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Isolation and Respiratory Measurements of Mitochondria from Arabidopsis thaliana
Published on: January 5, 2018
Respiratory Properties of Mitochondria from Rice Seedlings Germinated under Water and Their Changes during Air
1Laboratory for Plant Ecological Studies, Faculty of Science, Kyoto University, Kyoto 606, Japan.
Abstract:
Respiratory activities were compared among rice seedlings germinated in air for 6 days (aerobic seedlings), those germinated under water for 5 days (submerged seedlings), and those grown in air for 1 day after 5 days' submerged germination (air-adapted seedlings). The respiratory activity of the submerged seedlings increased rapidly on transfer to air and reached a plateau at 16 hours in air. Respiration of the submerged seedlings was as sensitive to cyanide as those of aerobic and air-adapted seedlings. 2,4-Dinitrophenol had no effect on the respiration of the submerged seedlings, but stimulated those of the other two types of seedlings. Mitochondria from three types of seedlings did not differ in the ADP/O ratio and the respiratory control ratio (RCR) when succinate was oxidized. However, mitochondria from submerged seedlings (submerged mitochondria) showed poor RCR of about unity when malate was oxidized. Both the rate of succinate oxidation and succinate dehydrogenase activity were low in submerged mitochondria, but increased during air adaptation. Although submerged mitochondria oxidized malate very slowly, this activity increased after exposure to air without any increase in malate dehydrogenase activity. When NAD(+) was added to submerged mitochondria, oxidation of malate was restored to the level of the aerobic controls. Addition of NAD(+) enhanced the state 3 rate in submerged mitochondria, and RCR recovered to nearly the same value as that of the aerobic controls. Similar effects of NAD(+) on 2-oxoglutarate oxidation were observed. All these defects in submerged mitochondria were repaired during air adaptation. These results suggest that NAD(+)-linked substrate oxidation was low in submerged mitochondria because of NAD(+) deficiency, and that the oxidation increased with an increasing level of NAD(+) during air adaptation.
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