저수온 스트레스에 의한 북방전복, Haliotis discus hannai의 생리학적 반응

박철지,민병화,김관석,이장욱,노재구,김현철,박종원,명정인 한국패류학회 2011 The Korean Journal of Malacology Vol.30 No.4

This study was performed to obtain the basic data on physiological responses of low water temperature stress of the cultured Pacific abalone, Haliotis discus hannai. Abalones were exposed at low water temperatures of 7℃ and 4℃. We have investigated survival rate, superoxide dismutase (SOD) activity and total protein (TP) in the abalone by the exposure times (0, 3, 6, 12, 24, 72, 120, 168 and 216 hours). Survival rate of the abalone at 7℃ experiment was 90.8%, whereas at 4℃ experiment was 0% after exposure 10 days. SOD activity was significantly increased until 12 hours after exposure to 4℃, and then was recovered to starting level after 24 hours. However, there was no significant difference between control (12℃) and 7℃ experiments. TP was significantly increased until 216 hours after 24 hours at 4℃ experiment, but 7℃ experiment showed no significant differences compared to control (12℃) experiment. Therefore, H. discus hannai was acclimated in low water temperature stress at 7℃, but at 4℃, all abalone died possibly because they exceed the limits of defense ability to too low temperature. 본 연구는 북방전복 (H. discus hannai) 의 겨울철 저수온으로 발생하는 폐사원인 규명을 위한 생리학적인 반응을 조사하기 위하여 수온 12℃를 대조구로 7℃ 및 4℃의 실험구를 설정하고 10일간에 걸쳐 생존율, 항산화효소의 활성 및 총 단백질 농도를 측정하였다. 그 결과 7℃ 저수온 실험구에서는SOD 활성 및 총 단백질 농도가 대조구와 차이를 나타내지 않았으며 생존율은 90.8%를 나타내었다. 반면에 4℃ 저수온 실험구에서는 SOD 활성은 노출직후에 급격히 상승하여 12시간째까지 대조구보다 유의적으로 높게 나타났다. 또한 총 단백질농도에 있어서는 24시간째부터 실험종료 시까지 대조구보다유의적으로 높게 나타났으며, 생존율은 3일째 10마리의 폐사를 시작으로 10일째 전 개체가 페사하였다. 이상의 결과로 북방전복 (H. discus hannai) 은 7℃의 저수온 실험구에서는다양한 생리적 기작을 이용하여 생존이 가능한 반면에 4℃의저수온 실험구에서는 생리적 순응반응을 나타내고 있으나 방어기작의 한계를 벗어나 생존이 불가능하다고 추정된다.

교배실험을 통한 북방전복 (Haliotis discus hannai) 의 패각색 변이에 대한 유전적 지배

박철지,남원식,이명석,강지윤,김경길 한국패류학회 2014 The Korean Journal of Malacology Vol.30 No.4

Purple-colored shell individuals were discovered among normal green-colored shell individuals in artificial seed of Pacific abalone, Haliotis discus hannai, reared on an ordinary type of diatom and artificial diet. In the present study, factorial mating experiments were designed to clarify the genetic control of the variant (purple type) and normal (green type) of shell color. The parental population of purple type and green type individuals were derived from a single family between a female and male of each type of coloration. The all mating families were reared in same tank for the same breed environment. The individual of 4 type families were distinguished by paternity test using microsatellite DNA. In factorial mating experiments, all individuals offspring of GG (green type female and green type male), GP (green type female and purple type male) and PG (purple type female and green type male) mating types appeared to green type. In only PP (purple type female and purple type male) mating type, all individuals offspring appeared to purple type. The results suggested that the purple shell color is controlled by recessive purple type allele and a dominant green type allele at a single locus. 본 연구는 북방전복의 보라색패각색의 유전적 지배를 명확히 파악하는 것을 목적으로 하였다. 환경적 요인과 유전적 요인을 명확히 구분하기 위하여 모든 실험구는 같은 사료를 공급하여 실험 하였으며, 또한 동시에 생산된 4가계를 혼합사육한후 microsatellite DNA를 이용하여 친자확인으로 가계를 분리하여 가계간의 패각색의 분리를 확인하였다. 그 결과, 교배형태가 ♀Gr × ♂Gr (GG type), ♀Pu × ♂Gr (PG type) 및 ♀Gr × ♂Pu (GP type) 의 경우 모든 자식개체의 패각색은 녹색으로 나타난 반면, ♀Pu × ♂Pu (PP type) 의 교배구의 모든 자식개체는 보라색패각형으로 나타났다. 따라서 패각색 형태는 녹색이 우성, 보라색이 열성형질로 가정할 경우 패각색의 표현형 분리의 설명이 가능하며, 보라색패각색의 유전지배는 1개의 유전자좌에 대하여 2개의 대립유전자에 의해 결정된다고 생각된다.

북방전복 (Haliotis discus hannai) 삼배체 유도를 위한 저온자극 처리 조건의 개선

박철지,권민수,김은정,남윤권 한국패류학회 2018 The Korean Journal of Malacology Vol.34 No.4

Optimized protocols of cold shock treatment for the induction of triploidy were developed in Pacific abalone Haliotis discus hannai. Time windows of polar body extrusions were analyzed and various technical parameters including initial treatment times, treatment temperatures (-1ºC, 1ºC and 3ºC) and treatment durations (10 to 24 min) were tested in order to develop the best combination(s) among parameters enabling the efficient blocking the polar bodies. Based on a series of tests regarding hatchability of embryos, normality of larvae and induction efficiency of triploidy, the most appropriate protocol to block the first polar body was proven to be the cold shock treatment at 1ºC for 14 min with an initial treatment at 11 min post insemination. On the other hand, the most efficient condition for blocking the second polar body was the cold shock at 1ºC for 14 min with an initial treatment at 25-27 min post insemination. Under these treatment conditions, larval yields of cold-shock treated groups relative to non-treated control groups were up to 70% with a fairly consistent triploidy rate up to near 100%. Efficacy and reproducibility of triploid induction were further validated under scaled-up conditions; however, the yield of triploid larvae was highly influenced depending on the gamete quality used. Results from this study could be a useful basis for developing the practical protocol for mass production of triploid seedlings in this abalone species.

북방전복 (Haliotis discus hannai) 의 성별에 따른 성장형질 및 유전모수 추정

박철지,박종원,김보라,정규현,김영진,손윤석,김경길 한국패류학회 2016 The Korean Journal of Malacology Vol.32 No.4

The purpose of this study is to estimate genetic parameter and growth traits by sex of Pacific abalone, Haliotis discus hannai. The abalone 10 family produced using 1:1 mating system of male and female for analyses of sex ratio and growth traits (shell length, shell width, and total weight) by sex. Overall mean in phenotypic traits at 30-month-old showed 85.01 mm of shell length, 57.49 mm of shell width and 73.34 g of total weight respectively. The sex ratio (female : male) was 1:0.93 (n = 191:177). The values of growth traits by sex showed significant difference (P < 0.05). The each value of female growth traits were shown to be higher than the values of male growth traits. The heritability of growth traits by sex were estimated that the heritability of female growth traits are higher than male that. The results suggest a possibility of improving the growth of cultured abalone using selection breeding by sex.

북방전복 (Haliotis discus hannai) 의 mitochondrial DNA 영역별 유전적 변이성 분석

박철지,남원식,이정호,노재구,김현철,박종원,황인준,김성연 한국패류학회 2013 The Korean Journal of Malacology Vol.29 No.4

The seven mitochondrial DNA regions (ND2, ND5, ND4, ND4L, ND6, ND1 and 12SrRNA) of Haliotis discus hannai were examined to estimate the availability as a genetic marker for the study of population genetic. The region with the highest genetic variation was ND4 (Haplotype diversity = 1.0000, Nucleotide diversity = 0.0108). On the other hand, ND2 and ND1 regions have significantly appeared genetic divergence between clusters (divergence of 90% and 87%). Also, pairwise FST between clusters within ND2 and ND1 regions showed high values; 0.4061 (P = 0.0000), 0.4805 (P = 0.0000) respectively. Therefore we can infer that it is the most efficient and accurate way to analyze the region of ND4 with the highest variation in addition to the regions of ND2 and ND1, which formed clusters with high bootstrap value, for study of population genetic structure in this species. 본 연구는 우리나라의 주요양식 품종인 북방전복을 대상으로 지금까지 전복류에서는 사용되지 않았던 mtDNA의protein coding 영역 ND2, ND5, ND4, ND4L, ND6, ND1의 6개영역과 protein noncoding 영역인 12SrRNA(ribosomal RNA) 을 포함해 총 7개 영역을 이용하여 각 영역의 유전적 변이성 및 개체간 유전적 유연관계 등을 분석하여각 영역별 특성을 파악하고 이러한 특성을 고려하여 유전학적분석에 적합한 분자유전마커를 개발하였다. 유전적 변이성은ND4 영역 (Haplotype diversity = 1.000, Nucleotidediversity = 0.010823) 이 가장 높게 나타났으며, 개체간의유전적 차이는 ND2 및 ND1 영역이 각각 90% 및 87%로 유의적으로 명확히 구분할 수 있었다. 따라서 유전적 변이성이가장 높은 ND4 영역과 영역내의 클러스터 간의 유전적 차이가 명확한 ND2 및 ND1 영역을 복합적으로 활용할 경우 북방전복의 집단유전학 및 계통분류학 분석에 유용한 분자유전마커로 사용할 수 있을 것이라 생각된다.

독도연안에 서식하는 전복의 유전학적 특성

박철지,이정호,노재구,김현철,민병화,명정인 한국패류학회 2009 The Korean Journal of Malacology Vol.25 No.3

This study was conducted to investigate the genetic characteristics of wild population of Pacific abalone, Haliotis discus hannai in Dokdo island. We used six polymorphic microsatellite marker to investigate the genetic diversity and population structure. The loci Hdh1321 and Hdh512 had the highest number of allele (34 and 22 respectively) and loci Hdh145 and Awb083 had the lowest (5 and 7 respectively). The mean number of allele per locus was 14.8. The average observed and expected heterozygosities were 0.664 and 0.824 respectively, and the average FIS was 0.195. We compared the population genetic parameters of Dokdo population with previously published data of the same species. At the result, the parewise FST test showed significant difference between the Dokdo population and six populations (published data), suggesting that the genetic relationship of Dokdo population was separated from six populations.

이배체 및 삼배체 전복(Haliotis discus hannai) 치패에서 주요 열충격 단백질 유전자들(heat shock protein genes)의 발현 특징

박철지,김은정,남윤권 한국수산과학회 2020 한국수산과학회지 Vol.53 No.4

Basal and heat shock-induced mRNA expression patterns of major heat shock protein (HSP) genes, including those encoding heat shock protein (HSP) 90, HSP70, HSP70-12A, heat shock inducible protein 70 (HSIP70), heat shock binding protein 1 (HSPBP1), HSP60, and HSP40 were examined in the gill and hepatopancreas of 1-year-old diploid and triploid abalone Haliotis discus hannai juveniles. Under non-stimulated conditions at 19ºC, triploid abalones displayed, in general, higher mRNA levels of various HSPs (HSP70, HSIP70, HSPBP1, HSP70-12A, and HSP60 in the gill and HSIP70, HSPBP1, and HSP60 in the hepatopancreas) than did communally cultured diploids. Conversely, only the hepatopancreatic expression of HSP70-12A was higher in diploids than in triploids. However, the fold changes in gene expression in response to an acute thermal challenge (elevation from 19 to 30ºC) were generally greater in diploids than in triploids, such that the difference in basal expression was diminished, weakened, or even reversed after heat shock treatment. However, unlike other HSP genes, the basal expression of HSP60 (higher in 3N) was more pronounced after heat shock treatment. Collectively, the results of this study suggest that triploid abalones have different capacities for not only basal expression but also the heat-induced expression of HSPs in an HSP member-dependent manner.

RFLP Analysis of the mtDNA COI Region in Four Abalone Species

박철지,Akihiro Kijima 한국수산과학회 2006 Fisheries and Aquatic Sciences Vol.9 No.3

북방전복 (Haliotis discus hannai) 의 선발육종 연구를 위한 microsatellite multiplex PCR법 개발

박철지,남원식,이명석,강지윤,김경길 한국패류학회 2014 The Korean Journal of Malacology Vol.30 No.4

The multiplex PCR system including six microsatellites from Haliotis discus hannai, consisting of dinucleotide and trinucleotide repeat units, is developed. The six loci were coamplified in a single reaction employing dye-labeled primers. Alleles from these loci were sized using an internal standard by automated sample processing in an ABI3100 Genetic Analyser. Amplified alleles in profiles containing selected microsatellites were typed clearly, providing easily interpretable results. In this results suggest that the presented multiplex PCR system may be a useful tool in a selective breeding program of H. discus hannai in which genetic identification will allow different genotypes to be reared together from fertilization. This should have a great impact as it will make selective breeding more efficient. Moreover, it will be useful in a variety of applications, including strain and hybrid identification, parentage assignment, pedigree reconstruction, estimating genetic diversity and/or inbreeding. 북방전복 선발육종에 필요한 친자확인 및 가계분석을 효율적으로 실험하기 위하여 microsatellite multiplex PCR 기술을 개발하였다. 개발한 mutiplex PCR 기술은 6개microsatellite locus Hdh145, Hdh512, Hdh1321, Awb017, Awb083 및 Awb098을 한번의 PCR 증폭으로 다중분석이 가능하다. 이 기술은 높은 친자확인 성공률 및 가계분석에 있어서도 모두 멘델의 분리법칙을 따르고 있다. 더욱이대량의 시료처리를 필요로 하는 경우에 있어서도 시간절약 및비용 절감뿐만 아니라 샘플 처리과정의 간소화가 가능하여handling errors를 줄일 수 있다. 따라서 본 연구에서 개발된multiplex PCR은 친자확인, 가계분석, 집단유전학 및 계통분류학 분석에 유용하게 사용할 수 있을 것이라 생각된다.

선발육종기술을 이용한 북방전복의 성장

박철지,이정호,노재구,김현철,박종원,황인준,김성연 한국패류학회 2012 The Korean Journal of Malacology Vol.28 No.4

속성장을 위한 선발 1세대 전복 (Selected abalone : SA)의 성장률을 조사하기 위하여 동일 사육 환경 조건에서 대조구전복 (control abalone : CA) 과 비교실험을 하였다. 그 결과각 성장형질 (각장, 각폭 및 중량) 은 SA구가 CA구보다 유의하게 빠른 것으로 나타났다. 45일째 및 90일째의 평균 각장에있어 SA구가 CA구 보다 각각 31.7% 및 17.8% 빠르게 나타났으며, 중량에 있어서도 각각 43.7% 및 25.6% 빠른 것으로나타났다. 더욱이 90일째의 각장과 중량의 상대성장 비교에있어서도 SA구가 CA구 보다 유의적으로 높은 상관관계를 나타내었다. 이러한 결과는 선발육종기술을 이용하여 양식전복의 성장률을 증대할 수 있는 가능성을 시사하고 있다. The growth rates of offspring (SA) of selected abalone, Haliotis discus hannai, population for rapid growth were compared to those of offsping (CA) of the control abalone at bred under the same condition. The growth traits (shell length, shell breadth and total weight) of SA were significantly faster than those of CA at this experiment. At the 45th day and 90th day, the growth rates in shell length of SA were estimated 31.7% and 17.8% faster than those of CA, total weight of SA were estimated 43.7% and 25.6%, respectively. Moreover, the relative growth rate between shell length and total weight of SA showed significant differences higher than one in CA at 90th day. The results suggest a possibility of improving the growth rate of cultured abalone using selection techniques.