OPTIMIZATION OF HORMONE COMPOSITION OF NUTRIENT MEDIUM FOR TRITICUM AESTIVUM L. CULTIVATION IN VITRO
E.D. Nikitina, V.N. Mukhin
Altai Research Institute of Agriculture, Nauchny gorodok, 35, Barnaul, 656910, Russia
Summary. The investigations were carried out to determine the optimal content of hormones in the differential nutrient medium providing the realization of morphogenetic potencies of immature embryos culture of four spring soft wheat varieties Skala, Spektr, Zarnitsa and Zhnitsa. Donor plants were grown in breeding greenhouses of arched type during winter-spring vegetation period. For callus induction the explants 1.5…1.7 mm in size were used, which were subsequently passed to Linsmaier and Skoog medium, possessing 0.8 % of agar, 3 % of sucrose and 2 mg/l of 2,4-dichlorophenoxyacetic acid (2,4-D). Cell cultures were incubated in darkness at the temperature 26 ± 1degrees. In 30…35 days in accordance with the scheme of complete factorial experiment of 3E2 type calluses were passed to differential medium supplemented with 2,4-D at the levels of 0.5, 2.5, 4.0 mg/l and with kinetin (6-furfurylaminopurine) at the levels of 0.5, 2.25 and 4.0 mg/l. Number of replications for each of 9 variants is four. As a result 20 mathematic models (4 varieties x 5 stages of regeneration) designed as polynomial equations of second order were obtained. The optimal values of factors are not equal for cultures with genotypes analyzed and for different regeneration stages. For callus tissues of varieties Skala and Spektr the optimum on kinetin for all regeneration stages was 0.5 ml/l except for the morphogenesis. Optimal values of 2,4-D for the same varieties, except for rhizogenesis of Spectr variety, were within 2.3…3.2 mg/l. For cell cultures of Zarnitsa and Zhnitsa they were 0.5…1.9 mg/l. Recommended concentration intervals are 0.5..1.9 mg/l of kinetin and 1.9…3.2 of 2,4-D. The level of exogenic hormones necessary for stem differentiation is lower than the one for root formation. When the hormone concentration was 0.5 mg/l the frequency of embryoido-hemmorhizogenesis was 87.7 %, of rhizogenesis – 12.3 %.
Keywords: spring soft wheat, immature embryos culture, morphogenesis, plants regeneration, growth regulators, complete factorial experiment of 3E2 type.
For citation: Nikitina E.D., Mukhin V.N. Optimization of hormone composition of nutrient medium for Triticum Aestivum l. cultivation in vitro // Dostizheniya nauki i tekhniki APK. 2015. V. 29. № 6. pp. 44-48 (In Russ)