• Animal Bioscience
• /
• 제35권9호
• /
• pp.1327-1339
• /
• 2022

Objective: Fat deposition in poultry is an important factor in production performance and meat quality research. miRNAs also play important roles in regulating adipocyte differentiation process. This study was to investigate the expression patterns of miRNAs in duck adipocytes after differentiation and explore the role of miR-214 in regulating carnitine palmitoyltransferases 2 (CPT2) gene expression during duck adipocyte differentiation. Methods: Successful systems for the isolation, culture, and induction of duck primary fat cells was developed in the experiment. Using Illumina next-generation sequencing, the miRNAs libraries of duck adipocytes were established. miRanda was used to predict differentially expressed (DE) miRNAs and their target genes. The expression patterns of miR-214 and CPT2 during the differentiation were verified by quantitative real-time polymerase chain reaction and western blot. Luciferase reporter assays were used to explore the specific regions of CPT2 targeted by miR-214. We used a miR-214 over-expression strategy in vitro to further investigate its effect on differentiation process and CPT2 gene transcription. Results: There were 481 miRNAs identified in duck adipocytes, included 57 DE miRNA candidates. And the 1,046 targets genes of DE miRNAs were mainly involved in p53 signaling, FoxO signaling, and fatty acid metabolism pathways. miR-214 and CPT2 showed contrasting expression patterns before and after differentiation, and they were selected for further research. The expression of miR-214 was decreased during the first 3 days of duck adipocytes differentiation, and then increased, while the expression of CPT2 increased both in the transcriptional and protein level. The luciferase assay suggested that miR-214 targets the 3'untranslated region of CPT2. Overexpression of miR-214 not only promoted the formation of lipid droplets but also decreased the protein abundance of CPT2. Conclusion: Current study reports the expression profile of miRNAs in duck adipocytes differentiated for 4 days. And miR-214 has been proved to have the regulator potential for fat deposition in duck.

• 한국자원식물학회지
• /
• 제17권3호
• /
• pp.257-264
• /
• 2004

Selection of stress-tolerant ginseng lines in fields is very difficult because it is almost impossible to control properly the environmental conditions of soil. On the contrary, it can be studied with ease to search for stress-tolerant ginseng lines through in vitro culture because of easy manipulation of stress conditions. This study was conducted for the selection of ginseng pure lines tolerant to salt stress. Murashige ＆amp; Skoog(MS) media with 2.5 folds of KNO$_3$, NH$_4$NO$_3$, MgSO$_4$.7$H_2O$, KH$_2$PO$_4$, and CaC1$_2$.2$H_2O$ was established for the selection of ginseng pure lines tolerant to salt stress in vitro. Among 88 ginseng pure lines bred by Korea Ginseng and Tobacco Research Institute, Punggi Hwangsuk, 78093, 82886, 78135, 86024 and KG104 lines was tolerant to salt stress. For the stable production of quality Korean ginseng, genetic tolerance to salt stress is one of important factors since relatively high salt concentrations in the ginseng nursery soil environment of Korea. Ginseng inbred pure lines were tested for their tolerance to salt stress through in vitro culture technique.

• 한국산림과학회지
• /
• 제86권2호
• /
• pp.128-134
• /
• 1997

Populus davidiana는 종자의 활력이 짧고(약 3주), 삼목시 발근율이 저조하여 전통적인 유성 무성번식법의 어려움이 있어서 효과적인 기내증식법을 연구하였다. 10가지의 기본 배지를 이용하여, 1-2개의 액아를 포함한 잎이 붙어 있는 줄기절편체를 0.2mg/l의 BAP가 첨가된 배지에서 배양한 결과 MS 배지와 LP배지에서 가장 많은 신초를 생산하였다. 두 배지에서 5주간 배양한 결과 절편체당 9개 이상씩 신초를 생산하였다. 신초생산과 신초의 생장에 있어 클론간의 상당한 차이를 보였다. 몇 클론은 생장조절물질을 첨가하지 않아도 신초를 생산하였다. 기내배양된 신초 중 60% 이상이 0.2mg/l의 IBA가 첨가된 1/2 GD배지에서 발근이 가능하였다.

• Animal cells and systems
• /
• 제1권1호
• /
• pp.99-105
• /
• 1997

Using three species of Korean frogs (Rana dybowskii, R. rugosa and R. nigromaculata), the annual spermatogenic pattern, the seasonal changes in the steroidogenic competence, and responsiveness of testis to gonadotropins in terms of testosterone secretion in vitro were examined. The spermatogenic pattern of R. dybowskii was classified as a discontinuous type since spermatogenesis stops completely after spawning in late winter (February) until mid-summer (July). In contrast, the pattern of R. nigromaculata and R. rugosa was classified as a potent continuous type since sperm was always present in the seminiferous tubules all year round. In all three species, the levels of testicular testosterone and that of testosterone secreted by testis following in vitro culture were very low in late summer (August), but increased thereafter until winter (hibernation period). Interestingly, responsiveness of testis in vitro to gonadotropins in terms of testosterone secretion increased markedly in November (early hibernation period). Specifically, bullfrog LH was more effective than FSH in stimulating the secretion of testosterone by frog testis in vitro during hibernation period. This fact suggests that testosterone secretion by testis during hibernation is at least regulated by the pituitary gonadotropin rather than environmental factors. Taken together, the data presented here suggest that testicular cycles of three species of Korean frogs are closely linked to their females breeding cycles, and are eventually controlled by various environmental cues.

• 한국식물생명공학회:학술대회논문집
• /
• 한국식물생명공학회 2005년도 춘계학술대회 및 국제심포지움 초록집
• /
• pp.158-158
• /
• 2005

• 한국식물생명공학회:학술대회논문집
• /
• 한국식물생명공학회 2004년도 생명공학 실용화를 위한 비젼
• /
• pp.164-164
• /
• 2004

• Asian-Australasian Journal of Animal Sciences
• /
• 제26권4호
• /
• pp.501-508
• /
• 2013

The effects of amino acids and dipeptides on in vitro production of porcine embryos and accumulation of ammonia in culture medium during developmental stages were examined in this study. The maturation, fertilization and development of embryonic cultures were performed in modified Tissue culture medium (mTCM)-199 supplemented with 10% (v/v) porcine follicular fluid, modified Tyrode's albumin lactate pyruvate (mTALP) medium, and modified North Carolina State University (mNCSU)-23 medium, respectively. In addition, amino acids and dipeptides of different concentrations and combinations were used to treat the embryos. The addition of L-alanyl-L-glutamine (AlnGln)+L-glycyl-L-glutamine (GlyGln) significantly (p<0.05) improved oocyte maturation, fertilization and the incorporation and oxidation of 14C(U)-glucose when compared to the control group and other treatment groups. Additionally, 2-4 cell, 8-16 cell, morula and blastocyst development increased significantly (p<0.05) following treatment with AlnGln+GlyGln when compared to the control group and other treatment groups, while this treatment reduced the accumulation of ammonia. Taken together, these findings suggest that treatment with AlnGln+GlyGln may play an important role in increasing the rate of porcine oocyte maturation, fertilization and embryonic development by reducing the level of accumulated ammonia measured in the culture media.

• 한국자원식물학회지
• /
• 제24권2호
• /
• pp.208-213
• /
• 2011

둥근마 기내 식물체의 증식을 위해서는 고체배지보다 액체배지에서 액아수, 초장, 생체중, 괴경형성의 수가 높아 기내식물체 증식에 효과적이었다. 배지의 무기염류 농도는 MS 기본 염류의 배지에서 액아수와 생체중이 각각 28.8개, 3.6 g로 기본 염류의 양을 반으로 줄이거나 두 배로 늘인 배지에서보다 높았다. 당원으로는 sucrose가 glucose 보다 더 효과적이었고, sucrose의 농도는 60 $g{\cdot}L^{-1}$에서 액아수와 괴경형성수가 가장 높아 효율적이었다. 질소원은 $NH_4NO_3$와 $KNO_3$의 조성 비율이 $1{\times}{\frac{1}{2}}$인 것이 다른 처리에 비해 생육이 우수한 것으로 나타났다. 배지내 수소이온 농도는 pH 4.8 ~ 6.8 범위내에서 pH간에 뚜렷한 유의 차이를 보이지 않았다. 생장시기별 생장량에 있어서 엽수, 마디수 및 초장은 접종후 2주부터 5주까지 급격히 증가하고 12주까지는 서서히 증가하는 유사한 경향을 보였다. 반면, 생체중은 접종 3주에서 11주까지 꾸준하게 증가하여 최고치에 달했다. 괴경은 접종 2주부터 형성되기 시작하여, 5~8주 사이에 높은 증가율을 가지며 11주에 최고 형성수가 되었다.

• 한국육종학회지
• /
• 제41권4호
• /
• pp.385-390
• /
• 2009

A resveratrol synthase (RS) gene was isolated from peanut (Arachis hypogaea, L. cv. Jinpoong) plants. This gene was placed under the control of the cauliflower mosaic virus 35S promoter (CaMV35S) and introduced into two Korean varieties of potato (Solanum tuberosum L. cvs. Jasim and Jowon) plants by Agrobacterium-mediated gene transfer. Putative transformants were screened by PCR with primers designed from CaMV 35S promoter, NOS terminator and RS gene. Most of selected transgenic potato plants showed the amplification of expected fragments by PCR of genomic DNA with gene-specific primers, while they were absent in untransformed control plants. Expression of the resveratrol synthase gene was also examined by northern blot analysis. The transformants showed a band which was lacking in the control plant, confirming that the introduced gene is transcribed into mRNA in the transformants. The strength of the band, which reflected the level of mRNA expression, differed among the individual transformants. Among the transformants obtained, the highest trans-resveratrol content in the transgenic young leaves of purple-fleshed "Jashim" was $2.11{\mu}gg^{-1}$ fresh weight and that in the microtubers in vitro of purple fleshed "Jashim" was $8.31{\mu}gg^{-1}$ fresh weight. This amount of resveratrol may have a positive biological effect on human health.

• The Plant Pathology Journal
• /
• 제22권4호
• /
• pp.386-390
• /
• 2006

Extensive research of the Potato virus X(PVX) has been performed in in vitro transcription system using the bacteriophage T7 promoter. We constructed an efficient T-DNA based binary vector, pSNU1, and modified vectors carrying PVX replicons. The suitability of the construct to transiently express PVX RNA using Agrobacterium tumefaciens was tested by analysis of infectivity in plants. The expressed PVX RNA was infectous and systemically spread in three plant species including Nicotiana benthamiana, N. tabacum cv. Xanthi-nc, and Capsicum annuum cv. Chilsungcho. The PVX full length construct, pSPVXp31, was caused severe mosaic symptoms on N. benthamiana, severe necrotic lesions on C. annuum while milder symptoms and delayed mosaic symptoms were appeared on the systemic leaves on N. tabaccum. RT-PCR analysis confirmed the presence of PVX RNAs on both inoculated and systemic leaves in all three plant species tested. Our results indicated that PVX replicons were efficiently expressed PVX RNA in at least three tested species. Further investigation win be needed to elucidate the mechanism of PVX replication, translation, movement and assembly/disassembly processes.