The characterization of the soybean polygalacturonase-inhibiting proteins (Pgip) gene family reveals that a single member is responsible for the activity detected in soybean tissues.
Polygalacturonase-inhibiting proteins (PGIPs) are leucine-rich repeat (LRR) proteins that inhibit fungal endopolygalacturonases (PGs). They are encoded by multigene families whose members show functional redundancy and subfunctionalization for recognition of fungal PGs. In order to expand the information on the structure and functional features of legume PGIP, we have isolated and characterized four members of the soybean Pgip gene family and determined the properties of the encoded protein products. Sequence analysis showed that these genes form two clusters: one cluster of about 5 kbp containing Gmpgip1 and Gmpgip2, and the other containing Gmpgip3 and Gmpgip4 within a 60 kb fragment of a separate BAC clone. Sequence diversification of the four members resides mainly in the xxLxLxx region that includes residues forming the beta-sheet B1. When compared with other legume Pgip genes, Gmpgip3 groups with the bean genes Pvpgip1 and Pvpgip2, suggesting that these genes are closer to the ancestral gene. At the protein level, only GmPGIP3 shows the capability to inhibit fungal PGs. The spectrum of inhibition of GmPGIP3 against eight different fungal PGs mirrors that of the PGIP purified from soybean tissues and is similar to that of the bean PvPGIP2, one of the most efficient inhibitors so far characterized. We also report that the four Gmpgip genes are differentially regulated after wounding or during infection with the fungal pathogen Sclerotinia sclerotiorum. Following fungal infection Gmpgip3 is up regulated promptly, while Gmpgip2 is delayed.
D'Ovidio
R
R
Dipartimento di Agrobiologia e Agrochimica, Università della Tuscia, via San Camillo De Lellis, s.n.c, 01100 Viterbo, Italy. dovidio@unitus.it
Roberti
S
S
Di Giovanni
M
M
Capodicasa
C
C
Melaragni
M
M
Sella
L
L
Tosi
P
P
Favaron
F
F
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000378:metabolism; D005658:Fungi / Q000378:metabolism / Q000502:physiology; D014644:Genetic Variation; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000502:physiology*; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011096:Polygalacturonase / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D013025:Glycine max / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology
D005656:Fungal Proteins; C081479:PGIP protein, plant; D010940:Plant Proteins; D011096:Polygalacturonase