The sweet cherry plant (Prunus avium L.) is primarily self-incompatible, with so-called S-alleles responsible for the inability of flowers to be pollinated not only by their own pollen grains but also by pollen from other cherries having the same S-alleles. This characteristic has wide-ranging impacts on commercial growing, harvesting, and breeding. However, mutations in S-alleles as well as changes in the expression of M locus-encoded glutathione-S-transferase (MGST) can lead to complete or partial self-compatibility, simplifying orchard management and reducing possible crop losses. Knowledge of S-alleles is important for growers and breeders, but current determination methods are challenging, requiring several PCR runs. Here we present a system for the identification of multiple S-alleles and MGST promoter variants in one-tube PCR, with subsequent fragment analysis on a capillary genetic analyzer. The assay was shown to unequivocally determine three MGST alleles, 14 self-incompatible S-alleles, and all three known self-compatible S-alleles (S3', S4', S5') in 55 combinations tested, and thus it is especially suitable for routine S-allele diagnostics and molecular marker-assisted breeding for self-compatible sweet cherries. In addition, we identified a previously unknown S-allele in the 'Techlovicka´ genotype (S54) and a new variant of the MGST promoter with an 8-bp deletion in the ´Kronio´ cultivar.

Research and Breeding Institute of Pomology Holovousy Ltd Holovousy 129 508 01 Hořice Czech Republic

Zobrazit více v PubMed

de Nettancourt D. Incompatibility in Angiosperms. 1st ed. Springer; Berlin/Heidelberg, Germany: 1977. pp. 1–232. DOI

Muñoz-Sanz J.V., Zuriaga E., Cruz-García F., McClure B., Romero C. Self-(In)compatibility Systems: Target Traits for Crop-Production, Plant Breeding, and Biotechnology. Front. Plant Sci. 2020;11:195. doi: 10.3389/fpls.2020.00195. PubMed DOI PMC

Crane M.B., Brown A.G. Incompatibility and Sterility in the Sweet Cherry, Prunus avium L. J. Pomol. Hortic. Sci. 1938;15:2–116. doi: 10.1080/03683621.1938.11513493. DOI

Tao R., Yamane H., Sugiura A., Murayama H., Sassa H., Mori H. Molecular Typing of S-alleles through Identification, Characterization and cDNA Cloning for S-RNases in Sweet Cherry. J. Am. Soc. Hortic. Sci. 1999;124:224–233. doi: 10.21273/JASHS.124.3.224. DOI

Yamane H., Ikeda K., Ushijima K., Sassa H., Tao R. A Pollen-Expressed Gene for a Novel Protein with an F-box Motif that is Very Tightly Linked to a Gene for S-RNase in Two Species of Cherry, Prunus cerasus and P. avium. Plant Cell Physiol. 2003;44:764–769. doi: 10.1093/pcp/pcg088. PubMed DOI

Schröpfer S., Schuster M., Flachowsky H. Detection of S-alleles in a sweet cherry collection from Azerbaijan and Turkey using next generation sequencing. Acta Hortic. 2022;1342:269–276. doi: 10.17660/ActaHortic.2022.1342.38. DOI

Matsumoto D., Tao R. Distinct Self-recognition in the Prunus S-RNase-based Gametophytic Self-incompatibility System. Hortic. J. 2016;85:289–305. doi: 10.2503/hortj.mi-ir06. DOI

Matsumoto D., Tao R. Recognition of a wide-range of S-RNases by S locus F-box like 2, a general-inhibitor candidate in the Prunus-specific S-RNase-based self-incompatibility system. Plant Mol. Biol. 2016;91:459–469. doi: 10.1007/s11103-016-0479-2. PubMed DOI

Matsumoto D., Tao R. Recognition of S-RNases by an S locus F-box like protein and an S haplotype-specific F-box like protein in the Prunus-specific self-incompatibility system. Plant Mol. Biol. 2019;100:367–378. doi: 10.1007/s11103-019-00860-8. PubMed DOI

Lisek A., Kucharska D., Głowacka A., Rozpara E. Identification of S-haplotypes of European cultivars of sour cherry. J. Hortic. Sci. Biotechnol. 2017;92:484–492. doi: 10.1080/14620316.2017.1289071. DOI

Lewis D., Crowe L.K. Structure of the incompatibility gene. Heredity. 1954;8:357–363. doi: 10.1038/hdy.1954.38. DOI

Sonneveld T., Tobutt K.R., Vaughan S.P., Robbins T.P. Loss of Pollen-S Function in Two Self-Compatible Selections of Prunus avium Is Associated with Deletion/Mutation of an S Haplotype–Specific F-Box Gene. Plant Cell. 2004;17:37–51. doi: 10.1105/tpc.104.026963. PubMed DOI PMC

Marchese A., Bošković R.I., Caruso T., Raimondo A., Cutuli M., Tobutt K.R. A new self-compatibility haplotype in the sweet cherry ’Kronio’, S5’, attributable to a pollen-part mutation in the SFB gene. J. Exp. Bot. 2007;58:4347–4356. doi: 10.1093/jxb/erm322. PubMed DOI

Cachi A.M., Wünsch A. Characterization and mapping of non-S gametophytic self-compatibility in sweet cherry (Prunus avium L.) J. Exp. Bot. 2011;62:1847–1856. doi: 10.1093/jxb/erq374. PubMed DOI

Ono K., Akagi T., Morimoto T., W�Nsch A., Tao R., Wünsch A. Genome Re-Sequencing of Diverse Sweet Cherry (Prunus avium) Individuals Reveals a Modifier Gene Mutation Conferring Pollen-Part Self-Compatibility. Plant Cell Physiol. 2018;59:1265–1275. doi: 10.1093/pcp/pcy068. PubMed DOI

Bošković R., Tobutt K.R. Correlation of stylar ribonuclease zymograms with incompatibility alleles in sweet cherry. Euphytica. 1996;90:245–250. doi: 10.1007/BF00023865. DOI

Bošković R., Russell K., Tobutt K. Inheritance of stylar ribonucleases in cherry progenies, and reassignment of incompatibility alleles to two incompatibility groups. Euphytica. 1997;95:221–228. doi: 10.1023/A:1002945529157. DOI

Sonneveld T., Robbins T.P., Bošković R., Tobutt K.R. Cloning of six cherry self-incompatibility alleles and development of allele-specific PCR detection. Theor. Appl. Genet. 2001;102:1046–1055. doi: 10.1007/s001220000525. DOI

Sonneveld T., Tobutt K.R., Robbins T.P. Allele-specific PCR detection of sweet cherry self-incompatibility (S) alleles S1 to S16 using consensus and allele-specific primers. Theor. Appl. Genet. 2003;107:1059–1070. doi: 10.1007/s00122-003-1274-4. PubMed DOI

Sonneveld T., Robbins T.P., Tobutt K.R. Improved discrimination of self-incompatibility S-RNase alleles in cherry and high throughput genotyping by automated sizing of first intron polymerase chain reaction products. Plant Breed. 2006;125:305–307. doi: 10.1111/j.1439-0523.2006.01205.x. DOI

Vaughan S.P., Russell K., Sargent D.J., Tobutt K.R. Isolation of S-locus F-box alleles in Prunus avium and their application in a novel method to determine self-incompatibility genotype. Theor. Appl. Genet. 2006;112:856–866. doi: 10.1007/s00122-005-0187-9. PubMed DOI

Cachi A.M., Wünsch A., Vilanova A., Guàrdia M., Ciordia M., Aletà N. S-locus diversity and cross-compatibility of wild Prunus avium for timber breeding. Plant Breed. 2017;136:126–131. doi: 10.1111/pbr.12450. DOI

Lisek A., Rozpara E., Glowacka A., Kucharska D., Zawadzka M. Identification of S-genotypes of sweet cherry cultivars from Central and Eastern Europe. Hort. Sci. 2015;42:13–21. doi: 10.17221/103/2014-HORTSCI. DOI

Szikriszt B., Doǧan A., Ercisli S., Akcay M.E., Hegedüs A., Halász J. Molecular typing of the self-incompatibility locus of Turkish sweet cherry genotypes reflects phylogenetic relationships among cherries and other Prunus species. Tree Genet. Genomes. 2013;9:155–165. doi: 10.1007/s11295-012-0543-2. DOI

Wünsch A., Hormaza J.I. Cloning and characterization of genomic DNA sequences of four self-incompatibility alleles in sweet cherry (Prunus avium L.) Theor. Appl. Genet. 2004;108:299–305. doi: 10.1007/s00122-003-1418-6. PubMed DOI

Vaughan S.P., Bošković R.I., Gisbert-Climent A., Russell K., Tobutt K.R. Characterisation of novel S-alleles from cherry (Prunus avium L.) Tree Genet. Genomes. 2008;4:531–541. doi: 10.1007/s11295-007-0129-6. DOI

Ikeda K., Watari A., Ushijima K., Yamane H., Hauck N.R., Iezzoni A.F., Tao R. Molecular Markers for the Self-compatible S4’-haplotype, a Pollen-part Mutant in Sweet Cherry (Prunus avium L.) J. Amer. Soc. Hort. Sci. 2004;129:724–728. doi: 10.21273/JASHS.129.5.0724. DOI

Muñoz-Espinoza C., Espinosa E., Bascuñán R., Tapia S., Meneses C., Almeida A. Development of a molecular marker for self-compatible S4′ haplotype in sweet cherry (Prunus avium L.) using high-resolution melting. Plant Breed. 2017;136:987–993. doi: 10.1111/pbr.12546. DOI

Schuster M. Self-incompatibility (S) genotypes of cultivated sweet cherries—An overview update 2020. Open Agrar. Repos. 2020 doi: 10.5073/20201016-141600. DOI

Lapins K. Stella, a self-fruitful sweet cherry. Can. J. Plant Sci. 1971;51:252–253. doi: 10.4141/cjps71-051. DOI

Kou X., Zhang L., Yang S., Li G., Ye J. Selection and validation of reference genes for quantitative RT-PCR analysis in peach fruit under different experimental conditions. Sci. Hortic. 2017;225:195–203. doi: 10.1016/j.scienta.2017.07.004. DOI

Cmejlova J., Rejlova M., Paprstein F., Cmejla R. A new one-tube reaction kit for the SSR genotyping of apple (Malus × domestica Borkh.) Plant Sci. 2021;303:110768. doi: 10.1016/j.plantsci.2020.110768. PubMed DOI

Holušová K., Čmejlová J., Suran P., Čmejla R., Sedlák J., Zelený L., Bartoš J. High-resolution genome-wide association study of a large Czech collection of sweet cherry (Prunus avium L.) on fruit maturity and quality traits. Hortic Res. 2022;10:uhac233. doi: 10.1093/hr/uhac233. PubMed DOI PMC

Peace C.P. DNA-informed breeding of rosaceous crops: Promises, progress and prospects. Hortic. Res. 2017;4:17006. doi: 10.1038/hortres.2017.6. PubMed DOI PMC

Sebolt A.M. Breeder profile: Nnadozie Oraguzie. RosBREED Newslett. 2011;2:5–6.

Patzak J., Henychová A., Paprštein F., Sedlák J. Molecular S-genotyping of sweet cherry (Prunus avium L.) genetic resources. Hort. Sci. 2019;46:146–152. doi: 10.17221/245/2017-HORTSCI. DOI