Theoretical and Applied Genetics

, Volume 106, Issue 6, pp 971–978 | Cite as

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

  • H. S. ParisEmail author
  • N. Yonash
  • V. Portnoy
  • N. Mozes-Daube
  • G. Tzuri
  • N. Katzir


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.


Pumpkin Squash Gourd Diversity Taxonomy Genetic distance AFLP ISSR SSR 



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.


  1. Andres TC (1987) Cucurbita fraterna, the closest wild relative and progenitor of C. pepo. Cucurbit Genet Coop Rep 10:69–71Google 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 23:73–77Google 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 44:111–119CrossRefPubMedGoogle Scholar
  4. Brown RN, Myers JR (2000) Searching for molecular markers linked to ZYMV resistance in squash. Cucurbit Genet Coop Rep 23:69–70Google Scholar
  5. Buntjer JB (2000) Phylogenetic computer tools, version 1.2. Wageningen University, The NetherlandsGoogle Scholar
  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 21:25–28Google Scholar
  7. Danin-Poleg Y, Reis N, Tzuri G, Katzir N (2001) Development and characterization of microsatellite markers in Cucumis. Theor Appl Genet 102:61–72Google Scholar
  8. Decker DS (1988) Origin(s), evolution, and systematics of Cucurbita pepo (Cucurbitaceae). Econ Bot 42:4–15Google Scholar
  9. Decker DS, Wilson HD (1987) Allozyme variation in the Cucurbita pepo complex: C. pepo var. ovifera vs C. texana. Syst Bot 12:263–273Google 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 27:19–28Google Scholar
  11. Duchesne AN (1786) Essai sur l'histoire naturelle des courges. Panckoucke, ParisGoogle Scholar
  12. Felsenstein J (1993) PHYLIP (Phylogeny Inference Package), version 3.5c. Department of Genetics, University of Washington, SeattleGoogle Scholar
  13. Ferriol M, Picó B, Nuez F (2001) Genetic variability in pumpkin (Cucurbita maxima) using RAPD markers. Cucurbit Genet Coop Rep 24:94–96Google 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 101:860–864CrossRefGoogle Scholar
  15. Gwanama C, Labuschagne MT, Botha AM (2000) Analysis of genetic variation in Cucurbita moschata by random amplified polymorphic DNA (RAPD) markers. Euphytica 113:19–24CrossRefGoogle Scholar
  16. Ignart F, Weeden NF (1984) Allozyme variation in cultivars of Cucurbita pepo L. Euphytica 33:779–785Google 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 93:1282–1290CrossRefGoogle 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 331–335Google Scholar
  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 510:433–439Google Scholar
  20. Mantel N (1967) The detection of disease clustering and a generalized regression approach. Cancer Res 27:209–220PubMedGoogle Scholar
  21. Naudin C (1856) Nouvelles recherches sur les caractères spécifiques et les variétés des plantes du genre Cucurbita. Ann Sci Natur Bot IV 6:5–73Google Scholar
  22. Nee M (1990) The domestication of Cucurbita (Cucurbitaceae). Econ Bot 44 (supplement):56–68Google Scholar
  23. Nei M, Li WH (1979) Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Natl Acad Sci USA 76:5269–5273PubMedGoogle Scholar
  24. Page RDM (1996) TREEVIEW: an application to display phylogenetic trees on personal computers. Comput Appl Biosci 12:357–358PubMedGoogle Scholar
  25. Paris HS (1986) A proposed subspecific classification for Cucurbita pepo. Phytologia 61:133–138Google Scholar
  26. Paris HS (1989) Historical records, origins, and development of the edible cultivar groups of Cucurbita pepo (Cucurbitaceae). Econ Bot 43:423–443Google 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 YorkGoogle Scholar
  28. Paris HS (2001) Characterization of the Cucurbita pepo collection at the Newe Ya'ar Research Center, Israel. Plant Genet Res Newslett 126:41–45Google Scholar
  29. Paris HS, Nerson H (1986) Genes for intense fruit pigmentation of squash. J Hered 77:403–409Google 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 510:367–374Google 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 3:225–238Google 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 99:535–540CrossRefPubMedGoogle 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 GenevaGoogle Scholar
  34. Sneath PHA, Sokal RR (1973) Numerical taxonomy: the principles and practice of numerical classification. W.H. Freeman Co., San Francisco, CaliforniaGoogle Scholar
  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 21:48–50Google Scholar
  36. Staub JE, Serquen FC, Gupta M (1996) Genetic markers, map construction, and their application in plant breeding. HortScience 31:729–741Google 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 115:225–241CrossRefGoogle 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 53:11–17Google 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 23:4407–4414PubMedGoogle Scholar
  40. Whitaker TW, Bemis WP (1964) Evolution in the genus Cucurbita. Evolution 18:553–559Google Scholar
  41. Wilson HD, Doebley J, Duvall M (1992) Chloroplast DNA diversity among wild and cultivated members of Cucurbita (Cucurbitaceae). Theor Appl Genet 84:859–865Google Scholar
  42. Zietkiewicz E, Rafalski A, Labuda D (1994) Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20:176–183CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • H. S. Paris
    • 1
    Email author
  • N. Yonash
    • 2
  • V. Portnoy
    • 1
  • N. Mozes-Daube
    • 1
  • G. Tzuri
    • 1
  • N. Katzir
    • 1
  1. 1.Department of Vegetable Crops, Agricultural Research Organization, Newe Ya'ar Research Center, P. O. Box 1021, Ramat Yishay 30-095, Israel
  2. 2.The Israel Gene Bank for Agricultural Crops, Agricultural Research Organization, Volcani Center, P. O. Box 6, Bet Dagan 50-250, Israel

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