Pigment Analyses

In the 2004 field and greenhouse experiments, leaf pigments were determined on the same leaflet used in the gs (n = 6). Ten leaf discs (total leaf area of 5 cm2) were cut using a puncher from a single leaf of individual control plants and plants treated with UV-B, then macerated with a few drops of 80% acetone in a pestle and mortar as described by Apostol and Zwiazek (2004). The extract was centrifuged at 1,500 g for 5 minutes. The supernatant was diluted with 10 mL of 80% acetone. Leaf pigments were estimated from absorbance measurements of the supernatant made on a Lambda 1 UV/Visible Spectrophotometer (Perkin-Elmer Corp., Wilton, CT). The chlorophyll concentrations were determined from absorbance at 664 nm and 647 nm (Graan and Ort, 1984). Carotenoid concentrations were determined from absorbance at 483, 663 and 638 nm following the procedure of Davies (1976). Anthocyanins were estimated from absorbance measurements at 534, 643, and 661 nm, as previously described by Lichtenthaler (1987).

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