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Comparative analysis of genetic diversity in pea assessed by RFLP- and PCR-based methods

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Abstract

DNA-based molecular-marker techniques have been proven powerful in genetic diversity estimations. Among them, RFLP was the first and is still the most commonly used in the estimation of genetic diversity of eukaryotic species. The recently developed PCR-based multiple-loci marker techniques, which include RAPD, AFLP, Microsatellite-AFLP and inter-SSR PCR, are playing increasingly important roles in this type of research. Despite the wide application of these techniques, no direct comparison of these methods in the estimation of genetic diversity has been carried out. Here we report a direct comparison of DNA-based RFLP with various PCR-based techniques regarding their informativeness and applicability for genetic diversity analysis. Among ten pea genotypes studied, all the PCR-based methods were much more informative than cDNA-RFLP. Genetic diversity trees were derived from each marker technique, and compared using Mantel's test. By this criterion, all trees derived from the various molecular marker techniques, except for the tree derived from inter-SSR PCR, were significantly correlated, suggesting that these PCR-based techniques could replace RFLP in the estimation of genetic diversity. On the basis of this result, AFLP analysis was applied to assess the genetic diversity of a sample of accessions representing the various species and subspecies within the genus Pisum.

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References

  • Amos B, Pemberton J (1992) DNA fingerprinting in a non-human population. Curr Opin Genet Dev 2:857–860

    Google Scholar 

  • Arens P, Odinot P, Heusden AW van, Lindhout P, Vosman B (1995) GATA- and GACA-repeats are not evenly distributed throughout the tomato genome. Genome 38:84–90

    Google Scholar 

  • Arnau J, Housego AP, Oliver RP (1994) The use of RAPD markers in the genetic analysis of the plant pathogenic fungus Cladosporium fulvum. Curr Genet 25:438–444

    Google Scholar 

  • Beckman JS, Soller M (1986) Restriction fragment length polymorphisms and genetic improvement of agricultural species. Euphytica 35:111–124

    Google Scholar 

  • Ben-Ze'ev N, Zohary D (1973) Species relationships in the genus Pisum L. Israel J Bot 22:73–91

    Google Scholar 

  • Beyermann B, Nurnberg P, Weihe A, Meixner M, Epplen JT, Borner T (1992) Fingerprinting plant genomes with oligonucleotide probes specific for simple repetitive DNA sequences. Theor Appl Genet 83:691–694

    Google Scholar 

  • Blixt S (1972) Mutation genetics in Pisum. Agric Hort Genet 30:1–293

    Google Scholar 

  • Botstein D, White RL, Skolnick MH, Davies RW (1980) Construction of a genetic map in man using restriction fragment length polymorphism. Am J Hum Genet 32:314–331

    CAS  PubMed  Google Scholar 

  • Brown PTH (1992) DNA fingerprinting in plant breeding. Agro-Food-Industry Hi-Tech November/December

  • Condit R, Hubbell SP (1991) Abundance and DNA sequence of twobase repeat regions in a tropical tree genome. Genome 34:66–71

    Google Scholar 

  • Dallas JF (1988) Detection of DNA “fingerprints” of cultivated rice by hybridization with a human minisatellite DNA probe. Proc Natl Acad Sci USA 85:6831–6835

    Google Scholar 

  • Delsney M, Grellet F, Tremousaygue D, Raynal M, Panabieres F (1988) Structure, evolution et expression de l'ADN nucleaire. Bull Soc Bot Fr 135:23–38

    Google Scholar 

  • Dos Santos JB, Nienhuis J, Skroch P, Tivang J, and Slocum MK (1994) Comparison of RAPD and RFLP genetic markers in determining genetic similarity among Brassica oleracea L. genotypes. Theor Appl Genet 87:909–915

    Google Scholar 

  • Ellis THN (1994) Approaches to the genetic mapping of pea. Mod Meth Plant Anal 16:117–160

    Google Scholar 

  • Ellis THN, Turner L, Hellens RP, Lee D, Harker CL, Enard C, Domoney C, Davies DR (1992) Linkage maps in pea. Genetics 130:649–663

    CAS  PubMed  Google Scholar 

  • Epplen JT (1988) On simple GAT/CA sequences in animal genomes: a critical reappraisal. J Hered 79:409–417

    Google Scholar 

  • Gebhardt C, Ritter E, Debener T, Schachtschabel U, Walkemeier B (1989) RFLP analysis and linkage mapping in Solanum tuberosum. Theor Appl Genet 78:65–75

    Google Scholar 

  • Hamann A, Zink D, Nagl W (1995) Microsatellite fingerprinting in the genus Phaseolus. Genome 38:507–515

    Google Scholar 

  • Havey MJ, Muehlbauer FJ (1989) Variability for restriction fragment lengths and phylogenies in lentil. Theor Appl Genet 77:839–843

    Google Scholar 

  • Jeffreys AJ, Wilson V, Thein SL (1985) Individual-specific “fingerprints” of human DNA. Nature 316:76–79

    CAS  PubMed  Google Scholar 

  • Jung C, Pillen K, Frese L, Fahr S, Melchinger AE (1993) Phylogenetic relationships between cultivated and wild species of the genus Beta revealed by DNA “fingerprinting”. Theor Appl Genet 86:449–457

    Google Scholar 

  • Kesseli R, Ochoa O, Michelmore R (1991) Variation at RFLP loci in Lactuca spp. and origin of cultivated lettuce (L. sativa). Genome 34:430–436

    Google Scholar 

  • Kesseli RV, Paran I, Michelmore RW (1994) Analysis of a detailed genetic linkage map of Lactuca sativa (Lettuce) constructed from RFLP and RAPD markers. Genetics 136:1435–1446

    Google Scholar 

  • Lapitan NLV (1992) Organization and evolution of higher plant nuclei genomes. Genome 35:171–181

    CAS  Google Scholar 

  • Leberg PL (1992) Effects of population bottlenecks on genetic diversity as measured by allozyme electrophoresis. Evolution 46:477–494

    Google Scholar 

  • Manly BFJ (1985) The statistics of natural selection. Chapman and Hall, London

    Google Scholar 

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

    CAS  PubMed  Google Scholar 

  • Marx GA (1987) A suite of mutants that modify pattern formation in pea leaves. Plant Mol Biol Rep 5:311–335

    Google Scholar 

  • Messmer MM, Melchinger AE, Woodman WL, Lee EA, Lamkey KR (1991) Genetic diversity among progenitors and elite lines from the Iowa Stiff Stalk Synthetic (BSSS) maize population: comparison of allozyme and RFLP data. Theor Appl Genet 83:97–107

    Google Scholar 

  • Miller JC, Tanksley SD (1990) RFLP analysis of phylogenetic relationships and genetic variation in the genus Lycopersicon. Theor Appl Genet 80:437–448

    CAS  Google Scholar 

  • Monckton DG, Jeffreys AJ (1993) DNA profiling. Curr Opin Biotechnol 4:660–664

    Google Scholar 

  • Moser H, Lee M (1994) RFLP variation and genealogical distance, multivariate distance, heterosis, and genetic variation in oats. Theor Appl Genet 87:947–956

    Google Scholar 

  • Murray MG, Thompson WF (1982) Repeat sequence interspersion in coding DNA of peas does not reflect that in total pea DNA. Plant Mol Biol 1:143–153

    Google Scholar 

  • Nei M (1987) Molecular evolutionary genetics. Columbia University Press, New York

    Google Scholar 

  • N'goran JAK, Laurent V, Risterucci AM, Lanaud C (1994) Comparative genetic diversity studies of Theobroma cacao L. using RFLP and RAPD markers. Heredity 73:589–597

    Google Scholar 

  • Nybom H (1990) Genetic variation in ornamental apple trees and their seedlings (Malus Rosaceae) revealed by DNA “fingerprinting” with M13 repeat probe. Hereditas 133:17–28

    Google Scholar 

  • Perry BC, Mahinur SA, Arvind AB, Lavi U, Jiang R (1994) Length polymorphisms of simple sequence repeat (SSR) DNA as molecular markers in plants. In: Gresshoff PM (ed) Plant genome analysis. CRC Press, Boca Raton, London, pp 47–56

    Google Scholar 

  • Poulsen GB, Kahl G, Weising K (1994) Differential abundance of simple repetitive sequences in species of Brassica and related Brassicaceae. Plant Syst Evol 190:21–30

    Google Scholar 

  • Rogstad SH, Patton JC, Schaal BA (1988) M13 repeat probe detects DNA minisatellite-like sequences in gymnosperms and angiosperms. Proc Natl Acad Sci USA 85:9176–9178

    Google Scholar 

  • Rus-Kortekaas W, Smulders MJM, Arens P, Vosman B (1994) Direct comparison of levels of genetic variation in tomato detected by a GACA-containing microsatellite probe and by random amplified polymorphic DNA. Genome 37:375–381

    Google Scholar 

  • Saitou N, Nei M (1987) The neighbour-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  PubMed  Google Scholar 

  • Schmidt T, Boblenz K, Metzlaff M, Kaemmer D, Weising K, Kahl G (1993) DNA fingerprinting in sugar beet (Beta vulgaris)-identification of double haploid breeding lines. Theor Appl Genet 85:653–657

    Google Scholar 

  • Sharma (1996). Theor Appl Genet (in press)

  • Smith OS, Smith JSC, Bowen SL, Tenborg RA (1992) Numbers of RFLP probes necessary to show associations between lines. Maize Genet Coop Newslett 66:66

    Google Scholar 

  • Tanksley SD, Young ND, Paterson AH, Bonierbale MW (1989) RFLP mapping in plant breeding — new tools for an old science. Bio Technol 7:257–264

    Google Scholar 

  • Tinker NA, Fortin MG, Mather DE (1993) Random amplified polymorphic DNA and pedigree relationships in spring barley. Theor Appl Genet 85:976–984

    Google Scholar 

  • Tivang JG, Nienhuis J, Smith OS (1994) Estimation of sampling variance of molecular-marker data using the bootstrap procedure. Theor Appl Genet 89:259–264

    Google Scholar 

  • Tzuri G, Hillel J, Lavi U, Harberfeld A, Vainstein A (1991) DNA fingerprint analysis of ornamental plants. Plant Sci 76:91–97

    Google Scholar 

  • Vedel F, Delseny M (1987) Repetitive and variability of higher-plant genomes. Pl Physiol Biochem 25:191–210

    Google Scholar 

  • Vogel JM (1995) A microsatellite-based multiplexed genome assay (abtracts). In: Plant Genome. Scharago International, New York, p 84

    Google Scholar 

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

    CAS  PubMed  Google Scholar 

  • Welsh J, McClelland M (1990) Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 18:7213–7218

    CAS  PubMed  Google Scholar 

  • Williams JGK, Kubelik AR, Livak K, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535

    CAS  PubMed  Google Scholar 

  • Zabeau M, Vos P (1993) Selective restriction fragment amplification: a general method for DNA fingerprinting. European Patent Application 924026 29.7 (publication number: 0534 858 A1)

  • Zhou Z, Gustafson JP (1995) Genetic variation detected by DNA fingerprinting with a rice minisatellite probe in Oryza sativa. Theor Appl Genet 91:481–488

    Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

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Communicated by G. E. Hart

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Lu, J., Knox, M.R., Ambrose, M.J. et al. Comparative analysis of genetic diversity in pea assessed by RFLP- and PCR-based methods. Theoret. Appl. Genetics 93, 1103–1111 (1996). https://doi.org/10.1007/BF00230132

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  • DOI: https://doi.org/10.1007/BF00230132

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