W14/15 esterase gene haplotype can be a genetic landmark of cultivars and species of the genus Gentiana L

Abstract

We have identified multiple alleles for a single gene termed W14/15. This gene encodes closely related but not identical proteins W14 and W15 that accumulate in overwinter buds of Gentiana triflora (Takahashi et al. in Breed Sci 56:39-46, 2006; Hikage et al. in Mol Genet Genomics 278:95-104, 2007). In this study, structural analysis of the W14/15 gene was carried out for 21 different gentian lines/cultivars consisting of 5 different species, to survey species- or line/cultivar-specific haplotypes. Within the samples examined, multiple variant forms were found. Those were categorized into seven major types (type I-VII) and ten subtypes based on the presence of three short insertion/deletion sites, three RFLP sites, and several SNP sites. Each line/cultivar had a distinct set of W14/15 gene variants for an allelic pair. Phylogenetic analysis showed that the W14/15 alleles cluster into groups that are characteristic of gentian species, i.e., G. triflora, G. scabra, G. pneumonanthe, G. septemfida and an unknown species other than the former four. In addition, within the same gentian species, different sets of haplotypes were found. Thus, the W14/15 alleles provide useful landmarks to resolve phylogenies of the genus or section Gentiana, as well as to analyze pedigree and breeding history of the cultivars derived from those Gentiana sp.

Development of Simple Sequence Repeat Markers for Identification of Japanese Gentian Cultivars

“…synthetic genes were lower than those detected with the SSR markers developed in this study (Shimada et al, 2009). W14/W15 showed high allelic variability among Japanese gentian cultivars, but it was difficult to increase the polymorphic information because they are coded by a single locus (Hikage et al, 2011); therefore, our SSR markers have advantages over the markers developed previously and are useful to protect Japanese gentian breeders' rights.…”

“…'Momokorin' and 'Aokorin' were sib-cultivars from common parents and showed the closest genetic relationship. 'Maerchen Ashiro' and 'Shineblue Ashiro' were classified in the same cluster, as both have a similar genetic background (Hikage et al, 2011). The white-flowered cultivars 'Polarno White', 'Pure White', and 'Crystal Ashiro' also showed high genetic similarity.…”

“…Thus, the SSR markers could estimate genetic similarity among gentian cultivars. Hikage et al (2011) undertook pedigree analysis of Gentiana cultivars and species using haplotypes of the W14/15 esterase gene. SSR markers have been utilized as a tool for pedigree analysis in maize (Smith et al, 1997) and olive (Díaz et al, 2006).…”

“…More recently, Shimada et al (2009) reported that sequence characterized amplified region (SCAR) markers based on length polymorphisms in intron regions of flavonoid biosynthetic genes could distinguish vegetatively propagated gentian cultivars. Phylogenetic and pedigree analysis of Gentiana was conducted using single nucleotide polymorphisms (SNPs) of the W14/ W15 alleles, encoding a member of the α/β-hydrolase fold superfamily (Hikage et al, 2011). Mishiba et al (2009) …”

Development of DNA markers that discriminate between white- and blue-flowers in Japanese gentian plants

“…The haplotypes of W14/ 15 alleles also provided data to resolve phylogenetic relationships in the genus or section Gentiana, and to analyze the pedigree and breeding history of the cultivars derived from those Gentiana spp. (Hikage et al 2011). Although some useful DNA markers have been developed for Japanese gentian, none have been developed for identification of the white flower trait, which is another important flower color variation.…”