Nature of enhanced respiration during sprouting of aged potato seed-tubers

Respiration of 18-month-old Solarium tuberosum L. tubers was about 53% greater than that of 6-month-old tubers during sprouting at 23°C; yet, a significant loss of sprout vigor in the older tubers was apparent. Involvement of alternative oxidase (AO) in the age-induced difference in tuber respiration was assessed. AO was only detected in immunoblots if tissue disks from tubers were pre-incubated for 24 h prior to isolation of submitochondrial membrane particles (SMPs). No AO¹ was detected in SMPs from nonincubated tuber tissue of either age, indicating that it was not contributing to tuber respiration during sprouting as previously thought. Respiratory control and ADP/O ratios indicated that oxidative phosphorylation was fully coupled to electron transport in mitochondria isolated from 6- and 18-month-old tubers. Cytochrome c oxidase (EC 1.9.3.1) activities of intact mitochondria were also not affected by tuber age. The difference in respiration during sprouting was unique to whole tubers, as oxygen consumption by mitochondria from young and oid tubers was equal on a milligram protein basis. Sprouting 18-month-old tubers had 15% more mitochondrial protein per gram fresh weight than did 6-month-old tubers. Older tubers also produced more ATP than younger tubers prior to and during sprouting, through a fully coupled, Cyt-mediated respiratory pathway, reduced sprout vigor notwithstanding. From 5 to 10 days of sprouting, coinciding with development of the age-induced difference in whole-tuber respiration, ATP concentration in 18-month-old tubers increased to become 52% higher than that in 6-month-old tubers. ATP synthase (EC 3.6.1.34), assessed by SDS-PAGE and immunoblots of β- and oligomycin-sensitivity conferring protein-subunits, also increased as a proportion of SMP protein in older tubers during this period. Relative to 6-month-old tubers, the increased respiration and associated oxidative phosphorylation of 18-rnonth-old tubers during sprouting were probably in response to a lower adenylate energy charge (AEC) prior to sprouting (from 0 fo 5 days). From 5 to 10 days of sprouting, AEC of 18-rnonth-old tubers increased to equal that of 6-month-old tubers and the two tuber ages maintained the same AEC for the remainder of the 20-day sprouting interval. Higher respiration and lower AEC of older tubers in storage at 4°C, along with the fact that older tubers respired at a higher rate to achieve the same AEC as younger tubers during sprouting, indicate greater utilization of ATP by older tubers.