New Phytol 2001, 151:743–751 CrossRef 38 Fravel D: Role of antib

New Phytol 2001, 151:743–751.CrossRef 38. Fravel D: Role of antibiosis in the biocontrol of plant diseases. Annu Rev Phytopathol 1988, 26:75–91.CrossRef 39. Whipps JM: Effect of media on growth

and interactions between a range of soil-borne glasshouse pathogens and antagonistic fungi. New Phytol 1987, 107:127–142.CrossRef 40. Doyle JJ, Doyle JL: Isolation of plant DNA from fresh tissue. Focus 1990, 12:13–15. 41. White TJ, Bruns T, Lee S, Taylor JW: Amplification and direct sequencing of TPX-0005 purchase fungal ribosomal RNA genes for phylogenetics. In PCR protocols: a guide to methods and applications. Edited by: Innis MA, Gelfand DH, Sninsky JJ, White TJ. Academic, New York; 1990:315–322. 42. Abarenkov K, Henrik Nilsson INK1197 mouse R, Larsson KH, Alexander IJ, Eberhardt U, Erland S, Høiland K, Kjøller R, Larsson E, Pennanen T, Sen R, Taylor AF, Tedersoo L, Ursing BM, Vrålstad T, Liimatainen K, Peintner U, Kõljalg U: The UNITE database for molecular identification of fungi–recent updates and future perspectives. New Phytol 2010, 186:281–285.PubMedCrossRef 43. Nonomura H, Hayakawa M, et al.: New methods for the selective isolation of soil actinomycets. In Biology of Actinomycetes’88. Edited by: Okami Y. Japan Scientific

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Wheat Roots. Appl Environ Microbiol 2003, 69:5603–5608.PubMedCrossRef 47. Debaud JC, Gay G: In vitro fruiting under controlled conditions of ectomycorrhizal fungus Hebeloma cylindrosporum associated with Pinus pinaster. New Phytol 1987, 105:429–435.CrossRef 48. Di Battista C, Selosse MA, Bouchard D, Stenström E, Le Tacon F: Variations in symbiotic efficiency, Phloretin phenotypic characters and ploidy level among different isolates of the ectomycorrhizal basidiomycete Laccaria bicolor strain S238. Mycol Res 1996, 100:1315–1324.CrossRef 49. Molina R, Palmer JG: Isolation, maintenance and pure culture manipulation of ectomycorrhizal fungi. In Methods and principles of mycorrhizal research. Edited by: Schenk NC. The American Phytopathological Society, St. Paul, MN, USA; 1982:115–129. 50. Davis BD, Mingioli ES: Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol 1950, 60:17–28.PubMed 51. Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 1962, 15:467–472.CrossRef 52. Epple P, Apel K, Bohlmann H: Overexpression of an endogenous thionin enhances resistance of Arabidopsis against Fusarium oxysporum. Plant Cell 1997, 9:509–520.PubMed Competing interests The authors declare to have no competing interests.

Pharm World Sci 23:148–152PubMedCrossRef 39 Lorefalt B, Toss G,

Pharm World Sci 23:148–152PubMedCrossRef 39. Lorefalt B, Toss G, Granerus AK (2007) Bone mass in elderly patients with Parkinson’s

disease. Acta Neurol Scand 116:248–254PubMedCrossRef 40. Cauley JA, Fullman RL, Stone KL, Zmuda JM, Bauer DC, Barrett-Connor E, Ensrud K, Lau EM, Orwoll ES (2005) Factors associated with the lumbar spine and proximal femur bone mineral density in older men. Osteoporos Int 16:1525–1537PubMedCrossRef 41. Kanis JA, Johnell O, Oden A, Johansson H, De Laet C, Eisman JA, Fujiwara S, Kroger H, McCloskey EV, Mellstrom D, Melton LJ, Pols H, Reeve J, Silman A, Tenenhouse A (2005) Smoking and fracture risk: a meta-analysis. Osteoporos Int 16:155–162PubMedCrossRef 42. Powers KM, Kay DM, Factor SA, Zabetian CP, Higgins DS, Samii A, Nutt JG, Griffith A, Leis B, Roberts JW, Martinez ED, Montimurro JS, Checkoway H, Payami H (2008) Combined effects of smoking, RepSox mw coffee, and NSAIDs on Parkinson’s disease risk. Mov Disord 23:88–95PubMedCrossRef”
“Calcium supplements have been used for decades in the prevention

and, as an adjuvant, for treatment of osteoporosis because low calcium intakes are frequent and have negative effects on bone health. There is an abundant literature showing the beneficial effects of an adequate calcium intake on the maintenance of bone mineral density (BMD) in adults, and on the slowing of the loss of BMD in the elderly. There is even some evidence that it has a moderate effect on fracture risk. In other words, the prescription of calcium supplements in the prevention of osteoporosis has its place, in so far as it causes no Selleck Alpelisib harm. Although a very high intake of 3–4 g per day is not recommended,

there is no proof that such intakes are harmful. Hypercalciuria in kidney stone formers and gastrointestinal intolerance are the only well-known contraindications. Fractional calcium absorption decreases with higher intakes and protects the body from excess intake, at least in part. Indeed, calcium supplementation had no safety restrictions. The negative effects of calcium supplements listed in the ADAM7 recent report of the Institute of Medicine of the US [1] include kidney stones, milk-alkali syndrome and hypercalcemia with its various consequences. But the risk of renal stones is not confirmed [2], that of hypercalcemia is not documented, and as for provoking the rare milk-alkali syndrome, it needs more than just a calcium supplement. If the same strict scientific parameters were applied for assessing the upper tolerable intake level of calcium (or the buy Tozasertib lowest observed adverse effect level), as for assessing the positive effects of calcium supplements on bone, it would be impossible to define an upper safety limit. New information on the possibility of negative cardiovascular effects puts a cloud in the so far quiet sky of calcium supplementation. In the paper by I. Reid et al.

PubMedCrossRef 19 Petersen C, Moller LB: Control of copper homeo

selleck chemical PubMedCrossRef 19. Petersen C, Moller LB: Control of copper homeostasis in Escherichia coli by a P-type ATPase, CopA, and a MerR-like transcriptional activator, CopR. Gene 2000, 261:289–298.PubMedCrossRef 20. Stoyanov JV, Hobman JL, Brown NL: CueR (YbbI) of Escherichia coli is a MerR family regulator controlling expression of the copper exporter CopA. Mol Microbiol

2001, 39:502–511.PubMedCrossRef 21. Reeve WG, Tiwari RP, Kale NB, Dilworth MJ, Glenn AR: ActP controls copper homeostasis in Rhizobium leguminosarum bv. viciae and Sinorhizobium melliloti preventing low-pH induced copper toxicity. Mol Microbiol 2002, 43:981–991.PubMedCrossRef 22. Kim JS, Kim MH, find more Joe MH, Song SS, Lee IS, Choi SY: The SctR of Salmonella enterica serovar Typhimurium encoding a homologue of the MerR protein is involved in the copper-responsive regulation of cuiD. FEMS Microbiol Lett 2002, 210:99–103.PubMedCrossRef 23. Brocklehurst KR, Hobman JL, Lawley B, Blank L, Marshall SJ, Brown NL, Morby AP: ZntR is a Zn(II)-responsive MerR-like transcriptional regulator of zntA in Escherichia coli. Mol Microbiol 1999, RGFP966 purchase 31:893–902.PubMedCrossRef 24. Outten CE, Outten FW, O’Halloran TV: DNA distortion mechanism for transcriptional activation by ZntR, a Zn(II)-responsive MerR homologue in Escherichia coli. J Biol Chem 1999, 274:37517–37524.PubMedCrossRef 25. Kidd SP, Brown NL: ZccR- a MerR-like regulator from Bordetella pertussis, which responds

to zinc, cadmium and cobalt. Biochem Biophys Res Comm 2003, 302:697–702.PubMedCrossRef 26. Checa SK, Espariz M, Perez Audero ME, Botta PE, Spinelli SV, Soncini FC: Bacterial sensing of and resistance to gold salts. Mol Microbiol 2007, 63:1307–1318.PubMedCrossRef 27. Changela A, Chen K, Xue Anidulafungin (LY303366) Y, Holschen J, Outten CE, O’Halloran TV, Mondragon A: Molecular basis of metal-ion selectivity and zeptomolar sensitivity by CueR. Science 2003, 301:1383–1387.PubMedCrossRef 28. Helmann JD, Ballard BT, Walsh CT: The MerR Metalloregulatory Protein Binds Mercuric

Ion as a Tricoordinate, Metal-Bridged Dimer. Science 1990, 248:946–948.CrossRef 29. Shewchuk LM, Verdine GL, Nash H, Walsh CT: Mutagenesis of the cysteines in the metalloregulatory protein MerR indicates that a metal-bridged dimer activates transcription. Biochemistry 1989, 28:6140–6145.PubMedCrossRef 30. Sambrook J, Fritsch EF, Maniatis T: Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York; 1989. 31. Gibson T: Studies on the Eppstein-Barr virus genome. University of Cambridge, Cambridge, U.K; 1984. 32. Stanssens P, Opsomer C, McKeown YM, Kramer W, Zabeau M, Fritz HJ: Efficient oligonucleotide-directed construction of mutations in expression vectors by the gapped duplex DNA method using alternating selectable markers. Nucleic Acid Res 1989, 17:4441–4454.PubMedCrossRef 33. Praszkier J, Wilson IW, Pittard AJ: Mutations affecting translational coupling between the rep genes of an IncB miniplasmid. J Bacteriol 1992, 174:2376–2383.PubMed 34.

Infect Immun 2000, 68:953–955 CrossRefPubMed 8 Sahly H, Podschun

Infect Immun 2000, 68:953–955.CrossRefPubMed 8. Sahly H, Podschun R, Oelschlaeger TA, Greiwe M, Parolis H, Hasty D, Kekow J, Ullmann U, Ofek I, Sela S: Capsule impedes adhesion Ulixertinib supplier to and invasion of epithelial cells by Klebsiella pneumoniae. Infect Immun 2000, 68:6744–6749.CrossRefPubMed 9. Schembri MA, CH5183284 Dalsgaard D, Klemm P: Capsule shields the function of short bacterial adhesins. J Bacteriol 2004, 186:1249–1257.CrossRefPubMed 10. Schembri MA, Blom J, Krogfelt KA, Klemm P: Capsule and fimbria interaction in Klebsiella pneumoniae. Infect Immun 2005, 73:4626–4633.CrossRefPubMed 11. Campos MA, Vargas MA, Regueiro V, Llompart CM, Albertí S, Bengoechea JA: Capsule

polysaccharide mediates bacterial resistance to antimicrobial peptides. Infect Immun 2004, 72:7107–7114.CrossRefPubMed 12. Llobet E, Tomás JM, Bengoechea JA: Capsule polysaccharide is a bacterial decoy for antimicrobial peptides. Microbiology 2008, 154:3877–3886.CrossRefPubMed 13.

Regueiro V, Campos MA, Pons J, Albertí S, Bengoechea JA: The uptake of a Klebsiella pneumoniae capsule polysaccharide Ro 61-8048 ic50 mutant triggers an inflammatory response by human airway epithelial cells. Microbiology 2006, 152:555–566.CrossRefPubMed 14. Regueiro V, Moranta D, Campos MA, Margareto J, Garmendia J, Bengoechea JA:Klebsiella pneumoniae increases the levels of Toll-like receptors 2 and 4 in human airway epithelial cells. Infect Immun 2009, 77:714–724.CrossRefPubMed Phosphoribosylglycinamide formyltransferase 15. Cortés G, Álvarez D, Saus C, Albertí S: Role of lung epithelial cells in defense against Klebsiella pneumoniae

pneumonia. Infect Immun 2002, 70:1075–1080.CrossRefPubMed 16. Cortés G, Borrell N, de Astorza B, Gómez C, Sauleda J, Albertí S: Molecular analysis of the contribution of the capsular polysaccharide and the lipopolysaccharide O side chain to the virulence of Klebsiella pneumoniae in a murine model of pneumonia. Infect Immun 2002, 70:2583–2590.CrossRefPubMed 17. Westphal O, Jann K: Bacterial lipopolysaccharides extraction with phenol-water and further applications of the procedure. Meth Carbohydrate Chem 1963, 5:83–91. 18. Hirschfeld M, Ma Y, Weis JH, Vogel SN, Weis JJ: Cutting edge: repurification of lipopolysaccharide eliminates signaling through both human and murine toll-like receptor 2. J Immunol 2000, 165:618–622.PubMed 19. Manthey CL, Perera PY, Henricson BE, Hamilton TA, Qureshi N, Vogel SN: Endotoxin-induced early gene expression in C3H/HeJ (Lpsd) macrophages. J Immunol 1994, 153:2653–2663.PubMed 20. Bitter T, Muir HM: A modified uronic acid carbazole reaction. Anal Biochem 1962, 4:330–334.CrossRefPubMed 21. Rahn A, Whitfield C: Transcriptional organization and regulation of the Escherichia coli K30 group 1 capsule biosynthesis ( cps ) gene cluster. Mol Microbiol 2003, 47:1045–1060.CrossRefPubMed 22.

(A) ATP levels in the

(A) ATP levels in the culture supernatant. ATP concentrations were determined and plotted against the incubation period. (B) ATP levels in the bacterial pellet. Total ATP levels in the Selleck GS-9973 bacterial pellet were normalized against OD600nm of each culture and plotted against the incubation time period. (C) Ratio of quantity of ATP in the culture supernatant to that of the bacterial cells. Acinetobacter junii cultures were spun down and separated into culture supernatant and cell pellet. ATP levels in each fraction were determined. The ratio of ATP from supernatant to that of bacterial cells from the same volumes

of cultures was plotted against the incubation period. Results are the average of 4 experiments and error bars represent standard deviations. Discussion We report here that ATP can be detected Dactolisib mw in the culture supernatant of a wide variety of bacterial species including both Gram-positive and Gram-negative bacteria of laboratory and clinical strains (Figure 2 and Table 5). The concentrations of extracellular ATP (from several nanomolar to several hundred nanomolar) were

much lower than the 1–5 mM reported for intracellular ATP [6–9], and total extracellular ATP represents up to 3 to 5% of that in bacterial culture (Figure 4). One LOXO-101 research buy noticeable exception is Acinetobacter junii AJ4970 that had ratios of extracellular to intracellular ATP > 0.5 (Figure 7C), suggesting that a significant portion of total ATP was present in the culture supernatant of this

bacterial strain. The extracellular ATP is unlikely an artifact due to any contamination of culture supernatant by bacterial cells since filtration did not reduce the ATP level (Figure 1). However, Adenosine triphosphate we have yet to establish the mechanism of how ATP was released into the culture medium. The simplest explanation is that ATP was released from dead and lysed bacteria. This explanation is plausible for low extracellular ATP levels when total extracellular ATP is less than 5% of the intracellular ATP levels; however, it cannot explain the high extracellular ATP levels observed with AJ4970 which has comparable quantities of extracellular ATP compared to the intracellular ATP (Figure 7C). In addition we have shown that live bacteria of both E. coli and Salmonella (but not dead bacteria or culture supernatant) are able to actively deplete ATP at approximately 5 μM/hr or 83 nM/min (Figure 5A and B) – a very high rate compared to the peak extracellular ATP concentration of 15 nM to 35 nM/OD600nm in E. coli and Salmonella cultures (Figure 4). Thus the quantity of ATP released into culture supernatant is likely to be much higher than that detected in the supernatant. Genetic analysis showed that ATP release is linked to cytochrome bo oxidases and thus argues against the bacterial cell death and lysis as the sole source of the extracellular ATP (Figure 4).


Ikeja 7-Cl-O-Nec1 ic50 1 – - – 1 – 1Cstr S. Ilala 2 – 1 – 2 – 1Sstr S. Kaapstad – 4 1 – 5 – 1Pstr, 1Sstr S. Kalamu 1 – - – 1 – - S. Kalina 2 – - – 2 – 1Cstr S. Kingston 2 3 – - 5 – 1Pstr, 1Cstr S. Kokomlemle 2 1 – - 3 – 1Pstr, 1Cstr S. Korlebu 2 – - – 2 2Cstr – S. Lagos 4 2 – - 6 2Pstr 1Ptet, 2Cstr S. Moero 1 – - – 1 – - S. Monschaui 1 1 – 3 5 3Hstr 1Pstr S. Muenster 17 6 3 11 37 1Camp, 1Cstr, 1Pnal, 1Hsul, 1Hstr 5Hstr, 6Cstr, 4Pstr, 2Sstr, 1Htet S. Nima 3 – - – 3 – - S. Nottingham 2 1 – - 3 – 1Pstr-tet S. Depsipeptide chemical structure Oranienburg 1 – - – 1 – 1Cstr S. Othmarschen 1 – - – 1 1Cstr – S. Ouakam – - 1 – 1 – 1Sstr S. Poona 2 1 – - 3 – 1Pstr, 2Cstr S. Rissen

1 – - – 1 – - S. Ruiru 8 – - – 8 1Cstr, 1Cstr-tet 3Cstr S. Saintpaul – 1 – - 1 – 1Ptet S. Salford 1 – - – 1 – - S. Schwarzengrund 1 3 – - 4 – 1Cstr , 3Pstr S. Senftenberg – 8 – 2 10 – 4Pstr, 2Pstr-tet, 1Pstr-sul-tet S.

Shangani – 1 – - 1 – 1Pstr -sul S. Soumbedioune 4 – - – 4 – 3Cstr S. Stanley – - – 1 1 – 1Hstr S. Stanleyville – 1 – - 1 – 1Pstr-tet S.Tennessee 3 – - – 3 – 1Cstr S. Trachau 1 1 – - 2 1Cstr 1Pstr S. Typhi – 1 – - 1 1Pstr – S. Typhimurium 3 4 – - 7 4Pamp-chl-str-sul-tmp, 3Cstr – S. Umbadah 1 – - – 1 – - S. Umbilo 1 – - – 1 – 1Cstr S. Urbana 13 1 2 – 16 1Cchl-tmp-nal-mec 4Cstr, 1Cstr-ftx, 2Cstr-tet, 1Cstr-cip, 1Pstr, 1Sstr S. Virchow 1 – - – 1 – 1Cstr S. Waycross 2 1 – - 3 1Cstr 1Cstr, 1Pcip Afatinib order S. Yoruba 1 – - – 1 – 1Cstr S. group B 4,5,12:-:- 1   – - 1 1Cstr-tet – S. group C 6,7,14:d:- 1 9 – - 10 – 5Pstr-sul, 4Pstr, 1Cstr S. group E 3,10:e,h:- 1 5 – - 6 – 1Pstr-sul-tet, 1Pstr, 1Cstr S. group G 13,22:z:- – - – 1 1 – 1Hstr Salmonella

enterica ssp. salamae 1 – - – 1 – - Total 159 192 8 24 383 52 247 (52%) (55%) (16%) (96%) (53%) (7%) (34%) aFor example, entry 7Pstr-tet, means that 7 isolates Oxalosuccinic acid from poultry feces were resistant/intermediate to streptomycin and tetracycline. Abbreviations: C, cattle feces; P, poultry feces; S, swine feces; H, hedgehog feces, amp, ampicillin; chl, chloramphenicol; str, streptomycin; sul, sulphonamides; tmp, trimethoprim; tet, tetracycline; nal, nalidixic acid; cip, ciprofloxacin; ftx, cefotaxime; mec, mecillinam. Figure 1 Pulsed-field gel analysis with Xba I (A) and Bln I (B) to assess the genetic similarity of the Salmonella isolates from animal and human feces from Burkina Faso. Fifty Salmonella strains belonging to serotypes Muenster (n = 20), Typhimurium (n = 17), Typhimurium var. Copenhagen (n = 3), Albany (n = 4), Virchow (n = 3) and Ouakam (n = 3) were analysed. FT = phage type. Antimicrobial resistance On the whole, 52 (14%) of the 383 Salmonella isolates were resistant to one or more antimicrobials tested: 23 of these were from the cattle, 23 from the poultry and 6 from the hedgehog feces (Table 1). The salmonella isolates from the swine feces were susceptible to the tested antimicrobials. Six isolates were multiresistant: 4 S. Typhimurium isolates from the poultry feces (ampicillin, chloramphenicol, streptomycin, sulfonamides and trimethoprim), 1 S.

J Antimicrob Chemother 2007,59(5):1001–1004 PubMedCrossRef 9 Vil

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Adv Drug Delivery Rev 2011, 63:730–747 CrossRef 25 Lvov Y, Deche

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Increases in body water were similar to the placebo and creatine

Increases in body water were similar to the placebo and MS-275 nmr creatine monohydrate groups. The vast majority of the improvement observed in the present study can most likely be attributed to the training protocol itself, rather than the supplementation. Since creatine ethyl ester supplementation showed a large increase

in serum creatinine levels throughout the study with no significant increase in serum and total muscle creatine content, it can be concluded that a large portion of the creatine ethyl ester was being degraded Evofosfamide price within the GI tract after ingestion. Furthermore, it appears that the skeletal muscle uptake of creatine ethyl ester uptake was not significant enough to increase skeletal muscle creatine levels without significant degradation to creatinine occurring. Acknowledgements We would like to thank the individuals that participated as subjects in this study. This study was supported by supplement donations from Labrada Nutritionals (Houston, TX) and AST Sport Science (Colorado Springs, CO) to Baylor University. Written consent for participation was obtained from all subjects. All researchers involved independently collected, analyzed, and interpreted the results from this study and have no financial interests concerning the outcome of the investigation. References 1. Greenhaff Blasticidin S molecular weight P: The nutritional biochemistry

of creatine. J Nutr Biochem 1997, 8:610–8.CrossRef 2. Bemben M, Lamont H: Creatine supplementation tetracosactide and exercise performance: Recent findings. Sports Med 2005, 35:107–25.CrossRefPubMed 3. Demant T, Rhodes E: Effects of creatine supplementation on exercise performance. Sports Med 1999, 28:49–60.CrossRefPubMed

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Photoperiod was 12 h with 350 μmol m−2 s−1 PPFD and temperature w

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. see more 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 Vismodegib 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, Oxymatrine 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 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.