Влияние мелатонина и обезвоживания на уровень пол и дыхание зародышей семян гороха, рост проростков и окислительную активность митохондрий эпикотилей

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Исследовали влияние предобработки 0.1 мкМ мелатонином и кратковременного обезвоживания набухающих семян на водный дефицит, уровень ПОЛ и дыхание зародышей, а также последующий рост эпикотилей проростков гороха (Pisum sativum L.) и окислительную активность выделенных из них митохондрий. Две группы зародышей одинакового возраста, но отличающихся по весу и по стадии развития исследовали отдельно: набухшие зародыши, но не прорвавшие оболочки семени (группа 1) и проклюнувшиеся зародыши (группа 2). Уровень водного дефицита зародышей 1 и 2 группы в отсутствие доступа воды повышался на 8.7 и 6.9% соответственно, что оказалось чувствительным для дыхательной активности зародышей. Было показано, что зародыши группы 2, прорвавшие оболочку семени и вступившие в контакт с кислородом воздуха, переживали резкий подъем дыхательной активности, который был сопряжен с повышением уровня ПОЛ, регистрируемого по накоплению продуктов, реагирующих с тиобарбитуровой кислотой. Кратковременное обезвоживание тормозило дыхание зародышей этой группы, связанное с активностью цитохромного пути, но не сопровождалось возникновением окислительного стресса, а напротив, снижало уровень ПОЛ. У набухших, но не прорвавших оболочку семени зародышей группы 1 также отсутствовал окислительный стресс при обезвоживании. Предобработка мелатонином существенно повышала активность цитохромного пути дыхания у обеих групп зародышей, как в контроле, так и у подвергнутых обезвоживанию семян. У митохондрий, выделенных из эпикотилей 5-дневных проростков, выросших из семян, предобработанных мелатонином и подвергнутых обезвоживанию, наблюдалась активация окисления НАД-зависимого субстрата (малата), как в контрольных, так и в стрессовых условиях. Также было показано, что предобработка мелатонином стимулировала рост эпикотилей проростков группы 2 во всех вариантах опытов, тогда как у проростков группы 1 только после действия обезвоживания. Сделан вывод, что предобработка семян мелатонином стимулировала процесс дыхания зародышей за счет активации цитохромного пути, а также активировала окисление малатамитохондриями эпикотилей проростков и действовала как стимулятор роста эпикотилей после обезвоживания. Впервые показано, что кратковременное обезвоживание тканей зародыша может не сопровождаться окислительным стрессом.

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作者简介

И. Генерозова

Федеральное Государственное бюджетное учреждение науки Институт физиологии растений им. К.А. Тимирязева Российской академии наук

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Email: igenerozova@mail.ru
俄罗斯联邦, Москва

С. Васильев

Федеральное Государственное бюджетное учреждение науки Институт физиологии растений им. К.А. Тимирязева Российской академии наук

Email: igenerozova@mail.ru
俄罗斯联邦, Москва

П. Буцанец

Федеральное Государственное бюджетное учреждение науки Институт физиологии растений им. К.А. Тимирязева Российской академии наук

Email: igenerozova@mail.ru
俄罗斯联邦, Москва

А. Шугаев

Федеральное Государственное бюджетное учреждение науки Институт физиологии растений им. К.А. Тимирязева Российской академии наук

Email: igenerozova@mail.ru
俄罗斯联邦, Москва

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2. Fig. 1. The effect of dehydration and melatonin treatment on the water deficiency of germ tissues of group 1 (1) and group 2 (2) 18-hour pea seeds. Experimental options: I – control seeds, II – seeds after treatment with melatonin, III – seeds after dehydration, IV – seeds after dehydration and treatment with melatonin. The figures show the arithmetic averages and their standard errors. Statistically significant differences between the mean values were determined using the Raglan test in the ANOVA program, the results marked with different letters are statistically significant at P < 0.05.

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3. Fig. 2. The effect of dehydration and melatonin treatment on the content of TBK-RP in the tissues of embryos of group 1 (1) and group 2 (2) of 18-hour pea seeds. Experimental options: I – control seeds, II – seeds after treatment with melatonin, III – seeds after dehydration, IV – seeds after dehydration and treatment with melatonin. The figures show the arithmetic averages and their standard errors. Statistically significant differences between the mean values were determined using the Raglan test in the ANOVA program, the results marked with different letters are statistically significant at P < 0.05.

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4. Fig. 3. The effect of dehydration and melatonin treatment on the rate of oxygen uptake by embryos of group 1 of 18-hour seeds: (1) – the rate of oxygen uptake by embryos (Vc); (2) – activity of the cytochrome respiratory pathway; (3) – maximum activity of the alternative respiratory pathway; (4) – residual respiration. Experimental options: I – control seeds, II – seeds after treatment with melatonin, III – seeds after dehydration, IV – seeds after dehydration and treatment with melatonin. The figures show the arithmetic averages and their standard errors. Statistically significant differences between the average values of the experimental and control samples were determined using the Tukey test in the ANOVA program, the results related to the corresponding substrate (1 or 2), marked with different letters, are statistically significant at P < 0.05.

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5. Fig. 4. The effect of dehydration and melatonin treatment on the rate of oxygen uptake by embryos of group 2 of 18-hour seeds: (1) – the rate of oxygen uptake by embryos (Vc); (2) – activity of the cytochrome respiratory pathway; (3) – maximum activity of the alternative respiratory pathway; (4) – residual respiration. Experimental options: I – control seeds, II–seeds after treatment with melatonin, III – seeds after dehydration, IV – seeds after dehydration and treatment with melatonin. The figures show the arithmetic averages and their standard errors. Statistically significant differences between the average values of the experimental and control samples were determined using the Tukey test in the ANOVA program, the results related to the corresponding substrate (1 or 2), marked with different letters, are statistically significant at P < 0.05.

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6. Fig. 5. The effect of dehydration and melatonin treatment on the epicotyl length of 5-day pea seedlings grown from seeds of group (1) and group 2 embryos (2). Experimental options: I – control seeds, II – seeds after treatment with melatonin, III – seeds after dehydration, IV – seeds after dehydration and melatonin treatments. The figures show the arithmetic averages and their standard errors. Statistically significant differences between the mean values were determined using the Raglan test in the ANOVA program, the results marked with different letters are statistically significant at P < 0.05.

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7. Fig. 6. The effect of dehydration and melatonin treatment on respiration and activity of various oxidation pathways of malate (a) and succinate (b) by the mitochondria of epicotyles of 5-day pea seedlings: (1) – the rate of oxidation of substrates in state 3 (V3); (2) – the rate of oxidation of substrates along the cytochrome pathway (Vcit); (3) – the rate of oxidation of substrates by an alternative pathway (Valt); (4) is the rate of residual respiration (Vost). Experimental options: I – control seeds, II–seeds after treatment with melatonin, III – seeds after dehydration, IV – seeds after dehydration and treatment with melatonin. The figures show the arithmetic averages and their standard errors. Statistically significant differences between the average values of the experimental and control samples were determined using the Tukey test in the ANOVA program, the results related to the corresponding substrate (1 or 2), marked with different letters, are statistically significant at P < 0.05.

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