Department of Agronomy, University of Wisconsin, Madison, Wisconsin 53706.
Plant Physiol. 1977 Nov;60(5):716-22. doi: 10.1104/pp.60.5.716.
The influence of low temperature on soybean (Glycine max [L.] Merr. cv. Wells) energy transduction via mitochondrial respiration and dehydrogenases was investigated in this study during imbibition and germination. Mitochondria were isolated from embryonic axes of seeds treated at 10 and 23 C (control) by submergence in H(2)O for 6 hours and maintenance for an additional 42 hours in a moist environment. Arrhenius plots of initial respiration rates revealed that those from cold-treated axes had respiratory control (RC) ratios of near 1.0 above an inflection in the plot at 8 C. Arrhenius plots of control axes mitochondrial respiration showed RC ratios of 2.8 above and 5.0 below an inflection temperature of 12.5 C. Energies of activation for mitochondrial respiration between 20 and 30 C for the cold and control treatments were 7.8 and 15.6 kcal/mole, respectively. These data indicate possible differences in mitochondrial membranes, degree of mitochondrial integrity, and mitochondrial enzyme complement between the two treatments.Glutamate dehydrogenase (GDH), malate dehydrogenase (MDH), alcohol dehydrogenase (ADH), glucose-6-phosphate dehydrogenase (G6P-DH), and NADP-isocitrate dehydrogenase (NADP-ICDH) were assayed from whole seeds and axes (after germination) during the 48 hours of temperature treatments. Activity of these dehydrogenases decreased during the first 6 hours with the exception of MDH. After germination at 23 C (48 hours) all five dehydrogenases increased in activity. Arrhenius plots of cotyledon dehydrogenase activities indicated that one inflection temperature between 6 and 18 C was present for each enzyme assayed. Differences were seen in Arrhenius plots of axes dehydrogenase activities with the two temperature treatments in the cases of GDH and MDH from mitochondrial pellets and with differences in enzyme extraction media. These data suggest that the temperature treatments yield differences in mitochondrial enzyme complement. There were no detectable inflection temperatures for the activities of G6P-DH and ADH extracted from axes. Arrhenius plots of NADP-ICDH activity indicated extreme cold sensitivity. The slopes of the plots for axes NADP-ICDH were very similar to those for mitochondrial respiration (23 C treatment) suggesting that this enzyme may limit mitochondrial respiration at low temperature in soybean tissues grown at moderate temperatures.
本研究探讨了低温对大豆(Glycine max [L.] Merr. cv. Wells)在吸胀和萌发过程中线粒体呼吸和脱氢酶能量传递的影响。将种子的胚轴在 10 和 23°C(对照)下分别在水中浸泡 6 小时,并在潮湿环境中再维持 42 小时,然后从这些种子中分离出线粒体。初始呼吸速率的 Arrhenius 图表明,在 8°C 处出现拐点时,冷处理轴的呼吸控制(RC)比接近 1.0。对照轴线粒体呼吸的 Arrhenius 图显示,在 12.5°C 的拐点温度以上和以下,RC 比分别为 2.8 和 5.0。冷处理和对照处理下线粒体呼吸在 20 至 30°C 之间的活化能分别为 7.8 和 15.6 千卡/摩尔。这些数据表明,两种处理之间可能存在线粒体膜、线粒体完整性程度和线粒体酶组成的差异。谷氨酸脱氢酶(GDH)、苹果酸脱氢酶(MDH)、醇脱氢酶(ADH)、葡萄糖-6-磷酸脱氢酶(G6P-DH)和 NADP-异柠檬酸脱氢酶(NADP-ICDH)从整个种子和轴(萌发后)在 48 小时的温度处理中进行了测定。除 MDH 外,这些脱氢酶的活性在最初的 6 小时内下降。在 23°C(48 小时)下萌发后,所有五种脱氢酶的活性都增加。子叶脱氢酶活性的 Arrhenius 图表明,每种酶的拐点温度都在 6 至 18°C 之间。在 GDH 和 MDH 来自线粒体沉淀的情况下,以及在酶提取介质方面,在轴的脱氢酶活性的 Arrhenius 图中可以看到两种温度处理之间的差异。这些数据表明,温度处理导致线粒体酶组成的差异。从轴上提取的 G6P-DH 和 ADH 的活性没有检测到拐点温度。NADP-ICDH 活性的 Arrhenius 图表明其对低温极为敏感。轴 NADP-ICDH 图的斜率与线粒体呼吸(23°C 处理)非常相似,这表明该酶可能在中温下生长的大豆组织中限制低温下的线粒体呼吸。