Journal Article

Semi-Automatic Laser Beam Microdissection of the Y Chromosome and Analysis of Y Chromosome DNA in a Dioecious Plant, <i>Silene latifolia</i>

Sachihiro Matsunaga, Shigeyuki Kawano, Takeshi Michimoto, Tetsuya Higashiyama, Shunsuke Nakao, Atsushi Sakai and Tsuneyoshi Kuroiwa

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 40, issue 1, pages 60-68
Published in print January 1999 | ISSN: 0032-0781
Published online January 1999 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/oxfordjournals.pcp.a029475
Semi-Automatic Laser Beam Microdissection of the Y Chromosome and Analysis of Y Chromosome DNA in a Dioecious Plant, Silene latifolia

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Silene latifolia has heteromorphic sex chromosomes, the X and Y chromosomes. The Y chromosome, which is thought to carry the male determining gene, was isolated by UV laser microdissection and amplified by degenerate oligonucleotide-primed PCR. In situ chromosome suppression of the amplified Y chromosome DNA in the presence of female genomic DNA as a competitor showed that the microdissected Y chromosome DNA did not specifically hybridize to the Y chromosome, but hybridized to all chromosomes. This result suggests that the Y chromosome does not contain Y chromosome-enriched repetitive sequences. A repetitive sequence in the microdissected Y chromosome, RMY1, was isolated while screening repetitive sequences in the amplified Y chromosome. Part of the nucleotide sequence shared a similarity to that of X-43.1, which was isolated from microdissected X chromosomes. Since fluorescence in situ hybridization analysis with RMY1 demonstrated that RMY1 was localized at the ends of the chromosome, RMY1 may be a subtelomeric repetitive sequence. Regarding the sex chromosomes, RMY1 was detected at both ends of the X chromosome and at one end near the pseudoautosomal region of the Y chromosome. The different localization of RMY1 on the sex chromosomes provides a clue to the problem of how the sex chromosomes arose from autosomes.

Keywords: Dioecious plant; Microdissection; Sex chromosome; Subtelomeric sequence

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Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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