title: Elevated atmospheric CO2 and strain of rhizobium alter freezing tolerance and cold-induced molecular changes in alfalfa (Medicago sativa).
abstract: OBJECTIVE
  The objective of the study was to assess the impact of elevated CO2 in interaction with rhizobial strains on freezing tolerance and cold-induced molecular changes in alfalfa.
  
  
  METHODS
  Alfalfa inoculated with two different strains of rhizobium (A2 and NRG34) was grown and cold acclimated (2 weeks at 2 degrees C) under either 400 (ambient) or 800 micromol mol(-1) (elevated) CO2.
  
  
  RESULTS
  Plants acclimated under 400 micromol mol(-1) CO2 were more freezing tolerant than those maintained under 800 micromol mol(-1). Cryoprotective sugars typically linked with the acquisition of freezing tolerance such as sucrose, stachyose and raffinose increased in roots in response to low temperature but did not differ between CO2 treatments. Similarly high CO2 did not alter the expression of many cold-regulated (COR) genes although it significantly increased the level of transcripts encoding a COR gene homologous to glyceraldehyde-3-phosphate-dehydrogenase (GAPDH). A significant effect of rhizobial strain was observed on both freezing tolerance and gene expression. Plants of alfalfa inoculated with strain A2 were more freezing tolerant than those inoculated with strain NRG34. Transcripts of COR genes homologous to a pathogenesis-related protein (PR-10) and to a nuclear-targeted protein were markedly enhanced in roots of alfalfa inoculated with strain A2 as compared with strain NRG34. Transcripts encoding the vegetative storage proteins (VSPs) beta-amylase and chitinase were more abundant in roots of non-acclimated plants inoculated with strain NRG34 than with strain A2.
  
  
  CONCLUSIONS
  Taken together, the results suggest that elevated CO2 stimulates plant growth and reduces freezing tolerance. The acquisition of cold tolerance is also influenced by the rhizobial strain, as indicated by lower levels of expression of COR genes and sustained accumulation of VSP-encoding transcripts in alfalfa inoculated with strain NRG34 as compared with strain A2.
authors.affiliation: Agriculture and Agri-Food Canada, 2560, Soils and Crops Research and Development Centre, 2560 Hochelaga Boulevard, Quebec, Quebec, G1V 2J3, Canada. bertranda@agr.gc.ca
authors.forename: Annick
authors.initials: A
authors.lastname: Bertrand
authors.forename: Danielle
authors.initials: D
authors.lastname: Prévost
authors.forename: Francine J
authors.initials: FJ
authors.lastname: Bigras
authors.forename: Yves
authors.initials: Y
authors.lastname: Castonguay
mesh_terms: D000064:Acclimatization / Q000235:genetics / Q000502:physiology*; D001272:Atmosphere / Q000737:chemistry*; D050260:Carbohydrate Metabolism; D002245:Carbon Dioxide / Q000032:analysis* / Q000494:pharmacology*; D005615:Freezing; D018506:Gene Expression Regulation, Plant; D000455:Medicago sativa / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism / Q000382:microbiology*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000382:microbiology; D016962:Sinorhizobium meliloti / Q000145:classification* / Q000502:physiology*
chemical_list: D010940:Plant Proteins; D002245:Carbon Dioxide