title: Recovery from heat, salt and osmotic stress in Physcomitrella patens requires a functional small heat shock protein PpHsp16.4.
abstract: BACKGROUND
  Plant small heat shock proteins (sHsps) accumulate in response to various environmental stresses, including heat, drought, salt and oxidative stress. Numerous studies suggest a role for these proteins in stress tolerance by preventing stress-induced protein aggregation as well as by facilitating protein refolding by other chaperones. However, in vivo evidence for the involvement of sHsps in tolerance to different stress factors is still missing, mainly due to the lack of appropriate mutants in specific sHsp genes.
  
  
  RESULTS
  In this study we characterized the function of a sHsp in abiotic stress tolerance in the moss Physcomitrella patens, a model for primitive land plants. Using suppression subtractive hybridization, we isolated an abscisic acid-upregulated gene from P. patens encoding a 16.4 kDa cytosolic class II sHsp. PpHsp16.4 was also induced by salicylic acid, dithiothreitol (DTT) and by exposure to various stimuli, including osmotic and salt stress, but not by oxidative stress-inducing compounds. Expression of the gene was maintained upon stress relief, suggesting a role for this protein in the recovery stage. PpHsp16.4 is encoded by two identical genes arranged in tandem in the genome. Targeted disruption of both genes resulted in the inability of plants to recover from heat, salt and osmotic stress. In vivo localization studies revealed that PpHsp16.4 localized in cytosolic granules in the vicinity of chloroplasts under non stress conditions, suggesting possible distinct roles for this protein under stress and optimal growth.
  
  
  CONCLUSIONS
  We identified a member of the class II sHsp family that showed hormonal and abiotic stress gene regulation. Induction of the gene by DTT treatment suggests that damaged proteins may act as signals for the stress-induction of PpHsp16.4. The product of this gene was shown to localize in cytosolic granules near the chloroplasts, suggesting a role for the protein in association with these organelles. Our study provides the first direct genetic evidence for a role of a sHsp in osmotic and salt stress tolerance, and supports a function for this protein particularly during the stress recovery stage of P. patens.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Ruibal
authors.forename: Alexandra
authors.initials: A
authors.lastname: Castro
authors.forename: Valentina
authors.initials: V
authors.lastname: Carballo
authors.forename: László
authors.initials: L
authors.lastname: Szabados
authors.affiliation: Laboratorio de Biología Molecular Vegetal, Facultad de Ciencias, Universidad de la República, Iguá 4225, CP 11400 Montevideo, Uruguay. svidal@fcien.edu.uy.
authors.forename: Sabina
authors.initials: S
authors.lastname: Vidal
mesh_terms: D000040:Abscisic Acid / Q000494:pharmacology; D017360:Arabidopsis / Q000187:drug effects / Q000378:metabolism; D019068:Bryopsida / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000378:metabolism; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D017353:Gene Deletion; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D055786:Gene Knockout Techniques; D018390:Gene Targeting; D017343:Genes, Plant; D050888:Heat-Shock Proteins, Small / Q000235:genetics / Q000378:metabolism; D006358:Hot Temperature; D008970:Molecular Weight; D012275:Oryza / Q000187:drug effects / Q000378:metabolism; D009997:Osmotic Pressure / Q000187:drug effects*; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D012965:Sodium Chloride / Q000494:pharmacology*; D013312:Stress, Physiological / Q000187:drug effects*; D013997:Time Factors
chemical_list: D050888:Heat-Shock Proteins, Small; D010940:Plant Proteins; D012965:Sodium Chloride; D000040:Abscisic Acid