Figure 4.

Fluorescence in situ hybridization of centromeric TCAST satDNA and TR families determined in this work by TRF analysis. Chromosomes are counter-stained with DAPI. FISH showing centromeric TCAST satDNA (TCAST subf3 as a probe; red signals) on T. castaneum chromosomes in meiotic pro metaphase (A). Arrows point to chromosomes CH2, CH3, CH4 and Yp. Chromosomes were named according to the karyotype analysis provided by Stuart and Mocelin.²⁹ Two-coloured FISH performed on chromosomes in mitotic pro metaphase show localization of new TR families (green): Cast1 (B), Cast2 (C), Cast3 (D), Cast4 (E), Cast5 (F), Cast6 (G), Cast7 (H), Cast8 (I) and Cast9 (J) with respect to centromeric regions marked with TCAST satDNA (red). The bar represents 1 μm. Aside to chromosome spreads are shown Southern blot analyses of genomic DNA digested with REs and hybridized with Cast1 (B), Cast2 (C), Cast4 (E), Cast 5 (F) and Cast6 (G). Only TR families with >0.5% of genomic DNA are presented.