Molecular characteristic of stable and unstable white gene alleles in highly mutable lines from natural Drosophila melanogaster populations

Abstract

Mutations in the white locus emerged in highly mutable isofemale Drosophila melanogaster lines from the populations of Novosibirsk 2013 (NS3 line), Nalchik 2014 (N119 line), and Sakhalin Island 2014 (S46 line). A single white-eyed male found in the NS3 line was sterile. Phenotypically mutant derivatives (white gene alleles) differing in eye color (pure white, different shades of yellow (honey), orange (apricot), cherry, and red (wild type)) emerged during the N119 and S46 line breeding in the laboratory. Molecular genetic study of the structure of wild type white locus in initial lines and white-mutant derivatives de novo emerging from them, as well as other white lines from the fund of the Laboratory of Population Genetics of the Institute of Cytology and Genetics (Siberian Branch, Russian Academy of Sciences), was conducted. The pairs of primers flanking different white gene regions were selected. Six such pairs overlapped the coding part of the gene. Molecular genetic analysis demonstrated that most DNA defects were limited to the region which includes the first exon (34 lines). Among them, four mutant events were accompanied by an insertion of DNA fragments of approximately 800 bp; one mutation event was accompanied by a deletion of approximately 200 bp; in 29 cases, no PCR product was obtained (this can indicate that as a minimum one of the primer binding sites is damaged). The inserted DNA fragments have no homology with known D. melanogaster sequences presented in the NCBI database. The complete white gene deletion with the manifestation of mutant “white eyes” phenotype was registered in four cases (and only in the N119 line derivatives). Normal PCR product was obtained in 22 cases for all six DNA fragments. Among them, there are both alleles phenotypically mutant by the eye color (white, cherry, or orange) and revertants to the wild type (red). The abundance of defects in the beginning of the gene can indicate a multiplicity of mobile genetic element insertion sites in this part of the white gene in D. melanogaster.