Microdissection and chromosome painting of X and B chromosomes in the grasshopper Eyprepocnemis plorans.
The relative location of 2 repetitive DNAs, i.e. ribosomal (rDNA) and a tandemly repeated satellite DNA (satDNA), with respect to the centromere, suggested that B chromosomes in the grasshopper Eyprepocnemis plorans derived intraspecifically from the X chromosome. To test this hypothesis, we microdissected X and B chromosomes and amplified the obtained DNA by 2 different procedures, the conventional DOP-PCR method and the single-cell whole-genome amplification GenomePlex method. We then generated DNA probes to perform chromosome painting. Our results have confirmed that X and B chromosomes share many DNA sequences between them and with most of the autosomes, especially at locations where the satDNA and rDNA reside, in consistency with previous information. This supports the hypothesis of an intraspecific origin of B chromosomes in E. plorans. Nevertheless, the present results did not help to clarify whether Bs were derived from the X chromosome or else from 1 or more autosomes.
Departamento de Genética, Universidad de Granada, Granada, Spain.
M
M
Teruel
J
J
Cabrero
F
F
Perfectti
M J
MJ
Acosta
A
A
Sánchez
J P M
JP
Camacho
D000818:Animals; D020223:Chromosome Painting / Q000379:methods*; D002875:Chromosomes / Q000737:chemistry*; D004247:DNA / Q000032:analysis / Q000235:genetics; D015342:DNA Probes / Q000737:chemistry; D004275:DNA, Ribosomal / Q000032:analysis / Q000235:genetics; D004276:DNA, Satellite / Q000032:analysis / Q000235:genetics; D004625:Embryo, Nonmammalian; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism; D005843:Geography; D006110:Grasshoppers / Q000196:embryology / Q000235:genetics*; D017404:In Situ Hybridization, Fluorescence; D007211:Indoles / Q000378:metabolism; D008297:Male; D008677:Metaphase; D042282:Microdissection / Q000379:methods*; D008938:Mitosis; D017422:Sequence Analysis, DNA; D013090:Spermatocytes / Q000378:metabolism; D014960:X Chromosome / Q000737:chemistry*
D015342:DNA Probes; D004275:DNA, Ribosomal; D004276:DNA, Satellite; D005456:Fluorescent Dyes; D007211:Indoles; C007293:DAPI; D004247:DNA