Abstract
BackgroundThe wheat mutant line three-pistil (TP) exhibits three pistils per floret. As TP normally has two or three seeds in each of the florets on the same spike, there is the possibility of increasing the number of grains per spike. Therefore, TP is a highly valuable mutant for breeding and for the study of floral development in wheat. To map the three-pistil gene (Pis1), genotyping-by-sequencing single-nucleotide polymorphism (GBS-SNP) data from an F2 mapping population (CM28 × CM28TP) was used to construct a genetic map that is of significant value.ResultsIn the present study, a high-density genetic map of wheat containing 2917 GBS-SNP markers was constructed. Twenty-one linkage groups were resolved, with a total length of 2371.40 cM. The individual chromosomes range from 2.64 cM to 454.55 cM with an average marker density of 0.81 cM. The Pis1 gene was mapped using this high-resolution map, and two flanking SNP markers tightly linked to the gene, M70 and M71, were identified. The Pis1 is 3.00 cM from M70 and 1.10 cM from M71. In bread wheat genome, M70 and M71 were found to delimit a physical distance of 3.40 Mb, which encompasses 127 protein-coding genes. To validate the GBS-generated genotypic data and to eliminate missing marker data in the Pis1 region, five Kompetitive Allele-Specific PCR (KASP) assays were designed from corresponding GBS sequences, which harbor SNPs that surround Pis1. Three KASP-SNP markers, KM70, KM71, and KM75, were remapped to the Pis1 gene region.ConclusionsThis work not only lays the foundation for the map-based cloning of Pis1 but can also serve as a valuable tool for studying marker-trait association of important traits and marker-assisted breeding in wheat.
Highlights
The wheat mutant line three-pistil (TP) exhibits three pistils per floret
This work lays the foundation for the map-based cloning of Pis1 but can serve as a valuable tool for studying marker-trait association of important traits and marker-assisted breeding in wheat
To verify the GBS-generated genotypic data and to eliminate missing marker data in the Pis1 region, five Kompetitive allele-specific PCR (KASP) markers were designed from corresponding GBS sequences harboring Singlenucleotide polymorphism (SNP) surrounding Pis1 (Table 2)
Summary
As TP normally has two or three seeds in each of the florets on the same spike, there is the possibility of increasing the number of grains per spike. One effective method of improving wheat yield is to increase the number of grains per spike [1, 2], which can be achieved through a wide-range of genetic variations in the morphological framework of wheat, including superfluous spikelets and multiple spikelets [3] as well as multiple rows of spikes [4]. Three-pistil (TP) mutant wheat has normal spike morphology but exhibits three pistils per floret [5]. Previous studies have shown that the three-pistil trait is controlled by the Pis gene, a dominant gene located on chromosome arm 2DL [5, 6] between simple sequence repeat (SSR) markers Xgwm539 and Xgwm349 [7].
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