Allele-specific PCR detection of sweet cherry self-incompatibility alleles S₃, S₄ and S₉ using consensus and allele-specific primers in the Czech Republic

Abstract

Sharma K., Sedlák P., Zeka D., Vejl P., Soukup J., 2014. Allele-specific PCR detection of sweet cherry selfincompatibility alleles S 3 , S 4 and S 9 using consensus and allele-specific primers in the Czech Republic. Hort. Sci. (Prague), 41: 153-159.Prunus avium species of the Rosaceae family exhibit gametophytic self-incompatibility. Determination of the selfincompatibility genotype of individuals is essential for genetic studies and the development of informed management strategies. The PCR-based detection of S-allele helps to promote and speed up traditional breeding activity and hence molecular analysis of the perspective genotypes has become more intensive in all cherry growing countries. The alleles S 3 , S 4 and S 9 from 34 accessions of Czech collections were determined using the polymerase chain reaction (PCR) method. Initially, DNA extracts were amplified with consensus primers that amplify across the first, second, or both introns of the S-ribonuclease gene which shows a considerable length polymorphism. The new allele specific primers were designed with the goal to overcome some occurring difficulties in the detection of expected alleles by previously published allele specific primers. S-alleles fragments of standard cultivars used in this study were PCR amplified, sequenced to validate the designed primers. The study demonstrates the advantage of newly designed primers application in testing of sweet cherry genotypes. Keywords: Prunus avium; S-alleles; allele-specific primers; consensus primersSweet cherries Prunus avium (L.) are usually diploid (2n = 2× = 16) trees (Webster 1996;Iezzoni 2008) which are characterized by mighty growth and a pyramidal tree top. Gametophytic self-incompatibility (GSI) in cherry is controlled by the multi-allelic S-locus, which prevents self-fertilization (de Nettancourt 2001;Franklin-Tong, Franklin 2003). The S-genotype is agronomically important because cultivars with the same S-genotype are cross incompatible.The stylar component of GSI was determined to be a ribonuclease (S-RNase) gene in the Rosaceae (Bošković, Tobutt 1996;Sassa et al. 1996). In the past, several P. avium S-alleles were identified as a result of cross pollination experiments and progeny testing in sweet cherry (Crane, Brown 1937) and later correlated with stylar ribonucleases (Bošković, Tobutt 1996). These S-RNases were also characterized at the molecular level (Tao et al. 1999;

Development of a molecular marker for self‐compatible S4′ haplotype in sweet cherry (Prunus avium L.) using high‐resolution melting

“…Normal pollen tube growth was observed in varieties 'Lapins' and 'Staccato,' which agreed with their self-compatible S4 0 haplotype (Schuster, 2012;Sharma, Sedl ak, Zeka, Vejl, & Soukup, 2014). Images of the pollen tube elongation raising pistils in the varieties 'Lapins'…”

“… S ‐allele genotypes previously reported for varieties, incompatibility groups and origin were included. (1) Bošković & Tobutt, ; (2) Choi, Tao, & Anderson, ; (3) Hauck, Iezzoni, Yamane, & Tao, ; (4) Lacis, Kaufman, Rashal, Trajkovski, & Iezzoni, ; (5) Schuster, ; (6) Schuster, Flachowsky, & Köhler, ; (7) Sharma et al., ; (8) Sonneveld et al., ; (9) Sonneveld et al., ; (10) Tao et al., ; (11) Wiersma et al., ; (12) Wünsch & Hormaza, . SI, self‐incompatible; SC, self‐compatible; O, universal donor.…”

TheR2R3MYBtranscription factorPavMYB10.1involves in anthocyanin biosynthesis and determines fruit skin colour in sweet cherry (Prunus aviumL.)

“…To identify the molecular mechanism underlying fruit skin colour in sweet cherry, we constructed an F1 population. Because Prunus avium exhibits gametophytic self-incompatibility, the dark-red variety 'Lapins' and the dark-red variety 'Wanhongzhu' were selected as cross-parents (Sharma et al, 2014). Analyses of the F1 hybrids showed that the 415 dark-red and 150 blush fruit skin phenotypes segregated in a 3 : 1 ratio.…”

Studying on the strictly self-compatibility mechanism of ‘Liuyefeitao’ peach (Prunus persica L.)

“…The S-RNase play an important role in GSI, which is necessary and sufficient for pollen recognition [ 23 ]. Lots of S-RNase have been isolated in Rosaceae, such as: Malus pumila [ 24 ], Pyrus serotina [ 25 ], Prunus avium [ 26 ], Prunus dulcis [ 27 ], Prunus salicina [ 28 ], Eriobotrya japonica [ 29 ] and Prunus armeniaca [ 30 ]. We found a highly expressed gene (PRUPE_ppa018459mg) in ‘Liuyefeitao’ which is homologous to the S-RNase gene of Prunus dulcis ( S2 Table ).…”