The ditelosomic line for the long arm of chromosome 4B (4BL) of Chinese Spring (CS) wheat is not available because it is completely male sterile. Since all deletions in the 4B short arm (4BS) cause male sterility in the homozygous condition, a male-fertility gene should be located in a distal region of 4BS. Among the selfed progeny of a hybrid between a male-sterile 4BS deletion plant (4BS-8) and a Japanese common wheat cultivar Norin 61 (N61), we obtained self-fertile 4BS-8 homozygous deletion plants. We also found fertile nullisomic-4B plants among the selfed progeny of a hybrid between a monosomic line 4B of N61 and CS. This fact suggested that N61 has a novel male-fertility gene on a chromosome other than 4B. We established a self-fertile ditelosomic 4BL line after backcrossing a fertile 4BS-8 plant to CS monotelodisomic 4BL. By cytological observation and PCR analysis using 16 SSR markers, we verified that this ditelosomic line lacked the entire 4BS arm and found that three of the markers were on 4BS. We conducted deletion mapping using three 4BS homozygous deletion lines and three 4BS-specific markers.