• Journal of Ginseng Research
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• 제43권1호
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• pp.38-48
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• 2019

Background: Interspecific ginseng hybrid, Panax ginseng ${\times}$ Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with $GA_3$ to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L $GA_3$ for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The $F_1$ hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides $F_{11}$ in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.

• 한국산림과학회지
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• 제108권3호
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• pp.302-310
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• 2019

정확하고 신속한 품종식별은 효율적인 육종과 육종가의 권리 보호를 위하여 필수적이다. 대추나무 품종을 구분하는 전통적인 방법은 형태적 특성을 근거로 하지만, 시기적 제약과 환경의 영향으로 형태적 형질만을 이용하여 구별하기에는 어려움이 있다. 이에 본 연구에서는 ISSR 분석으로 얻어진 단편을 복제하여 안정적인 식별을 위한 SCAR 표지를 개발하였다. 대리조와 보은대추 품종에서 특이성을 보이는 ISSR 밴드를 대상으로 정제, 복제, 염기서열 분석을 수행함으로써 827Dalizao550, 827Boeun750, 846Boeun700, 847Dalizao850로 명명된 4개의 클론을 확인하였다. 대추나무 품종 특이성 분석을 위한 4쌍의 SCAR 프라이머를 제작하였으며, 대추나무와 묏대추나무를 포함하는 32개체에 대하여 PCR 분석을 수행하였다. 827Dalizao550 SCAR 표지의 PCR 결과 6개체(대리조, 산동이조, 동조, 원령 신 2호, 묏대추나무 2, 묏대추나무 4)에서 550 bp의 특이적인 밴드가 증폭되었고, 나머지 개체에서는 예기치 않은 밴드(490 bp)가 증폭되었다. 또한 847Dalizao850 SCAR 표지의 PCR 결과 대리조, 산동이조, 동조에서 850 bp의 밴드가 나타났고 예기치 않은 900 bp의 밴드가 5개체(파조, 묏대추나무 1, 묏대추나무 2, 묏대추나무 3, 묏대추나무 4)에서 증폭되었다. 이상의 결과로 보아 새롭게 개발된 표지들은 대추나무 품종식별을 위한 빠르고 신뢰성 있는 수단으로 이용될 수 있을 것으로 생각된다. 하지만 보다 다양한 품종들의 식별을 위해서는 다형성 정보의 추가와 SCAR 표지의 개발이 필요하다.

• 식물분류학회지
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• 제49권2호
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• pp.118-126
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• 2019

The taxonomic rank of the tiny-leaved terrestrial orchid Cephalanthera subaphylla Miyabe & $Kud{\hat{o}}$ has been somewhat controversial, as it has been treated as a species or as an infraspecific taxon, under C. erecta (Thunb.) Blume [C. erecta var. subaphylla (Miyabe & $Kud{\hat{o}}$) Ohwi and C. erecta f. subaphylla (Miyabe & $Kud{\hat{o}}$) M. Hiro]. Allozyme markers, traditionally employed for delimiting species boundaries, are used here to gain information for determining the taxonomic status of C. subaphylla. To do this, we sampled three populations of five taxa (a total of 15 populations) of Cephalanthera native to the Korean Peninsula [C. erecta, C. falcata (Thunb.) Blume, C. longibracteata Blume, C. longifolia (L.) Fritsch, and C. subaphylla]. Among 20 putative loci resolved, three were monomorphic (Dia-2, Pgi-1, and Tpi-1) across the five species. Apart from C. longibracteata, there was no allozyme variation within the remaining four species. Of the 51 alleles harbored by these 17 polymorphic loci, each of the 27 alleles at 14 loci was unique to a single species. Accordingly, we found low average values of Nei's genetic identities (I) between ten species pairs (from I = 0.250 for C. erecta versus C. longifolia to I = 0.603 for C. falcata vs. C. longibracteata), with C. subaphylla being genetically clearly differentiated from the other species (from I = 0.349 for C. subaphylla vs. C. longifolia to 0.400 for C. subaphylla vs. C. falcata). These results clearly indicate that C. subaphylla is not genetically related to any of the other taxa of Cephalanthera that are native to the Korean Peninsula, including C. erecta. In a principal coordinate analysis (PCoA), C. subaphylla was positioned distant not only from C. falcata, C. longibracteata, and C. longifolia, but also from C. erecta. Finally, K = 5 was the best clustering scheme using a Bayesian approach, with five clusters precisely corresponding to the five taxa. Thus, our allozyme results strongly suggest that C. subaphylla merits the rank of species.

• 한방비만학회지
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• 제18권2호
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• pp.106-114
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• 2018

Objectives: We investigated about the microbial properties and changes in the efficacy of the Codonopsis lanceolata (CL) by natural fermentation. Methods: CL was fermented for four weeks in a well-ventilated place with 2.5% salt. pH, total sugar, total polyphenol, and total flavonoid were measured to determine fermentation characteristics according to fermentation period and salt treatment. Polymerase chain reaction denaturing gradient gel electrophoresis and random amplification of polymorphic DNA-polymerase chain reaction were carried out for microbial analysis during fermentation. In addition, HepG2 cell was cultured to check the lipid accumulation through oil red O staining and the glucose uptake was analyzed by measuring the 2-NBDG at C2C12 cell. Results: The pH level and the total sugar decreased with the CL fermentation. Total polyphenol and flavonoid increased after CL fermentation. It was confirmed that Leuconostoc mesenteroides were maintained continuously during fermentation. In the salt treatment CL, there was a sharp increase in Rahnella aquatilis. Lactobacillus plantarum matrix was observed in fermented CL. In addition, Lactococcus lactis, Weissella koreensis, R. aquatilis, L. plantarum, Leu. mesenteroides have been added to the salt treatment. Glucose uptake were significantly increased after fermentation with salt for four weeks. Lipid accumulation in the HepG2 cells was observed that there was difference (P<0.01) between free fatty acid group (100%) and decreased 4 weeks after fermentation (90.38%) at $800{\mu}g/mL$. Conclusions: Total polyphenol and flavonoid were increased after CL fermentation. Especially, percentage of the glucose uptake and lipid accumulation inhibition increased in CL fermentation with salt. It is expected that fermentation of salt treated CL will be more effective in diabetes and fatty liver.

• Asian-Australasian Journal of Animal Sciences
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• 제32권7호
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• pp.913-921
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• 2019

Objective: The objectives of this study were to compare identified informative regions through two genome-wide association study (GWAS) approaches and determine the accuracy and bias of the direct genomic value (DGV) for milk production traits in Korean Holstein cattle, using two genomic prediction approaches: single-step genomic best linear unbiased prediction (ss-GBLUP) and Bayesian Bayes-B. Methods: Records on production traits such as adjusted 305-day milk (MY305), fat (FY305), and protein (PY305) yields were collected from 265,271 first parity cows. After quality control, 50,765 single-nucleotide polymorphic genotypes were available for analysis. In GWAS for ss-GBLUP (ssGWAS) and Bayes-B (BayesGWAS), the proportion of genetic variance for each 1-Mb genomic window was calculated and used to identify informative genomic regions. Accuracy of the DGV was estimated by a five-fold cross-validation with random clustering. As a measure of accuracy for DGV, we also assessed the correlation between DGV and deregressed-estimated breeding value (DEBV). The bias of DGV for each method was obtained by determining regression coefficients. Results: A total of nine and five significant windows (1 Mb) were identified for MY305 using ssGWAS and BayesGWAS, respectively. Using ssGWAS and BayesGWAS, we also detected multiple significant regions for FY305 (12 and 7) and PY305 (14 and 2), respectively. Both single-step DGV and Bayes DGV also showed somewhat moderate accuracy ranges for MY305 (0.32 to 0.34), FY305 (0.37 to 0.39), and PY305 (0.35 to 0.36) traits, respectively. The mean biases of DGVs determined using the single-step and Bayesian methods were $1.50{\pm}0.21$ and $1.18{\pm}0.26$ for MY305, $1.75{\pm}0.33$ and $1.14{\pm}0.20$ for FY305, and $1.59{\pm}0.20$ and $1.14{\pm}0.15$ for PY305, respectively. Conclusion: From the bias perspective, we believe that genomic selection based on the application of Bayesian approaches would be more suitable than application of ss-GBLUP in Korean Holstein populations.

• Laboraroty Animal Research
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• 제34권4호
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• pp.176-184
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• 2018

In this study, we observed chronological changes in the immunoreactivity and expression level of myelin basic protein (MBP), one of the most abundant proteins in the central nervous system, in the hippocampus of Zucker diabetic fatty (ZDF) rats and their control littermates (Zucker lean control; ZLC). In the ZLC group, body weight steadily increased with age; the body weight of the ZDF group, however, peaked at 30 weeks of age, and subsequently decreased. Based on the changes of body weight, animals were divided into the following six groups: early (12-week), middle (30-week), and chronic (52-week) diabetic groups and their controls. MBP immunoreactivity was found in the alveus, strata pyramidale, and lacunosum-moleculare of the CA1 region, strata pyramidale and radiatum of the CA3 region, and subgranular zone, polymorphic layer, and molecular layer of the dentate gyrus. MBP immunoreactivity was lowest in the hippocampus of 12-week-old rats in the ZLC group, and highest in 12-week-old rats in the ZDF group. Diabetes increased MBP levels in the 12-week-old group, while MBP immunoreactivity decreased in the 30-week-old group. In the 52-week-old ZLC and ZDF groups, MBP immunoreactivity was detected in the hippocampus, similar to the 30-week-old ZDF group. Western blot results corroborated with immunohistochemical results. These results suggested that changes in the immunoreactivity and expression of MBP in the hippocampus might be a compensatory response to aging, while the sustained levels of MBP in diabetic animals could be attributed to a loss of compensatory responses in oligodendrocytes.

• 생명과학회지
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• 제31권1호
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• pp.52-58
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• 2021

키위에 세균성 궤양병을 일으키는 Pseudomonas syringae pv. actinidiae는 유전적 특성과 생산하는 독소에 따라 5개의 biovar (1, 2, 3, 5, 6)로 나누어진다. 그중 최근 전 세계적으로 유행하고 있는 biovar 3는 2011년부터 국내에서 분리되고 있다. RAPD 분석을 바탕으로 국내에서 분리된 biovar 3 균주는 6개의 subgroup (I, IV, V, VI, VII, VIII)으로 나누어진 바 있다. 본 연구에서는 차등되는 RAPD 밴드의 염기서열로부터 6개 subgroup 각각에 특이적인 SCAR primers를 개발하였다. 각 subgroup에 특이적인 이들 primers를 사용하여 2011-2017에 국내에서 분리한 biovar 3 균주들의 subgroup 분포를 조사하였다. 조사된 54개 균주 중 35개(64.8%)가 subgroup V에, 9개(16.7%) 균주가 subgroup IV, 4개(7.4%)가 subgroup VI, 3개(5.6%) 균주가 subgroup VII, 2개(3.7%)가 subgroup VIII, 그리고 1개(1.9%) 균주가 subgroup I에 속하였다. Subgroups IV, V 및 VI에 속하는 균주들은 각각 중국, 뉴질랜드, 칠레 균주와 연관이 있었다. 이 연구에 따르면 우리나라의 biovar 3 균주들은 유전적으로 다양하며 꽃가루를 통해 외국으로부터 유입된 것으로 추정된다.

• Animal Bioscience
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• 제34권7호
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• pp.1105-1115
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• 2021

Objective: To conserve and utilize the genetic resources of a traditional Chinese indigenous pig breed, Liangshan pig, we assessed the genetic diversity, genetic structure, and genetic distance in this study. Methods: We used 50K single nucleotide polymorphism (SNP) chip for SNP detection of 139 individuals in the Liangshan Pig Conservation Farm. Results: The genetically closed conserved population consisted of five overlapping generations, and the total effective content of the population (Ne) was 15. The whole population was divided into five boar families and one non-boar family. Among them, the effective size of each generation subpopulation continuously decreased. However, the proportion of polymorphic markers (P_N) first decreased and then increased. The average genetic distance of these 139 Liangshan pigs was 0.2823±0.0259, and the average genetic distance of the 14 boars was 0.2723±0.0384. Thus, it can be deduced that the genetic distance changed from generation to generation. In the conserved population, 983 runs of homozygosity (ROH) were detected, and the majority of ROH (80%) were within 100 Mb. The inbreeding coefficient calculated based on ROH showed an average value of 0.026 for the whole population. In addition, the inbreeding coefficient of each generation subpopulation initially increased and then decreased. In the pedigree of the whole conserved population, the error rate of paternal information was more than 11.35% while the maternal information was more than 2.13%. Conclusion: This molecular study of the population genetic structure of Liangshan pig showed loss of genetic diversity during the closed cross-generation reproduction process. It is necessary to improve the mating plan or introduce new outside blood to ensure long-term preservation of Liangshan pig.

• 한국버섯학회지
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• 제19권1호
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• pp.76-80
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• 2021

느타리 품종구분을 위한 마커의 개발을 위하여 곤지7호의 어버이 일핵 균사중의 하나인 MT07156-97의 전체 유전자 염기서열을 바탕으로 제작한 251개의 SSR 프라이머를 제작하였다. 우선적으로 수한1호, 곤지7호, 흑타리 품종에 다형성 여부를 관찰하여 20개의 SSR을 선발하고, 이를 10개 품종에 적용하였다. 단일의 프라이머로는 일부 품종이 구분되지 않았으므로, 선발된 프라이머 간의 다양한 조합(multiplex 방식)을 적용한 결과 모든 품종을 판별할 수 있는 분자마커 다형성을 보인 프라이머 "166+115" 조합을 선발하였다. 별도로 프라이머 115와 166가 만들어 낸 산술적인 유전자좌(loci) 31개보다 12개 많은 40개의 유전자좌가 증폭되어 다양한 품종에 특이적인 분자마커를 제공할 수 있었다. 개발된 분자마커는 종균의 품질관리, 품종의 판별, 신품종 보호에 활용될 수 있을 것이다.

• 생명과학회지
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• 제31권6호
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• pp.537-542
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• 2021

자웅이주 식물로 알려진 Schisandra nigra Max. (흑오미자)는 우리나라 제주도에 자생하고 있으며, 열매를 얻기 위해 일부 농가에서 재배되고 있다. 열매 생산을 위해서는 수그루에 비해 암그루의 가치가 더 높기 때문에 묘목단계에서 일찍 성별을 아는 것은 중요하다. 이 연구에서는 S. nigra의 유전체에서 암그루에 특이적인 부위에 관련된 SCAR 마커를 개발하였다. 120개의 무작위로 구성된 RAPD 프라이머 중에서 OPB-03 프라이머가 암그루에서 749 bp의 밴드를 안정적으로 증폭시켰다. 암그루 특이적인 PCR 산물을 분리하여 클로닝한 뒤 염기서열을 결정하였다. 이 암그루 특이적인 절편을 탐침으로 사용한 Southern hybridization에서 암그루에서만 양성반응이 나타나고 수그루에서는 잡종화가 일어나지 않았다. 이러한 결과는 749 bp의 DNA 절편이 암그루의 유전체에는 존재하지만, 수그루에는 없음을 시사하였다. RAPD 마커로부터 암그루에서만 436 bp를 증폭시키는 SCAR 프라이머를 설계하였다. 이 프라이머 쌍은 암그루와 자생지에서 수집한 4개의 자웅동주 식물에서만 예상되었던 크기의 DNA 절편을 증폭하였다. 이 연구에서 개발된 SCAR 마커는 묘목 단계에서 암꽃이 피는 개체를 선발하는데 사용될 수 있을 것이다.