Purification and molecular characterization of a soybean polygalacturonase-inhibiting protein.
A polygalacturonase-inhibiting protein (PGIP) was detected in soybean (Glycine max (L.) Merr.) seedlings. The protein was purified from germinating seeds and appeared to consist of at least three components with very close molecular weights (between 37 and 40 kDa) but each showing a unique N-terminal sequence. Primers specific for N-terminal and C-terminal nucleotide sequences of field bean (Phaseolus vulgaris L.) PGIP were used in a polymerase chain reaction (PCR) on soybean DNA, and only one amplification band was obtained. The amplified product was cloned and one of the PCR clones was sequenced. The nucleotide sequence comprises 942 bp with a single open reading frame which encodes a polypeptide of 313 amino-acid residues with a predicted molecular weight of 33984 Daltons and an isoelectric point of 8.21. Analysis of genome organization showed a single gene copy of PGIP with few related sequences, and wounding of soybean hypocotyls showed a strong induction of expression of the PGIP gene. The PGIP showed different activities toward three purified fungal endo-polygalacturonases (endo-PGs) (two endo-PGs from Sclerotinia sclerotiorum and one endo-PG from Aspergillus niger). A possible involvement of soybean PGIP in plant defence against fungal pathogens is discussed.
Favaron
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F
Istituto di Patologia vegetale, Padova, Italy.
D'Ovidio
R
R
Porceddu
E
E
Alghisi
P
P
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002852:Chromatography, Ion Exchange; D018744:DNA, Plant; D004591:Electrophoresis, Polyacrylamide Gel; D007887:Fabaceae / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics / Q000302:isolation & purification*; D010946:Plants, Medicinal; D011096:Polygalacturonase / Q000037:antagonists & inhibitors*; D017386:Sequence Homology, Amino Acid; D013025:Glycine max / Q000737:chemistry* / Q000235:genetics
D018744:DNA, Plant; C081479:PGIP protein, plant; D010940:Plant Proteins; D011096:Polygalacturonase