Flow cytometric sorting of maize chromosome 9 from an oat-maize chromosome addition line

L. J. Li, K. Arumuganathan, H. W. Rines, R. L. Phillips, O. Riera-Lizarazu, D. Sandhu, Y. Zhou, K. S. Gill

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

Large numbers of maize chromosome 9 can be collected with high purity by flow cytometric sorting of chromosomes isolated from a disomic maize chromosome addition line of oat. Metaphase chromosome suspensions were prepared from highly synchronized seedling root-tips of an oat-maize chromosome-9 addition line (OM9) and its parental oat and maize lines. Chromosomes were stained with propidium iodide for flow cytometric analysis and sorting. Flow-karyotypes of the oat-maize addition line showed an extra peak not present in the parental oat line. This peak is due to the presence of a maize chromosome-9 pair within the genome of OM9. Separation of maize chromosome 9 by flow cytometric sorting of a chromosome preparation from a normal maize line was not possible because of its size similarity (DNA content) to maize chromosomes 6, 7 and 8. However, it is possible to separate maize chromosome 9 from oat chromosomes and chromatids. An average of about 6×103 chromosomes of maize chromosome 9 can be collected by flow-sorting from chromosomes isolated from 30 root tips (ten seedlings) of the oat-maize addition line. Purity of the maize chromosome 9, sorted from the oat-maize chromosome addition line, was estimated to be more than 90% based on genomic in situ hybridization analysis. Sorting of individual chromosomes provides valuable genomic tools for physical mapping, library construction, and gene isolation.

Original languageEnglish (US)
Pages (from-to)658-663
Number of pages6
JournalTheoretical and Applied Genetics
Volume102
Issue number5
DOIs
StatePublished - 2001

Keywords

  • Genomic in situ hybridization
  • Maize
  • Maize chromosome-9 sorting
  • Oat
  • Oat-maize chromosome-9 addition line

ASJC Scopus subject areas

  • Biotechnology
  • Agronomy and Crop Science
  • Genetics

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