STUDIES ON RESPIRATORY ENZYMES AND OXYGEN UPTAKE BY MITOCHONDRIA OF CMS SYSTEMS OF SORGHUM

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Date
2002
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Mahatma Phule Krishi Vidyapeeth, Rahuri.
Abstract
The mam objective of the present investigation was to study some biochemical parameters associated with CMS phenotype m sorghum using isolated mitochondria Mitochondria were isolated from the etiolated seedlings of the two CMS systems of sorghum viz, 1049A, 1049B, RSV 33R, SPH-1229 and 104A, 104B, RS-585, CSH-15R The assays of soluble proteins and respiratory enzymes viz, cytochrome c oxidase and succinate dehydrogenase were earned out from isolated mitochondria Further, the total respiration, cyanide-sensitive, cyanide-insensitive and residual respiration were measured from the isolated mitochondria and etiolated seedlings of these lines The protocol for the isolation and punfication of mitochondiral DNA was also standardized. The isolated mitochondna from both the CMS lmes, 1049 A and 104A, contained higher soluble protems than their respective isonuclear mamtamer, restorer and hybnd lmes The cyanide-sensitive respiration was measured by using SHAM, a potential inhibitor of alternative oxidase, while cyanide-insensitive respiration was measured by using KCN, a potential inhibitor of cytochrome c oxidase TUe cyanide-sensitive respiration was higher, whereas cyanide-insensitive respiration was lower in isolated mitochondria and etiolated seedlings of CMS lines than their corresponding mamtamer, restorer and hybrid lines When both inhibitors were added simultaneously in the reaction media, residual respiration was observed in isolated mitochondria and etiolated seedlings of both the CMS systems The cytochrome c oxidase and succinate dehydrogenase activities in both the CMS lines were lower than their respective isonuclear mamtamer lines and also restorer and hybrid lines The CMS system 104A, 104B, RS-585 and CSH-15R showed comparatively higher cytochrome c oxidase and succinate dehydrogenase activities than the other CMS system, 1049A, 1049B,RSV-33Rand SPH-1229 The higher protein content, cyanide-sensitive respiration and lower cytochrome c oxidase activity in CMS lines than their isonuclear lines indicates that mutation might have occurred in the mitochondrial DNA which probably resulted in the synthesis of new proteinĀ® Specifically the respiratory complex IV might have been mutated because some of the subuntis of this complex are encoded by the mitochondrial genome, reflecting the alteration in the cytochrome c oxidase activity and cyamde sensitive respiration The lower cyanide-insensitive respiration and succinate dehydrogenase activity in CMS lines than their corresponding isonuclear maintamer lines suggested that it might be the result of the CMS character and not the cause of CMS, because alternative oxidase and succinate dehydrogenase are encoded by the nuclear DNA The lower alternate oxidase activity m CMS than its isonuclear lme may perhaps be correlated to the higher yield or yield contributing characters That's why the hybrid CSH-15R yields more than the hybrid SPH-1229.
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