31Wheat (Triticum aestivum L.) is a self-pollinating crop whose hybrids offer the potential to provide a major boost 32 in yield. Male sterility induced by the cytoplasm of Triticum timopheevii is a powerful method for hybrid seed 33 production. Hybrids produced by this method are often partially sterile and full fertility restoration is crucial for 34 wheat production using hybrid cultivars. To identify genetic loci controlling fertility restoration in wheat, we 35 produced two CMS-based backcross (BC1) mapping populations. The restorer lines Gerek 79 and 71R1203 were 36 used to pollinate the male-sterile winter wheat line CMS-Sperber. Seed set and numbers of sterile spikelets per 37 spike were evaluated in 340 and 206 individuals of the populations derived from Gerek 79 and 71R1203, 38 respectively. Genetic maps were constructed using 930 and 994 SNPs, spanning 2,160 and 2,328 cM over 21 39 linkage groups in the two populations, respectively. Twelve quantitative trait loci (QTL) controlled fertility 40 restoration in both BC1 populations, including a novel restorer-of-fertility (Rf) locus flanked by the single 41 nucleotide polymorphism (SNP) markers IWB72413 and IWB1550 on chromosome 6AS. The locus was mapped 42 as a qualitative trait in the BC1 Gerek 79 population and was designated Rf9. Ninety-three putative candidate genes 43 were predicted for the QTL region on chromosome 6AS. Among them were genes encoding tetratricopeptide and 44 pentatricopeptide repeat-containing proteins in rice known to be associated with fertility restoration. This finding 45 is a promising step to better understand the functions of genes for improving hybrid wheat. 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 Since the discovery of male sterility and restoration systems in the 1960s, hybrid wheat triggered attention due to 62 its potential for improved grain and straw productivity and yield stability particularly under harsh and marginal 63 environments (Longin et al. 2012; Whitford et al. 2013). The major gains of hybrid versus line varieties are 64 improved trait values due to heterosis (Castillo et al. 2014). Heterosis or hybrid vigor is the phenomenon by which 65 the progeny derived from a cross of two inbred lines outperform the parent lines and has provided considerable 66 economic benefits in worldwide crop production (Chen and Liu 2014). Heterosis in winter wheat has been reported 67 to provide uniform plant establishment and tolerance against frost, lodging, and diseases such as leaf rust, stripe 68 rust, Septoria tritici blotch and powdery mildew (Gupta et al. 2019).
69To harness yield gains associated with hybrid vigor, the cytoplasmic male sterility (CMS) system provides a cost-70 effective tool for efficient hybrid seed production (Chen and Liu 2014). CMS in plants is based on rearrangements 71 of mitochondrial DNA that lead to chimaeric genes and a condition under which a plant is unable to produce fertile 72 pollen (Whitford et al. 2013). CMS evades the need for manual removal of anthers, thus facilitating a technolo...