Skip to main content
Log in

HPLC analyses of seed storage proteins reveal polymorphism in Italian common bean (Phaseolus vulgaris L.) ecotypes

  • Published:
Euphytica Aims and scope Submit manuscript

Abstract

Phaseolin (PHAS) and phytohemagglutinin (PHA) represent the main seed protein fractions in common bean. Currently, several ecotypes of common bean cultivated in marginal areas exist, above all in Southern Italy regions. Such germplasm could be lost due to the genetic uniformity of new cultivars. Moreover, the first step to obtain community recognition and denomination requires the full characterization of the available germplasm. In the present study, PHAS and PHA extracted in saline solutions from nine ecotypes of common bean ‘di Controne’ (Campania region) were analysed by anionic and cation exchange-high performance liquid chromatography (AE- and CE-HPLC), SDS-PAGE and IEF. The results obtained by AE-HPLC have allowed us either to standardize a high resolution, cost-effective and time-saving biochemical methodology or to compare the studied ecotypes with some cultivars of common bean most widely grown in Italy. The elution profiles of the nine ecotypes, already analyzed at the morpho-physiological and molecular levels, allowed us to differentiate the ecotypes either from a qualitative point of view (presence/absence of one or more peaks) or a quantitative one. For instance, the ecotypes of common bean ‘di Controne’ could be clearly differentiated from some commercial cultivars of dwarf and/or climbing bean. In contrast, the analysis of the electrophoresis patterns supplied less information.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Avitabile, A., A. Pentangelo, I. Giordano, A. Di Mauro & R. Rao, 1999. Caratterizzazione bioagronomica dell'ecotipo ‘Fagiolo di Controne’. Atti V Convegno Nazionale Biodiversità, ‘Biodiversità e sistemi ecocompatibili’, pp. 471-8. Caserta 9-10 Settembre 1999.

  • Bonetti, A., A. Miggiano, G. Dinelli & A. Lovato, 1995. Identification of bean (Phaseolus vulgaris L.) cultivars grown in Italy by field and electrophoresis tests: a comparative study. Seed Sci & Technol 23: 69-84.

    Google Scholar 

  • Bradford, M., 1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein dye binding. Analyt Biochem 72: 248-54.

    Article  PubMed  CAS  Google Scholar 

  • Brown, J.W.S., T.C. Osborn, F.A. Bliss & T.C. Hall, 1982. Bean lectins Part 2: Relationship between qualitative lectin variation in Phaseolus vulgaris L. and previous observations on purified bean lectins. Theor Appl Genet 62: 361-7.

    CAS  Google Scholar 

  • Cooke, R.J., 1995. Varietal identification of crop plants. In: J.H. Skerritt & R. Appels (Eds.), New Diagnostics in Crop Sciences, pp. 33-63, CAB INTERNATIONAL, Wallingford, UK.

    Google Scholar 

  • Gepts, P., T.C. Osborn, K. Rashka & F.A. Bliss, 1986. Phaseolin protein variability in wild forms and landraces of the common bean (Phaseolus vulgaris L.): evidence for multiple centers of domestications. Econ Bot 40: 451-68.

    CAS  Google Scholar 

  • Gepts, P. & F.A. Bliss, 1988. Dissemination pathways of common bean (Phaseolus vulgaris, Fabaceae) deduced from phaseolin electrophoretic variability: II Europe and Africa. Econ Bot 42: 86-104.

    Google Scholar 

  • Hubert, L.J., 1987. Assignment Methods in Combinational Data Analysis. Marcell Dekker, New York, p. 326.

    Google Scholar 

  • Laemmli, U.K., 1970. Cleavage of structure proteins during assembly of the head of Bacteriophage T4. Nature 227: 680-5.

    Article  PubMed  CAS  Google Scholar 

  • Limongelli, G.M., G. Laghetti, P. Perrino & A.R. Piergiovanni, 1996. Variation of seed storage proteins in landraces of common bean (Phaseolus vulgaris L.) from Basilicata, Southern Italy. Euphytica 92: 393-399.

    Article  CAS  Google Scholar 

  • Lioi, L., 1989a. Variation of the storage protein phaseolin in common bean (Phaseolus vulgaris L.) from the Mediterranean area. Euphytica 44: 151-155.

    Article  CAS  Google Scholar 

  • Lioi, L., 1989b. Geographical variation of phaseolin patterns in an old world collection of Phaseolus vulgaris. Seed Sci & Technol 17: 317-324.

    Google Scholar 

  • Lioi, L., 1991. Electrophoretic variation and geographical distribution of the seed protein phytohemagglutinin in cultivated Phaseolus vulgaris L. J Genet & Breed 45: 97-102.

    CAS  Google Scholar 

  • Mennella, G., M. Bianchi, V. Onofaro Sanajà, A. Tonini & V. Magnifico, 1995. Caratterizzazione biochimica e morfo-fisiologica di cultivar di Brassica oleracea L. Sementi Elette 6: 9-16.

    Google Scholar 

  • Mennella, G., A. Iori, V. Onofaro Sanajà & V. Magnifico, 1996. Broccoli and cauliflower cultivar identification through IE-HPLC seed protein analysis. Acta Horticulturae 407: 115-121.

    CAS  Google Scholar 

  • Mennella, G., A. D'Alessandro, V. Onofaro Sanajà, & V. Magnifico, 1998. Caratterizzazione biochimica di pomodoro San Marzano mediante tecniche elettroforetiche e cromatografiche.Italus Hortus 4: 36-41.

    Google Scholar 

  • Mennella, G., V. Onofaro Sanajà, A. Tonini, & V. Magnifico, 1999. Seed storage protein characterization of Solanum species and of cultivars and androgenetic lines of S. melongena L. by SDS-PAGE and AE-HPLC. Seed Sci & Technol 27: 23-35.

    Google Scholar 

  • Mennella, G., V. Onofaro Sanajà, A. D'Alessandro, R. Coppola & I. Poma, 2001. Tomato ecotype characterization by anionic exchange-high performance liquid chromatography analysis of endosperm seed proteins. Acta Horticulturae 546: 453-457.

    CAS  Google Scholar 

  • Osborn, T.C., M. Burow & H.A. Bliss, 1988. Purification and characterization of arcelin seed protein from common bean. Plant Physiol 86: 399-405.

    PubMed  CAS  Google Scholar 

  • Pustzai, A., E.M.W. Clarke, T.P. King & J.C. Stewart, 1979. Nutritional evaluation of kidney beans (Phaseolus vulgaris): chemical composition, lectin content, and nutritional value of selected cultivars. J Sci Food Agric 30: 843-848.

    Google Scholar 

  • Rohlf, F.J., 1993. NTSYS-pc Numerical Taxonomy and Multivariate Analysis System. Exeter Publication, Setauker, NY, p. 326.

    Google Scholar 

  • Romero Andreas, J., B.S. Yandell & F.A. Bliss, 1986. Bean arcelin I. Inheritance of a novel seed protein of Phaseolus vulgaris L. and its effect on seed composition. Theor Appl Genet 72: 123-128.

    Article  Google Scholar 

  • Singh, S.P., P. Gepts & D.G. Debouck, 1991. Races of common bean (Phaseolus vulgaris, Fabaceae). Econ Bot 45: 379-396.

    Google Scholar 

  • Smith, J.A. & S.L. Desborough, 1987. The endosperm seed protein solin: biochemical characterization, induction by ABA and species-specific subunits. Theor Appl Genet 74: 739-745.

    Article  CAS  Google Scholar 

  • Smith, J.S.C. & O.S. Smith, 1992. Fingerprinting crop varieties. Adv in Agron 47: 85-140.

    Article  CAS  Google Scholar 

  • Sokal, R. & C. Michener, 1958. A statistical method for evaluating systematic relationship. Univ Kan Sci Bull 38: 1409-1438.

    Google Scholar 

  • Thorne, J., Ch.O. Toro, J. Vargas & D.G. Debouck, 1995. Variability in Andean nuña common beans (Phaseolus vulgaris, Fabaceae). Econ Bot 49: 78-95.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mennella, G., Onofaro Sanajà, V., D'Alessandro, A. et al. HPLC analyses of seed storage proteins reveal polymorphism in Italian common bean (Phaseolus vulgaris L.) ecotypes. Euphytica 134, 85–95 (2003). https://doi.org/10.1023/A:1026127004224

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1026127004224

Navigation