Theoretical and Applied Genetics

, Volume 107, Issue 5, pp 814–822 | Cite as

Lectin-related resistance factors against bruchids evolved through a number of duplication events

  • L. LioiEmail author
  • F. Sparvoli
  • I. Galasso
  • C. Lanave
  • R. Bollini


Abundant lectin-related proteins found in common beans (Phaseolus vulgaris L.) have been shown to confer resistance against the larvae of a number of bruchid species. Genes encoding for these proteins are members of the lectin multigene family, the most representative components being arcelins, phytohemagglutinins and α-amylase inhibitors. Arcelins have been described in seven variants, some of which are resistance factors against the Mexican bean weevil (Zabrotes subfasciatus), a major bean predator. In this study the isolation and sequencing of arcelin genes from wild P. vulgaris genotypes, containing Arc3 and Arc7 variants, is reported, and similarities and evolutionary relationships among the seven known arcelins are described. The evolutionary analysis shows that arcelins 3 and 4 cluster together and are the most-ancient variants. A duplication event gave rise to two additional clusters, one comprising arcelins 1, 2 and 6 and separated from the cluster of arcelins 5 and 7. A multiple number of arcelin genes were found in arcelin 3 and 4 genotypes indicating that more than one type of arcelin gene may be present in the same locus. Some of these sequences are reminiscent of ancient duplication events in arcelin evolution demonstrating that arcelins have evolved through multiple duplications. A further aim of this paper was to better understand and describe the evolution of the entire lectin multigene family. Beside arcelins, a number of other types of sequences, such as putative lectins and sequences not easily classifiable, were found in genotypes containing Arc3 and Arc4. These results, together with the evolutionary analysis, indicate that lectin loci are quite complex and confirm their origin by multiple duplication events.


α-Amylase inhibitor Arcelin Evolution Phytohemagglutinin Seed storage proteins 



The authors are grateful to Dr. A. Marina Torres, Genetic Resources Unit, CIAT, Cali, Colombia, for providing the P. vulgaris accessions used in this study. This work was partially supported by grants of the MiUR Italy project: Cluster C03-legge 488–92.


  1. Adachi J, Hasewaga M (1994) MOLPHY programs for molecular phylogenetics, ver. 2.1.2. Institute of Statistical Mathematics, TokyoGoogle Scholar
  2. Acosta-Gallegos JA, Quintero C, Vargas J, Toro O, Tohme J, Cardona C (1998) A new variant of arcelin in wild common bean, Phaseolus vulgaris L., from southern Mexico. Genet Res Crop Evol 45:235–242CrossRefGoogle Scholar
  3. Bollini R, Chrispeels MJ (1978) Characterization and subcellular localization of vicilin and phytohemagglutinin, the two major reserve proteins of Phaseolus vulgaris L. Planta 142:291–298Google Scholar
  4. Bollini R, Vitale A, Chrispeels MJ (1983) In vivo and in vitro processing of seed reserve protein in the endoplasmic reticulum: evidence for two glycosylation steps. J Cell Biol 96:999–1007PubMedGoogle Scholar
  5. Bompard-Gilles C, Rousseau P, Rougé P, Payan F (1996) Substrate mimicry in the active center of a mammalian α-amylase: structural analysis of an enzyme-inhibitor complex. Curr Biol 4:1441–1452Google Scholar
  6. Burnette WN (1981) "Western Blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 112:195–203PubMedGoogle Scholar
  7. Cardona C, Kornegay J, Posso CE, Morales F, Ramirez H (1990) Comparative value of four arcelin variants in the development of dry bean lines resistant to the Mexican bean weevil. Entomol Exp Appl 56:197–206Google Scholar
  8. Chrispeels MJ, Raikhel NV (1991) Lectins, lectin genes, and their role in plant defence. Plant Cell 3:1–9Google Scholar
  9. Debouck DG (2000) Biodiversity, ecology and genetic resources of Phaseolus beans – seven answered and unanswered questions. Proc 7th MAFF Int Workshop Genet Resour, Part 1 Wild Legumes, AFFRC and NIAR, JapanGoogle Scholar
  10. Finardi-Filho F, Mirkov E, Chrispeels MJ (1996) A putative precursor protein in the evolution of the bean α-amylase inhibitor. Phytochemistry 43:57–62CrossRefPubMedGoogle Scholar
  11. Galtier N, Gouy M, Gautier C (1996) SEAVIEW and PHYLO_WIN: two graphic tools for sequence alignment and molecular phylogeny. Comput Appl Biosci 12:543–548PubMedGoogle Scholar
  12. Gerhardt IR, Paes NS, Block Jr C, Mendes PAM, Leite A, Chrispeels MJ, Grossi de Sa MF (2000) Molecular characterization of a new arcelin-5 gene. Biochim Biophys Acta 1490:87–98CrossRefPubMedGoogle Scholar
  13. Goossens A, Geremia R, Bauw G, Van Montagu M, Angenon G (1994) Isolation and characterisation of arcelin-5 proteins and cDNAs. Eur J Biochem 225:787–795PubMedGoogle Scholar
  14. Hartweck LM, Vogelzang RD, Osborn TC (1991) Characterization and comparison of arcelin seed protein variants from common bean. Plant Physiol 97:204–211Google Scholar
  15. Huesing JE, Shade RE, Chrispeels MJ, Murdock LL (1991) α-Amylase inhibitor, not phytohemagglutinin, explains resistance of common bean seeds to cowpea weevil. Plant Physiol 96:993–996Google Scholar
  16. Ishimoto M, Sato T, Chrispeels MJ, Kitamura K (1996) Bruchid resistance of transgenic azuki bean expressing seed α-amylase inhibitor in common bean. Entomol Exp Appl 79:309–315Google Scholar
  17. Lanave C, Preparata G, Saccone C, Serio G (1984) A new method for calculating evolutionary substitution rates. J Mol Evol 20:86–93PubMedGoogle Scholar
  18. Lioi L, Bollini R (1989) Identification of a new arcelin variant in wild bean seeds. Annu Rep Bean Improv Coop 32:28Google Scholar
  19. Loris R, Hamerlick T, Bouckaert J, Wyns L (1998) Legume lectin structure. Biochim Biophys Acta 1383:9–36CrossRefPubMedGoogle Scholar
  20. Minney PHB, Gatehouse MRA, Dobie P, Dendy J, Cardona C, Gatehouse AJ (1990) Biochemical bases of seed resistance to Zabrotes subfasciatus (bean weevil) in Phaseolus vulgaris (common bean); a mechanism for arcelin toxicity. J Insect Physiol 36:757–767Google Scholar
  21. Mirkov TE, Wahlstrom JM, Hagiwara K, Finardi-Filho F, Kjemtrup S, Chrispeels MJ (1994) Evolutionary relationships among proteins in the phytohemagglutinin-arcelin-α-amylase inhibitor family of the common bean and its relatives. Plant Mol Biol 26:1103–1113PubMedGoogle Scholar
  22. Mirkov TE, Evans SV, Wahlstrom J, Gomez L, Young NM, Chrispeels MJ (1995) Location of the active site of the bean α-amylase inhibitor and involvement of a Trp, Arg, Tyr triad. Glycobiology 3:45–50Google Scholar
  23. Mourey L, Pédelacq JD, Birck C, Fabres C, Rougé P, Samama JP (1998) Crystal structure of the arcelin-1 dimer from Phaseolus vulgaris at 1.9 Å resolution. J Biol Chem 273:12914–12922CrossRefGoogle Scholar
  24. Nodari RO, Tsai SM, Gilbertson RL, Gepts P (1993) Towards an integrated linkage map of common bean. 2. Development of an RFLP-based linkage map. Theor Appl Genet 85:513–520Google Scholar
  25. Osborn TC, Blake T, Gepts P, Bliss FA (1986) Bean arcelin 2. Genetic variation, inheritance and linkage relationships of a novel seed protein of Phaseolus vulgaris L. Theor Appl Genet 71:847–855Google Scholar
  26. Osborn TC, Alexander DC, Sun SSM, Cardona C, Bliss FA (1988) Insecticidal activity and lectin homology of arcelin seed protein. Science 240:207–210Google Scholar
  27. Paes NS, Gerhardt IR, Coutinho MV, Yokoyama M, Santana E, Harris N, Chrispeels MJ, Grossi de Sa MF (2000) The effect of arcelin-1 on the structure of the midgut of bruchid larvae and immunolocalization of the arcelin protein. J Insect Physiol 46:393–402CrossRefPubMedGoogle Scholar
  28. Peumans WJ, Van Damme EJM (1995) The role of lectins in plant defense. Plant Physiol 109:347–352PubMedGoogle Scholar
  29. Rougé P, Barre A, Causse H, Chatelain C, Porthé G (1993) Arcelin and α-amylase inhibitor from seeds of common bean (Phaseolus vulgaris) are truncated lectins. Biochem Syst Ecol 21:695–703CrossRefGoogle Scholar
  30. Saccone C, Lanave C, Pesole G, Preparata G (1990) Influence of base composition on quantitative estimates of gene evolution. Methods Enzymol 183:570–583PubMedGoogle Scholar
  31. Santino A, Valsasina B, Lioi L, Vitale A, Bollini R (1991) Bean (Phaseolus vulgaris L.) seed lectins: a novel electrophoretic variant of arcelin. Plant Physiol (Life Sci Adv) 10:7–11Google Scholar
  32. Sharon N, Lis H (1990) Legume lectins – a large family of homologous proteins. FASEB J 4:3198–3208PubMedGoogle Scholar
  33. Sparvoli F, Bollini R (1998) Arcelin in wild bean (Phaseolus vulgaris L.) seeds: sequence of arcelin 6 shows it is a member of the arcelins 1 and 2 subfamily. Genet Res Crop Evol 45:383–388CrossRefGoogle Scholar
  34. Sparvoli F, Daminati MG, Lioi L, Bollini R (1996) In vivo endoproteolytically cleaved phaseolin is stable and accumulates in developing Phaseolus lunatus L. seeds. Biochim Biophys Acta 1292:15–22CrossRefPubMedGoogle Scholar
  35. Sparvoli F, Lanave C, Santucci A, Bollini R, Lioi L (2001) Lectin and lectin related proteins in Lima bean (Phaseolus lunatus L.) seeds: biochemical and evolutionary studies. Plant Mol Biol 45:587–597CrossRefPubMedGoogle Scholar
  36. Staswick P, Chrispeels MJ (1984) Expression of lectin genes during seed development in normal and phytoemagglutinin-deficient cultivars of Phaseolus vulgaris. J Mol Appl Genet 2:525–535PubMedGoogle Scholar
  37. Swofford DL (1998) PAUP*. Phylogenetic analysis using parsimony (*and other Methods). Version 4 Sinauer Associates, Sunderland, MassachusettsGoogle Scholar
  38. Tohme J, Gonzalez DO, Beebe S, Duque MC (1996) AFLP analysis of gene pools of a wild bean core collection. Crop Sci 36:1375–1384Google Scholar
  39. Tonelli C, Consonni SF, Dolfini SL, Dellaporta A, Viotti A, Gavazzi G (1991) Genetic and molecular analysis of Sn, a light-inducible, tissue-specific regulatory gene in maize. Mol Gen Genet 225:401–410PubMedGoogle Scholar
  40. Van Damme EJM, Barre A, Bemer V, Rougé P, Van Leuven, Peumans WJ (1995) A lectin and a lectin-related protein are the two most prominent proteins in the bark of yellow wood (Cladrastis lutea). Plant Mol Biol 29:579–598PubMedGoogle Scholar
  41. Van Damme EJM, Peumans WJ, Barre A, Rougé P (1998) Plant lectins: a composite of several families of structurally and evolutionary related proteins with diverse biological roles. Crit Rev Plant Sci 17:575–692CrossRefGoogle Scholar
  42. Vitale A, Ceriotti A, Bollini R (1985) Molecular analysis of a phytohemagglutinin-defective cultivar of Phaseolus vulgaris L. Planta 166:201–207Google Scholar
  43. Young NM, Thibault P, Watson DC, Chrispeels MJ (1999) Post-traslational processing of two α-amylase inhibitors and an arcelin from the common bean, Phaseolus vulgaris. FEBS Lett 446:203–206CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • L. Lioi
    • 1
    Email author
  • F. Sparvoli
    • 2
  • I. Galasso
    • 1
  • C. Lanave
    • 3
  • R. Bollini
    • 2
  1. 1.Istituto di Genetica Vegetale, CNR, Via Amendola 165/A, 70126 Bari, Italy
  2. 2.Istituto di Biologia e Biotecnologia Agraria, CNR, Via Bassini 15, 20133 Milano, Italy
  3. 3.Istituto di Tecnologie Biomediche, Sezione di Bioinformatica e Genomica, CNR, Via Amendola 168/5, 70126 Bari, Italy

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