Abstract Sterile and semi-fertile F1 plants were obtained by intergeneric sexual hybridization between paternal Brassica oleracea var. alboglabra (genome CC, 2n=18) and maternal Sinapis alba (genome SS, 2n=24), BC1 plants were obtained by backcrossing between paternal B. oleracea and maternal semi-fertile F1 plants. Genomic in situ hybridization (GISH) combined with dual-colour fluorescence in situ hybridization (dcFlSH) showed that sterile F1 plants contained 21 chromosomes consisting of one B. oleracea chromosome set and one S. alba chromosome set, belonging to expected hybrids, and semi-fertile F1 plants contained 30 chromosomes consisting of two B. oleracea chromosome sets and one S. alba chromosome set. It is obvious that the semi-fertile F1 plants belong to unexpected hybrids. 1 -3 trivalents were detected at meiotic metaphase I of semi-fertile F1 pollen mother cells (PMCs). Different separation ratios of S chromosomes were detected at anaphase I. A monosomic alien addition line (MAAL) was identified by GISH-dcFISH from BC1 plants; it contained 19 chromosomes consisting of 18 C chromosomes and 1 S chromosome. At meiotic metaphase 1, 9 divalents from B. oleracea and one univalent from S. alba could be detected. Sometimes, one putative C-S trivalent could also be detected. The achievement of B. oleracea-S. alba monosomic alien addition lines lays a foundation for gene introgression, location and cloning. [ABSTRACT FROM AUTHOR]