APPLICATION OF RAPD TECHNIQUE FOR IDENTIFICATION OF INTERSPECIFIC HYBRIDS FROM GENUS Capsicum
Beata Sikora
University of Technology and Life Sciences in BydgoszczPaweł Nowaczyk
University of Technology and Life Sciences in BydgoszczAbstract
Molecular biology techniques based on DNA analysis are being increasingly used in modern plant breeding. In most cases DNA amplification methods using the PCR are being applied. The aim of this study was to assess the applicability of RAPD technique in identification of interspecific hybrids from genus Capsicum. The material consisted of selected interspecific hybrids (C. frutescens × C. annuum), (C. frutescens × C. chinense) and (C. frutescens × C. baccatum) and their respective parental breeding lines. The research was conducted with fifteen decamer primers. The size of amplified products ranged from 122–2127 bp, and their number from 3 to 17 per primer. All 15 primers made it possible to analyze 143 loci, of which 111 were polymorphic. The analysis of electrophoretograms allowed the identification of all studied genotypes and confirmation of hybrid origin of all tested hybrids. Two RAPD markers were obtained for the C. frutescens × C. annuum hybrid, two markers for C. frutescens × C. baccatum hybrid and five markers for C. frutescens × C. chinense. The data obtained were used to calculate the genetic distance between the investigated genotypes and construction of dendrograms.
Keywords:
molecular markers, PCR, hybrid seed testing, pepper, CapsicumReferences
Ballester J., de Vicente M.C., 1998. Determination of F1 hybrid seed purity in pepper using PCR-based markers. Euphytica 103, 223–226.
Caranta C., Pflieger S., Lefebvre V., Daubèze A. M., Thabuis A., Palloix A., 2002. QTLs involved in the restriction of cucumber mosaic virus (CMV) long-distance movement in pepper. Theor. Appl. Genet. 104, 586–591.
Costa F.R., Pereira T.N.S., Vitoria A.P., de Campos K.P., Rodrigues R., da Silva D.H., et al., 2006. Genetic diversity among Capsicum accessions using RAPD markers. Crop Breed. Appl. Biot. 6, 18–23.
DeWitt D., Bosland P.W., 1996. Peppers of the World: An Identification Guide. CAB International, Wallinford.
Djian-Caporalino C., Lefebvre V., Sage-Daubèze A.M., Palloix A., 2006. Capsicum. In: Genetic resources, chromosome engineering, and crop improvement: Vegetable crops, Vol. 3, R.J. Singh (ed.). CRC Press, Taylor & Francis, Boca Raton.
Eshbaugh W.H., 1993. History and exploitation of a serendipitous new crop discovery. In: New Crops, J. Janick, J.E. Simon (eds). New York, Wiley, 132–139.
FAOSTAT data, http://faostat.fao.org, 2011 (accessed on 11th February 2013).
Greenleaf W.H., 1956. Inheritance of resistance to tobacco-etch virus in Capsicum frutescens and in Capsicum annuum. Phytopathology 46, 371–375.
Heiser C.B., Smith P.G., 1953. The cultivated Capsicum peppers. Econ. Bot. 7, 214–227.
Holmes F.O., 1937. Inheritance of resistance to tobacco mosaic disease in the pepper. Phytopathology 27, 637–642.
Ilbi H., 2003. RAPD markers assisted varietal identification and genetic purity test in pepper, Capsicum annuum. Sci. Hortic. 97, 211–218.
Ince A.G., Karaca M., Onus A.N., 2009. Development and utilization of diagnostic DAMD-PCR markers for Capsicum accessions. Genet. Resour. Crop Ev. 56, 211–221.
Ince A.G., Karaca M., Onus A.N., 2010. Genetic relationships within and between Capsicum species. Biochem. Genet. 48, 83–95.
Jang I., Moon J.H., Yoon J.B., Yoo J.H., Yang T.J., Kim Y.J., Park H.G., 2004. Application of RAPD and SCAR markers for purity testing of F1 hybrid seed in chili pepper (Capsicum annuum). Mol. Cells 18, 295–299.
Jensen R.J., McLeod M.J., Eshbaugh W.H., Guttaman S.I., 1979. Numerical taxonomic analysis of allozymic variation in Capsicum. Taxon 28, 315–327.
Kochieva E.Z., Ryzhova N.N., van Dooijeweert W., Boukema I.W., Arens P., 2004. Assessment of genetic relationships in the genus Capsicum using different DNA marker systems w EUCARPIA: XIIth meeting on genetics and breeding of Capsicum and Eggplant. Wageningen, Plant Research International, 44–50.
Kumar P., Gupta V.K., Misra A.K., Modi D.R., Pandey B.K., 2009. Potential of molecular markers in plant biotechnology. Plant Omics J. 2, 141–162.
Lanteri S., Acquadro A., Quagliotti L., Portis E., 2003. RAPD and AFLP assessment of genetic variation in a landrace of pepper (Capsicum annuum L.), grown in North-west Italy. Genet. Res. Crop Evol. 50, 723–735.
Lefebvre V., 2004. Molecular markers for genetic and breeding: Development and use in pepper (Capsicum spp.). In: Molecular marker systems in plant breeding and crop improvement-Biotechnology in agriculture and forestry No. 55., Lörz H., Wenzel G. Springer, Verlag, Berlin, 189–214
Lefebvre V., Palloix A., Rivers M., 1993. Nuclear RFLP between cultivars (Capsicum annuum L.). Euphytica 71, 189–199.
Livingstone K.D., Lackney V.K., Blauth J.R., Van Wijk R., Jahn M.K., 1999. Genome mapping in Capsicum and the evolution of structure in the Solanaceae. Genetics 152, 1183–1202.
Livneh O., Nagler Y., Tal Y., Gafni S.B., Beckmann J.S., Sela J., 1990. RFLP analysis of a hybrid cultivar of pepper (Capsicum annuum) and its use in distinguishing between parental lines and in hybrid identification. Seed Sci. Technol. 18, 209–21.
Loaiza-Figueroa F., Ritland K., Laborde Cancino J.A., Tanksley S.D., 1989. Patterns of genetic variation of the genus Capsicum (Solanaceae) in Mexico. Plant System. Evol. 165, 159–188.
Mongkolporn O., Dokmaihom Y., Kanchana-Udomkan C., Pakdeevaraporn P., 2004. Genetic purity test of F1 hybrid Capsicum using molecular analysis. J. Hortic. Sci. Biotech. 79, 449–451.
Nei M., Li W.-H., 1979. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc. Natl. Acad. Sci. USA 76, 5269–5273.
Panda R.C., Kumar O.A., Rao K.G.R., 1986. The use of seed protein electrophoresis in the study of phylogenetic relationships in chili pepper (Capsicum L.). Theor. Applied Genet. 72, 665–670.
Paran I., Aftergoot E., Shifriss C., 1998. Variation in Capsicum annum reavealed by RAPD and AFLP markers. Euphytica 99, 167–173.
Pickersgill B., 1971. Relationships between weedy and cultivated forms in some species of chili peppers (genus Capsicum). Evolution 25, 683–691.
Pickersgill B., 1991. Cytogenetics and evolution of Capsicum L. In: Chromosome engineering in plants: genetics, breeding, evolution. Part B. Tsuchiya T., Gupta P.K. (eds.). Amsterdam, Elsevier, 139–160.
Pickersgill B., 1997. Genetic resources and breeding of Capsicum spp. Euphytica 96, 129–133.
Prince J.P., Lackney V.K., Angeles C., Blauth J.R., Kyle M.M., 1995. A survey of DNA polymorphism within the genus Capsicum and the fingerprinting of pepper cultivars. Genome 38, 244–251.
Rajesh M.K., Jerarda B.A., Preethia P., Thomasb R.J., Fayasa T.P., Rachanaa K.E., Karuna A., 2013. Development of a RAPD-derived SCAR marker associated with tall-type palm traitin coconut. Sci. Hort. 150, 312–316.
Rodriguez J.M., Berke T., Engle L., Nienhuis J., 1999. Variation among and within Capsicum species revealed by RAPD markers. Theor. Appl. Genet. 99, 147–156.
Stachowiak A., Świerczyński S., 2012. Phenological, morphological and genetic variability of 15 clones of rootstocks for apple. Acta Sci. Pol., Hortorum Cultus 11, 183–192.
Stoffella P.J., Locascio S.J., Howe T.K., Olson S.M., Shuler K.D., Vavrina C.S., 1995. Yield and fruit size stability differs among bell pepper cultivars. J. Amer. Soc. For. Hort. Sci. 120, 325–328.
Soller M., Beckmann J.S., 1983. Genetic polymorphism in varietal identification and genetic improvement. Theor. Appl. Gen. 67, 25–33.
Tanksley S.D., Iglesias-Olivas J., 1984. Inheritance and transfer of multiple-flower character from Capsicum chinense into Capsicum annuum. Euphytica 33, 769–777.
Todorova V., 2007. Fruit characterization and influence of variation factors in pepper Kapiya type varieties and breeding lines (Capsicum annuum L.). Bulg. J. Agric. 13, 309–315.
Van de Peer Y., De Wachter Y., 1994. TREECON for Windows: a software package for the construction and drawing of evolutionary trees for the Microsoft Windows environment. Comput. Applic. Biosci. 10, 569–70.
Zewdie Y., Bosland P.W., 2000. Evaluation of genotype, environment and genotype-byenvironment interaction for capsaicinoids in Capsicum annuum L. Euphytica 111, 185–190.
Caranta C., Pflieger S., Lefebvre V., Daubèze A. M., Thabuis A., Palloix A., 2002. QTLs involved in the restriction of cucumber mosaic virus (CMV) long-distance movement in pepper. Theor. Appl. Genet. 104, 586–591.
Costa F.R., Pereira T.N.S., Vitoria A.P., de Campos K.P., Rodrigues R., da Silva D.H., et al., 2006. Genetic diversity among Capsicum accessions using RAPD markers. Crop Breed. Appl. Biot. 6, 18–23.
DeWitt D., Bosland P.W., 1996. Peppers of the World: An Identification Guide. CAB International, Wallinford.
Djian-Caporalino C., Lefebvre V., Sage-Daubèze A.M., Palloix A., 2006. Capsicum. In: Genetic resources, chromosome engineering, and crop improvement: Vegetable crops, Vol. 3, R.J. Singh (ed.). CRC Press, Taylor & Francis, Boca Raton.
Eshbaugh W.H., 1993. History and exploitation of a serendipitous new crop discovery. In: New Crops, J. Janick, J.E. Simon (eds). New York, Wiley, 132–139.
FAOSTAT data, http://faostat.fao.org, 2011 (accessed on 11th February 2013).
Greenleaf W.H., 1956. Inheritance of resistance to tobacco-etch virus in Capsicum frutescens and in Capsicum annuum. Phytopathology 46, 371–375.
Heiser C.B., Smith P.G., 1953. The cultivated Capsicum peppers. Econ. Bot. 7, 214–227.
Holmes F.O., 1937. Inheritance of resistance to tobacco mosaic disease in the pepper. Phytopathology 27, 637–642.
Ilbi H., 2003. RAPD markers assisted varietal identification and genetic purity test in pepper, Capsicum annuum. Sci. Hortic. 97, 211–218.
Ince A.G., Karaca M., Onus A.N., 2009. Development and utilization of diagnostic DAMD-PCR markers for Capsicum accessions. Genet. Resour. Crop Ev. 56, 211–221.
Ince A.G., Karaca M., Onus A.N., 2010. Genetic relationships within and between Capsicum species. Biochem. Genet. 48, 83–95.
Jang I., Moon J.H., Yoon J.B., Yoo J.H., Yang T.J., Kim Y.J., Park H.G., 2004. Application of RAPD and SCAR markers for purity testing of F1 hybrid seed in chili pepper (Capsicum annuum). Mol. Cells 18, 295–299.
Jensen R.J., McLeod M.J., Eshbaugh W.H., Guttaman S.I., 1979. Numerical taxonomic analysis of allozymic variation in Capsicum. Taxon 28, 315–327.
Kochieva E.Z., Ryzhova N.N., van Dooijeweert W., Boukema I.W., Arens P., 2004. Assessment of genetic relationships in the genus Capsicum using different DNA marker systems w EUCARPIA: XIIth meeting on genetics and breeding of Capsicum and Eggplant. Wageningen, Plant Research International, 44–50.
Kumar P., Gupta V.K., Misra A.K., Modi D.R., Pandey B.K., 2009. Potential of molecular markers in plant biotechnology. Plant Omics J. 2, 141–162.
Lanteri S., Acquadro A., Quagliotti L., Portis E., 2003. RAPD and AFLP assessment of genetic variation in a landrace of pepper (Capsicum annuum L.), grown in North-west Italy. Genet. Res. Crop Evol. 50, 723–735.
Lefebvre V., 2004. Molecular markers for genetic and breeding: Development and use in pepper (Capsicum spp.). In: Molecular marker systems in plant breeding and crop improvement-Biotechnology in agriculture and forestry No. 55., Lörz H., Wenzel G. Springer, Verlag, Berlin, 189–214
Lefebvre V., Palloix A., Rivers M., 1993. Nuclear RFLP between cultivars (Capsicum annuum L.). Euphytica 71, 189–199.
Livingstone K.D., Lackney V.K., Blauth J.R., Van Wijk R., Jahn M.K., 1999. Genome mapping in Capsicum and the evolution of structure in the Solanaceae. Genetics 152, 1183–1202.
Livneh O., Nagler Y., Tal Y., Gafni S.B., Beckmann J.S., Sela J., 1990. RFLP analysis of a hybrid cultivar of pepper (Capsicum annuum) and its use in distinguishing between parental lines and in hybrid identification. Seed Sci. Technol. 18, 209–21.
Loaiza-Figueroa F., Ritland K., Laborde Cancino J.A., Tanksley S.D., 1989. Patterns of genetic variation of the genus Capsicum (Solanaceae) in Mexico. Plant System. Evol. 165, 159–188.
Mongkolporn O., Dokmaihom Y., Kanchana-Udomkan C., Pakdeevaraporn P., 2004. Genetic purity test of F1 hybrid Capsicum using molecular analysis. J. Hortic. Sci. Biotech. 79, 449–451.
Nei M., Li W.-H., 1979. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc. Natl. Acad. Sci. USA 76, 5269–5273.
Panda R.C., Kumar O.A., Rao K.G.R., 1986. The use of seed protein electrophoresis in the study of phylogenetic relationships in chili pepper (Capsicum L.). Theor. Applied Genet. 72, 665–670.
Paran I., Aftergoot E., Shifriss C., 1998. Variation in Capsicum annum reavealed by RAPD and AFLP markers. Euphytica 99, 167–173.
Pickersgill B., 1971. Relationships between weedy and cultivated forms in some species of chili peppers (genus Capsicum). Evolution 25, 683–691.
Pickersgill B., 1991. Cytogenetics and evolution of Capsicum L. In: Chromosome engineering in plants: genetics, breeding, evolution. Part B. Tsuchiya T., Gupta P.K. (eds.). Amsterdam, Elsevier, 139–160.
Pickersgill B., 1997. Genetic resources and breeding of Capsicum spp. Euphytica 96, 129–133.
Prince J.P., Lackney V.K., Angeles C., Blauth J.R., Kyle M.M., 1995. A survey of DNA polymorphism within the genus Capsicum and the fingerprinting of pepper cultivars. Genome 38, 244–251.
Rajesh M.K., Jerarda B.A., Preethia P., Thomasb R.J., Fayasa T.P., Rachanaa K.E., Karuna A., 2013. Development of a RAPD-derived SCAR marker associated with tall-type palm traitin coconut. Sci. Hort. 150, 312–316.
Rodriguez J.M., Berke T., Engle L., Nienhuis J., 1999. Variation among and within Capsicum species revealed by RAPD markers. Theor. Appl. Genet. 99, 147–156.
Stachowiak A., Świerczyński S., 2012. Phenological, morphological and genetic variability of 15 clones of rootstocks for apple. Acta Sci. Pol., Hortorum Cultus 11, 183–192.
Stoffella P.J., Locascio S.J., Howe T.K., Olson S.M., Shuler K.D., Vavrina C.S., 1995. Yield and fruit size stability differs among bell pepper cultivars. J. Amer. Soc. For. Hort. Sci. 120, 325–328.
Soller M., Beckmann J.S., 1983. Genetic polymorphism in varietal identification and genetic improvement. Theor. Appl. Gen. 67, 25–33.
Tanksley S.D., Iglesias-Olivas J., 1984. Inheritance and transfer of multiple-flower character from Capsicum chinense into Capsicum annuum. Euphytica 33, 769–777.
Todorova V., 2007. Fruit characterization and influence of variation factors in pepper Kapiya type varieties and breeding lines (Capsicum annuum L.). Bulg. J. Agric. 13, 309–315.
Van de Peer Y., De Wachter Y., 1994. TREECON for Windows: a software package for the construction and drawing of evolutionary trees for the Microsoft Windows environment. Comput. Applic. Biosci. 10, 569–70.
Zewdie Y., Bosland P.W., 2000. Evaluation of genotype, environment and genotype-byenvironment interaction for capsaicinoids in Capsicum annuum L. Euphytica 111, 185–190.
Beata Sikora
University of Technology and Life Sciences in Bydgoszcz
University of Technology and Life Sciences in Bydgoszcz
Paweł Nowaczyk
University of Technology and Life Sciences in Bydgoszcz
University of Technology and Life Sciences in Bydgoszcz
License
Articles are made available under the conditions CC BY 4.0 (until 2020 under the conditions CC BY-NC-ND 4.0).
Submission of the paper implies that it has not been published previously, that it is not under consideration for publication elsewhere.
The author signs a statement of the originality of the work, the contribution of individuals, and source of funding.
