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Purification and molecular characterization of a soybean polygalacturonase-inhibiting protein

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Abstract

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 endoPGs from Sclerotinia sclerotiorum and one endo-PG from Aspergillus niger). A possible involvement of soybean PGIP in plant defence against fungal pathogens is discussed.

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Abbreviations

IEF:

isoelectrofocusing

PCR:

polymerase chain reaction

PG:

polygalacturonase

PGIP:

polygalacturonase-inhibiting protein

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Authors and Affiliations

  1. Istituto di Patoiogia vegetale, via Gradenigo 6, I-35131, Padova, Italy

    F. Favaron & P. Alghisi

  2. Dipartimento di, Agrobiologia e Agrochimica, via S. Camille de Lellis, I-01100, Viterbo, Italy

    R. D'Ovidio & E. Porceddu

Authors
  1. F. Favaron
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  2. R. D'Ovidio
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  3. E. Porceddu
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  4. P. Alghisi
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Additional information

The nucleotide sequence data of soybean PGIP is registered in EMBL GeneBank and DDBJ nucleotide-sequence data bases under the accession number X78274

The authors gratefully acknowledge the assistance of Dr. C. Castiglioni (Istituto di Patologia vegetale, Padova University, Italy) for PGIP purification and Dr. G. Gay (Dipartimento di Agrobiologia e Agrochimica, Viterbo University, Italy) for reading the manuscript. This research was supported by National Research Council of Italy, special project RAISA, paper no. 1542.

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Favaron, F., D'Ovidio, R., Porceddu, E. et al. Purification and molecular characterization of a soybean polygalacturonase-inhibiting protein. Planta 195, 80–87 (1994). https://doi.org/10.1007/BF00206295

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  • DOI: https://doi.org/10.1007/BF00206295

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