Photoperiod was 12 h with 350 μmol m−2 s−1 PPFD and temperature was cycled 23/20 °C (light/dark). Instantaneous whole-canopy gas exchange rate was measured using a LI-6400 (Li-Cor Inc., Lincoln, NE, USA) with a custom-made whole-shoot Arabidopsis cuvette (Fig. 1). Cuvette PPFD was maintained at 350 μmol m−2 s−1

PPFD, CO2 was maintained at 400 μmol mol−1, and temperature and relative humidity were set to growth chamber conditions. Each block was measured on a different day, 28–31 days after sowing. this website Following measurements for each plant, leaf area was determined from digital photographs of the rosette using Scion Image (Scion Corporation, Frederick, MD, USA). Fig. 1 Cuvette used for whole-plant gas exchange measurements. The cuvette is mounted on the LI-6400 IRGA and cuvette control system (gold-plated panel, fan and aluminum box, upper photograph). This system allows accurate, rapid measurement of CO2 (A) and H2O (E) exchange of whole shoots of Arabidopsis plants. The whole-plant cuvette incorporates a leaf temperature thermocouple that interfaces directly with the LI-6400. Intrinsic WUE (A/g s), stomatal conductance (g s), internal CO2 concentration (C i), and other variables can be calculated from

these measurements. All interior surfaces are Teflon coated or Ni-plated, the cuvette has extremely Selleck Belinostat low leak rates when operated in lab conditions with high external CO2, and the circular design provides excellent mixing using the LI-6400 fans. Plants can be rapidly changed using multiple inserts (lower photo) A:C i responses were measured for three accessions (Tsu-1, SQ-8, and Kas-1) which differed in A and δ13C. Cuvette conditions were the same as above, pheromone but light was increased to

1,000 μmol m−2 s−1 PPFD. Photosynthetic carbon dioxide response curves were measured on four rosettes of each accession. The number of replications of A:C i measurements were limited by chamber environment equilibration time at each CO2 set point. The least squares iterative curve-fitting procedure (Sharkey et al. 2007) model was used to fit Farquhar et al.’s (1980) biochemical model of photosynthesis and obtain DNA Damage inhibitor maximal carboxylation rate (V cmax) and maximal photosynthetic electron transport rate (Jmax). Leaf water content (Experiment 3) 39 natural accessions from the native range of Arabidopsis previously used in Mckay et al. (2003) were measured for LWC and leaf δ13C. Four replicates of each ecotype were grown in a greenhouse at UC Davis in a randomized block design. Seeds were sown in 250-mL pots in peat-based potting mix with slow-release fertilizer and vernalized at 4 °C for 5 days. Day length was extended to 16 h using supplemental lighting at 350 μmol m−2 s−1 PPFD. Greenhouse mean relative humidity and air temperature were 44 % and 23 °C, respectively.