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ENVIRONMENTAL STRESS AND ADAPTATION

Cold Tolerance of C₄ photosynthesis in Miscanthus x giganteus: Adaptation in Amounts and Sequence of C₄ Photosynthetic Enzymes¹

Department of Crop Sciences, University of Illinois, Urbana, Illinois 61801–4730 (S.L.N., S.P.M., S.P.L.); and Department of Biological Sciences, University of Essex, Colchester, CO4 3SQ, United Kingdom (A.K.A.-S., C.A.R.)

Field-grown Miscanthus x giganteus maintains high photosynthetic quantum yields and biomass productivity in cool temperate climates. It is related to maize (Zea mays) and uses the same NADP-malic enzyme C₄ pathway. This study tests the hypothesis that M. x giganteus, in contrast to maize, forms photosynthetically competent leaves at low temperatures with altered amounts of pyruvate orthophosphate dikinase (PPDK) and Rubisco or altered properties of PPDK. Both species were grown at 25°C/20°C or 14°C/11°C (day/night), and leaf photosynthesis was measured from 5°C to 38°C. Protein and steady-state transcript levels for Rubisco, PPDK, and phosphoenolpyruvate carboxylase were assessed and the sequence of C₄-PPDK from M. x giganteus was compared with other C₄ species. Low temperature growth had no effect on photosynthesis in M. x giganteus, but decreased rates by 80% at all measurement temperatures in maize. Amounts and expression of phosphoenolpyruvate carboxylase were affected little by growth temperature in either species. However, PPDK and Rubisco large subunit decreased >50% and >30%, respectively, in cold-grown maize, whereas these levels remained unaffected by temperature in M. x giganteus. Differences in protein content in maize were not explained by differences in steady-state transcript levels. Several different M. x giganteus C₄-PPDK cDNA sequences were found, but putative translated protein sequences did not show conservation of amino acids contributing to cold stability in Flaveria brownii C₄-PPDK. The maintenance of PPDK and Rubisco large subunit amounts in M. x giganteus is consistent with the hypothesis that these proteins are critical to maintaining high rates of C₄ photosynthesis at low temperature.

¹ This work was supported in part by the U.S. Department of Agriculture National Research Initiative Competitive Grants Program (grant nos. 2000–35100–9057 and 2002–35100–12424 to S.P.L. and S.P.M.).

² Present address: King Abdul Aziz University, Education College, PO Box 1450, Jeddah, Saudi Arabia.

^* Corresponding author; e-mail stevel{at}life.uiuc.edu; fax 217–244–7563.

Received February 3, 2003; returned for revision February 27, 2003; accepted April 7, 2003.

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