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MtMKK5 inhibits nitrogen-fixing nodule development by enhancing defense signaling

  • Hojin Ryu
    • Journal of Plant Biotechnology
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    • v.49 no.4
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    • pp.300-306
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    • 2022
  • The mitogen-activated protein kinase (MAPK) signaling cascade is essential for a wide range of cellular responses in plants, including defense responses, responses to abiotic stress, hormone signaling, and developmental processes. Recent investigations have shown that the stress, ethylene, and MAPK signaling pathways negatively affect the formation of nitrogen-fixing nodules by directly modulating the symbiotic signaling components. However, the molecular mechanisms underlying the defense responses mediated by MAPK signaling in the organogenesis of nitrogen-fixing nodules remain unclear. In the present study, I demonstrate that the Medicago truncatula mitogen-activated protein kinase kinase 5 (MtMKK5)-Medicago truncatula mitogen-activated protein kinase 3/6 (MtMPK3/6) signaling module, expressed specifically in the symbiotic nodules, promotes defense signaling, but not ethylene signaling pathways, thereby inhibiting nodule development in M. truncatula. U0126 treatment resulted in increased cell division in the nodule meristem zone due to the inhibition of MAPK signaling. The phosphorylated TEY motif in the activation domain of MtMPK3/6 was the target domain associated with specific interactions with MtMKK5. I have confirmed the physical interactions between M. truncatula nodule inception (MtNIN) and MtMPK3/6. In the presence of high expression levels of the defense-related genes FRK1 and WRKY29, MtMKK5a overexpression significantly enhanced the defense responses of Arabidopsis against Pseudomonas syringae pv. tomato DC3000 (Pst DC3000). Overall, my data show that the negative regulation of symbiotic nitrogen-fixing nodule organogenesis by defense signaling pathways is mediated by the MtMKK5-MtMPK3/6 module.

Investigation of the efficacy of mycotoxin-detoxifying additive on health and growth of newly-weaned pigs under deoxynivalenol challenges

  • Holanda, Debora Muratori;Kim, Sung Woo
    • Animal Bioscience
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    • v.34 no.3_spc
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    • pp.405-416
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    • 2021
  • Objective: This study evaluated the effects of feeding diets naturally contaminated with deoxynivalenol (supplemental 2 mg/kg) on health, growth, and the effects of a mycotoxin-detoxifying additive in newly-weaned pigs. Methods: Thirty-six pigs (27 day-old) were housed individually and assigned to 3 treatments for 5 weeks: CON (diet containing minimal deoxynivalenol), MT (diet with supplemental 1.9 mg/kg of deoxynivalenol), and MT+D (MT + mycotoxin-detoxifying additive, 0.2%, MegaFix, ICC, São Paulo, Brazil). The mycotoxin-detoxifying additive included bentonite, algae, enzymes, and yeast. Blood was taken at week 2 and 5. Jejunal tissue were taken at week 5. Data were analyzed using the MIXED procedure of SAS. Results: Pigs fed MT+D tended to have decreased (p = 0.056) averaged daily feed intake during week 1 than MT. At week 2, serum aspartate aminotransferase/alanine aminotransferase in MT tended to be lower (p = 0.059) than CON, whereas it was increased (p<0.05) for MT+D than MT, indicating hepatic damages in MT and recovery in MT+D. Pigs fed MT had lower (p<0.05) blood urea nitrogen/creatinine than CON, supporting hepatic damage. At week 5, pigs fed MT tended to have reduced (p = 0.079) glucose than CON, whereas it was increased (p<0.05) for MT+D than MT, indicating impaired intestinal glucose absorption in MT, which was improved in MT+D. Pigs fed CON tended to have increased (p = 0.057) total glutathione in jejunum than MT, indicating oxidative stress in MT. Pigs fed MT+D had a reduced (p<0.05) proportion of Ki-67-positive cells in jejunum than MT, indicating lower enterocyte proliferation in MT+D. Conclusion: Feeding supplemental 1.9 mg/kg of deoxynivalenol reduced growth and debilitated hepatic health of pigs, as seen in leakage of hepatic enzymes, impaired nitrogen metabolism, and increase in oxidative stress. The mycotoxin-detoxifying enhanced hepatic health and glucose levels, and attenuated gut damage in pigs fed deoxynivalenol contaminated diets.

Analysis of Benthic Macroinvertebrate Community and Biological Evaluation of Water Quality in the Gapyung Region of the Upper North Han River, Korea (한강 상류수계(가평)의 저서성 대형무척추동물 군집분석 및 생물학적 수질평가)

  • 배경석;유승성;원두희;김민영;신재영
    • Journal of Environmental Health Sciences
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    • v.28 no.2
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    • pp.149-160
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    • 2002
  • Analysis of benthic macroinvertebrate community structure and biological estimation of water quality by the benthic macroinvertebrates from the Mt. Myungji(1267m), Mt. Kalbong(840m), Mt. Daegeum(704m) and Mt. Jubal(489m) region of Gapyung-gun, Kyunggi-do were investigated from April to November, 2001. Total taxa of benthic macroinvertebrates were 105 species, 38 families, 13 orders, 5 classes in 4 phyla. Aquatic insects were 33 species in ephemeroptera, 5 species in odonata, 13 species in plecoptera, 2 species in hemiptera, 1 species in megaloptera, 27 species in trichoptera, 3 species in coleoptera and 13 species in diptera, respectively. Non-insects were 1 species in platyhelminthes, 5 species in gastropoda, 1 species in oligochaeta, 1 species in crustacea, respectively. Ephemeroptera, trichoptera and plecoptera as indicators in clean water were very abundantly. Occurrence species at each survey area was 67 species at Mt. Myungji and 69 species at Mt. Kalbong, 48 species at Mt. Daege.um and 47 species at Mt. Jubal, respectively. Mean species diversity indices at Mt. Myungji, Mt. Kalbong, Mt. Daegeum and Mt. Jubal were 3.30, 3.33, 2.59 and 3.43, respectively. According to the saprobic system based on the species diversity indices of benthic macroinvertebrates, Mt. Myungji, Mt. Kalbong and Mt. Jubal are determined as Limnosaprobic area, respectively, but Mt. Daegeum is determined as $\beta$-mesosaprobic area.

Analysis of Benthic Macroinvertebrate Community and Biological Estimation of Water Quality at Creeks in the Cheonwang-bong of National Park of Mt. Jiri and Its Nearby Area (지리산 천왕봉 일대의 저서무척추동물 군집분석 및 생물학적 수질평가)

  • 배경석;길혜경;유병태
    • Journal of environmental and Sanitary engineering
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    • v.18 no.2
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    • pp.16-26
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    • 2003
  • Analysis of benthic macroinvertebrate community and biological estimation of water quality was conducted at the national park of Mt. Jiri and its nearby area in Gurye, Sancheong and Hadong-gun from June to November, 2002. Total taxa of benthic macroinvertebrates were 124 species, 45 families, 15 orders, 5 classes in 3 phyla. Aquatic insects were 33 species in ephemeroptera, 28 species in Trichoptera, 21 species in plecoptera, 13 species in Diptera, 8 species in odonata, 5 species in hemiptera and 5 species in coleoptera, respectively. Non-insects were 5 species in Mollusca, 2 species in Hirudinea, 1 species in oligochaeta and 1 species in crustacea, respectively. Ephemeroptera, trichoptera and plecoptera as indicators in clean water were very abundant. Occurrence species at each survey area was 68 species at Mt. 1025-goji, 59 species at Mt. Eungseok-bong, 57 species at Mt. Wangdeung-jae, 50 species at Mt. Cheonwang-bong and 39 species at Mt. 645-goji, respectively. Mean species diversity indices at Mt. Cheonwang-bong, Mt. 1025-goji, Mt. Wangdeung-jae, Mt. Eungseok-bong and Mt. 645-goji were 3.33, 3.03, 3.41, 3.02 and 2.91, respectively, According to the saprobic system based on the species diversity indices of benthic macroinvertebrates, most survey areas except some sites are determined as Limnosaprobic area.

Effect of Seeding Date on Forage Yield and Chemical Composition of Kenaf in Jeju (제주지역에서 파종기에 따른 양마의 사료수량 및 조성분 변화)

  • 조남기;송창길;조영일;고지병
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.6
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    • pp.439-442
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    • 2001
  • This study has been conducted from April 25, 1999 to June 25, 1999 in Jeju Province in order to determine the influence of seeding date on growth, yield and chemical composition of Kenaf (Hibiscus cannabinus L.) Delaying the seeding date from April 25 to June 25 decreased the plant height of Kenaf from 278.5 cm to 205.7 cm. As for the number of leaves, number of withering leaves, number of branches, and stem diameter decreased as the seeding date was delayed. As the seeding date was delayed, fresh forage yield decreased from 98.5MT/㏊ to 45.9MT/㏊, dry matter yield from 20.7MT/㏊ to 8.2MT/㏊, crude protein (CP) yield from 2.9MT/ha to 1.3MT/ha and total digestible nutrients (TDN) yield form 1l.6MT/㏊ to 5MT/㏊. However, both leaf yield and stem yield were nearly the same tendency. Delaying the seeding date from April 25 to June 25 increased crude protein contents of leaves from 21.5% to 24.4%, crude fat contents from 5.2% to 6.1%, nitrogen free extract (NFE) contents form 39.9% to 41.2% and TDN contents from 64.3% to 69.7%. However, delaying seeding date decreased crude fiber contents from 20.8% to 17.5%, and crude ash contents from 7.9% to 7.0%. Based on the these findings, optimum seeding date for forage production of Kenaf seems to be about 25 April in atmospheric phenomena and volcanic ash soils of Jeju island.

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Effects of Split Nitrogen Application on Agronomic Characteristics, Forage Yield, and Chemical Composition of Kenaf in Jeju (제주지역에서 질소분시 횟수에 따른 양마의 생육특성, 사료수량 및 조성분 변화)

  • 조남기;강영길;송창길;조영일;정재수;고미라;오은경
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.22 no.1
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    • pp.9-14
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    • 2002
  • Evegradies 71' kenaf (Hibiscus cannabinus L.) was grown at 160,000 plants per ha in a volcanic ash soil in Jeju island to determine the optimum frequency of split N application fer forage production. Nitrogen fertilizer was applied with 240 kg/ha, and frequencies of the split N applications were 1, 2, 3, 4 and 5 times. Plant height averaged across two harvests increased from 187 to 201cm as N was split-applied from one to three applications and then decreased to 201cm in five applications. This pattern held for the number of branches and leaves per plant stem diameter and weight of plant per plant. Fresh forage yield increased from 91.8 to 114.2 MT/ha. dry matter yield from 12.70 to 16.6 MT/ha, crude protein yield from 1.75 to 2.48MT/ha, and total digestible nutrients (TDN) yield from 5.39 to 7.63 MT/ha as N was split-applied from one to four applications, and then decreased to 106.6, 15.0, 2.32, and 7.22 MT/ha in five applications, respectively As N was split-applied from one to five applications, crude protein content increased from 13.8 to 15.4%. ether extract content from 4.1 to 5.9%, and TDN content from 42.4 to 48.1%. but crude fiber decreased from 38.2 to 37.1% and crude ash content from 11.5 to 8.6%. Nitrogen free extract content was about 19.0% regardless of cutting height.

Effects of Phosphate Application Rate on Growth Characters, Forage Yield and Feed Value of Jeju Native Soybean (제주 재래대두의 인산시비량 차이가 생육형질, 수량 및 사료가치에 미치는 영향)

  • 조남기;강영길;송창길;윤상태;조영일;김동현
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.23 no.3
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    • pp.143-150
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    • 2003
  • This study was conducted to determine the response of main growth characters, yield and chemical composition of Jeju native soybean based on the difference between phosphate rates (0, 50, 100, 150, 200, 250kg/ha) from May 11 to September 10 in 2002 in Jeju island. Days to flowering was delayed from 92 days to 97 days as the increasing of phosphate rate. Also plant height grew longer from 109cm to 124cm. The number of branches and leaves, stem diameter, root length and weight of root grew powerful as phosphate rate increased from 0 to 250kg/ha. Fresh forage yield was 26.5MT/ha in the nonphosphate plot and then was 36.9MT/ha as phosphate rate increased to 250kg/ha. And the difference between 200kg/ha and 250kg/ha in phosphate rate was not significant. Dry matter, crude protein and TDN yield increased 6.0∼7.9MT/ha, 0.9∼1.4MT/ha and 3.4∼4.9MT/ha respectively, as the increasing of phosphate rate. Also crude protein, crude fat, NFE and TDN content increased 15.5∼18.3%, 2.2∼3.3%, 42.5∼43.5%, and 56.7∼61.9% respectively. In contrast with this, crude fiber and crude ash decreased 32.1∼28.1% and 7.7∼6.8% respectively. To reach the climax of forage yield was estimated optimum phosphate rate to be 200kg/ha.

Molecular Characterization of Metallothionein Gene of the Korean Bitterling Acheilognathus signifer (Cyprinidae) (묵납자루 (Acheilognathus signifer; Cyprinidae) metallothionein 유전자의 클로닝 및 특징 분석)

  • Lee, Sang-Yoon;Bang, In-Chul;Nam, Yoon-Kwon
    • Korean Journal of Ichthyology
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    • v.23 no.1
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    • pp.10-20
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    • 2011
  • Genetic determinant for metallothionein (MT), a cysteine-rich protein playing essential roles in metal detoxification and homeostasis, was characterized in the Korean bitterling (Acheilognathus signifer, Cyprinidae), an endemic fish species. The full-length A. signifer MT (AsMT) cDNA (551 bp) is composed of a single open-reading frame (ORF) to encode a polypeptide of 60 amino acids containing 20 cysteine residues whose positions are conserved in most cypriniform MTs. At the genomic level, the AsMT (2,593 bp spanning the 5'-flanking region to the 3'-untranslated region) represented a conserved tripartite (three exons interrupted by two introns) structure with AT-rich introns. The upstream regulatory region (-1,914 bp from the ATG initiation codon) of AsMT displayed various sites and motifs for transcription factors involved in the metal-mediated regulation and stress/immune responses. The AsMT transcript was ubiquitously detected in various organs with variable expression levels, where the ovary and intestine showed the highest expression, while the heart and skeletal muscle represented the lowest level. During an exposure to copper (immersion in $0.5\;{\mu}M$ Cu for 48 h), the levels of AsMT transcripts were significantly elevated in the liver (more than 3.5-fold), moderately in the gill, kidney, and spleen (ranging from 1.5- to 2.5-fold), and barely in the brain and intestine. Results of this study could form a useful basis to explore the metal-related stress physiology of this endangered fish species.

Development and Evaluation of an Oral Controlled Release Delivery System for Melatonin

  • Lee, Beom-Jin;Parrott, Keith A.;Sack, Robert L.;Ayres, James W.
    • Journal of Pharmaceutical Investigation
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    • v.23 no.3
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    • pp.9-18
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    • 1993
  • Sugar spheres loaded with melatonin (MT) were coated with $Aquacoat^{\circledR}$ to control the release rate of MT over 8 hours. A zero-order release pattern over 8 hours was obtained with 20% coating on 8-10 mesh beads in USP basket dissolution studies. MT in 20% coated beads was quite stable at room temperature with less than 5% MT degraded during 6 months' storage. Dissolution profiles were also unchanged after 6 months. An oral preparation containing MT-loaded uncoated beads for immediate release and 20% coated beads with $Aquacoat^{\circledR}$ for controlled release over 8 hours was evaluated in six human subjects. When total 0.5 mg MT as low dose (immediate release portion of MT, 0.1 mg) was administered to four subjects, average peak plasma MT concentration was reached at about 600 pg/ml and maintained at about 10 pg/ml over 8 hours. Plasma MT concentration-time profiles were similar in shape to computer-simulated profiles. However, maximal plasma MT concentrations were three times greater compared to computer simulated curve. These results suggest that MT dose, ratio of immediate and controlled release MT, and pharmacokinetic parameters selected are adjusted to mimic endogenous MT concentration-time curve. In another study, 0.2 mg MT having 10% of immediate release portion and 80% controlled release portion produced plasma MT concentration-time curve which is more similar to endogenous profiles. A low bioavailability (<20%) may result from extensive first pass metabolism and remaining amounts of MT from controlled beads. A good correlation between plasma MT concentration and urinary excretion rate of 6-sulphatoxymelatonin (6-STMT), a major metabolite of MT was observed. As plasma MT concentration increased, urinary excretion rate of 6-STMT increased concomitantly. The linear relation between plasma MT and urinary excretion rate of 6-STMT was statistically significant. This result suggests that urinary 6-STMT may be used as an index of circadian rhythms of MT in humans.

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Hyperproduction of L-Threonine by Adding Sodium Citrate as Carbon Source in Transformed Escherichia coli Mutant. (형질전환된 Escherichia coli변이주에서 Sodium citrate를 이용한 고농도 L-Threonine 생산)

  • 이만효;김병진;정월규;최선욱;박해룡;황용일
    • Journal of Life Science
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    • v.14 no.5
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    • pp.868-873
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    • 2004
  • The efficient fermentative production of L-threonine fermentation was achieved by using Escherichia coli MT201, transformed a plasmid carrying pyruvate carboxylase gene. It is an attempt to supply oxaloacetate to the L-threonine biosynthetic pathway. In order to improve the L-threonine productivity of E. coli MT201, a plasmid pPYC which is an expression vector of the pyruvate carboxylase gene of Coryne-bacterium glutamicum, was introduced. When E. coli MT/pPYC was incubated with medium containing only glucose as a carbon source, both the cell growth and L-threonine production were reduced, compared to the results from fermentation of E. coli MT201. In order to circumvent this effect, we attempted the addition of a mixed carbon source, composed of glucose and sodium citrate at a ratio of 1.5:3.5. It was shown that L-threonine production and cell growth (OD660) with E. coli MT/pPYC reached up to 75.7 g/l and 48, respectively, at incubation for 75 hr under fed-batch fermentation conditions. It is assumed that overproduction of L-threonine by anaplerotic pathway leads unbalance of TCA cycle and sodium citrate might playa role to recover normal TCA cycle.