doi: 10.15389/agrobiology.2024.3.525eng

UDC: 633.1:58.04:58.085



N.A. Lagmetova1 , Z.M. Alieva1, K.U. Kurkiev2,
M.Kh. Gadjimagomedova2

1Dagestan State University, 43а, ul. Magometa Gadzhieva, Makhachkala, Republic of Dagestan, 367000 Russia, e-mail (✉ corresponding author),;
2Dagestan Experimental Station — Branch of the Federal Research Center Vavilov All-Russian Institute of Plant Genetic Resources, s. Vavilovo, Derbent District, Republic of Dagestan, 368600 Russia, e-mail,

Lagmetova N.A.
Kurkiev K.U.
Alieva Z.M.
Gadjimagomedova M.Kh.

Final revision received November 14, 2023
Accepted  March 28, 2024

Studies of the salt resistance of × Triticosecale Wittmack and the identification of optimal cultivars in this regard are relevant due to the widespread saline soils in the areas where this crop is grown. Biotechnological methods attract attention among the methods for assessing salt resistance, however, they have practically not been developed for triticale, and the varietal specificity of its response to salinity in vitro has not been sufficiently studied. This work for the first time submits data on the dependence of callus formation, crude and dry biomass, the accumulation of proline in callus tissues and the intensity of lipid peroxidation on the 0.5, 0.75 and 1 % NaCl in the nutrient medium. In addition, we revealed the varietal specific changes in these indicators under salinization in winter triticale samples. The aim of the work was to study the effect of different levels of NaCl salinity on morphophysiological and biochemical changes in in vitro calluses of five winter triticale varieties. Mature embryos of grain samples of winter triticale Triskell, Sotnik, PRAG530l-1934, Timbo and Almaz (the collection of the Dagestan Experimental Station, a branch of the FRC Vavilov All-Russian Institute of Plant Genetic Resources) were used. To initiate in vitro culture, grains were sequentially sterilized for 30 s in 96 % ethanol and 20 min in the commercial preparation Belizna, and then washed 3 times for 5 min with sterile distilled water. The embryos isolated from grains were placed with a cut down on a nutrient medium to form a callus. Four variants of the Murashige-Skuga nutrient medium (MS) differed in NaCl concentration were MS added with 2,5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 0,5 mg/l 6-benzylaminopurine (BAP) (MS1, control); MS + 2,4-D + BAP + 0.5 % NaCl (MS2); MS + 2,4-D + BAP + 0.75 % NaCl (MS3); MS + 2,4-D + BAP + 1 % NaCl (MS4). Mature embryos were cultures on nutrient media for 30 days in a climatic chamber (MLR-352H, Sanyo, Japan, a 16-hour photoperiod, 24±1 °C, illumination of 3000 lux and humidity of 80 %). The size of calluses, the growth of raw and dry biomass, the accumulation of free proline, and the intensity of lipid peroxidation (LP) were determined. In the experiments, the Timbo and Almaz cultivars showed the greatest sensitivity to salinity in vitro and produced calli only at a low NaCl concentration (0.5 %). The cultivars Sotnik and PRAG530l-1934 withstood an average salinity level (0.75 % NaCl). The Triskell sample turned out to be resistant, since callus formation occurred even at 1% NaCl). Sodium chloride added at a concentration of 0.75 % led to a decrease in the callus size of the Triskell, Sotnik and PRAG530l-1934 cultivars by 1.8, 1.9 and 2 times, respectively, vs. the control (р < 0.05). On the MS4 nutrient medium, the size of the callus in the Triskell sample decreased 2.9 times (р < 0.05). On MS2, the lowest decrease in crude callus biomass occurred in Triskell samples (1.6-fold vs. control, р < 0.05)) and Almaz (1.7-fold vs. control, р < 0.05), and the largest was in Timbo (3-fold vs. control, р < 0.05). On MS3, Triskell, PRAG530l-1934 and Sotnik cultivars showed a decrease in crude biomass vs. control by 2.1, 2.5 and 2.5 times (р < 0.05), respectively. High rates dry biomass accumulation on MS2 were characteristic of the Triskell and Sotnik samples, 62 and 57 % vs. control, р < 0.05). On MS3, the largest increase was found in Triskell (43 %, р < 0.05), the smallest in PRAG530l-1934 (23 %, р < 0.05) and Sotnik (25 %, р < 0.05). On MC4, the dry callus biomass of the Triskell was 21 % vs. control. Proline accumulation in calli was revealed on nutrient media with an excessive NaCl content, the most intense in a resistant Triskell sample. With 0.5, 0.75 and 1 % NaCl, prolin concentrations increased by 4.9, 6.2 and 6.9 times (р < 0.05), respectively. In sensitive Almaz and Timbo cultivars, there was an increase in lipid peroxidation. The most sustainable Triskell cultivar was characterized by generally high growth rates, intensive accumulation of proline and lower LP activity.

Keywords: Triticosecale Wittmack, triticale, callus, salt tolerance, chloride salinization, in vitro.



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