Several Influencing Factors on Fluorescent in situ Hybridization Experimental System Applied to Dendranthema spp.

Fluorescent in situ hybridization (FISH) was applied to investigate the phylogenetic relationship among Dendranthema spp. Genomic DNA of wild species which was used as probe did not give specific signals, while 18S-26S rDNA from Arabidopsis, which was used as control probe, showed the loci on the target chromosomes clearly. Satisfied results of FISH were gotten when denaturing digoxingenen-labeled probe and chromosome together in oven at 80℃ for 1015min. There is little influence on the result by the stringency of washing when rDNA was used as probe. The result also indicates the limitation of genomic in situ hybridization (GISH) when used as an approach to analyze the phylogenetic relationship among Dendranthema spp. and the origin of cultivated chrysanthemum.