Supplementary Material for: Cryptic Introgression of <i>Dasypyrum villosum</i> Parental DNA in Wheat Lines Derived from Intergeneric Hybridization

Cytogenetic and DNA molecular analyses have been carried out in 3 wheat introgression lines (ILs; CS×V58, CS×V59, and CS×V60) derived from <i>Triticum aestivum</i> cv. ‘Chinese Spring’ (CS) × <i>Dasypyrum villosum</i><i>(Dv)</i> intergeneric hybridization. All lines, which showed several phenotypic differences compared to CS, had the same chromosome number (2n = 42) and structure as CS, and neither chromosomes nor chromatin from <i>Dv</i> were apparently added to their complement. However, Feulgen/DNA cytophotometry showed that there was more nuclear DNA in the lines than in the parental wheat (by 1.85%, 2.76%, and 1.26% in CS×V58, CS×V59, and CS×V60, respectively). Molecular investigation indicated the presence of <i>Dv</i> DNA in the ILs. AFLP analysis of genomic DNA from the ILs, CS, and <i>Dv</i> detected a total of 120 polymorphic bands, of which 7 (5.8%) were present in some or all the ILs and <i>Dv</i> but were absent in CS. PCR amplification, sequence analysis of amplicons, and Southern blot hybridization confirmed the presence of <i>Dv</i>-specific sequences in each of the ILs. These results indicate cryptic introgression of <i>Dv</i> DNA sequences into the genome of the ILs. Some implications of this finding are discussed.