A promising QTL QSns.sau-MC-3D.1 likely superior to WAPO1 for wheat spikelet number per spike shows no adverse effects on yield-related traits

Spikelet number per spike (SNS) is one of the crucial factors determining wheat yield. Thus, improving our understanding of the genes that regulate SNS could help develop higher-yielding wheat varieties. A genetic linkage map constructed using the GenoBaits Wheat 16K Panel and the 660K SNP array contained 5991 polymorphic SNP markers spanning 2813.26 cM. A total of twelve QTL for SNS were detected in the recombinant inbred line (RIL) population msf × Chuannong 16 (MC), and two of them, i.e., QSns.sau-MC-3D.1 and QSns.sau-MC-7A , were stably expressed. QSns.sau-MC-3D.1 had high LOD values ranging from 4.99 to 11.06 and explained 9.71-16.75% of the phenotypic variation. Comparison of QSns.sau-MC-3D.1 with previously reported SNS QTL suggested that it is likely a novel one. A kompetitive allele-specific PCR (KASP) marker, KASP-10, tightly linked to QSns.sau-MC-3D.1 was developed to successfully validate its effect in three segregated populations and a natural population. Genetic analysis indicated that WHEAT ORTHOLOG OFAPO1 ( WAPO1 ) was a candidate gene for QSns.sau-MC-7A . The combined additive effect of QSns.sau-MC-3D.1 and WAP01 had a great additive effect increasing SNS by 7.10%. In addition, our results suggested that SNS is not affected by 1BL/1RS translocations in the MC RIL population. Correlation analysis between two major QTL and other agronomic traits showed that QSns.sau-MC-3D.1 was likely independent of these agronomic traits. However, the H2 haplotype of WAPO1 may affect effective tiller number and plant height. This indicated that the breeding potential of QSns.sau-MC-3D.1 is better than that of WAPO1 . The geographical distribution of QSns.nsau-MC-3D.1 showed that QSns.sau-MC-3D.1 positive allele frequency was dominant in most wheat-producing regions of China and it has been positively selected among modern cultivars released in China since the 1940s. Two genes, TraesCS3D03G0222600 and TraesCS3D03G0216800 , associated with SNS development were predicted in the physical interval of QSns.sau-MC-3D.1 . qRT-PCR results of the two genes showed that only the expression level of TraesCS3D03G0216800 was significantly different between msf and CN16. These results enrich our understanding of the genetic basis of wheat SNS and will be useful for fine mapping and cloning of genes underlying QSns.sau-MC-3D.1 , and provide a basis for marker-assisted selection breeding. Author summary In this study, we identified two major QTL ( QSns.sau-MC-3D.1 and QSns.sau-MC-7A ) in a RIL population. WAPO1 was demonstrated to be the candidate gene for QSns.sau-MC-7A. QSns.sau-MC-3D.1 was a novel and stably expressed QTL, and further confirmed in different genetic backgrounds. Our results further demonstrate that QSns.sau-MC-3D.1 has better breeding potential because of its no adverse effect on other agronomic traits than WAPO1 , and it has been positively selected during Chinese breeding programs since the 1940s. Taken together, the identification of QSns.sau-MC-3D.1 offers a promising resource to further increase wheat yields. ### Competing Interest Statement The authors have declared that no competing interests exist.