Abstract
Sunflower (Helianthus annuus L.) production is challenged by different biotic and abiotic stresses, among which downy mildew (DM) is a severe biotic stress that is detrimental to sunflower yield and quality in many sunflower-growing regions worldwide. Resistance against its infestation in sunflower is commonly regulated by single dominant genes. Pl17 and Pl19 are two broad-spectrum DM resistance genes that have been previously mapped to a gene cluster spanning a 3.2 Mb region at the upper end of sunflower chromosome 4. Using a whole-genome resequencing approach combined with a reference sequence-based chromosome walking strategy and high-density mapping populations, we narrowed down Pl17 to a 15-kb region flanked by SNP markers C4_5711524 and SPB0001. A prospective candidate gene HanXRQChr04g0095641 for Pl17 was identified, encoding a typical TNL resistance gene protein. Pl19 was delimited to a 35-kb region and was approximately 1 Mb away from Pl17, flanked by SNP markers C4_6676629 and C4_6711381. The only gene present within the delineated Pl19 locus in the reference genome, HanXRQChr04g0095951, was predicted to encode an RNA methyltransferase family protein. Six and eight SNP markers diagnostic for Pl17 and Pl19, respectively, were identified upon evaluation of 96 diverse sunflower lines, providing a very useful tool for marker-assisted selection in sunflower breeding programs.
Highlights
Sunflower (Helianthus annuus L.) production is challenged by different biotic and abiotic stresses, among which downy mildew (DM) is a severe biotic stress that is detrimental to sunflower yield and quality in many sunflower-growing regions worldwide
We report the use of reference sequence-based chromosome walking toward the target genes, Pl17 and Pl19, identify candidate genes, and develop user-friendly single nucleotide polymorphism (SNP) markers diagnostic for Pl17 and Pl19
The genome sequence of chromosome 4 was extracted from the HA412-HO reference assembly from 3,621,089 to 6,852,749 bp and the XRQ assembly from 5,662,479 to 5,707,598 bp, which covers the Pl17 and Pl19 loci reported in previous studies[16,17]
Summary
Sunflower (Helianthus annuus L.) production is challenged by different biotic and abiotic stresses, among which downy mildew (DM) is a severe biotic stress that is detrimental to sunflower yield and quality in many sunflower-growing regions worldwide. Resistance against its infestation in sunflower is commonly regulated by single dominant genes. Using a whole-genome resequencing approach combined with a reference sequence-based chromosome walking strategy and high-density mapping populations, we narrowed down Pl17 to a 15-kb region flanked by SNP markers C4_5711524 and SPB0001. A prospective candidate gene HanXRQChr04g0095641 for Pl17 was identified, encoding a typical TNL resistance gene protein. Downy mildew (DM), incited by the oomycete Plasmopara halstedii (Farl.) Berlese & de Toni, is one of the most severe biotic factors affecting sunflower production worldwide, in Europe and North America[1,2]. Motif ga — — — ag ag gt ct tg ac ca tg gaa Sequence Resource HA412-HO HA412-HO HA412-HO HA412-HO HA412-HO HA412-HO HA412-HO HA412-HO HA412-HO XRQ XRQ XRQ XRQ
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