부세(Larimichthys crocea)의 유전적 다양성 분석을 위한 차세대염기서열 분석법 기반의 Microsatellite 마커 개발

Abstract

Demand for large yellow croker (Larimichthys crocea), which is used as a raw material for Gulbi of high economic value, has been increased. In this study, a microsatellite marker was developed using a next-generation sequencing method for efficient gene management of broodstock used for artificial seed production, and for the genetic analysis of the breeding populations. More than 15G bases of total reads were generated through Next-generation sequencing (NGS) with a quality score of Q20 of 99% reliability for 94.5% of the total. From microsatellite regions selected through the calculated results, 192 markers were selected. Out of 75 sets of markers selected through PCR screening, 24 markers were selected after confirming amplification and diversity by performing genotyping by synthesizing dye labeled primers. Based on the range according to the dye, we developed 2 sets of multiplex PCRs in which 8 loci are 1 set. Genetic analysis of large yellow crocker seeds and broodstock groups using the developed marker indicated that the average number of alleles for the total 144 individuals was 13.66 by applying allele richness. This was similar to the number of alleles (K) with an average of 13.62. The average expected heterozygous rate (He) was 0.857, and the average observed heterozygous rate (Ho) was 0.770. The polymorphism information index (PIC) showed an average of 0.835, which confirmed that genetic diversity was well maintained and that there was no linkage imbalance between markers. In addition, the evaluation of the exclusion power of 16 markers showed a high discrimination power of 7.248 X 10-16. The microsatellite marker developed in this study is expected to be useful for genetic analysis and genetic diversity identification of large yellow crocker populations in the future.