Активність та вміст фітогормонів-антагоністів у первинних калюсах ізогенних ліній сої з контрастною фотоперіодичною реакцією

Abstract

У роботі представлено результати дослідження вмісту та активності фітогормонів – антагоністів ІОК та АБК в первинних калюсах ізогенних за генами Е-серії ліній сої Glycine max (L.) Merr, які різняться за ступенем фотоперіодичної реакції. Виявлено, що на процес первинного калюсогенезу впливає рівень активності та вмісту рістконтролюючих фітогормонів ІОК та АБК. Показник відношення ІОК/АБК, який відображає фітогормональний баланс, корелює із частотою калюсогенезу та залежить від генотипу і фотоперіодичної реакції ізолінії.

В работе представлены результаты исследования содержания и активности фитогормонов – антагонистов ИУК и АБК в первичных каллусах изогенных по генам Е-серии линий сои Glycine max (L.) Merr., которые различаются по степени фотопериодической реакции. Выявлено, что на процесс первичного каллусогенеза влияет уровень активности и содержания ростконтролирующих фитогормонов ИУК и АБК. Показатель отношения ИУК/АБК, который отражает фитогормональный баланс, коррелирует с частотой каллусогенеза и зависит от генотипа и фотопериодической реакции изолинии.

Aim. The aim of work was to study the activity and content of main phytohormones that regulate auxin and ABA growth in primary callus isogenic lines of soybean Glycine max (L.) Merr. (NILs), differing in photoperiodic response. Five NILs genotypes, which differ on loci E-genes and photoperiodic response were the material of the study. Methods. In the paper, there were used standard biotechnological methods for getting primary callus culture, thin layer chromatography (TLC) methods for distribution and determination of phytohormones (IAA and ABA) and biotesting methods for research activity phytohormones. Results. The results of the study of frequency primary callusogenesis in soybean isogenic lines with contrasting photoperiodic response showed the difference in frequency callusogenesis between isolines is observed only during 4-7 days in the first week of cultivation. By morphological features, callus tissue was a compact, watered, yellowish, rapidly growing callus. Determination of activity phytohormones-antagonists in primary callus NILs soybean showed that the activity of IAA was higher than the activity of the hormone-antagonist ABA in all isolines regardless of their photoperiodic response. Maximum activity IAA was found in line Clark with shortday reaction, and the minimum - in the isoline L 94-1110 with photoperiodic neutral reaction. As the culture medium doesn’t contain native auxins we can assume that endogenous auxin, which is synthesized by primary callus tissues, determines their level of activity. It is a higher level of activity of auxin in shortday isolines, determines their faster rate of the callusogenesis. ABA activity in the callus is slightly lower than auxin in all investigated NILs. Though the connection between photoperiodic response isolines and ABA activity was not found. The results of the study of hormones antagonistic content in primary callus showed that the IAA content far exceeds ABA growth inhibition hormone. Growing, watered, not morphogenic callus characterized by intense growth callus cells both by intense proliferation, and by the expense a specific form of plant cell growth - "Growth stretching," which is determined by IAA physiological effect. It is established, those callus varieties Clark and isolines L 63-3016, L 65-3366, which show shortday photoperiodic response, are characterized by the maximum content of IAA. Isolines with photoperiodic neutral reaction are characterized by lower content IAA in callus. By the content of ABA investigated isolines don’t differ significantly, with the exception of Clark, which has a maximum content of both IAA and ABA. The phytohormonal balance indicator of IAA / ABA content for hormone-antagonists is clearly correlated with the photoperiodic response: SD isolines are characterized by higher value comparing to photoperiodic neutral genotypes. The results suggest that E-series genes can be involved in the callusogenesis control indirectly through their phytohormonal balance determination. Conclusions. Thus, in the course of the studies, we found that IAA activity in primary callus of isogenic soybean lines is higher than the active hormone antagonist - ABA. It is shown that growth stimulation phytohormon content is significantly higher than growth inhibition content in the rapidly growing, watered, not morphogenic callus of all investigated isogenic lines. The ratio of IAA / ABA reflects the phytohormonal balance in primary callus tissues and correlates with the frequency callusogenesis, depending on genotype and photoperiodic response isoline. NILs with a shortday photoperiodic response is characterized by more intense callusogenesis and high rate of IAA / ABA comparing to photoperiodic neutral isolines.