Варіанти гену LEAFY в геномах українських зразків інвазійного виду Reynoutria sachalinensis

Abstract

Highly variable nuclear genes, such as LEAFY, are an effective tool for identifying hybrid/heterozygous forms. However, the widely used method of direct sequencing of PCR products is not suitable for detecting possible intragenomic variants of these genes. Alternative approaches, such as Illumina sequencing of amplicons, make LEAFY genes promising markers for barcoding hybrid and polyploid plants. Important objects of such studies are invasive species of the Reynoutria genus, which include polyploid forms of hybrid origin. Aim. To characterize the polymorphism of the second intron of the LEAFY gene in Ukrainian samples of R. sachalinensis. Methods. PCR amplification, amplicon sequencing using the Illumina method, bioinformatic analysis. Results. Six haplotypes were identified in the genomes of three R. sachalinensis samples. One of these, H2, is common, while the others are specific to individual samples. Haplotypes H1 and H2 correspond to variants S1 and S2 previously identified in North American specimens. Haplotypes H3–H6 contain a mutation present in certain populations of the species primary range, on Hokkaido and Sakhalin, but is absent in North American specimens. Conclusions. Analysis of the second intron sequences of the LEAFY gene revealed intragenomic polymorphism in this region in Ukrainian R. sachalinensis specimens. The use of Illumina amplicon sequencing of the second intron of the LEAFY gene is a promising method for barcoding invasive populations of Reynoutria species.