Carotenoids in roots indicated the level of stress induced by mannitol and sodium azide treatment during the early stages of maize germination
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.
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