Horticulture, Environment, and Biotechnology

, Volume 57, Issue 1, pp 61–68 | Cite as

Development of RAPD and SCAR markers related to watermelon mosaic virus and zucchini yellow mosaic virus resistance in Cucurbita moschata

  • Dae Kook Kim
  • Sang-Gi Seo
  • Soon Bae Kwon
  • Young-Doo Park
Research Report Genetics and Breeding


Squash (Cucurbita spp.) is a major vegetable crop in the Cucurbitaceae family, and has high economic value. Squash is an important foodstuff and has possible health benefits because of its antioxidant, anti-diabetic, anti-carcinogenic, and anti-microbial potential. However, the highly virulent watermelon mosaic virus (WMV) and zucchini yellow mosaic virus (ZYMV) are serious problems for squash worldwide. Single or mixed infections of WMV and ZYMV can destroy entire squash plants. Therefore, breeding squash cultivars with resistance to WMV and ZYMV is of major importance for squash cultivation. In this study, random amplified polymorphic DNA (RAPD) and sequence-characterized amplified region (SCAR) markers were developed using C. moschata lines resistant or susceptible to WMV and ZYMV. A total of 100 random primers were tested for their ability to discriminate between resistant and susceptible lines in RAPD analysis, and 4 RAPD markers (OPF10, OPF19, OPF20, and OPL19) related to WMV and ZYMV resistance were selected. Based on the RAPD results, genetic similarities were calculated and cluster analysis was conducted using unweighted pair group method with arithmetic mean (UPGMA) method. C. moschata lines were clearly segregated into resistant and susceptible lines by cluster analysis. One of the RAPD markers was successfully converted into a SCAR marker (VirSq-F19). This marker could be used as a tool for selecting WMV- and ZYMV-resistant squash in the early selection stages of a practical squash breeding program.

Additional key words

breeding program polymorphism pumpkin virus-resistant squash 


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Literature Cited

  1. Agarwal, M., N. Shrivastava, and H. Padh. 2008. Advances in molecular marker techniques and their applications in plant sciences. Plant Cell Rep. 27;617–631.CrossRefPubMedGoogle Scholar
  2. Ardiel, G.S., T.S. Grewal, P. Deberdt, B.G. Rossnagel, and G.J. Scoles. 2002. Inheritance of resistance to covered smut in barley and development of a tightly linked SCAR marker. Theor. Appl. Genet. 104;457–464.CrossRefPubMedGoogle Scholar
  3. Bolanos-Herrera, A. and A.R. Valdivia Torres. 1994. Inheritance of virus resistance in Cucurbita moschata and C. maxima (Doctoral dissertation, MS thesis. Dept of Plant Breeding, Cornell Univ., Ithaca, NY). ISO 690. Cornell University, Ithaca, NY (EUA), & CATIE, Turrialba (Costa Rica).Google Scholar
  4. Clough, G.H. and P.B. Hamm. 1995. Coat protein transgenic resistance to watermelon mosaic and zucchini yellows mosaic virus in squash and cantaloupe. Plant Dis. 79;1107–1109.CrossRefGoogle Scholar
  5. Deng, Z., S. Xiao, S. Huang, and F.G. Gmitter. 1997. Development and characterization of SCAR markers linked to the Citrus tristeza virus resistance gene from Poncirus trifoliate. Genome 40;697–704.CrossRefPubMedGoogle Scholar
  6. Doyle, J.J. and J.L. Doyle. 1990. Isolation of plant DNA from fresh tissue. Focus 12;13–15.Google Scholar
  7. Fletcher, J.D., A.R. Wallace, and B.T. Rogers. 2000. Potyviruses in New Zealand buttercup squash (Cucurbits maxima Duch.): Yield and quality effects of ZYMV and WMV 2 virus infections. N. Z. J. Crop Hortic. Sci. 28;17–26.CrossRefGoogle Scholar
  8. Fuchs, M., D.M. Tricoli, K.J. Carney, M. Schesser, J.R. McFerson, and D. Gonsalves. 1998. Comparative virus resistance and fruit yield of transgenic squash with single and multiple coat protein genes. Plant Dis. 82;1350–1356.CrossRefGoogle Scholar
  9. Gilbert-Albertini, F., H. Lecoq, M. Pitrat, and J.L. Nicolet. 1993. Resistance of Cucurbita moschata to Watermelon mosaic virus type 2 and its genetic relation to resistance to zucchini yellow mosaic virus. Euphytica 69;231–237.CrossRefGoogle Scholar
  10. Hadrys, H., M. Balick, and B. Schierwater. 1992. Applications of random amplified polymorphic DNA (RAPD) in molecular ecology. Mol. Ecol. 1;55–63.CrossRefPubMedGoogle Scholar
  11. Harris, K.R., K.S. Ling, W.P. Wechter, and A. Levi. 2009. Identification and utility of markers linked to the zucchini yellow mosaic virus resistance gene in watermelon. J. Am. Soc. Hortic. Sci. 134;529–534.Google Scholar
  12. Kim, J.S., S.H. Lee, H.S. Choi, M.K. Kim, H.R. Kwak, J.S. Kim, M. Nam, D.J. Cho, I.S. Cho, and G.S. Choi. 2012. 2007-2011 Characteristics of plant virus infections on crop samples submitted from agricultural places. Res. Plant Dis. 18;277–289.CrossRefGoogle Scholar
  13. Lodhi, M.A., G.N. Ye, N.F. Weeden, and B.I. Resich. 1994. A simple and efficient method for DNA extraction from grapevine cultivars and Vitis species. Plant Mol. Biol. Rep. 12;6–13.CrossRefGoogle Scholar
  14. McDermott, J.M., U. Brandle, F. Dutly, U.A. Haemmerli, S. Keller, K.E. Muller, and M.S. Wolf. 1994. Genetic variation in powdery mildew of barley: Development of RAPD, SCAR and VNTR markers. Phytopathology 84;1316–1321.CrossRefGoogle Scholar
  15. Munger H.M. and R. Provvidenti. 1987. Inheritance of resistance to zucchini yellow mosaic virus in Cucurbita moschata. Cucurbit Genet. Coop. Rep. 10;80–81.Google Scholar
  16. Pachner, M and T. Lelley. 2004. Different genes for resistance to zucchini yellow mosaic virus (ZYMV) in Cucurbita moschata, p. 237–243. In: A. Lebeda and H.S. Paris (eds.). Progress in Cucurbit Genetics and Breeding Research: Proceedings of Cucurbitaceae. Palacky Univ. Olomouc, Czech Republic.Google Scholar
  17. Pachner, M., H.S. Paris, and T. Lelley. 2011. Genes for resistance to zucchini yellow mosaic in tropical pumpkin. J. Hered. 102;330–335.CrossRefPubMedGoogle Scholar
  18. Paran, I. and R.W. Michelmore. 1993. Development of reliable PCR-based markers linked to downy mildew resistance genes in lettuce. Theor. Appl. Genet. 85;985–993.CrossRefPubMedGoogle Scholar
  19. Paris, H.S., S. Cohen, Y. Burger, and R. Yoseph. 1988. Single-gene resistance to zucchini yellow mosaic virus in Cucurbita moschata. Euphytica 37;27–29.CrossRefGoogle Scholar
  20. Paris, H.S. and R.N. Brown. 2005. The genes of pumpkin and squash. HortScience 40;1620–1630.Google Scholar
  21. Park, Y., N. Katzir, Y. Brotman, J. King, F. Bertrand, and M. Havey. 2004. Comparative mapping of ZYMV resistances in cucumber (Cucumis sativus L.) and melon (Cucumis melo L.). Theor. Appl. Genet. 109;707–712.CrossRefPubMedGoogle Scholar
  22. Provvidenti, R., R.W. Robinson, and H.M. Munger. 1978. Multiple virus resistance in Cucurbita species. Cucurbit Genet. Coop. 1;26–27.Google Scholar
  23. Staniaszek, M. and H. Habdas. 2006. RAPD technique application for intraline evaluation of androgenic carrot plants. Floia Hortic. 2;87–97Google Scholar
  24. Van Die, I.M., H.E. Bergmans, and W.P. Hoekstra. 1983. Transformation in Escherichia coli: studies on the role of the heat shock in induction of competence. J. Gen. Microbiol. 129;663–670.PubMedGoogle Scholar
  25. Williams, J.G.K., A.R. Kubelik, K.J. Livak, J.A. Rafalski, and S.V. Tingey. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18;6531–6535.CrossRefPubMedPubMedCentralGoogle Scholar
  26. Yadav, M., S. Jain, R. Tomar, G.B. Prasad, and H. Yadav. 2010. Medicinal and biological potential of pumpkin: an updated review. Nutr. Res. Rev. 23;184–190.CrossRefPubMedGoogle Scholar
  27. Zhang. Y. and J.R. Stommel. 2000. Development of SCAR and CAPS markers linked to the beta gene in tomato. Crop Sci. 41;1602–1608.CrossRefGoogle Scholar
  28. Zheng. C., R. Chang, L. Qiu, P. Chen, X. Wu, and S. Chen. 2003. Identification and characterization of a RAPD/SCAR marker linked to a resistance gene for soybean mosaic virus in soybean. Euphytica 132;199–210.CrossRefGoogle Scholar
  29. Zitter, T.A., D.L. Hopkins, and C.E. Thomas. 1996. Compendium of Cucurbit Diseases. APS Press, St. Paul, MN, USA.Google Scholar

Copyright information

© Korean Society for Horticultural Science and Springer-Verlag GmbH 2016

Authors and Affiliations

  • Dae Kook Kim
    • 1
  • Sang-Gi Seo
    • 2
  • Soon Bae Kwon
    • 3
  • Young-Doo Park
    • 1
  1. 1.Department of Horticultural BiotechnologyKyung Hee University, Seocheon-dong, Giheung-gu, YonginGyeonggiKorea
  2. 2.Breeding Research InstituteHongik Bio Co., Anjeong-ri, Paengseong-eup, PyeongtaekGyeonggiKorea
  3. 3.Gangwondo Agricultural Research and Extension ServicesGangwon, ChunchonKorea

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