Pea dehydrins: identification, characterisation and expression.
An antiserum raised against dehydrin from maize (Zea mays) recognised several polypeptides in extracts of pea (Pisum sativum) cotyledons. A cDNA expression library was prepared from mRNA of developing cotyledons, screened with the antiserum and positive clones were purified and characterised. The nucleotide sequence of one such clone, pPsB12, contained an open reading frame which would encode a polypeptide with regions of significant amino acid sequence similarity to dehydrins from other plant species. The deduced amino acid sequence of the pea dehydrin encoded by B12 is 197 amino acids in length, has a high glycine content (25.9%), lacks tryptophan and is highly hydrophilic. The polypeptide has an estimated molecular mass of 20.4 kDa and pI = 6.4. An in vitro synthesised product from the clone comigrates with one of the in vivo proteins recognised by the antiserum. A comparison of the pea dehydrin sequence with sequences from other species revealed conserved amino acid regions: an N-terminal DEYGNP and a lysine-rich block (KIKEKLPG), both of which are present in two copies. Unexpectedly, pea dehydrin lacks a stretch of serine residues which is conserved in other dehydrins. B12 mRNA and dehydrin proteins accumulated in dehydration-stressed seedlings, associated with elevated levels of endogenous abscisic acid (ABA). Applied ABA induced expression of dehydrins in unstressed seedlings. Dehydrin expression was rapidly reversed when seedlings were removed from the stress or from treatment with ABA and placed in water. During pea cotyledon development, dehydrin mRNA and proteins accumulated in mid to late embryogenesis. Dehydrin proteins were some of the most actively synthesised at about the time of maximum fresh weight and represent about 2% of protein in mature cotyledons.
Roberton
M
M
Division of Plant Industry, CSIRO, Camberra, ACT, Austria.
Chandler
P M
PM
D000040:Abscisic Acid / Q000494:pharmacology; D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015180:Electrophoresis, Gel, Two-Dimensional; D007887:Fabaceae / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D005786:Gene Expression Regulation / Q000187:drug effects; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D010946:Plants, Medicinal; D012689:Sequence Homology, Nucleic Acid
D010940:Plant Proteins; C070556:dehydrin proteins, plant; D000040:Abscisic Acid