Display options
Share it on

Theor Appl Genet. 1992 Apr;83(6):684-90. doi: 10.1007/BF00226685.

Use of the polymerase chain reaction to detect spacer size heterogeneity in plant 5S-rRNA gene clusters and to locate such clusters in wheat (Triticum aestivum L.).

TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik

A V Cox, M D Bennett, T A Dyer

Affiliations

  1. Jodrell Laboratory, Royal Botanic Gardens, TW9 3DS, Surrey, Kew, Richmond, UK.

PMID: 24202741 DOI: 10.1007/BF00226685

Abstract

We have used the polymerase chain reaction to analyse variation in the size of individual 5S-ribosomal gene spacer sequences. This reaction can be used to demonstrate inter- and intraspecific variation in spacer size, and combined with DNA sequencing it may thus be a valuable taxonomic tool. Two sets of nested polymerase chain reaction primers were designed to amplify the nontranscribed spacer DNA between repeated 5S-rRNA genes. These "universal" primers were used to generate fragments from the genomic DNA from several unrelated monocotyledonous plants. Ribosomal RNA spacer sequences generated in these experiments could also be used to locate 5S-rRNA gene clusters on specific chromosomes in hexaploid wheat (Triticum aestivum). Three distinct spacer sizes were observed after amplification. These were assigned locations on chromosomes by analysing amplification products of genomic DNA from nullisomic/tetrasomic and ditelosomic wheat stocks. "Large" 508-bp 5S repeats are located on the short arm of chromosome 5B and "short" 416-bp and 425-bp repeat unit variants are located on the short arms of chromosomes 1B and 1D, respectively. No other loci were detected. The spacer fragments were cloned, sequenced, and shown to be homologous to wheat 5S-rRNA spacers previously identified. Spacers of uniform size but with some sequence heterogeneity were shown to be located at each locus.

References

  1. Nature. 1989 Jun 15;339(6225):501-5 - PubMed
  2. Proc Natl Acad Sci U S A. 1984 Dec;81(24):8014-8 - PubMed
  3. Annu Rev Biochem. 1980;49:727-64 - PubMed
  4. Science. 1988 Jan 29;239(4839):487-91 - PubMed
  5. Plant Mol Biol. 1986 Sep;6(5):339-45 - PubMed
  6. Nucleic Acids Res. 1987 Jan 26;15(2):529-42 - PubMed
  7. Cell. 1990 Jan 26;60(2):235-45 - PubMed
  8. Nucleic Acids Res. 1988 Jun 10;16(11):5198 - PubMed
  9. Nucleic Acids Res. 1986 Jun 11;14(11):4605-16 - PubMed
  10. Evolution. 1990 Dec;44(8):2089-2105 - PubMed
  11. Biochemistry. 1991 Mar 19;30(11):2735-47 - PubMed
  12. Nucleic Acids Res. 1980 Nov 11;8(21):4851-65 - PubMed
  13. Annu Rev Biochem. 1988;57:873-914 - PubMed
  14. Cell. 1986 May 9;45(3):325-6 - PubMed
  15. Nucleic Acids Res. 1988;16 Suppl:r1-70 - PubMed

Publication Types