PubMed · 7550639
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G Fiskesjö. 1995. Allium test.. https://doi.org/10.1385/0-89603-282-5%3A119
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Plasma membranes purified from onion roots contain two distinct NAD(P)H-dehydrogenases of 27 and 31 kDa that differ in their physicochemical properties, substrate specificities and inhibitors sensitivities. The 27-kDa enzyme used both NADH and NADPH as electron donors. The 31-kDa enzyme was fully specific for NADH and accounted for the bulk of NADH-ferricyanide oxidoreductase. We have used NADPH- and NADH-ferricyanide oxidoreductase activities as markers for investigating the orientation of the 27- and 31-kDa enzymes at the plasma membrane, respectively. These activities were assayed in right-side-out vesicles isolated by two-phase partition, inside-out vesicles obtained by treatment with the detergent Brij 58 and membranes permeabilized with Triton X-100. Upon addition of Brij 58 to right-side-out plasma membrane vesicles, both NADPH- and NADH-ferricyanide oxidoreductases were activated to the same degree as the plasma membrane H(+)-ATPase. Redox activities were similar when measured in the presence of either Brij 58 or Triton X-100. Our results demonstrate that both enzymes expose their catalytic sites toward the cytoplasmic side of the plasma membrane.
Outer epidermal cells from the basal, equatorial, near-apical, and apical regions of the third turgid onion (Allium cepa L. var. yellow, sweet Spanish) leaf base were treated (3 and 6 h in the dark = T3 and T6, respectively) with brassinolide (Br, a brassinosteroid plant growth regulator; effects on excised pieces compared with those in water controls: there were no statistical differences between the T3 and T6 results). Br induced increases in the volume and changes in morphologies of the major nucleoli to a greater extent than observed for major nucleoli in basal through near-apical controls. No major nucleoli were activated in control or Br-treated apical tissue. Minor nucleolar organizer regions in control and Br-treated tissue remained inactive in all locations. We propose that Br is a major ribosomal cistron regulator.