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극동산 뱀장어(Anguilla japonica)의 친자확인을 위한 유전자 마커 개발
노은수,신은하,박경현,김은미,김영옥,유용운,김신권,남보혜 한국수산과학회 2022 한국수산과학회지 Vol.55 No.5
The Japanese eel Anguilla japonica is a highly valued research object that is important for aquaculture in Asia, including the Republic of Korea. However, few studies have been conducted analyzing parentage using microsatellite markers derived from the Japanese eel. We acquired Japanese eel genome data using next generation sequencing technology, and constructed a draft genome comprising 1,087 Mbp. Using the Simple Sequence Repeat Identification Tool program, 444,724 microsatellites were identified. Of these, 1,842 microsatellites located in the 3' untranslated region, which are stably inherited, were finally selected. Ninety-six primers were selected to validate polymorphism at these microsatellites, and 9 primers were finally identified for multiplex analysis. Using multiplex polymerase chain reaction with three different fluorescence chemistries, we performed parentage analysis of an artificial Japanese eel population. CERVUS software was used to calculate the logarithm of the odds (LOD) scores and the confidence of the parentage assignments. The results presented here show that 83 out of 85 paternity cases were assigned at 95% confidence to a candidate father and mother with LOD scores ranging from 4.79 to 28.2. This study provided a microsatellite marker-based assay for parentage analysis of Japanese eels, which will be useful for selective breeding and genetic diversity studies.
문흥규,노은운,하유미,심경구,Moon, Heung-Kyu,Noh, Eun-Woon,Ha, Yoo-Mi,Shim, Kyung-Ku 한국식물생명공학회 2002 식물생명공학회지 Vol.29 No.2
히어리나무 1년생 및 10년생을 재료로 액아배양을 통한 기내번식을 시험하였다. 줄기의 증식은 zeatin과 BA의 공조처리가 효과적이었으며 MS배지에 zeatin 0.5∼3.0 mg/L, 0.2 mg/L BA 처리시 주효하였다. 1년생이 10년생보다 전반적으로 증식 및 생장이 양호하였으며, 배양 6개월 후에는 10년생에서도 매월 절편 당 3개의 줄기유도가 가능하였다. 기내 줄기의 발근은 1년생은 97%, 10년생은 62%를 나타내었고, 토양이식시 1 년생 유래 배양묘는 67%, 10년생은 48%생존되어 모수령에 따른 차이를 나타냈다. 본 실험결과 히어리나무의 액아배양으로 유시, 성숙목의 대량번식이 가능함을 보여주었으나 선발개체의 효율적인 기내번식을 위해서는 재유령화의 기술개발과 토양순화율을 증진시켜야 하는 것으로 나타났다. We have developed an in vitro micropropagation system via shoot formation from axillary buds using nodal segments of Corylopsis coreana. Explants from both juvenile tree (one-year-old greenhouse stock seedlings) and mature tree (ten-years-old tree in nursery) were compared with regard to propagation efficiency. Combined treatment of both BA and zeatin were effective on shoot proliferation since the best result was obtained on MS medium supplemented with 0.5∼3.0 mg/L zeatin and 0.2 mg/L BA. Generally, juvenile explants were better in both shoot proliferation and growth than mature explants. However, as the duration of in vitro culture was proceed to 6 months, explants from mature tree also produced three shoots per explant. Distinctive differences in rooting and adaptability to soil of shoots obtained from mother trees. Whereas shoots originated from juvenile explants rooted as high as 97%, those from adult explants showed 62% rooting. Similar result was also observed in soil acclimatization. The plantlets derived from juvenile plants survived 67%, while only 48% of those from adult trees survived. The results showed a possibility of the micropropagation of Corylopsis coreana through shoot formation from axillary buds. In addition, the advance of the research still remain to enhance the frequency of acclimatization of plantlets from mature trees for practical application.
NPTII 및 HPT 유전자가 삽입된 현사시의 항생제에 대한 저항성 분석
이은정,노은운,박재인 한국식물생명공학회 2001 식물생명공학회지 Vol.28 No.5
본 실험은 nptII 및 hpt 유전자가 삽입된 현사시나무를 이용하여 각 항생제 저항성 유전자로 형질전환된 세포의 효율적인 선발 조건을 규명하기 위하여 수행되었다. 액아가 포함된 줄기절편과 잎절편 조직의 생장, 발근 유도, 캘러스 유도로 항생제에 대한 감수성에 대한 효과를 검정하였다. 형질전환되지 않은 대조식물체의 잎절편을 이용한 경우 50 mg/L의 kanamycin이나 2 mg/L hygromycin으로 캘러스 유도 및 생장을 억제할 수 있었으나 형질전환된 식물은 100 mg/L kanamycin, 50 mg/L hygromycin에서도 왕성한 생장을 나타냈다. 절간조직의 개아의 경우 100 mg/L kanamycin, 5 mg/L hygromycin으로 줄기신장을 완전히 억제 가능하였으며, 뿌리유도는 50 mg/L kanamycin, 5 mg/L hygromycin에서 선발이 가능하였다. 형질전환체는 모두 이보다 높은 농도에서 왕성한 생장을 보였다. 형질전환이 되지 않은 세포의 생장을 억제하는 데는 hygromycin이 kanamycin보다 더 효율이 좋은 것으로 나타났다. 따라서 hpt유전자가 npt II 유전자보다 훨씬 강력한 선발표지임이 확인되었으며 엽절편의 캘러스 유도와 생장과 절간조직의 발근유도 모두 항생제에 예민하게 작용하여 형질전환체의 식별에 효과적으로 사용할 수 있음을 알 수 있었다. This study was peformed to find out the optimal conditions for the selection of transformed cells using already established transgenic plants. Several transgenic poplar (Populus alba$\times$P giandulosa) lines carrying npt II or hpt gene as a selectable marker were tested against kanamycin or hygromycin. Two culture explants, leaf discs and nodes, were compared regarding their sensitivity to the antibiotics. When leaf discs of untransformed control plants were cultured on callus inducing media in the presence of varying levels of kanamycin or hygromycin, only those cultured on the media containing lower than 50 mg/L kanamycin or 2 mg/L hygromycin formed callus. However, much higher concentration of kanamycin was needed to suppress the growth of axillary buds of untransformed plants. On the other hand, hygromycin at the concentration of 5 mg/L effectively suppressed shoot growth of untransformed plants. Root induction from untransformed plants could also be suppressed at the concentration of 50 mg/L kanamycin or 5 mg/L hygromycin. The transgenic plants showed resistance to 100 mg/L kanamycin or 50 mg/L hygromycin in the growth of callus, shoots, and roots. Hygromycin appeared to be more efficient in selecting untransformed cells than kanamycin.