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Identifying Mutations by High Resolution Melting in a TILLING Population of Rice
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DNA fingerprinting to detect genetic variation in rice using hypervariable DNA sequences.

W Ramakrishna1, K V Chowdari, M D Lagu

  • 1Plant Molecular Biology Unit, Division of Biochemical Sciences, National Chemical Laboratory, 411 008, Pune, India.

TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik
|November 1, 2013
PubMed
Summary

Researchers explored DNA fingerprinting in rice using mini- and microsatellite probes. The R18.1 probe demonstrated high polymorphism, capable of identifying numerous rice genotypes and enabling Mendelian segregation analysis.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Plant Science

Background:

  • DNA fingerprinting is crucial for plant variety identification and genetic studies.
  • Mini- and microsatellite DNA sequences offer potential for high-resolution genetic markers.

Purpose of the Study:

  • To evaluate the efficacy of various mini- and microsatellite probes for generating DNA fingerprints in rice.
  • To assess the polymorphism level and discriminatory power of these markers for rice cultivar and species identification.

Main Methods:

  • Testing of multiple DNA probes including R18.1 (cattle-derived), M13 repeat, pV47 (human minisatellite), and other repeat sequences (Per gene, telomere, chi, apolipoprotein B 3' hypervariable region).
  • Analysis of polymorphism in rice cultivars and wild species.
  • Investigation of Mendelian segregation in an F2 population using the R18.1 probe.
  • Assessment of DNA fingerprint stability across somatic tissues and within cultivars.

Main Results:

  • Probes R18.1, pV47, and M13 repeat yielded high levels of polymorphism, enabling identification of all tested rice cultivars and wild species.
  • The R18.1 probe exhibited the highest polymorphism, with an estimated capacity to distinguish 2.5×10^20 rice genotypes.
  • Loci detected by R18.1 segregated in a Mendelian fashion in an F2 population.
  • Generated DNA fingerprints were somatically stable, with consistent hybridization patterns across plants of the same cultivar.

Conclusions:

  • Mini- and microsatellite DNA sequences, particularly the R18.1 probe, are highly effective for generating reliable DNA fingerprints in rice.
  • These molecular markers are valuable tools for rice genotype identification, genetic analysis, and cultivar characterization.
  • This study represents the first report on the application of these specific molecular genetic markers for rice genotype identification.