Use of SNP markers by KASP assay for MAS studies in sunflower against Plasmopara halstedii

Use of SNP markers by KASP assay for MAS studies in sunflower against Plasmopara halstedii

Downy mildew is a fungal disease caused by Plasmopara halstedii and leads to loss of yield up to 100% in sunflower. Diseasecontrol is performed mostly through chemical seed treatment and breeding. Due to the time consuming nature of conventionalbreeding, it is supported by biotechnological approaches. Marker-assisted selection (MAS) is a strategic approach in molecular breedingusing molecular markers. Single nucleotide polymorphisms (SNPs) such as insertions, deletions, and base-pair substitutions are moreadvantageous than other molecular markers. The abundance and biallelic nature of SNPs in a genome provide flexibility in the choosingof SNPs at the desired loci. Competitive allele-specific PCR (KASP) is a genotyping technology for screening of trait-specific SNPmarkers. In this study, SNP markers (NSA002867, NSA006138; NSA000052, NSA000354; NSA002220, NSA002251) linked with thedowny mildew resistance genes Plarg, Pl13, and Pl8, respectively, were analyzed via KASP in three parental crosses (RHA-419 × Colombi,RHA-419 × P64LC53, RHA-419 × Oliva) for Plarg, one parental cross (HA-R5 × P64LC53) for Pl13, one parental cross (P64LC53 × HA-89)for Pl8, and 140 F2 individuals. According to the allelic discrimination results, NSA002867 and NSA006138 markers were discriminativein all crosses for Plarg, NSA000354 marker was discriminative for Pl13, and NSA002220 and NSA002251 markers were discriminativefor Pl8. This study has revealed the potential use of SNP markers in combination with KASP assay for MAS studies, particularly downymildew resistance in sunflower.

___

  • Apaydın A (2008). Zirai Mücadele Teknik Talimatları. Volume II. Republic of Turkey General Directorate of Agricultural Research and Policies. Ankara, Turkey: TAGEM. pp. 8-11.
  • Chhetri M, Bariana H, Wong D, Sohail Y, Hayden M, Bansal U (2017). Development of robust molecular markers for marker-assisted selection of leaf rust resistance gene Lr23 in common and durum wheat breeding programs. Mol Breeding 37: 21.
  • Doyle JJ, Doyle JL (1987). A rapid DNA isolation procedure from small quantities of fresh leaf tissues. Phytochem Bull 19: 11-15.
  • Dußle CM, Hahn V, Knapp SJ, Bauer E (2004). PlArg from Helianthus argophyllus is unlinked to other known downy mildew resistance genes in sunflower. Theor Appl Genet 109: 1083-1086.
  • Force EM, Dunford NT, Salas JJ (2015). Sunflower Chemistry, Production, Processing, and Utilization. 1st Edition. Urbana, IL, USA: AOCS Press.
  • Forrest K, Pujol V, Bulli P, Pumphrey M, Wellings C, Herrera-Foessel S, Huerta-Espino J, Singh R, Lagudah E, Hayden M et al. (2014). Development of a SNP marker assay for the Lr67 gene of wheat using a genotyping by sequencing approach. Mol Breeding 34: 2109-2118.
  • Gascuel Q, Martinez Y, Vear F, Godiard L (2015). The sunflower downy mildew pathogen Plasmopara halstedii. Mol Plant Pathol 16: 109-113.
  • Gascuel Q, Bordat A, Sallet E, Pouilly N, Carrere S, Roux F, Vincourt P, Godiard L (2016). Effector polymorphisms of the sunflower downy mildew pathogen Plasmopara halstedii and their use to identify pathotypes from field isolates. PLOS ONE doi:10.1371/ journal.pone.0148513.
  • Gedil MA, Wye C, Berry ST, Seger B, Peleman J, Jones R, Leon A, Slabaugh MB, Knapp SJ (2001). An integrated restriction fragment length polymorphism - amplified fragment length polymorphism linkage map for cultivated sunflower. Genome 44: 213-221.
  • Gentzbittel L, Mestries E, Mouzeyar S, Mazeyrat F, Badaoui S, Vear F, De Labrouhe DT, Nicolas P (1999). A composite map of expressed sequences and phenotypic traits of the sunflower (Helianthus annuus L.) genome. Theor Appl Genet 99: 218-234.
  • He C, Holme J, Anthony J (2014). SNP genotyping: the KASP assay. Methods Mol Biol 1145: 75-86.
  • Hu J, Seiler G, Kole C (2010). Genetics, Genomics and Breeding of Sunflower. Boca Raton, FL, USA: CRC Press.
  • Jocić S, Cvejić S, Hladni N, Miladinović, D, Miklič V (2010). Development of sunflower genotypes resistant to downy mildew. Helia 33: 173-180.
  • Khera P, Upadhyaya HD, Pandey MK, Roorkiwal M, Sriswathi M, Janila P, Guo Y, McKain MR, Nagy ED, Knapp SJ et al. (2013). Single nucleotide polymorphism–based genetic diversity in the reference set of peanut (Arachis spp.) by developing and applying cost-effective competitive allele-specific polymerase chain reaction genotyping assays. Plant Genome 6. doi: 10.3835/ plantgenome2013.06.0019.
  • Kim C, Guo H, Kong W, Chandnani R, Shuang LS, Paterson AH (2016). Application of genotyping by sequencing technology to a variety of crop breeding programs. Plant Sci 242: 14-22.
  • Kursterer B, Rozynek B, Brahm L, Prufe M, Tsigos S, Horn R, Friedt W (2004). Construction of a genetic map and localization of major traits in sunflower (Helianthus annuus L.). Helia 40: 15- 24.
  • Lai Z, Livingstone K, Zou Y, Church SA, Knapp SJ, Andrews J, Rieseberg LH (2005). Identification and mapping of SNPs from ESTs in sunflower. Theor Appl Genet 111: 1532-1544.
  • Liu Z, Gulya TJ, Seiler GJ, Vick BA, Jan CC (2012). Molecular mapping of the Pl16 downy mildew resistance gene from HAR4 to facilitate marker-assisted selection in sunflower. Theor Appl Genet 125: 121-131.
  • Long YM, Chao WS, Ma GJ, Xu SS, Qi LL (2016). An innovative SNP genotyping method adapting multiple platforms and throughputs. Theor Appl Genet 130: 597-607.
  • Mouzeyar S, Roeckel-Drevet P, Gentzbittel L, Philippon J, De Labrouhe DT, Vear F, Nicolas P (1995). RFLP and RAPD mapping of the sunflower Pl1 locus for resistance to Plasmopara halstedii race 1. Theor Appl Genet 91: 733-737.
  • Mulpuri S, Liu Z, Feng J, Gulya T, Jan C (2009). Inheritance and molecular mapping of a downy mildew resistance gene, Pl13 in cultivated sunflower (Helianthus annuus L.). Theor Appl Genet 119: 795-803.
  • Qi LL, Talukder ZI, Hulke BS, Foley ME (2017). Development and dissection of diagnostic SNP markers for the downy mildew resistance genes PlArg and Pl8 and maker assisted gene pyramiding in sunflower (Helianthus annuus L.). Mol Genet Genomics doi: 10.1007/s00438-017-1290-8.
  • Radwan O, Bouzidi MF, Vear F, Philippon J, De Labrouhe TD, Nicolas P, Mouzeyar S (2003). Identification of non-TIR-NBSLRR markers linked to the Pl5 /Pl8 locus for resistance to downy mildew in sunflower. Theor Appl Genet 106: 1438-1446.
  • Roeckel-Drevet P, Gagne G, Mouzeyar S, Gentzbittel L, Philippon J, Nicolas P, De Labrouhe DT, Vear F (1996). Colocation of downy mildew (Plasmopara halstedii) resistance genes in sunflower (Helianthus annuus L.). Euphytica 91: 225-228.
  • Semagn K, Babu R, Hearne S, Olsen M (2014). Single nucleotide polymorphism genotyping using kompetitive allele-specific PCR (KASP): overview of the technology and its application in crop improvement. Mol Breeding 33: 1-14.
  • Shi Z, Liu S, Noe J, Arelli P, Meksem K, Li Z (2015). SNP identification and marker assay development for high-throughput selection of soybean cyst nematode resistance. BMC Genomics 16: 314.
  • Staton SE, Bakken BH, Blackman BK, Chapman MA, Kane NC, Tang S, Ungerer MC, Knapp SJ, Rieseberg LH, Burke JM (2012). The sunflower (Helianthus annuus L.) genome reflects a recent history of biased accumulation of transposable elements. Plant J 72: 142-53.
  • Talukder ZI, Gong L, Hulke BS, Pegadaraju V, Song QJ, Schultz Q, Qi LL (2014) A high-density SNP map of sunflower derived from RAD-sequencing facilitating fine-mapping of the rust resistance gene R12. PLOS ONE doi:10.1371/journal. pone.0098628.
  • Tan C-T, Assanga S, Zhang G, Rudd JC, Haley SD, Xue Q, Ibrahim A, Bai G, Zhang X, Byrne P, Fuentealba MP, Liu S (2017). Development and validation of KASP markers for wheat streak mosaic virus resistance gene Wsm2. Crop Sci 57: 340-349.
  • Thomson MJ (2014). High-throughput SNP genotyping to accelerate crop improvement. Plant Breed Biotechnol 2: 195-212.
  • Tozlu E (2008). Ayçiçeği’nde Sclerotinia sclerotiorum ve Sclerotinia minor’ın kültürel, biyolojik ve kimyasal mücadelesi. Atatürk University JAF 39: 281-286.
  • Umesha S, Dharmesh SM, Shetty SA, Krishnappa M, Shetty HS (1998). Biocontrol of downy mildew disease of pearl millet using Pseudomonas fluorescens. Crop Prot 17: 387-392.
  • Usatov AV, Klimenko AI, Azarin KV, Gorbachenko OF, Markin NV, Tikhobaeva VE, Kolosov YA, Usatova OA, Bakoev SY, Bibov MY et al. (2014). DNA-markers of sunflower resistance to the downy mildew (Plasmopara halstedii). Am J Biochem Biotechnol 10: 125-129.
  • Vear F, Gentzbittel L, Philippon J, Mouzeyar S, Mestries E, RoeckelDrevet P, De Labroube DT, Nicolas P (1997). The genetics of resistance to five races of downy mildew (Plasmopara halstedii) in sunflower (Helianthus annuus L.) Theor Appl Genet 95: 584- 589.
  • Yu JK, Tang S, Slabaugh MB, Heesacker A, Cole G, Herring MJ, Soper J, Han F, Chu WC, Webb DM et al. (2003). Towards a saturated molecular genetics linkage map for cultivated sunflower. CSSA 43:367-387.