Genetic Variation of Spongy Moth (Lymantria dispar) in Kazakhstan

The spongy moth (Lymantria dispar) is a significant defoliator of broadleaf forests and fruit trees, but its population genetic structure in Central Asia remains poorly defined. We analyzed 153 specimens collected from ten populations across Kazakhstan between 2023 and 2024, using a combination of diagnostic real-time PCR, RAPD genotyping, and mitochondrial COI sequencing. Subspecies identification and Asian introgression were initially assessed with the COI/FS1 TaqMan assay. All samples carried the European mitochondrial type L. dispar dispar, while the nuclear FS1 locus indicated Asian introgression in 92.8% of specimens, including eleven heterozygotes for the Asian allele. RAPD amplification with five polymorphic primers produced 107 variable bands, which we converted into a presence/absence matrix for population genetics analysis. Principal coordinate analysis showed weak but detectable population structure, and Bayesian clustering (STRUCTURE) supported the presence of two primary nuclear clusters. Populations from Ketpentau, Sumbe, Kazachka, Butakovka, and Pavlodar were assigned predominantly to one cluster, whereas most individuals from Almaty, Ile-Alatau, Koksu, North Kazakhstan, and Tekeli were assigned predominantly to the other, with limited admixture in several localities. COI sequences formed a single, minimally differentiated L. dispar clade, with no evidence of distinct mitochondrial lineages. Overall, these findings reveal extensive Asian nuclear introgression in populations retaining European mitochondrial haplotypes, supporting the interpretation of Kazakhstan as part of a broad contact zone within the larger dispar-asiatica gradient. The observed mito-nuclear discordance has important implications for quarantine diagnostics and highlights the need for multilocus molecular assays in regional biosurveillance.