Emeriewen Ofere Francis, Flachowsky Henryk, Peil Andreas
Julius Kühn-Institut (JKI), Federal Research Centre for Cultivated Plants, Institute for Breeding Research On Fruit Crops, Dresden, Germany.
BMC Res Notes. 2021 Jul 27;14(1):291. doi: 10.1186/s13104-021-05709-2.
The proposed candidate gene underlying the Malus fusca fire blight resistance locus on chromosome 10 was previously predicted to possess 880 amino acids and 8 exons. Eight base pair (8 bp) insertion/deletion in the first exon potentially distinguished resistant genotypes from susceptible ones. This study aimed at analyzing the candidate gene sequence in another set of original resistant and susceptible progeny, characterizing the sequence in a transgenic line transformed with the candidate gene under its own native promoter, as well as deciphering the potential genomic differences between this candidate gene and its homolog in the 'Golden Delicious' doubled haploid genome (GDDH13).
Sequences of amplicons of part of the candidate gene amplified in two progenies that showed resistant and susceptible fire blight phenotypes, confirmed the 8 bp insertion that distinguishes susceptible and resistant progenies. The transgenic line was positive for the candidate gene sequence, confirming a successful transfer into the background of apple cultivar 'Pinova', and possessed the same genomic sequence as the progeny with a resistant phenotype. Sequence analysis showed that the homolog gene on GDDH13 possesses a significant 18 bp deletion in exon 1 leading to a difference of 15 amino acid from the protein sequence of the candidate gene.
先前预测位于10号染色体上的苹果火疫病抗性位点的候选基因含有880个氨基酸和8个外显子。第一个外显子中的8个碱基对(8 bp)插入/缺失可能区分抗性基因型和敏感基因型。本研究旨在分析另一组原始抗性和敏感后代中的候选基因序列,表征在其自身天然启动子驱动下转入候选基因的转基因系中的序列,并解读该候选基因与其在‘金冠’双单倍体基因组(GDDH13)中的同源基因之间潜在的基因组差异。
在表现出抗性和敏感火疫病表型的两个后代中扩增的部分候选基因扩增子序列,证实了区分敏感和抗性后代的8 bp插入。转基因系的候选基因序列呈阳性,证实已成功转入苹果品种‘皮诺娃’的背景中,并且具有与具有抗性表型的后代相同的基因组序列。序列分析表明,GDDH13上的同源基因在第1外显子中存在18 bp的显著缺失,导致其蛋白质序列与候选基因的蛋白质序列有15个氨基酸的差异。