Purification, characterization, and molecular cloning of basic PR-1-type pathogenesis-related proteins from barley.
Partial amino acid sequences of two proteins, purified from barley leaves reacting hypersensitively to the powdery mildew fungus, showed a high degree of amino acid identity to the PR-1 proteins originally described in tobacco. The proteins, subsequently designated HvPR-1a and HvPR-1b, show apparent pI values of approximately 10.5 and 11, respectively and apparent M(r) 15,000. Independently, differential screening of a cDNA library prepared from barley leaves, exhibiting a compatible interaction with the powdery mildew fungus, resulted in isolation of cDNA species representing two PR-1 homologs. With the exception of one amino acid, the partial amino acid sequences of HvPR-1a and HvPR-1b are identical to internal sequences of the polypeptides derived from the two cDNA species. These derived polypeptides are each 164 amino acids long and both have putative N-terminal leader sequences of 24 amino acids. That these proposed leader sequences are functional is indicated by the observed occurrence of both proteins in the intercellular fluid. The proposed mature proteins (calculated M(r) 14,490 and 15,204) share 91% identical amino acids with each other and 56 to 74% with other PR-1 proteins. Northern blot hybridization and immunoblotting, respectively, show that both transcripts and both proteins accumulate following inoculation of susceptible and hypersensitivity resistant barley leaves with the powdery mildew fungus.
Bryngelsson
T
T
Department of Plant Breeding Research, Swedish University of Agricultural Sciences, Svalöv.
Sommer-Knudsen
J
J
Gregersen
P L
PL
Collinge
D B
DB
Ek
B
B
Thordal-Christensen
H
H
D000595:Amino Acid Sequence; D001203:Ascomycota / Q000502:physiology; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004247:DNA; D001467:Hordeum / Q000737:chemistry / Q000235:genetics*; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D010940:Plant Proteins; C053376:pathogenesis-related proteins, plant; D004247:DNA