and MNU. In: al. JJ-Ce (ed) Biotechnologies for plant mutation breeding. International Atomic Energy Agency, ViennaGoogle Scholar
- Szarejko I, Szurman-Zubrzycka M, Nawrot M, Marzec M, Gruszka D, Kurowska M, Chmielewska B, Zbieszczyk J, Jelonek J, Maluszynski M (2017b) In: al. JJ-Ce (ed) Biotechnologies for plant mutation breeding. International Atomic Energy Agency, ViennaGoogle Scholar