A chimeric open reading frame associated with cytoplasmic male sterility in alloplasmic wheat with Triticum timopheevi mitochondria is present in several Triticum and Aegilops species, barley, and rye.

Mitochondrial DNA from Triticum timopheevi has a chimeric gene, orf256, upstream of coxI. This gene is cotranscribed with coxI in cytoplasmic male sterile plants and produces a 7-kDa protein which is not produced in fertile or fertility-restored plants. T. aestivum, the nuclear donor in sterile plants, does not have orf256. Analysis by polymerase chain reaction of DNA from barley, rye, Aegilops bicornis, Ae. searsii, Ae. sharonensis, Ae. speltoides, Ae. tauschii, T. monococcum, and T. turgidum was done with oligonucleotide primers designed to detect orf256 or coxI sequences. Except for T. turgidum, these plants have various elements of the orf256 sequence over a 1-kb length of DNA immediately upstream of coxI in exactly the same arrangement as is found in the coxI region of T. timopheevi. Only T. timopheevi and Ae. speltoides have orf256 transcripts, and only cytoplasmic male-sterile plants involving these two species as maternal donors produce a protein from orf256. Part of an orf256-like sequence is present in T. turgidum but is at least slightly different in arrangement relative to coxI, as compared with the sequence in T. timopheevi. Neither maize nor sorghum have the orf256 sequence.