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

• Published in: Theoretical and applied genetics Vol. 102; no. 5; pp. 658 - 663
• Main Authors: LI, L. J, ARUMUGANATHAN, K, RINES, H. W, PHILLIPS, R. L, RIERA-LIZARAZU, O, SANDHU, D, ZHOU, Y, GILL, K. S
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer 01.04.2001; Berlin Springer Nature B.V
• Subjects: Agronomy. Soil science and plant productions; Avena sativa; Biological and medical sciences; Cell cycle; chromosome 9; Chromosomes; Corn; Cytogenetics; Fundamental and applied biological sciences. Psychology; Generalities. Genetics. Plant material; Genetics; Genetics and breeding of economic plants; Genomes; Hybridization; Soil sciences; Zea mays; United States > US; Genetic mapping; Flow cytometry; Monocotyledones; Addition line; Zea mays; Avena; Cereal crop; Gene; Gramineae; Chromosome 9; Angiospermae; Fluorescence in situ hybridization; Spermatophyta; Herbaceous plant; Genome
• ISSN: 0040-5752; 1432-2242
• DOI: 10.1007/s001220051694

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 6×10^sup 3^ 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.[PUBLICATION ABSTRACT]