Abstract:To explore the fertility restorer genes of C-type cytoplasmic male sterility (CMS-C) in maize, and analyze the gene functions, two F2 mapping populations were developed with restorer line K932R and two CMS-C allogeneic heterogeneous sterile lines K169S and K932S, respectively. Combined with SSR molecular markers and BSA-reseq technology, the fertility restorer gene in K932R was mapped. The target gene was cloned by PCR technology and gene expression analysis was performed. The results showed that the fertility restorer gene Rf 932 was mapped on the short arm of chromosome 8 by BSA-SSR molecular markers, between SSR markers 8-21 and 8-30, with the genetic distance of 2.3 cM and 7.7 cM, respectively. Rf 932 was located in a 0.38 Mb region on chromosome 8 by BSA-reseq technology, which contained 7 candidate genes. After the candidate genes being cloned in segments and aligned in sequences, Rf 932 was obtained with a full length of 2549 bp, consisting of 4 exons and 3 introns, encoding 385 amino acids; one of the key sites controlling fertility restoration, F187Y, had the same amino acid variation as Rf4, which was a new Rf4 allele; Rf 932 encoded a bHLH transcription factor that regulated the tapetum development. qRT-PCR analysis showed that there was no significant difference in the relative expression levels of Rf 932 in anthers between the sterile line and the restorer line at 3 developmental stages. The results enriched the restore gene bank of maize CMS-C, and laid a foundation for further analyzing the mechanism of fertility restoration.