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Nucleus-Selective Expression of Laccase Genes in the Dikaryotic Strain of Lentinula edodes

  • Ha, Byeongsuk;Lee, Sieun;Kim, Sinil;Kim, Minseek;Moon, Yoon Jung;Song, Yelin;Ro, Hyeon-Su
    • Mycobiology
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    • v.45 no.4
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    • pp.379-384
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    • 2017
  • In mating of Lentinula edodes, dikaryotic strains generated from certain monokaryotic strains such as the B2 used in this study tend to show better quality of fruiting bodies regardless of the mated monokaryotic strains. Unlike B2, dikaryotic strains generated from B16 generally show low yields, with deformed or underdeveloped fruiting bodies. This indicates that the two nuclei in the cytoplasm do not contribute equally to the physiology of dikaryotic L. edodes, suggesting an expression bias in the allelic genes of the two nuclei. To understand the role of each nucleus in dikaryotic strains, we investigated single nucleotide polymorphisms (SNPs) in laccase genes of monokaryotic strains to reveal nuclear origin of the expressed mRNAs in dikaryotic strain. We performed reverse transcription PCR (RT-PCR) analysis using total RNAs extracted from dikaryotic strains (A5B2, A18B2, and A2B16) as well as from compatible monokaryotic strains (A5, A18, and B2 for A5B2 and A18B2; A2 and B16 for A2B16). RT-PCR results revealed that Lcc1, Lcc2, Lcc4, Lcc7, and Lcc10 were the mainly expressed laccase genes in the L. edodes genome. To determine the nuclear origin of these laccase genes, the genomic DNA sequences in monokaryotic strains were analyzed, thereby revealing five SNPs in Lcc4 and two in Lcc7. Subsequent sequence analysis of laccase mRNAs expressed in dikaryotic strains revealed that these were almost exclusively expressed from B2-originated nuclei in A5B2 and A18B2 whereas B16 nucleus did not contribute to laccase expression in A2B16 strain. This suggests that B2 nucleus dominates the expression of allelic genes, thereby governing the physiology of dikaryons.

Effects of Hyeolbuchukeo-tang on the Estradiol Valerate-induced Polycystic Ovaries and the Conception of Rats (혈부축어탕(血府逐瘀湯)이 Estradiol Valerate로 유발된 흰쥐의 다낭성 난소 및 수태에 미치는 영향)

  • Bae, Sang-Jin;Kim, Hyung-Jun;Lee, Dong-Nyung
    • The Journal of Korean Obstetrics and Gynecology
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    • v.25 no.2
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    • pp.43-65
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    • 2012
  • Objectives: This study was designed to investigate the effects of Hyeolbuchukeotang( HCT) on the Estradiol Valerate(EV) induced Polycystic Ovaries and the Implantation of rats. Methods: PCO was induced by single intramuscular(IM) injection with EV in female rats. Normal group(n=8) were injected with sesame oil and orally administrated distilled water for 60 days. Control group(n=8) were injected with EV and orally administrated distilled water for 60 days. HCT treated group(n=8) were injected with EV and orally administrated HCT for 60 days. At the end day of experiment, we measured weights of body, ovaries, adrenal glands, uterus and contents of serum LH, FSH, ADD. The histomorphometrical changes of ovaries were also evaluated. And we observed the NGF and CRF expression by immunohistochemistry. Also we observed the mating and count the number of implantation. Results: - The weights(mg) of ovaries in HCT treated group($58.9{\pm}6.5$) were significantly increased (p<0.01) compared with control group ($42.3{\pm}8.5$). - The number of mature follicles in HCT treated group($9.8{\pm}2.6$) was significantly increased (p<0.01) compared with control group($6.1{\pm}2.1$). - The number of cystic follicles in HCT treated group($1.6{\pm}1.2$) was significantly decreased (p<0.01) compared with control group($3.8{\pm}1.5$). - The value of serum FSH(mIU/ml) in HCT treated group($3.870{\pm}2.151$) was significantly decreased(p<0.05) compared with control group($10.476{\pm}6.294$). - The expression of NGF-immunoreactive cells in the ovarian cells in HCT treated group was lesser observed than control group. - The number of implantation in HCT treated group($7.4{\pm}1.8$) was significantly increased (p<0.05) compared with control group($3.1{\pm}4.4$). Conclusions: Hyeolbuchukeo-tang(HCT) is effect on polycystic ovaries and the implantation of rats by EV-induced.

Isolation of Hypervirulent Agrobacterium spp from Korea and Application for Transformation of Tobacco (한국산 고감염 Agrobacterium spp의 분리 및 연초의 형질전환에 이용)

  • 양덕춘;정재훈;이정명
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.207-217
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    • 1998
  • Total of 78 strains were characterized based on the morphological characteristics of colonies isolated on Schroth, and New & Kerr's media for selection of hypervirulent wild-type Agrobacterium spp from galls, hairy root-like process and soil of Populus, Malus, Salix and Diopyros in Korea. Among them, 48 strains were able to induce tumors in carrot disc. Hypervirulent A. tumefaciens SP101 and SM042 were identified as biotype 1 and biotype 2, respectively, These strains formed fast growing, larger tumors as compared to those induced by other strains. The binary vector pGA643 with kanamycin resistant gene was mobilized from E. coli MC100 into A. tumefaciens strain SM042 isolated from soil, and/or disarmed vector PC2760 using a triparental mating method with E. coli HB101/pRK2013, and transconjugants, A. tumefaciens SM643 and PC643 were obtained in minimal media containing kanamycin and tetracycline. Tobacco tissues were cocultivated with conjugant Agrobacterium and then transferred to selective medium with 2,4-D and kanamycin to induce the transformants. Calli were formed more efficiently in cocultivation with A. tumefaciens SM643 than that with A. tumefaciens PC643. Most of calli transformed with A. tumefaciens PC643 were friable and regenerated into normal plantlets, while the calli transformed with A. tumefaciens SM643 were compact, hard, and mixed with friable calli. The friable calli formed normal shoots, while compact calli did not form shoots but only grew to typical compact tumor calli. When the shoots formed directly from tobacco stems without callus induction after transformation by A. tumefaciens SM643 with wild-type Ti-plasmid, normal transformed plants can be induced without using disarmed Ti-plasmid.

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Genetic Diversity and Population Structure of Liriope platyphylla (Liliaceae) in Korea (한국내 맥문동의 유전적 다양성과 집단 구조)

  • Huh, Hong-Wook;Choi, Joo-Soo;Lee, Bok-Kyu;Huh, Man-Kyu
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.328-333
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    • 2007
  • Genetic diversity and population structure of eleven Liriope platyphylla (Liliaceae) populations in Korea were determined using genetic variation at 20 allozyme loci. The percent of polymorphic loci within the enzymes was 55.9%. Genetic diversity at the species level and at the population level was high(Hes = 0.178; Hep = 0.168, respectively), whereas the extent of the population divergence was relatively low ($G_{ST}$ = 0.064). $F_{IS}$, a measure of the deviation from random mating within the 11 populations, was 0.311. Total genetic diversity values ($H_T$) varied between 0.0 and 0.535, giving an average over all polymorphic loci of 0.323. The interlocus variation in within population genetic diversity ($H_S$) was high (0.305). An indirect estimate of the number of migrants per generation (Nm = 3.66) indicates that gene flow is high among Korean populations of the species. In addition, analysis of fixation indices revealed a substantial heterozygosity deficiency in some populations and at some loci. Mean genetic identity between populations was 0.988. It is highly probable that directional toward genetic uniformity in a relatively the homogenous habitat is thought to be operated among Korean populations of L. platyphylla.

Current Status on the Occurrence and Management of Disease, Insect and Mite Pests in the Non-chemical or Organic Cultured Apple Orchards in Korea (무농약 유기재배 사과원의 병해충 발생과 관리 실태)

  • Choi, Kyung-Hee;Lee, Dong-Hyuk;Song, Yang-Yik;Nam, Jong-Chul;Lee, Soon-Won
    • Korean Journal of Organic Agriculture
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    • v.18 no.2
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    • pp.221-232
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    • 2010
  • During 2005~2009, current status on the occurrence and the management of the major disease, insect and mite pests were investigated in the non-chemical or organic cultured apple orchards in Korea. Numbers of certified organic or non-chemical apple orchards increased from 14 in 2005 to 78 in 2008. Severe damages on leaves and fruits were caused by the several diseases such as marssonina blotch, bitter rot, white rot, sooty blotch and flyspeck, and the several insect pests such as apple leaf-curling aphid, woolly apple aphid, oriental fruit moth and peach fruit moth on the almost certified organic or non-chemical pest control orchards. About 10 and 18 environmental-friendly materials were used to control diseases and insect or mite pests, respectively. But, lime sulfur and bordeaux mixture to diseases and machine oil, plant oil mixed with egg yolk, and pheromone mating disruptions to insect pests were effective under the adequate conditions.

Insulin Resistance in Late Pregnant Rats (임신 후반기 흰쥐의 인슐린 저항성과 그 기전)

  • Chun, Myung-Heup;Kim, Yong-Woon;Park, So-Young;Kim, Jong-Yeon;Lee, Suck-Kang
    • Journal of Yeungnam Medical Science
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    • v.12 no.2
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    • pp.319-330
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    • 1995
  • The influence of normal late pregnancy on insulin action and insulin secretion was studied in the Sprague-Dawley female rats. On 20th day after mating, intravenous glucose tolerance test(IVGTT) was performed in non pregnant control and pregnant rats. As results of IVGTT, glucose disappearance rate was not significantly different in both groups, but secretory response of insulin was significantly(p<0.05) increased in pregnant rat. And the ratio of insulin/glucose was significantly higher in pregnant rats, which means existence of insulin resistance. These insulin resistance was overcomed by increased secretory response of pancreatic insulin. Insulinogenic index(${\Delta}$ insulin/glucose - 5 min) was highly significantly (r=0.62, p<0.01) correlated with progesterone concentration. Glycogen level and amounts of $^{14}C$-glucose incorporated into glycogen after IVGTT were significantly(p<0.05) decreased in the liver, but were not changed significantly in soleus. Glycogen synthase activity of soleus and liver was not differ significantly in the both groups. Insulin binding at varying concentrations of insulin to crude membrane of pregnant liver was not significantly different from control. In conclusions, although these pregnant rats were normal glucose tolerance due to increased secretory response of insulin, that was correlated with progesterone concentration, pregnant rat had insulin resistance. The mechanisms of insulin resistance were not related to defect of insulin binding phase and glycogen synthase, but suggest pre-receptor and/or postreceptor phase.

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Detection of mcr-1 Plasmids in Enterobacteriaceae Isolates From Human Specimens: Comparison With Those in Escherichia coli Isolates From Livestock in Korea

  • Yoon, Eun-Jeong;Hong, Jun Sung;Yang, Ji Woo;Lee, Kwang Jun;Lee, Hyukmin;Jeong, Seok Hoon
    • Annals of Laboratory Medicine
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    • v.38 no.6
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    • pp.555-562
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    • 2018
  • Background: The emerging mobile colistin resistance gene, mcr-1, is an ongoing worldwide concern and an evaluation of clinical isolates harboring this gene is required in Korea. We investigated mcr-1-possessing Enterobacteriaceae among Enterobacteriaceae strains isolated in Korea, and compared the genetic details of the plasmids with those in Escherichia coli isolates from livestock. Methods: Among 9,396 Enterobacteriaceae clinical isolates collected between 2010 and 2015, 1,347 (14.3%) strains were resistant to colistin and those were screened for mcr-1 by PCR. Colistin minimum inhibitory concentrations (MICs) were determined by microdilution, and conjugal transfer of the mcr-1-harboring plasmids was assessed by direct mating. Whole genomes of three mcr-1-positive Enterobacteriaceae clinical isolates and 11 livestock-origin mcr-1-positive E. coli isolates were sequenced. Results: Two E. coli and one Enterobacter aerogenes clinical isolates carried carried IncI2 plasmids harboring mcr-1, which conferred colistin resistance (E. coli MIC, 4 mg/L; E. aerogenes MIC, 32 mg/L). The strains possessed the complete conjugal machinery except for E. aerogenes harboring a truncated prepilin peptidase. The E. coli plasmid transferred more efficiently to E. coli than to Klebsiella pneumoniae or Enterobacter cloacae recipients. Among the three bacterial hosts, the colistin MIC was the highest for E. coli owing to the higher mcr-1-plasmid copy number and mcr-1 expression levels. Ten mcr-1-positive chicken-origin E. coli strains also possessed mcr-1-harboring IncI2 plasmids closely related to that in the clinical E. aerogenes isolate, and the remaining one porcine-origin E. coli possessed an mcr-1-harboring IncX4 plasmid. Conclusions: mcr-1-harboring IncI2 plasmids were identified in clinical Enterobacteriaceae isolates. These plasmids were closely associated with those in chicken-origin E. coli strains in Korea, supporting the concept of mcr-1 dissemination between humans and livestock.

A Study on the Ovipositional Characteristics of Endangered Species Prosopocoilus astacoides blanchardi (Coleoptera: Lucanidae) for Indoor Mass-rearing (멸종위기종 두점박이사슴벌레의 실내 증식을 위한 산란 특성 연구)

  • Ko, Gyeong Hun;Kim, Dong-Soon
    • Korean journal of applied entomology
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    • v.60 no.4
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    • pp.423-429
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    • 2021
  • The ovipositional characteristics of the endangered species Prosopocoilus astacoides blanchardi was investigated for indoor mass-rearing. This insect required 3 weeks or more for mating after emergence when introduced at the 1:1 ratio of male to female. Among temperatures studied at 16, 20, 24, 28, and 32℃, the pre-ovipositional period of P. astacoides blanchardi was longest at 16℃ with an average of 109 days after emergence, while it was shortest at 32℃ with an average of 59.4 days. The number of eggs laid at 24℃ was highest with an average of 40.6 per female. The average lifespan of adults at 16℃ was 208.8 days for males and 263.9 days for females, while at 32℃ 96.1 days for males and 130.3 days for females. The number of laid eggs was dependent on insect rearing chamber size. The oviposition-trees did not significantly (P > 0.05) affect the number of laid eggs and ovipositional period. The results of this study could be used as rearing data for the restoration and mass propagation of the endangered species P. astacoides blanchardi.

Breeding of a New Cultivar, 'Sanggang' with Upright Stipe and Improved Shelf life in Agrocybe aegerita (직립형이고 저장성이 개선된 버들송이 신품종 '상강' 육성)

  • Jeon, Dae-Hoon;Ha, Tai-Moon;Choi, Jong-In;Ju, Young-Cheol;Yoo, Young-Bok
    • Korean Journal of Breeding Science
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    • v.42 no.6
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    • pp.711-716
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    • 2010
  • 'Sanggang', a new cultivar of Agrocybe aegerita, was bred with mating between monokaryotic strains isolated from 'GMAG45109' and 'GMAG45107' in Mushroom Research Station, Gyonggi Province A.R.E.S. in 2009. The optimum temperature for the mycelial growth of 'Sanggang' was 26 to 28$^{\circ}C$ on PDA(potato dextrose agar) medium. The optimum temperature for the primordia formation and fruiting body development of 'Sanggang', was 18 to 20$^{\circ}C$. In the bottle cultivation of 'Sanggang', the period of spawn running was around 38 days at 22 to 23$^{\circ}C$ and the period from scratching of inoculum to harvest was 12 days. These characteristics of 'Sanggang' were not different from those of 'Mihwang' (control). 'Sanggang' had brown-colored pileus, whereas 'Mihwang' had yellowish brown-colored one. In bottle cultivation, 'Sanggang' hada little smaller but stronger stipe and pilus than 'Mihwang', and 'Sanggang', as well as 'Mihwang', had upright stipe. And 'Sanggang' was shown to have lower veil opening ratio than that of 'Mihwang' at harvest time.The yield of fruiting bodies of 'Sanggang' was 134 g/850ml bottle, which was similar to that of 'Mihwang'. Resistance of'Sanggang' against Trichoderma spp., as well as 'Mihwang', was weak. Freshness of 'Sanggang' was maintained for 13 days under storage temperature 4$^{\circ}C$, while that of 'Mihwang' was maintained for 10 days.

Breeding of a new cultivar of Flammulina velutipes: 'Baeke' (팽이 신품종 '백이' 육성)

  • Im, Ji-Hoon;Jang, Kab-Yeul;Oh, Youn-Lee;Oh, MinJi;Lee, Seul-ki;Raman, Jegadeesh;Kong, Won-Sik
    • Journal of Mushroom
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    • v.16 no.4
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    • pp.299-303
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    • 2018
  • Flammulina velutipes, which is a white rot fungus, is a commercially important edible mushroom and is produced in large quantities with the help of an automated and mechanized cultivation system in Korea. F. velutipes has the lowest distribution rate among domestic cultivars, estimated at about 20 percent. As most white cultivars of F. velutipes produced and exported to Korea were introduced from Japan, farmers pay large amounts of royalties. Therefore, we have developed a new, purely domestic cultivar, "Baeke," to substitute for Japanese cultivars, which has improved storage characteristics for export. Baeke was bred by mating two monokaryotic strains isolated from ASI 4216-18 (Hansol) and ASI 4217-26 (Baekjung). Baeke showed faster mycelial growth and higher mycelial density upon incubation for seven days at $25^{\circ}C$ on PDA media than the control variety. The mycelial growth of Baeke was even maintained at $30^{\circ}C$ unlike the control. The lengths of pilei and stipes in Baeke harvested in the optimal stage were $11.2{\pm}0.5mm$ and $125{\pm}5.4mm$, respectively, and they were $11{\pm}0.5mm$ and $141.9{\pm}5.7mm$, respectively, in the control harvested in the optimal stage. The yields of Baeke ($257.4{\pm}13.5g$) and control ($270.7{\pm}17.8g$) per 1,100ml in bottle cultivation showed no significant difference. Overall, our results showed that Baeke was at par with foreign varieties of Flammulina velutipes in terms of quality and yield and had a uniformly shaped fruitbody, which added to its commercial value.