Identification and Development of a Species-Specific Molecular Marker for Discriminating Between Abies koreana and Abies nephrolepis

Abies koreana Wilson (Korean fir) and Abies nephrolepis (Trautv. Ex Maxim.) Maxim. (Khingan fir) are ecologically and economically significant coniferous species in East Asia. However, morphological similarities and hybridization complicate species identification, affecting conservation, forestry management, and commercial activities. Here, we developed a species-specific DNA marker using single nucleotide polymorphisms (SNPs) within the mitochondrial nad5 intron 1 region to discriminate between A. koreana and A. nephrolepis. In particular, PCR amplification and sequencing analyses of candidate drought and heat stress-responsive genes as well as mitochondrial nad5 intron 1 and nad5 intron 4 revealed a species-specific T-to-G substitution in nad5 intron 1. Allele-specific primers were designed, and competitive allele-specific labeled light emission technology was employed for SNP detection. The primers were validated using real-time PCR, achieving high specificity and reliability. Overall, the molecular diagnostic tool offers a practical solution for accurate species identification, aiding conservation efforts and sustainable resource management.