An under-methylated region in the spacer of ribosomal RNA genes of Lilium henryi

Abstract

Ribosomal RNA genes of Lilium henryi are almost completely methylated at CG and CNG sequences. A short under-methylated region was detected between 2.05 and 2.4 kbp upstream of the 18S sequences. It included the only sites of digestion by four methylation-sensitive restriction endonucleases - PstI, Hae II, Eco RII and Hpa II. Only about 15%-20% of rDNA repeats from shoot meristem are susceptible to each of the enzymes. The same repeats are apparently cut by all enzymes and occur in contiguous blocks. Because the region involved is likely to include regulatory sequences it may be that under-methylation occurs specifically in active rDNA repeats. To test this, rDNA was examined from pollen mother cells at pachytene where transcription has fallen to near zero. Under-methylation levels here were similar to those in shoot meristem tissue. Thus methylation of this region is not the agent responsible for rDNA gene inactivation in pachytene cells and it does not occur immediately genes become inactive. Even so, sequences in this region might be prevented from becoming methylated in transcribing repeats.