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This study presents representative images of germinating barley seeds subjected to sterilization using 0.1% H2O2 and 0.1% HClO4 for 60 minutes. The successful germination of seeds indicates the effectiveness of the sterilization process. Additionally, we confirm the absence of bacterial growth in plated mitochondrial aliquots, suggesting no copurification of bacteria with mitochondria. The apparent NOS activity in isolated mitochondria was assessed through DAF fluorescence, showcasing the potential metabolic processes influenced by substrates and inhibitors.
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1 A 1B Representative picture of germinating seeds which were sterilized with 0.1% H2O2 for 60 min Representative picturesof germinating seeds of barley on vertical plates which were sterilized with 0.1% H2O2and 0.1% HClO4 for 60 min 1C Plating of aliquots of mitochondria did not lead to bacterial growth confirming there is no bacteria (if present) copurified from mitochondria
Fluorescence (A.U.) Supplementary fig 1: Apparent NOS activity in mitochondria as indicated by DAF fluorescence. Mitochondria isolated from sterile roots were incubated for 30 min in the presence of DAF-2 10µM and substrates or inhibitors as indicated (500 µM cPTIO, or 1 mM NADPH, 2 mM L-Arg and NOS inhibitor 5 mM L-NMMA.
