Flower color in the weedy perennial Plantago lanceolata is phenotypically plastic. Darker flowers are produced at cooler ambient temperatures, and circumstantial evidence suggests that this is adaptive. The goal of this project was to investigate the chemical basis for the color plasticity. To test the hypothesis that increased anthocyanin production at low temperatures underlies the plasticity, extracts of P. lanceolata flowers produced at warm and cool temperatures were analyzed using UV/visible spectrophotometry coupled with mass spectrometry. Mass spectrometry allowed us to compare relative abundances of individual anthocyanins. Seventeen anthocyanins, derived from both cyanidin and delphinidin branches of the anthocyanin biosynthetic pathway, were detected. Most of these significantly increased in abundance under cool conditions. Genotypes differed significantly in anthocyanin levels and in their sensitivity to temperature change. Genotypes that showed greater floral color plasticity tended to show also greater temperature sensitivity with respect to anthocyanin production. Data suggest that the temperature regulation of the anthocyanin biosynthetic pathway occurs both upstream and downstream of the divergence of the cyanidin and delphinidin branches. The degree of temperature sensitivity, i.e. phenotypic plasticity, appears to be controlled downstream, whereas the overall temperature effect appears to be controlled upstream.

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