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Identification of chromosomal translocation causing inactivation of the gene encoding anthocyanidin synthase in white pomegranate (Punica granatum L.) and development of a molecular marker for genotypic selection of fruit colors
- Jeong, Hyeon-ju, Park, Moon-Young, Kim, Sunggil
- Horticulture, environment and biotechnology 2018 v.59 no.6 pp. 857-864
- Punica granatum, anthocyanidins, breeding programs, chromosome translocation, color, cultivars, fruits, genes, genetic markers, genotyping, messenger RNA, mutation, oxygenases, polymerase chain reaction, pomegranates, seedlings
- Previous studies have not detected transcripts of the gene encoding anthocyanidin synthase (ANS) in white pomegranates (Punica granatum L.) and suggest that a large-sized insertion in the coding region of the ANS gene might be the causal mutation. To elucidate the identity of the putative insertion, 3887-bp 5′ and 3392-bp 3′ partial sequences of the insertion site were obtained by genome walking and a gene coding for an expansin-like protein was identified in these genome-walked sequences. An identical protein (GenBank accession OWM71963) isolated from pomegranate was identified from BLAST search. Based on information of OWM71963, a 5.8-Mb scaffold sequence with genes coding for the expansin-like protein and ANS were identified. The scaffold sequence assembled from a red pomegranate cultivar also contained all genome-walked sequences. Analysis of positions and orientations of these genes and genome-walked sequences revealed that the 27,786-bp region, including the 88-bp 5′ partial sequences of the ANS gene, might be translocated into an approximately 22-kb upstream region in an inverted orientation. Borders of the translocated region were confirmed by PCR amplification and sequencing. Based on the translocation mutation, a simple PCR codominant marker was developed for efficient genotyping of the ANS gene. This molecular marker could serve as a useful tool for selecting desirable plants at young seedling stages in pomegranate breeding programs.