• Title/Summary/Keyword: germplasm

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The Effect of Simple Freezing Method on Viability of Frozen-thawed Primordial Germ Cells on the Chicken (간이 동결 방법이 닭 원시 생식 세포의 생존율에 미치는 영향)

  • Kim, Hyun;Cho, Young Moo;Han, Jae Yong;Choi, Sung Bok;Cho, Chang-Yeon;Suh, Sangwon;Ko, Yeoung-Gyu;Seong, Hwan-Hoo;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.41 no.4
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    • pp.261-270
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    • 2014
  • This study was conducted to establish the method for preserving chicken primordial germ cells (PGCs) that enables long-term storage in liquid nitrogen ($LN_2$) for developmental engineering or preservation of species. The purpose of this study is to clarify the effects of simple freeze-thaw treatment on viability of PGCs in chickens and to the optimal protocol for PGCs freezing. PGCs obtained from the germinal gonade of an early embryos of 5.5~6 day (stage 28) of Isa Brown, Korean Ogye (KO), White Leghorn and Commercial breeds, using the MACS method were suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG)). The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments : 2.5%, 5%, 10%, 15%, and 0% cryoprotectant (DMSO, EG, PG) as a control. Effects of exposure to simple freezing, with different concentrations of the cryoprotectant solution, were examined. After simple freezing, the viability of PGCs after freeze-thawing was significantly higher for Commercial breeds ($88.7{\pm}2.4%$) than KO ($85.1{\pm}0.4%$), Isa Brown ($84.6{\pm}0.2%$) and White Leghorn ($85.9{\pm}0.1%$) (p<0.05) using 10% EG cryoprotectant. Therefore, these systems may contribute in the improvement of cryopreservation for a scarce species in birds preservation. This study established a method for preserving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at a germplasm repository and ease of entry into a database.

The Evaluation of Various Conditions in the Cryopreservation of Primordial Germ Cells on Korean Native Chicken (Ogye) (한국재래닭(오계)의 원시 생식 세포의 냉동 보존에 있어서 여러 조건의 평가)

  • Kim, Hyun;Cho, Young Moo;Han, Jae Yong;Choi, Sung Bok;Byun, Mi Jeong;Kim, Young Sin;Ko, Yeoung-Gyu;Seong, Hwan-Hoo;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.41 no.4
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    • pp.249-259
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    • 2014
  • Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps and freezing media on the rates of viability of cryopreserved chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos of Korean Ogye (KO) and Commercial breeds (C), using the MACS method were suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG)). Gonads were harvested from stage 28 chick embryos and pooled in groups of 5, 10, 15, 20E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments : 2.5%, 5%, 10%, 15% and 0% cryoprotectant (DMSO, EG, PG) as a control. Effects of exposure to slow freezing and vitrification, with different concentrations of the cryoprotectant solution, were examined. After vitrification and slow freezing, survival rates of the frozen-thawed PGCs from the 10% EG plus FBS treatment were 85.63%, and 66.14% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05) (85.63% vs 66.81%) by vitrification. This study established a method for preserving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at a germplasm repository and ease of entry into a data base. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

Quantification of Protein and Amylose Contents by Near Infrared Reflectance Spectroscopy in Aroma Rice (근적외선 분광분석법을 이용한 향미벼의 아밀로스 및 단백질 정량분석)

  • Kim, Jeong-Soon;Song, Mi-Hee;Choi, Jae-Eul;Lee, Hee-Bong;Ahn, Sang-Nag
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.603-610
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    • 2008
  • The principal objective of current study was to evaluate the potential of near infrared reflectance spectroscopy (NIRS) as a non-destructive method for the prediction of the amylose and protein contents of un-hulled and brown rice in broad-based calibration models. The average amylose and protein content of 75 rice accessions were 20.3% and 7.1%, respectively. Additionally, the range of amylose and protein content were 16.6-24.5% and 3.8-9.3%, respectively. In total, 79 rice germplasms representing a wide range of chemical characteristics, variable physical properties, and origins were scanned via NIRS for calibration and validation equations. The un-hulled and brown rice samples evidenced distinctly different patterns in a wavelength range from 1,440 nm to 2,400 nm in the original NIR spectra. The optimal performance calibration model could be obtained by MPLS (modified partial least squares) using the first derivative method (1:4:4:1) for un-hulled rice and the second derivative method (2:4:4:1) for brown rice. The correlation coefficients $(r^2)$ and standard error of calibration (SEC) of protein and amylose contents for the un-hulled rice were 0.86, 2.48, and 0.84, 1.13, respectively. The $r^2$ and SEC of protein and amylose content for brown rice were 0.95, 1.09 and 0.94, 0.42, respectively. The results of this study suggest that the NIRS technique could be utilized as a routine procedure for the quantification of protein and amylose contents in large accessions of un-hulled rice germplasms.

Comparison Analysis of Aromatic Compounds in the Aromatic Rice Germplasm by Gas Chromatography and Mass Spectrometry (Gas Chromatography-Mass Spectrometry에 의한 향미 유전자원의 방향성 화합물 비교분석)

  • Kim, Jeong-Soon;Cho, Jum-Rae;Gwag, Jae-Gyun;Kim, Tae-San;Ahn, Sang-Nag;Lee, Sok-Young
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.54 no.1
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    • pp.88-103
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    • 2009
  • The volatile and semi-volatile compounds of 5 accessions of domestic scented and 25 accessions of introduced scented were extracted by solid phase microextraction (SPME) and analyzed by gas chromatographymass spectrum (GC-MS). A total of 156 volatile and semivolatile compounds were identified from 30 accessions of aromatic rice, including 32 alcohols, 25 acids, 25 ketones, 21 hydrocarbon, 18 esters, 16 aldehydes, 4 ethers, 5 amines, 2 phenols, 2 bases, and 8 miscellaneous compounds. By UPGMA/Neighbor-join tree analysis, the thirty accessions of aromatic rice could be classified into seven groups according to the major odor or aroma compounds. Group I included indica type of Basmati varieties. Group II and Group IV included japonica type introduced scented. Group III consisted only Hyangmibyeo1ho in domestic scented. Group V and Group VII included indica type of Basmati and non-Basmati varities. Group VI included four of domestic scented of seven accessions excepted Basmati6129, Basmati 6311, and Seratus Malam.

Variation of Growth Characteristics and Quality Related Components in Korean Indigenous Tea (Camellia sinensis) Germplasms (한국 재래종 차나무(Camellia sinensis)의 작물학적 특성 및 품질관련 성분 변이)

  • Lee, Min-Seuk;Lee, Jin-Ho;Lee, Jeong-Dae;Hyun, Jin-Wuk;Kim, Young-Gul;Hwang, Young-Sun;Lee, Hyeon-Jin;Choi, Su-San-Na;Lee, Su-Jin;Choung, Myoung-Gun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.3
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    • pp.333-338
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    • 2008
  • The tea has traditionally been used as a foodstuff by unique flavor, however recently not only the diversity of consumer demands but also the public interest in unique favorite and functional aspects have increased. It has been also reported that the main components contained in the leaves of tea (Camellia sinensis) include total nitrogen, free amino acids, polyphenols, and fiber, of which catechin has powerful bioactive effect such as anti-cancer, anti-aging, and anti-diabetic. (-)-Epigallocatechin gallate (EGCG) which is a major phenolic constituent of green tea extract has received considerable attention for a variety of important bioactivities. This study was carried out to obtain useful information for tea breeding programs, and to investigate the concentration of quality and functional related components in Korean indigenous tea germplasms. Korean indigenous tea lines were classified into three groups of sprout time, i.e, early, medium and late sprout time, and the ratio were 20%, 43% and 37%, respectively. There was a difference in characteristics among these Korean indigenous tea lines, leaf width of those ranged from 19.8 to 75 mm, leaf length was 35.5-160.0 mm, and leaf area was $660-8,400\;mm^2$. Experimental data on chlorophyll content (SPAD value) of Korean indigenous tea genetic resources ranged from 51.3 to 82.3. The concentrations of the total nitrogen, total free amino acids, and theanine were ranged 4.18-6.07%, 2.87-4.58%, and 1.64-2.66%, respectively. Also, catechin concentration showed from 11.54 to 15.07%, and concentration of caffeine was 2.82-4.23%. These results indicated indicated that it is possible to select elite lines with high concentration of quality related components and low concentration of caffeine from Korean domestic tea germplasms.

Optimal Extraction Condition of Anthocyanins in Soybean (Glycine max) with Black Seed Coats (검정콩 종피 함유 안토시아닌의 최적 추출조건)

  • Choung, Myoung-Gun;Hwang, Young-Sun;Lee, Hyeon-Jin;Choi, Su-San-Na;Lim, Jung-Dae;Kang, Sung-Taeg;Han, Won-Young;Baek, In-Youl;Kim, Hyeun-Kyeung
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.1
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    • pp.110-117
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    • 2008
  • Soybeans (Glycine max) with a black seed coat have been widely utilized as food source and as a medicinal herbs in Korea. The pigmentation in the seed coat of black soybean is due to accumulate anthocyanins in the epidermis palisade layer. The anthocyanin content and composition of the black soybean seed coat are considered as a standard. of quality evaluation of black soybean. The main objective of this study was to investigate the optimal condition for an extraction method of anthocyanins and compare anthocyanin quantity and composition within black soybean varieties and germplasms. In the test of extraction solvent, absorbance at 530 nm and Hunter's a value were increased as increasing the concentration of MeOH, but Hunter's Land b values were the exact opposite of absorbance and Hunter's a values. There was no significant difference for anthocyanin contents from 1% HCl - $H_2O$ to 1% HCl - 80% MeOH. In the aspects of anthocyanin contents and HPLC peak resolution, 1% HCl - 20% MeOH extraction solution was the most suitable solvent. Among the 5 kinds of extraction method using 1% HCl - 20% MeOH solution, the anthocyanin contents of room temperature extraction at 72 h was the highest among the methods. High extraction temperature, sonication and reflux method influenced on the decrease of anthocyanin contents because of breakdown of anthocyanins. There was no significant difference for extraction time between 12 h and 24 h. However, the optimal extraction condition were at room temperature for 12 h. The anthocyanin contents in seed coats of black soybean were determined on the basis of HPLC peak area at 530 nm. Ten black soybean varieties and germplasms were tested with optimal conditions founded in this study. On the basis of antocyanin component, these can be classified into three groups; C3G, C3G + D3G and C3G + D3G + Pt3G. The total anthocyanin content in seed coats ranged from 1.58 to 10.62 mg/g of seed. The total anthocyanin content of the variety "Geomjeongol" was about 7 times higher than that of variety "Heugchong". Information for extraction method and diversity in antocyanin of soybean seed coats can be used for future research for germplasm evaluation and development of high quality black soybean varieties.

Study on the Genetic Characteristics of Waterlogging Tolerant Pepper (Capsicum annuum L.) for Breeding Tolerant Varieties against Flooding Stress (내습성 고추 품종 육성을 위한 선발계통의 유전적 특성 구명)

  • Yang, Eun Young;Chae, Soo-Young;Hong, Jong-Pil;Lee, Hye-Eun;Park, Eun Joon;Moon, Ji-hye;Park, Tae-Sung;Roh, Mi-Young;Kim, Ok Rye;Kim, Sang Gyu;Kim, Dae Young;Lee, Sun Yi;Cho, Myeong Cheoul
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1111-1120
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    • 2017
  • This study was conducted to select pepper lines that were tolerant to excessive water injury among the pepper germplasm and investigate the genetic characteristics of those lines to contribute to the breeding of pepper cultivars with stable productivity in abnormal weather. Each of the tolerant and susceptible lines went through immersion treatment, and differentially expressed genes between them were analyzed. The tolerant line showed increased expression of the CA02g26670 gene, which is involved in the CONSTANS protein pathway and regulation of flowering by day length, but it exhibited decreased expressions of CA01g21450, CA01g22480, CA01g34470, CA02g00370 and CA02g00380. The susceptible line showed increased gene expressions of CA02g09720, CA02g21290, CA03g16520, CA07g 02110, and CA12g17910, which are involved in the inhibition of proteolytic enzyme activity, DNA binding, inhibition of cell wall-degrading enzyme, and inhibition of nodulin, respectively. Meanwhile the expressions of CA02g02820, CA03g21390, CA06g17700 and CA07g18230 decreased in the susceptible line, in relation to calcium-ion binding, high temperature, synthesis of phosphocholine and cold stress, respectively. The expressions of genes related to apoptosis and peroxidase increased, while that of CA02g16990, which functions as a nucleoside transporter, decreased in both the tolerant and susceptible lines. Based on the different gene expressions between the tolerant and susceptible lines, further studies are needed on breeding abiotic stress-tolerant lines.

Effect of Ethylene Glycol(EG) and Propylene Glycol(PG) on the Viability of Frozen-thawed Primordial Germ Cells(PGCs) on Korean Native Chicken(Ogye) by Vitrification (한국재래닭(오계)의 유리화 동결 시 생존율에 미치는 Ethylene Glycol(EG)과 Propylene Glycol(PG)의 영향)

  • Kim, Hyun;Kim, Dong Hun;Han, Jae Yong;Choi, Sung Bok;Ko, Yeoung Gyu;Do, Yoon Jung;Seong, Hwan-Hoo;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.197-205
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    • 2013
  • This study established a method for preserving chicken primordial germ cells (PGCs) that enables long-term storage in liquid nitrogen ($LN_2$) for preservation of the species. The purpose of this study was to compare the effects of Ethylene Glycol (EG) and Propylene Glycol (PG) on viability of cryopreserved PGCs with vitrification in Korean Native Chicken (Ogye), and to fine should be find or to the optimal protocol for PGCs freezing. One of the important components of cryopreservation process is cryopreservation medium that plays a vital role in preventing cellular injury during freeze-thawing. Cryoprotective agents have been known to improve cell viability after freeze-thawing. PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos, using the MACS method were suspended in a freezing medium containing a freezing and protecting agents. Gonads were harvested from stage 28 chick embryos and pooled in groups of 10E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments: 2.5% EG, 5% EG, 10% EG, 2.5% PG, 5% PG, 10% PG, and 0% cryoprotectant as a control. Effects of exposure to vitrification solution and vitrification, with different concentrations of the cryoprotectant solution, were examined. After freezing and thawing, survival rates of the frozen-thawed PGCs from the 0, 2.5, 5, 10 and 15% EG plus FBS treatment were 44.24%, 64.51%, 85.63%, 80.51% and 73.52% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05)(85.63% vs 66.81%). Therefore, these systems may contribute in the improvement of cryopreservation for a scarce species in birds preservation. This study established a method for preserving chicken PGC that enables systematic storage and labeling of cryopreserved PGCs in liquid N at a germplasm repository and ease of entry into a database. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

Yellowish Friable Embryogenic Callus (YFEC) Production and Plant Regeneration from Immature Embryo Cultures of Domestic Maize Cultivars and Genotypes (Zea may L.) (국내 옥수수 품종 및 계통의 미숙배 배양으로부터 Yellowish Friable Embryogenic 캘러스 (YFEC) 생산과 식물체 재생)

  • Cho Mi-Ae;Park Yun-Ok;Kim Jin-Suck;Park Ki-Jin;Min Hwang-Ki;Liu Jang-Ryol;Choi Pil-Son
    • Journal of Plant Biotechnology
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    • v.32 no.2
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    • pp.117-121
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    • 2005
  • Immature embryos of 3 cultivars (Du Me Chal, Mi Baek Chal, Heug Jeom Chal) and 5 genotypes (HW1, KL103, HW3, HW4, KW7) were cultured on medium containing MS salts, Eriksson's vitamins, 1 mg/L 2,4-dichlorophenoxyacetic acid, 25 mM proline, 100 mg/L casamino acid, 3 mM MES, 1.7 mg/L $AgNO_3$ and 20 g/L sucrose (SIM). Frequency of somatic embryo formation on explant of immature embryos showed in HW1 (45.20%), KL103 (5.75%), HW3 (37.20%), HW4 (30.10%), KW70 (55.20%), Mi Baek Chal (18.74%), Heug Jeom Chal (22.41%), Du Me Chal (36.72%) and Hi II type (<10%), respectively. Yellowish friable embryogenic callus (YFEC) such as type II callus of Hi II genotype only produced from the HW3 and Heug Jeom Chal, whereas other cultivars and genotypes were directly formed somatic embryos with late-embryonic stages or expanded yellowish compact somatic embryo with morphological abnormality. The yellowish friable embryogenic callus (YFEC) could be proliferated on the same medium, which were maintained embryogenic capacity for 6 months over. Upon transfer to first regeneration and second regeneration medium, somatic embryos converted to plantlets at a frequency of approximately 100%. However, the expanded somatic embryos with abnormal morphology were slowly proliferated when subcultured on the same medium, and some of them were degenerated or converted to plantlets at a frequency of approximately 25%. Accordingly, The Heug Jeom Chal and HW3 genotype will be further used for development of high frequency transformation system in domestic maize germplasm.

Variation of Anthocyanin Contents according to Collection Site and Maturity in Black Soybean (검정콩 수집지역과 성숙기에 따른 안토시아닌 함량 변이)

  • Yi, Eun-Seob;Lee, Yong-Sun;Kim, Hee-Dong;Kim, Yong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.53 no.4
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    • pp.376-381
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    • 2008
  • Anthocyanin contents of black soybean were analyzed for development of superior breeding lines with high anthocyanin contents. Total 292 genotypes of black soybean collected through the whole country were analyzed by HPLC in which C3G (Cyanidin-3-Glucoside), D3G (Delphinidin-3-glucoside), and Pt3G (Petunidin-3-Glucoside) were detected main anthocyanin pigments and each pigment contents were significantly different according to genotypes. C3G content showed the highest value in all materials and its variation was also wide, whereas D3G and Pt3G were not detected in 4 and 24 genotypes. Mean value of C3G, D3G, and Pt3G contents were $8.05{\pm}4.225$, $1.80{\pm}0.854$, and $1.15{\pm}0.781\;mg/g$, respectively. In case collected sites, genotypes collected in Chungnam region were higher the anthocyanin contents than other collections, which was $13.75{\pm}3.861\;mg/g$. It might be concluded that it takes more than 36 days for anthocyanin accumulation since beginning of seed-coat pigments formation, in that case it showed $13.09{\pm}4.190\;mg/g.$. Also total anthocyanin contents were present higher concentration in seed coat as maturation period was longer from flowering stage.