Assessment of genetic relationships in Cucurbita pepo (Cucurbitaceae) using DNA markers

Abstract.

Cucurbita pepo (pumpkin, squash, gourd), an economically important species of the Cucurbitaceae, is extremely variable in fruit characteristics. The objective of the present study was to clarify genetic relationships across a broad spectrum of the C. pepo gene pool, with emphasis on domesticates, using AFLP, ISSR and SSR markers. Forty-five accessions were compared for presence or absence of 448 AFLP, 147 ISSR, and 20 SSR bands, their genetic distances (GDs) were estimated and UPGMA cluster analysis was conducted. The results obtained from these three marker systems were highly correlated (P ≪ 0.001). Clustering was in accordance with the division of C. pepo into three subspecies, fraterna, texana and pepo, with the first two less distant to one another than to the last one. Within the clusters, sub-clustering occurred in accordance with fruit shape and size. The subsp. texana cluster consisted of six sub-clusters, one each for the representatives of its five cultivar-groups (Acorn, Crookneck, Scallop, Straightneck and Ovifera Gourd) and wild gourds. Within the subsp. pepo cluster, the representatives of two cultivar-groups (Zucchini and Orange Gourd) formed distinct sub-clusters and the representatives of two other groups (Cocozelle and Vegetable Marrow) tended to sub-cluster separately from one another but formed an assemblage with the representatives of the remaining group (Pumpkin). Within-group GDs were less than corresponding between-group GDs in nearly all comparisons. The smallest-fruited accession, 'Miniature Ball', appears to occupy a genetically central position within C. pepo.

This is a preview of subscription content, access via your institution.

Fig. 1.
Fig. 2.

References

  1. Andres TC (1987) Cucurbita fraterna, the closest wild relative and progenitor of C. pepo. Cucurbit Genet Coop Rep 10:69–71

    Google Scholar 

  2. Baranek M, Stift G, Vollmann J, Lelley T (2000) Genetic diversity within and between the species Cucurbita pepo, C. moschata and C. maxima as revealed by RAPD markers. Cucurbit Genet Coop Rep 23:73–77

    Google Scholar 

  3. Bradeen JM, Staub JE, Wye C, Antonise R, Peleman J (2001) Towards an expanded and integrated linkage map of cucumber (Cucumis sativus L.). Genome 44:111–119

    Article  CAS  PubMed  Google Scholar 

  4. Brown RN, Myers JR (2000) Searching for molecular markers linked to ZYMV resistance in squash. Cucurbit Genet Coop Rep 23:69–70

    Google Scholar 

  5. Buntjer JB (2000) Phylogenetic computer tools, version 1.2. Wageningen University, The Netherlands

  6. Danin-Poleg Y, Tzuri G, Reis N, Katzir N (1998) Application of inter-SSR markers in melon (Cucumis melo L.). Cucurbit Genet Coop Rep 21:25–28

    Google Scholar 

  7. Danin-Poleg Y, Reis N, Tzuri G, Katzir N (2001) Development and characterization of microsatellite markers in Cucumis. Theor Appl Genet 102:61–72

    CAS  Google Scholar 

  8. Decker DS (1988) Origin(s), evolution, and systematics of Cucurbita pepo (Cucurbitaceae). Econ Bot 42:4–15

    Google Scholar 

  9. Decker DS, Wilson HD (1987) Allozyme variation in the Cucurbita pepo complex: C. pepo var. ovifera vs C. texana. Syst Bot 12:263–273

    Google Scholar 

  10. Decker-Walters DS, Staub JE, Chung SM, Nakata E, Quemada HD (2002) Diversity in free-living populations of Cucurbita pepo (Cucurbitaceae) as assessed by random amplified polymorphic DNA. Syst Bot 27:19–28

    Google Scholar 

  11. Duchesne AN (1786) Essai sur l'histoire naturelle des courges. Panckoucke, Paris

  12. Felsenstein J (1993) PHYLIP (Phylogeny Inference Package), version 3.5c. Department of Genetics, University of Washington, Seattle

  13. Ferriol M, Picó B, Nuez F (2001) Genetic variability in pumpkin (Cucurbita maxima) using RAPD markers. Cucurbit Genet Coop Rep 24:94–96

    Google Scholar 

  14. Garcia-Mas J, Oliver M, Gomez-Paniagua H, Vicente MC de (2000) Comparing AFLP, RAPD and RFLP markers for measuring genetic diversity in melon. Theor Appl Genet 101:860–864

    Article  CAS  Google Scholar 

  15. Gwanama C, Labuschagne MT, Botha AM (2000) Analysis of genetic variation in Cucurbita moschata by random amplified polymorphic DNA (RAPD) markers. Euphytica 113:19–24

    Article  CAS  Google Scholar 

  16. Ignart F, Weeden NF (1984) Allozyme variation in cultivars of Cucurbita pepo L. Euphytica 33:779–785

    CAS  Google Scholar 

  17. Katzir N, Danin-Poleg Y, Tzuri G, Karchi Z, Lavi U, Cregan PB (1996) Length polymorphism and homologies of microsatellites in several Cucurbitaceae species. Theor Appl Genet 93:1282–1290

    Article  CAS  Google Scholar 

  18. Katzir N, Leshzeshen E, Tzuri G, Reis N, Danin-Poleg Y, Paris HS (1998) Relationships among accessions of Cucurbita pepo based on ISSR analysis. In: McCreight JD (ed) Proc Cucurbitaceae '98, ASHS, Alexandria, Virginia, pp 331–335

  19. Katzir N, Tadmor Y, Tzuri G, Leshzeshen E, Mozes-Daube N, Danin-Poleg Y, Paris HS (2000) Further ISSR and preliminary SSR analysis of relationships among accessions of Cucurbita pepo. In: Katzir N, Paris HS (eds) Proc Cucurbitaceae 2000. Acta Hort 510:433–439

    CAS  Google Scholar 

  20. Mantel N (1967) The detection of disease clustering and a generalized regression approach. Cancer Res 27:209–220

    CAS  PubMed  Google Scholar 

  21. Naudin C (1856) Nouvelles recherches sur les caractères spécifiques et les variétés des plantes du genre Cucurbita. Ann Sci Natur Bot IV 6:5–73

    Google Scholar 

  22. Nee M (1990) The domestication of Cucurbita (Cucurbitaceae). Econ Bot 44 (supplement):56–68

    Google Scholar 

  23. Nei M, Li WH (1979) Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Natl Acad Sci USA 76:5269–5273

    CAS  PubMed  Google Scholar 

  24. Page RDM (1996) TREEVIEW: an application to display phylogenetic trees on personal computers. Comput Appl Biosci 12:357–358

    CAS  PubMed  Google Scholar 

  25. Paris HS (1986) A proposed subspecific classification for Cucurbita pepo. Phytologia 61:133–138

    Google Scholar 

  26. Paris HS (1989) Historical records, origins, and development of the edible cultivar groups of Cucurbita pepo (Cucurbitaceae). Econ Bot 43:423–443

    Google Scholar 

  27. Paris HS (2000) History of the cultivar-groups of Cucurbita pepo. In: J. Janick (ed) Hort Rev 25 (2001):71–170. John Wiley, New York

    Google Scholar 

  28. Paris HS (2001) Characterization of the Cucurbita pepo collection at the Newe Ya'ar Research Center, Israel. Plant Genet Res Newslett 126:41–45

    Google Scholar 

  29. Paris HS, Nerson H (1986) Genes for intense fruit pigmentation of squash. J Hered 77:403–409

    Google Scholar 

  30. Périn C, Hagen L, Dogimont C, Conto V de, Lecomte L, Pitrat M (2000) Construction of a reference genetic map of melon. In: Katzir N, Paris HS (eds) Proc Cucurbitaceae 2000. Acta Hort 510:367–374

    CAS  Google Scholar 

  31. Powell W, Morgante M, Andre C, Hanafey M, Vogel J, Tingey S, Rafalaski A (1996) The comparison of RFLP, RAPD, AFLP and SSR (microsatellite) markers for germplasm analysis. Mol Breed 3:225–238

    Google Scholar 

  32. Sanjur OI, Piperno DR, Andres TC, Wessel-Beaver L (2002) Phylogenetic relationships among domesticated and wild species of Cucurbita (Cucurbitaceae) inferred from a mitochondrical gene: implications for crop plant evolution and areas of origin. Proc Natl Acad Sci USA 99:535–540

    Article  CAS  PubMed  Google Scholar 

  33. Schneider S, Roessli D, Excoffier L (2000) Arlequin: a software for population genetics data analysis. Ver 2.000. Genetics and Biometry Lab, Dept Anthropology, University of Geneva

  34. Sneath PHA, Sokal RR (1973) Numerical taxonomy: the principles and practice of numerical classification. W.H. Freeman Co., San Francisco, California

  35. Stachel M, Csanádi G, Vollmann J, Lelley T (1998) Genetic diversity in pumpkins (Cucurbita pepo L.) as revealed in inbred lines using RAPD markers. Cucurbit Genet Coop Rep 21:48–50

    Google Scholar 

  36. Staub JE, Serquen FC, Gupta M (1996) Genetic markers, map construction, and their application in plant breeding. HortScience 31:729–741

    CAS  Google Scholar 

  37. Staub JE, Danin-Poleg Y, Fazio G, Horejsi T, Reis N, Katzir N (2000) Comparative analysis of cultivated melon groups (Cucumis melo L.) using random amplified polymorphic DNA and simple sequence repeat markers. Euphytica 115:225–241

    Article  CAS  Google Scholar 

  38. Torres Ruiz RA, Hemleben V (1991) Use of ribosomal DNA spacer probes to distinguish cultivars of Cucurbita pepo L. and other Cucurbitaceae. Euphytica 53:11–17

    Google Scholar 

  39. Vos P, Hogers R, Bleeker M, Reijans M, Lee T van de, Hornes M, Frijters A, Pot J, Peleman J, Kuiper M, Zabeau M (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res 23:4407–4414

    CAS  PubMed  Google Scholar 

  40. Whitaker TW, Bemis WP (1964) Evolution in the genus Cucurbita. Evolution 18:553–559

    Google Scholar 

  41. Wilson HD, Doebley J, Duvall M (1992) Chloroplast DNA diversity among wild and cultivated members of Cucurbita (Cucurbitaceae). Theor Appl Genet 84:859–865

    CAS  Google Scholar 

  42. Zietkiewicz E, Rafalski A, Labuda D (1994) Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20:176–183

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements.

For providing seed samples gratis, we thank M. Manoah, A.R.O. Volcani Center, Bet Dagan, Israel; the United States North Central Regional Plant Introduction Station, Ames, Iowa; G. Santini, S.A.I.S., Cesena, Italy; G. P. Silvestri, C.C.S.A., Fidenza, Italy; Y. Asherov, Institute of Vegetable Seed Quality, Samarqand, Uzbekistan; H. D. Wilson, Texas A& M University, College Station; L. R. Oliver, University of Arkansas, Fayetteville; T. C. Andres, Bronx, New York; and P. S. Lichtenecker, Vienna, Austria. The financial support of M. E. Seeds of Qiryat Nordau, Netanya, Israel, is gratefully acknowledged. Contribution No. 141/01 from the Institute of Field and Garden Crops, Agricultural Research Organization, Bet Dagan, Israel.

Author information

Affiliations

Authors

Corresponding author

Correspondence to H. S. Paris.

Additional information

Communicated by H. Nybom

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Paris, H.S., Yonash, N., Portnoy, V. et al. Assessment of genetic relationships in Cucurbita pepo (Cucurbitaceae) using DNA markers. Theor Appl Genet 106, 971–978 (2003). https://doi.org/10.1007/s00122-002-1157-0

Download citation

Keywords.

  • Pumpkin
  • Squash
  • Gourd
  • Diversity
  • Taxonomy
  • Genetic distance
  • AFLP
  • ISSR
  • SSR