A novel mutant allele at the Cleistogamy 1 locus in barley.

key messageA chasmogamous mutant was induced by exposing a cleistogamous cultivar to sodium azide. The altered cly1 sequence in the mutant was not in the miR172 binding site, as is the case in other known cleistogamous alleles, but rather in a region encoding one of the gene product's two AP2 domains. The genetic basis of cleistogamy (in which pollination occurs before the flower opens) in barley is centered on the Cleistogamy 1 locus (cly1). The sequence of the microRNA (miR172)-targeting site in the gene, which belongs to the APETALA2 family, differs between cleistogamous and chasmogamous cultivars at a single nucleotide position, resulting in the differential ability of the lodicules to swell. Here, mutagenesis of the barley cultivar 'Misato Golden' (which carries the cly1.b allele), achieved using sodium azide, was used to induce a change from cleistogamy to chasmogamy (non-cleistogamous flowering). The cly1 coding sequence in the selected mutant differed from that of cly1.b by two non-synonymous mutations, one of which was responsible for an altered residue in one of the AP2 domains present in the Cly1 protein. Although there was no difference in the miR172 targeting site between cly1.b and the novel allele (designated cly1.b3), the mutant's lodicules' ability to swell was indistinguishable from that observed in cultivars carrying the chasmogamous allele Cly1.a. The phenotype of cly1.b3/cly1.b, cly1.b3/cly1.b2 and cly1.b3/cly1.c heterozygotes indicated that cly1.b3 is recessive or incompletely dominant with respect to these alleles.