On Methods of Isolation of Active, Tightly Coupled Mitochondria of Wheat Seedlings

Abstract

Tightly coupled mitochondria can be isolated from wheat seedlings 1) if young seedlings are used, 2) if the grinding medium is buffered and contains bovine serum albumin (BSA), 3) if the isolation procedure, including centrifugation is 10 to 11 minutes long. Mitochondria isolated in this manner oxidize α-ketoglutarate, malate and NADH. Respiratory control (R.C.) values are around 6, 3, and 10, respectively. ADP:O values with those substrates are 4.0, 2.9, and 1.7, respectively. Mitochondria are stable for 3 hours. It is suggested that the technique may be used for preparation of mitochondria for comparative studies since activities of these mitochondria are highly reproducible.