Optimization Of Hormone Composition Of Nutrient Medium For In Vitro Efficient Regeneration Of Bread Wheat
E. D. Nikitina, V. N. Mukhin, L. P. Khlebova, A. V. Matsyura, O. V. Bychkova
Optimal values of phytohormones in the differential nutrient medium providing the efficient realization of morphogenetic potencies of four spring bread wheat varieties (Skala, Spectr, Zarnitsa and Zhnitsa) from immature embryo cultures have been determined. For callus induction explants 1.5 – 1.7 mmin size were used, which were subsequently passed to the medium by Linsmaier&Skoog possessing 0.8 % of agar, 3 % of sucrose and 2.0 mg l-1 dichlorophenoxyacetic acid (2,4-D). Cell cultures were incubated in darkness at the temperature 26±1 °?. 30 – 35 days after in accordance with the scheme of complete factorial experiment of 32 type calli were passed to differential medium supplemented with 2,4-D at levels 0.5; 2.5; 4.0 mg l-1 and with kinetin (6-furfurylaminopurine) at levels 0.5; 2.25 and 4.0 mg l-1. Number of replications for each of 9 variants was four. As a result, 20 mathematic models (4 varieties × 5 stages of regeneration) designed as polynomial quadric equation were obtained. On the ground of the analysis of models it was established that optimal values for factors are not equal both for cultures of genotypes analyzed and for different regeneration stages. For callus tissues of Skala and Spectr an optimal value of kinetin for all regeneration stages was 0.5 mg l-1 except for the frequency of morphogenesis. Optimal values of 2,4-D for the same varieties were within 2.3 – 3.2 mg l-1. For cell cultures of Zarnitsa and Zhnitsa recommended concentration intervals made up 1.3 – 2.2 mg l-1 on kinetin except for the frequency of rhizogenesis, and 1.9 – 2.7 on 2,4-D. The level of exogenous phytohormones necessary for stem differentiation was lower than the one for root formation. The dependence of morphogenesis results on the hormonal status of the explant has been discussed.