PAG-II Plant Genome II Conference

Town & Country Conference Center, San Diego, CA, January, 1994.


PG-II: PRELIMINARY KARYOTYPE IN PINUS ELLIOTTII ENGELM. ELLIOTTII USING FLUORESCENCE IN SITU HYBRIDIZATION AND FLUOROCHROME BANDING PATTERNS

PRELIMINARY KARYOTYPE IN PINUS ELLIOTTII ENGELM. ELLIOTTII USING FLUORESCENCE IN SITU HYBRIDIZATION AND FLUOROCHROME BANDING PATTERNS.

R.L. Doudrick, C.D. Nelson, W.L. Nance, J.S. Heslop-Harrison, and T. Schwarzacher. R.L. Doudrick, C.D. Nelson, W.L. Nance, U.S.D.A. Forest Service, Southern Forest Experiment Station, Gulfport, MS, USA; J.S. Heslop-Harrison, T. Schwarzacher, John Innes Centre, Karyobiology Group, Norwich, U.K.


Idiograms and a preliminary karyotype were prepared from metaphase spreads of seedling root-tips in slash pine (Pinus elliottii Engelm. elliottii; 2n =24). Each metaphase had 11 pairs of long metacentric chromosomes and one pair of short submetacentrics. The long metacentric chromosomes were similar in shape and length and could only be distinguished individually by supplementing statistical analysis of characteristics with Qualitative analysis of patterns of probe hybridization and fluorochrome banding. Regions on chromosomes with homology to previously described 18S - 25S ribosomal DNA (RDNA) and 5S RDNA were located by fluorescence in situ hybridization. All cells observed had at least 12, and as many as 14 18S - 25S rDNA sites, on site present on each of seven separate pairs of chromosomes. One region of hybridization was observed at the centromeric region of an additional pair of chromosomes. A major site of 5S rDNA hybridization was localized on a ninth pair of chromosomes with minor sites on chromosomes with and without 18S - 25S sites. Chromosomes were stained with the DNA base-specific binding fluorochromes chromomycin A3 (CMA) and 4', 6-diamidino-2-phenylindole (DAPI). Bands of CMA appeared at interstitial and/or proximal regions of several pairs of chromosomes. The interstitial bands of CMA were mostly localized at secondary constrictions, the hybridization sites of 18 - 25S rDNA. Bands of DAPI appeared at interstitial and/or centromeric regions of nearly all chromosomes. A pair of bands of DAPI were observed near the centromeric region of one pair of chromosomes.


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