was verified by extended periods of cultivation both in the absence and presence of norflurazon. The pds and ble were amplified by PCR and partial region of pds including the mutation site and ble were subjected to DNA sequencing (see Figs. 3 and 5 for representative results). All the transformed norflurazon-resistant colonies revealed the DNA sequence of the donor plasmid derived from pPlat–pds. The point mutation conferring norflurazon resistance was identified in all transformed colonies (GC instead of TG in position 5185–5186 in the mutated pds) (Fig. 5A). Insertion of the foreign DNA was also confirmed by the presence of polymorphisms in the introns of the pds. The sequence of pds in pPlat–pds was derived from H. pluvialis strain NIES-144. While pds of transformed strains showed a sequence similar to that of the pPlat–pds, the WT strain (1844 em. Wille K-0084 (SCCAP)) pds (GenBank accession number: KP826910) and a spontaneous norflurazon-resistant strain showed a different sequence (Fig. 5B). Moreover, sequencing of partial PsaD 5' UTR, full ble CDS, and 3' UTR of PsaD verified insertion of PsaD-ble cassette in all tested colonies (data not shown).