• Title/Summary/Keyword: isozyme

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Effects of taurine and ginseng extracts on energy metabolism during exercise and their anti-fatigue properties in mice

  • Kim, Jisu;Beak, Suji;Ahn, Sanghyun;Moon, Byung Seok;Kim, Bom Sahn;Lee, Sang Ju;Oh, Seung Jun;Park, Hun-Young;Kwon, Seung Hae;Shin, Chul Ho;Lim, Kiwon;Lee, Kang Pa
    • Nutrition Research and Practice
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    • v.16 no.1
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    • pp.33-45
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    • 2022
  • BACKGROUND/OBJECTIVES: Ginseng extract (GSE) and taurine (TR) are widely used antifatigue resources in functional foods. However, the mechanism underlying the antifatigue effects of GSE and TR are still unclear. Hence, we investigated whether GSE and TR have synergistic effects against fatigue in mice. MATERIALS/METHODS: L6 cells were treated with different concentrations of TR and GSE, and cell viability was determined using 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium. Oxidative stress was analyzed by immunocytochemistry using MitoTrackerTM Red FM and an anti-8-oxoguanine antibody. Respiratory gas analysis was performed to investigate metabolism. Expression of an activated protein kinase was analyzed using immunohistochemistry. Gene expression of cluster of differentiation 36 and pyruvate dehydrogenase lipoamide kinase isozyme 4 was measured using reverse transcription-polymerase chain reaction. Mice were orally administered TR, GSE, or their combination for 30 days, and then fatigue-related parameters, including lactate, blood urea nitrogen, and glycogen, were measured after forced swimming. RESULTS: TR and GSE reduced oxidative stress levels in hydrogen peroxide-stimulated L6 cells and enhanced the oxygen uptake and lipid metabolism in mice after acute exercise. After oral administration of TR or GSE for 30 days, the fatigue-related parameters did not change in mice. However, the mice administered GSE (400 mg/kg/day) alone for 30 days could swim longer than those from the other groups. Further, no synergistic effect was observed after the swimming exercise in mice treated with the TR and GSE combination for 30 days. CONCLUSIONS: Taken together, our data suggest that TR and GSE may exert antifatigue effects in mice after acute exercise by enhancing oxygen uptake and lipid oxidation.

A comparative Study on the Activities of Glutamic Dehydrogenase and Glutamic Transaminase in Livers of the Crucian carp, Pigeon, and Rat (붕어, 비둘기, 흰쥐의 肝臟의 Glutamic Dehydrogenase 및 Glutamic Transaminase 의 活性에 관한 比較 硏究)

  • Kim, Yong Kyu;Nam, Sang Yul
    • The Korean Journal of Zoology
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    • v.12 no.2
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    • pp.50-56
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    • 1969
  • The present investigation involves a comparative study of enzymatic activities in various animals. The levels of the liver protein of rat (22.0$\pm$0.01mg/ml and pigeon (22.0$\pm$ 0.16) are twice as high as that of crucian carp (13.0$\pm$ 0.09)(p < 0.01). Generally, the specific acitivity (3.77 $\pm$0.18 unit/mg) of rat glutamic pyruvic transaminase (GPT) is highest, pigeon intermediate (1.93 $\pm$0.01), and crucian carp lowest (0.71$\pm$0.07). On the other hand, the specific activity (8.23$\pm$0.09 unit/mg)of rat glutamic oxaloacetic transaminase (GOT) is highest, pigeon intermediate (3.95$\pm$0.09), and crucian carp lowest (0.92$\pm$0.01) (p < 0.01). Ratios of GOT activity to GPT activity appear no remarkable difference from the levels of various animals. Specific activity of glutamic dehydrogenase(GDH) in pigeon tissue exceeds those of rat and crucian carp. In liver, rat GOT specific activity is greater than crucian carp and pigeon. On the other hand, pigeon GDH specific activity is greater than those of rat and crucian carp. This wouls seem to be in accord with protein metabolic intensity. The patterns for GDH isozyme were remarkably appeared in various animals. Glutamic dehydrogenase isozymes gave different electrophoretic mobilities in various animals. It is interesting that crucian carp, pigeon, and rat would show this difference, which may be indicative of an evolutional pattern. The fact that livers in various animals show quite different enzyme activities would suggest the existence of such a general phylogenetic relationship.

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Induction of Hepatic Microsomal Cytochrome P450 by N,N-dimethylformamide in Sprague-Dawley Rats (흰쥐에서 N,N-dimethylformamide에 의한 간장의 Microsomal Cytochrome P450의 유도)

  • Koh, Sang-Baek;Cha, Bong-Suk;Kang, Seung-Kyu;Joung, Hyo-Seok;Kim, Ki-Woong
    • Journal of Preventive Medicine and Public Health
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    • v.32 no.1
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    • pp.88-94
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    • 1999
  • Objectives. In order to gain a better understanding of the mechanism of DMF toxicity, recent studies have focused on hepatic drug metabolizing enzymes. In this study, we investigated the effects of DMF on the induction of P450 and the activities of other related enzymes in rat liver microsomes. Methods. DMF was administered to male Sprague Daweley rats by intraperitoneal injection at 0(control), 450(D1), 900(D2), 1,800(D3) mg DMF/kg body weight in olive oil once a day for three days. Hepatic P450 was measured by method of Omura and Sato. We evaluated selective assays for the three drug metabolizing cytochrome P450 isoenzymes 1A1, 2B1 and 2E1. Results. The content of microsomal protein, P450 and b5 were tended to be decreased in DMF treated group, but they were not statistically significant. The activity of NADPH-cytochrome P450 reductase was significantly increased dose dependently(p<0.01), but the activity of NADH-b5 reductase was decreased in the treated group(p<0.01). The activities of PROD and EROD were not significant between control and treated group. The activities of pNPH in the DMF treated groups were higher than that of the control group(p<0.01). When Western immunoblottings were carried out utilizing three monoclonal antibodies which were specific against P4501A1/1, P4502B1/2 and P4502E1, the strong density band corresponding to P4502E1 was observed with the microsomes obtained from the rats treated with DMF. But there were no significant increased in the P4501A1/2 and P4502B1/2 band densities in immunoblotting. Conclusions. These result suggested that P4502E1 was inducible by DMF and P4502E1 isozyme might be responsible for the hydroxylation of DMF to HMMF.

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Effects of Alcohol on Psychomotor Performance and Subjective Assessments In Normal Adults with Variation of Acetaldehyde Dehydrogenase I (정상성인에서 ALDH-I의 변이가 알콜의 정신운동성 수행과 주관적 평가에 미치는 영향)

  • Yoon, Bo Hyun;Yoon, Jin Sang
    • Korean Journal of Biological Psychiatry
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    • v.3 no.2
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    • pp.222-239
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    • 1996
  • The purpose of this study was to evaluate the effects of alcohol on the psychomotor performance and subjective assessment in healthy Korean adults with acetaldehyde dehydrogenase I(ALDH-I) isozyme variance. A total of 20 male subjects, half with active ALDH-I and the other half with inactive ALDH-I, were selected through both a self-reporting questionnaire examining alcohol sensitivity and the Higuchi's ethanol patch test detecting ALDH-I deficiency. In a doule-blind, placebo-controlled cross-over design, each subject consumed four doses of alcohol(0.25, 0.5, 0.75, 1.0g/kg) and placebo on five separate occasions at weekly intervals, Treatment order was fully balanced using a $5{\times}5$ Latin square, Psychomotor performance tests[coritical flicker fusion threshold(CFF) and choice reaction time(CRT)] and self-estimate questionnaires were conducted at baseline and at time points of 20, 40, 60, 90, 120, 150, 180 minutes after consuming the test drug for 20 minutes, Blood alcohol concentrations(BACs) using breath analyzer were measured at baseline and at time points of 10, 20, 30, 40, 50, 60, 75, 90, 105, 120, 150, 180 minutes after drinking, The BACs and the mean changes in the psychomotor performances and subjective assessments from pre-alcohol baseline, were compared between the two groups. The findings were summarized as follows : 1) BACs were tended to be higher in the inactive group than the active in all of the four alcohol doses. However significant group differences were only after the 0.5g/kg dose of alcohol. 2) The inactive group showed significant impairment in CFFT at most time points alter 0.75 and 1.0g/kg doses of alcohol. 3) In CRT, total reaction time(TRT) significantly prolonged in the inactive group than the active group at 20 minutes after 0.25 and 1.0g/kg doses of alcohol and at 40, 60, 90 minutes alter 0.75g/kg dose of alcohol. In the inactive group, recognition time component significantly increased at 20, 60, 90 minutes after 1.0g/kg dose of alcohol, while movement time component significantly increased at 40, 60 minutes after 0.75g/kg dose of alcohol. 4) Subjective evaluation of the effect of alcohol revealed that physical and mental conditions as well as a self-estimate of the effects of alcohol on performance were significantly worse in the inactive group than the active at some time points alter all of the lour alcohol doses, wihch were more pronounced after 0.75 and 1.0g/kg doses of alcohol. 5) Most of the group differences mentioned above, still remained statistically significant after BAC was entered as a covariate, These findings demonstrated that the alcohol sensitivity is higher in individuals with inactive ALDH-I than those with active ALDH-I both on the subjective assessments and the objective psychomotor performances. Furthermore, these results suggest thai the alcohol sensitivity may be determined by acetaldehyde concentration rather than BAC per se. In future studies, after more accurate genotyping for ALDH-I, the relationships between BAC, acetaldehyde concentration and alcohol sensitivities should be clearly defined.

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Difference in Electrophoretic Phenotypes of rice Cultivars Selected to Bensulfuron (Bensulfuron에 대(對)한 내성(耐性) 및 감수성(感受性) 수도품종(水稻品種)의 전기영동(電氣泳動) 표현형(表現型) 차이(差異))

  • Kuk, Y.I.;Guh, J.O.;Kim, Y.J.;Lee, D.J.
    • Korean Journal of Weed Science
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    • v.8 no.3
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    • pp.250-257
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    • 1988
  • The study was intended to know any relations between the rice tolerance to bensulfuron and varietal speciation in seed protein composition or any enzymatical allelies with or without chemical treatment. Rice varieties used were UCP-28, Chinsurah Boro II, Fukunohama, Fadehpur-2, IR 14252-13-2-2-5 as the tolerant group, and HP 93(3) FA, HP94(9) FA, Padilabou Alumbis, KH-17854, and IR 1846-2841-1 as the susceptible, respectively. Electrophoretic methods used were SDS-PAGE for seed protein, 7% PAGE for isozymes (acid phosphatase, peroxidase, malate dehydrogenase, and esterase from rice seedling) and variation in isoenzyme profiles (malate dehydrogenase, peroxidase, and esterase) as affected by different concentrations of bensulfuron(0, $10^{-6}$, $10^{-5}$ and $3{\times}10^{-5}M$) was also studied. The results are summarized as follows. -Among 16 bands separated in seed proteins, two different rice groups selected in terms of tolerance to bensulfuron were clustered in dissimilarity, which was based on relatively larger area in whole peaks and higher activities in N, O, P bands for the tolerant group. -Among isozymes obtained from rice seedlings without chemical treatments, the following specificities were obtained. The tolerant varieties had the relatively higher activity in D band out of 4 peroxidase bands. Malate dehydrogenase was separated into 3 bands and only tolerant varieties had A band and higher activities in Band C bands. Esterase was separated into 3-4 bands with higher activities in A and B bands for tolerant varieties. There were one major band accompanied by 2-3 minor bands for acid phosphatase in which only tolerant varieties had the B band. -The effect of Bensulfuron concentration on the isozyme activities showed that the activity of C band in peroxidase was not present in tolerant varieties which was contrary to the increased activities in susceptible varieties. However, D band was gradually disappeared only in susceptible varieties as the concentration of bensulfuron was increased. For malate dehydrogenase in the susceptible varieties, major bands D, E and F kept consistantly higher activities while minor bands A, B and C disappeared sensitively. Among 5 bands of esterase separated, D band was present only in the tolerant varieties while E band only in the susceptible. The activities in A, C, E bands were sharply decreased in the susceptible varieties as the concentration of bensulfuron was increased.

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Localization and characteristics of lactate and palate dehydrogenase in the sparganum and adult worm of Spirometra erinacei (만손열두조충에서 젖산탈수소효소와 말산탈수소효소의 분포와 특징)

  • Gwak, Gi-Hun;Jeon, Eun-U;Kim, Chang-Hwan
    • Parasites, Hosts and Diseases
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    • v.34 no.1
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    • pp.59-68
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    • 1996
  • This study investigated the enzyme histochemical localization and characteristics of lactate (LDH) and malate dehydrogenase (MDH) related with the oxidation-reduction metabolism in the sparganum and adult of 5. erinacei. By enzyme histochemical assay, activity of LDH was strong in the tegument and subtegumental muscle layers of the adult and sparganum. Activity of MDH was strong in the tegument of the sparganum and subtegumental muscle layers of the adult. However it was weak in the tegument of the adult. By electrophoresis, 45 kDa band was major and common in LDH of adults and spargana. The 150 kDa molecule was the major and common band in MDH of adults and r -spargana (from experimentally infected rats) . By isoelectrofocusing, isoelectric points (Pl) or 4 MDH isogyme from adult worm were 6.0.6.5, 6.7 and 7.1, respectively. Pl 6.0 was the major band. The active range of pH for MDH was about pH 6-8 and the optimum pH was pH 7 The effective temperature on the MDH was about $30^{\circ}C$$50^{\circ}C$ and the optimum temperature was about 40℃ in spargana md adult worm. In the stability against heat, when MDH was heated at 85℃ for 10 seconds, the activity was denatured perfectly. Maximum activity or MDH was 19.4 unit in the s-sparganum (from snakes), 24.5 unit in the r-sparganum (from rats) and 108.0 unit in the adult worm. The maximum activity was higher in adults than in spargana. The present result showed us that the nutrients absorbed through the tegument were transferred into inner tissues and were utilized as the source of metabolism. According to the habitat of the parasite, the isozymes of LDH and MDH are activated differently, and by this different activation the sparganum and adult can adapt themselves to parasitic circumstances.

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Silvicultural and Genetic Studies on Isozyme Patterns in Forest Trees - Inheritance of Leucine Aminopeptidase and Peroxidase Isozymes in ×Pinus taeda·rigida and P. densiflora - (유용임목(有用林木)의 동위효소(同位酵素)에 관(關)한 조림(造林) 및 유전학적(遺傳學的) 연구(硏究) - ×Pinus taeda·rigida와 P. densiflora에 있어서 Leucine Aminopeptidase와 Peroxidase 동위효소(同位酵素)의 유전(遺傳) -)

  • Park, Young Goo;Son, Won Ha
    • Journal of Korean Society of Forest Science
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    • v.43 no.1
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    • pp.39-50
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    • 1979
  • Twelve Pinus taeda were used as mother trees and one P. rigida plus tree was used as pollen tree for 12 cross combinations. Nine P. densiflora plus trees(4 mother and 7 pollen trees) were used as parents for 11 cross combinations. Those parents and 10 progenies were analyzed for LAP of ${\times}$P. taeda rigida hybrid and P. densiflora and for peroxidase of ${\times}$P. taeda rigida. The analysis, based on the banding patterns, indicate three alleles for LAP-A locus(A1, A2, A3) and two alleles for LAP-B locus (B1, B2) in ${\times}$P. taeda rigida hybrids. Chi-square test on the segregation for progenies did not show significant differences. The results indicated good agreement with monohybrid Mendelian inheritance. Independence test for occurrence frequency of 2 alleles(LAP-A3, LAP-B2) illustrated that there is neither linkage nor repulsion relationship between LAP-A3 and LAP-B2 alleles. Three band at LAP-A locus were always exhibited from all parents and their progenies of P. densiflora. However, the occurrence of two bands at LAP-B locus was variable, one bands assumed as homozygous alleles(B2/B2) and two bands as heterozygous alleles(B1/B2). The segregation ratio for progenies of P. densiflora suggested that LAP-B locus may be controlled by two alleles(B1 and B2). Three Peroxidase loci(Px-A, Px-B, Px-C) assumed to be controlled by allozyme in ${\times}$P. taeda rigida hybrid. The Px-B and Px-C loci could not find out the variations from banding patterns of parents and their progenies, while the Px-A locus showed the variations of occurrence frequency by two bands. The segregation ratio for A1/A2 at LAP-A locus suggest that the peroxidase allozymes of ${\times}$P. taeda rigida hybrid appeare to be monomeric products; that is, Px-A locus may be controlled by two alleles (A1 and A2).

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Stock Identification of Todarodes pacificus in Northwest Pacific (북서태평양에 서식하는 살오징어(Todarodes pacificus) 계군 분석에 대한 고찰)

  • Kim, Jeong-Yun;Moon, Chang-Ho;Yoon, Moon-Geun;Kang, Chang-Keun;Kim, Kyung-Ryul;Na, Taehee;Choy, Eun Jung;Lee, Chung Il
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.17 no.4
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    • pp.292-302
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    • 2012
  • This paper reviews comparison analysis of current and latest application for stock identification methods of Todarodes pacificus, and the pros and cons of each method and consideration of how to compensate for each other. Todarodes pacificus which migrates wide areas in western North Pacific is important fishery resource ecologically and commercially. Todarodes pacificus is also considered as 'biological indicator' of ocean environmental changes. And changes in its short and long term catch and distribution area occur along with environmental changes. For example, while the catch of pollack, a cold water fish, has dramatically decreased until today after the climate regime shift in 1987/1988, the catch of Todarodes pacificus has been dramatically increased. Regarding the decrease in pollack catch, overfishing and climate changes were considered as the main causes, but there has been no definite reason until today. One of the reasons why there is no definite answer is related with no proper analysis about ecological and environmental aspects based on stock identification. Subpopulation is a group sharing the same gene pool through sexual reproduction process within limited boundaries having similar ecological characteristics. Each individual with same stock might be affected by different environment in temporal and spatial during the process of spawning, recruitment and then reproduction. Thereby, accurate stock analysis about the species can play an efficient alternative to comply with effective resource management and rapid changes. Four main stock analysis were applied to Todarodes pacificus: Morphologic Method, Ecological Method, Tagging Method, Genetic Method. Ecological method is studies for analysis of differences in spawning grounds by analysing the individual ecological change, distribution, migration status, parasitic state of parasite, kinds of parasite and parasite infection rate etc. Currently the method has been studying lively can identify the group in the similar environment. However It is difficult to know to identify the same genetic group in each other. Tagging Method is direct method. It can analyse cohort's migration, distribution and location of spawning, but it is very difficult to recapture tagged squids and hard to tag juveniles. Genetic method, which is for useful fishery resource stock analysis has provided the basic information regarding resource management study. Genetic method for stock analysis is determined according to markers' sensitivity and need to select high multiform of genetic markers. For stock identification, isozyme multiform has been used for genetic markers. Recently there is increase in use of makers with high range variability among DNA sequencing like mitochondria, microsatellite. Even the current morphologic method, tagging method and ecological method played important rolls through finding Todarodes pacificus' life cycle, migration route and changes in spawning grounds, it is still difficult to analyze the stock of Todarodes pacificus as those are distributed in difference seas. Lately, by taking advantages of each stock analysis method, more complicated method is being applied. If based on such analysis and genetic method for improvement are played, there will be much advance in management system for the resource fluctuation of Todarodes pacificus.

Strain Improvement of the Genus Pleurotus by Protoplast Fusion (원형질체(原形質體) 융합(融合)에 의한 느타리버섯속(屬)의 품종개발(品種開發))

  • Yoo, Young-Bok;You, Chang-Hyun;Cha, Dong-Yeul
    • The Korean Journal of Mycology
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    • v.21 no.3
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    • pp.200-211
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    • 1993
  • Somatic hybrids of Pleurotus florida ASI 2016 and Pleurotus ostreatus ASI 2018 were obtained by protoplast fusion. The 40 fusants($P1{\sim}P40$) was examined for the yield on fermented and pasteurized rice straw in a tray. The carpophore yield of them were showed as the range of $27.0{\sim}155.2$, based on parental values of 100(ASI 2018), The pilei of fusants between orange white colored P. florida and dark grey colored P. ostreatus had mixed colors in the young stage. Other breeding programmes were performed to improve new varieties with high yield and good quality. A new oyster mushroom variety, Wonhyeongneutaribeosus(P72), was developed at the Agricultural Sciences Institute, Rural Development Administration in 1990. This P.florida-ostreatus-ostreatus hybrid P72 was selected from 38 protoplast fusion products($P41{\sim}P78$) between P.florida-ostreatus recombinant P5-M 43-arg rib and P. ostreatus ASI 2-13-0 2001-19-pro orn. The yield indexes of 38 hybrids ranged $40.5{\sim}152.7$ compared with the parental values of 100(ASI 2001). Hybrid P72 was characterized by the large fruiting bundle of semispherical shape with long stipe and by the small and circular pileus, resulting in lower harvesting cost. A significant increase in carpophore production was observed in somatic hybrids of protoplasts due to heterosis. A comparision of hybrid with parents P72 was made using isozyme analysis. The esterase banding patterns could be characterized by new bands in the hybrids. Seven fusion products of four crosses between P.florida ASI 2016 and P. ostreatus ASI 2018 were analysed with respect to the distribution of progenies and segregation of gene markers by random basidiospore analysis. Segregation of alleles should yield progeny of four genotypes in a Mendelian ratio of 1 : 1 : 1 : 1 for prototrophs, auxotrophs of one parental type, auxotrophs of the other parental type, and auxotrophic recombinants, respectively. However, five fusants of them did not detect one parental, P.ostreatus, type. Basidiospores could yield progeny of 16 genotypes in the cross of one of the recombinant P5-M43-arg $rib{\times}P. ostreatus$ ASI 2-13-pro orn but the segregants of three fusants were not detected clearly. The allele ratio of loci could be expected 1 : 1 : 1 : 1 for arg, rib, pro and orn. The ratio, however, would be changed to 4 : 1 : 1 : 1 with increasing proportion of argo In almost all the fusants, prototrophic recombinants were recovered in large numbers against auxotrophic markers. Parental genotypes were recovered with the recombinant progeny amounting to $38.68{\sim}99.56%$. The analysis provides proof of heterokaryosis and strong evidence for haploidy of vegetative nuclei, a sexual cycle consisting of nuclear fusion and meiosis.

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Germination and Proteome Profile Characteristics of Wheat Seeds Treated under Different Concentrations of Abscisic Acid (Abscisic acid 농도에 따른 밀 종자의 발아와 단백질체의 발현 특성)

  • Jeong, Jae-Hyeok;Kim, Dae-Wook;Hwang, Woon-Ha;An, Sung-Hyun;Jeong, Han-Yong;Lee, Hyeon-Seok;Choi, In-Bea;Choi, Kyung-Jin;Yun, Jong-Tak;Yun, Song Joong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.63 no.1
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    • pp.25-34
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    • 2018
  • This study was conducted to investigate the germination and proteome profile characteristics of wheat seeds treated under various concentrations of abscisic acid (ABA). After-ripening, the seeds of three wheat cultivars (Baegjoong, Keumkang, and Uri) showing different levels of dormancy were used. Germination index and germination rate of the cultivars was higher than 0.95% and 98%, respectively, and these were not significantly different under 0, 10, 30, and $50{\mu}M$ ABA at 7 d after germination. However, the growth of the shoot and radicle was significantly inhibited at 10, 30, and $50{\mu}M$ ABA compared to that at $0{\mu}M$ ABA. Mean ABA content of the embryos of seeds germinated at 0 and $50{\mu}M$ ABA for 7 d was 0.8 and $269.0ngmg^{-1}DW$, respectively. Proteins extracted from embryos germinated for 4 d were analyzed by two-dimensional gel electrophoresis, and proteins showing a difference of 1.5-fold or greater in their spot volume relative to that of $0{\mu}M$ ABA were identified. The expression of four protein spots increased at $50{\mu}M$ ABA and two protein spots were detected only at $50{\mu}M$ ABA; these six proteins were all identified as globulin types. Conversely, the expression of three protein spots decreased at $50{\mu}M$ ABA and were identified as cytosolic glutamine sysnthetase, isocitrate dehydrogenase, and S-adenosylmethionine synthetase 2. In conclusion, ABA did not inhibit the germination rate regardless of pre-harvest sprouting characteristics of the cultivars. However, the growth of the shoot and radicle was significantly inhibited by ABA, most likely through the down regulation of glutamine, methyl group donor, and polyamines biosynthesis, among others, while accompanied by globulin accumulation in the embryos.