Cloning and characterization of genomic DNA sequences of four self-incompatibility alleles in sweet cherry (Prunus avium L.)

• Published in: Theoretical and applied genetics Vol. 108; no. 2; pp. 299 - 305
• Main Authors: WÜNSCH, A, HORMAZA, J. I
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer 2004; Berlin Springer Nature B.V
• Subjects: 5' Untranslated Regions - genetics; Alleles; Allelomorphism; Amino Acid Sequence; Amino acids; Analysis; Base Sequence; Biological and medical sciences; Classical genetics, quantitative genetics, hybrids; Cloning; Cloning, Molecular; Cultivars; Deoxyribonucleic acid; DNA; DNA sequencing; DNA, Complementary; DNA, Plant - genetics; Ethylenediaminetetraacetic acid; Fundamental and applied biological sciences. Psychology; Genetic aspects; Genetic research; Genetics; Genetics of eukaryotes. Biological and molecular evolution; Molecular Sequence Data; Nucleotide sequencing; Prunus - enzymology; Prunus - genetics; Prunus avium; Pteridophyta, spermatophyta; Ribonuclease; Ribonucleases - chemistry; Ribonucleases - genetics; Sequence Homology, Amino Acid; Sequence Homology, Nucleic Acid; Species Specificity; Vegetals; Japan; Spain; Incompatibility; Cherry; Enzyme; Genomics; Rosaceae; Esterases; Self-incompatibility; Characterization; Allele; S-allele; Dicotyledones; DNA; Prunus avium; Angiospermae; Prunus; Hydrolases; Genetics; Spermatophyta; Agriculture; Pancreatic ribonuclease; RNase
• ISSN: 0040-5752; 1432-2242
• DOI: 10.1007/s00122-003-1418-6

Gametophytic self-incompatibility (GSI) in sweet cherry is determined by a locus S with multiple alleles. In the style, the S-locus codifies for an allele-specific ribonuclease ( S-RNase) that is involved in the rejection of pollen that carries the same S allele. In this work we report the cloning and genomic DNA sequence analysis including the 5' flanking regions of four S-RNases of sweet cherry ( Prunus avium L., Rosaceae). DNA from the cultivars Ferrovia, Pico Colorado, Taleguera Brillante and Vittoria was amplified through PCR using primers designed in the conserved sequences of sweet cherry S-RNases. Two alleles were amplified for each cultivar and three of them correspond to three new S-alleles named S23, S24 and S25 present in 'Pico Colorado', 'Vittoria' and 'Taleguera Brillante' respectively. To confirm the identity of the amplified fragments, the genomic DNA of these three putative S-RNases and the allele S12 amplified in the cultivar Ferrovia were cloned and sequenced. The nucleotide and deduced amino-acid sequences obtained contained the structural features of rosaceous S-RNases. The isolation of the 5'-flanking sequences of these four S-RNases revealed a conserved putative TATA box and high similarity among them downstream from that sequence. However, similarity was low compared with the 5'-flanking regions of S-RNases from the Maloideae. S6- and S24-RNase sequences are highly similar, and most amino-acid substitutions among these two RNases occur outside the rosaceous hypervariable region (RHV), but within another highly variable region. The confirmation of the different specificity of these two S-RNases would help elucidate which regions of the S-RNase sequences play a role in S-pollen specific recognition.