• Title/Summary/Keyword: desaturase-1

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Increase in Linolenate Contents by Expression of the fad3 Gene in Transgenic Tobacco Plants

  • Kang, Young-Hwi;Min, Bok-Kee;Park, Hee-Sung;Lim, Kyung-Jun;Huh, Tae-Lin;Lee, Se-Yong
    • BMB Reports
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    • v.29 no.4
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    • pp.308-313
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    • 1996
  • An 1.4 kb of the fad3 cDNA encoding microsomal linoleic acid desaturase catalyzing the conversion of linoleic acid (18:2, ${\omega}-6$) to linolenic acid (18:2, ${\omega}-3$) was introduced into tobacco plants by the Agrobacterium-mediated plant transformation, Among the transgenic tobacco plants conferring kanamycin resistance, five transformants showing increment in unsaturated fatty acid contents were selected and further analyzed for the transgenecity, In genomic Southern blot analyses, copy numbers of the integrated fad3 DNA in chromosomal DNA of the five transgenic tobacco plants were varied among the transgenic lines. By Northern blot analyses, the abundancy of the fad3 mRNA transcript directed by Cauliflower Mosaic Virus 35S promoter was consistent with the relative copy number of the fad3 DNA integrated in the chromosome of transgenic tobacco plants. When compared with the wild type, accumulation of linolenic acid in transgenic tobacco roots was elevated 3.7- to 4.7-fold showing a corresponding decrease in the linoleic acid contents; however, slight increments for linolenic acid were noticed in transgenic leaf tissues. These results indicated that the elevated level of fad3 expression is achieved in transgenic tobacco plants.

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Molecular Strategy for Development of Value-Added Sesame Variety

  • Chung, Chung-Han
    • Proceedings of the EASDL Conference
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    • 2004.10a
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    • pp.13-30
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    • 2004
  • There are two groups of significant functional constituents in sesame seeds on the whole; one is the vegetable oils and another is the anti-oxidative compounds. However, although high amounts of major fatty acids are synthesized in sesame seeds, their composition is unfavorable because the contents of alpha- and gamma-linolenic acid, the essential fatty acids, are very low or do not produced in sesame seeds. So, to increase these fatty acids in sesame seeds, one strategy is to overexpress their genes, ${\omega}$-3 fatty acid desaturase for alpha-linolenic acid and delta-6 fatty acid desaturase for gamma-linolenid acid, in them. Another molecular target is to enhance alpha-tocopherol, vitamin E, because its content is very low in sesame seeds. The enzyme, gamma-tocopherol methyltransferase, catalyzes the conversion of gamma-tocophero to alpha-tocopherol. Overexpression of this enzyme in sesame seeds could be also a good molecular breeding target. Reduction of phytic acid is also another molecular target in sesame seeds because phosphorus pollution may be caused by its high content in sesame seeds. Accordingly, to do so, one of target enzymes could be myo-inositol 1-phosphate synthase which is a key regulatory enzyme in the pathway of phytic aicd biosyntheses. In this lecture, a molecular strategy for development of value-added sesame crop is described in association with some results of our experiments involved in the molecular characterizations of the genes mentioned above.

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Genetics and Breeding for Modified Fatty Acid Profile in Soybean Seed Oil

  • Lee, Jeong-Dong;Bilyeu, Kristin D.;Shannon, James Grover
    • Journal of Crop Science and Biotechnology
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    • v.10 no.4
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    • pp.201-210
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    • 2007
  • Soybean [Glycine max(L.) Merr.] oil is versatile and used in many products. Modifying the fatty acid profile would make soy oil more functional in food and other products. The ideal oil with the most end uses would have saturates(palmitic + stearic acids) reduced from 15 to < 7%, oleic acid increased from 23 to > 55%, and linolenic acid reduced from 8 to < 3%. Reduced palmitic acid(16:0) is conditioned by three or more recessive alleles at the Fap locus. QTLs for reduced palmitic acid have mapped to linkage groups(LGs) A1, A2, B2, H, J, and L. Genes at the Fad locus control oleic acid content(18:1). Six QTLs($R^2$=4-25%) for increased 18:1 in N00-3350(50 to 60% 18:1) explained four to 25% of the phenotypic variation. M23, a Japanese mutant line with 40 to 50% 18:1 is controlled by a single recessive gene, ol. A candidate gene for FAD2-1A can be used in marker-assisted breeding for high 18:1 from M23. Low linolenic acid(18:3) is desirable in soy oil to reduce hydrogenation and trans-fat accumulation. Three independent recessive genes affecting omega-3 fatty acid desaturase enzyme activity are responsible for the lower 18:3 content in soybeans. Linolenic acid can be reduced from 8 to about 4, 2, and 1% from copies of one, two, or three genes, respectively. Using a candidate gene approach perfect markers for three microsomal omega-3 desaturase genes have been characterized and can readily be used in for marker assisted selection in breeding for low 18:3.

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Effects of Growth Hormone Gene Polymorphism on Lipogenic Gene Expression Levels in Diaphragm Tissues of Japanese Black Heifers

  • Ardiyanti, Astrid;Abe, Tsuyoshi;Tameoka, Nanae;Kobayashi, Eiji;Shoji, Noriaki;Ohtani, Yoshihisa;Suzuki, Keiichi;Roh, Sang-Gun;Katoh, Kazuo
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.8
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    • pp.1055-1062
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    • 2012
  • Two SNPs, i.e. L127V and T172M, of bovine growth hormone (GH) causing the presence of GH gene haplotypes A, B, and C was previously shown to alter intramuscular fatty acid (FA) composition in Japanese Black (JB) heifers. To determine the SNP effect on somatotropic hormone concentration and lipogenesis, we measured plasma GH, insulin, and insulin-like growth factor-1 (IGF-1) concentrations. We also measured mRNA levels of fatty acid synthase (FASN), stearoyl-coA desaturase (SCD), and sterol regulatory element binding proteins-1 (SREBP-1) and FA composition in diaphragm tissues. Heifers with genotype CC had the lowest plasma insulin concentration and FASN and SCD mRNA levels among genotypes. FASN mRNA levels in haplotype A tended to positively correlate with saturated FA (SFA) content and negatively correlated with C18:2 and unsaturated FA (USFA) contents. SCD mRNA levels in haplotype A positively correlated with monounsaturated FA (MUFA) contents and negatively correlated with C18:0 content. They also tended to positively correlate with C16:1, C18:1, and USFA contents and USFA/SFA ratio and negatively correlate with SFA content. Taken together, GH gene polymorphism affects the lipogenic genes expression levels and their relationships with fatty acid compositions in diaphragm tissues of JB heifers at 31 months of age.

Carrot Juice Administration Decreases Liver Stearoyl-CoA Desaturase 1 and Improves Docosahexaenoic Acid Levels, but Not Steatosis in High Fructose Diet-Fed Weanling Wistar Rats

  • Mahesh, Malleswarapu;Bharathi, Munugala;Reddy, Mooli Raja Gopal;Kumar, Manchiryala Sravan;Putcha, Uday Kumar;Vajreswari, Ayyalasomayajula;Jeyakumar, Shanmugam M.
    • Preventive Nutrition and Food Science
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    • v.21 no.3
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    • pp.171-180
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    • 2016
  • Non-alcoholic fatty liver disease (NAFLD) is one of the most prevalent liver diseases associated with an altered lifestyle, besides genetic factors. The control and management of NAFLD mostly depend on lifestyle modifications, due to the lack of a specific therapeutic approach. In this context, we assessed the effect of carrot juice on the development of high fructose-induced hepatic steatosis. For this purpose, male weanling Wistar rats were divided into 4 groups, fed either a control (Con) or high fructose (HFr) diet of AIN93G composition, with or without carrot juice (CJ) for 8 weeks. At the end of the experimental period, plasma biochemical markers, such as triglycerides, alanine aminotransferase, and ${\beta}$-hydroxy butyrate levels were comparable among the 4 groups. Although, the liver injury marker, aspartate aminotransferase, levels in plasma showed a reduction, hepatic triglycerides levels were not significantly reduced by carrot juice ingestion in the HFr diet-fed rats (HFr-CJ). On the other hand, the key triglyceride synthesis pathway enzyme, hepatic stearoyl-CoA desaturase 1 (SCD1), expression at mRNA level was augmented by carrot juice ingestion, while their protein levels showed a significant reduction, which corroborated with decreased monounsaturated fatty acids (MUFA), particularly palmitoleic (C16:1) and oleic (C18:1) acids. Notably, it also improved the long chain n-3 polyunsaturated fatty acid, docosahexaenoic acid (DHA; C22:6) content of the liver in HFr-CJ. In conclusion, carrot juice ingestion decreased the SCD1-mediated production of MUFA and improved DHA levels in liver, under high fructose diet-fed conditions. However, these changes did not significantly lower the hepatic triglyceride levels.

Application of a Reassortant Cucumber mosaic virus Vector for Gene Silencing in Tomato and Chili Pepper Plants

  • Hong, Jin-Sung;Rhee, Sun-Ju;Kim, Eun-Ji;Kim, Tae-Sung;Ryu, Ki-Hyun;Masuta, Chikara;Lee, Gung-Pyo
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.81-86
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    • 2012
  • We developed a reassortant RNA virus vector derived from $Cucumber$ $mosaic$ $virus$ (CMV), which has advantages of very wide host range and can efficiently induce gene silencing in a few model plants. Certain CMV isolates, however, show limited host ranges presumably because they naturally co-evolved with their own hosts. We used a reassortant comprised of two strains of CMV, Y-CMV and Gn-CMV, to broaden the host range and to develop a virus vector for virus-induced gene silencing (VIGS). Gn-CMV could infect chili pepper and tomato more efficiently than Y-CMV. Gn-CMV RNA1, 3 and Y-CMV RNA2-A1 vector were newly reconstructed, and the transcript mixture of RNA1 and 3 genomes of Gn-CMV and RNA2 genome of Y-CMV RNA2 containing portions of the endogenous phytoene desaturase (PDS) gene (CMV2A1::PDSs) was inoculated onto chili pepper (cv. Chung-yang), tomato (cvs. Bloody butcher, Tigerella, Silvery fir tree, and Czech bush) and $Nicotiana$ $benthamiana$. All the tested plants infected by the reassortant CMV vector showed typical photo-bleaching phenotypes and reduced expression levels of $PDS$ mRNA. These results suggest that the reassortant CMV vector would be a useful tool for the rapid induction of the RNA silencing of endogenous genes in chili pepper and tomato plants.

Effects of Hyaluronidase during In Vitro Maturation on Maturation and Developmental Competence in Porcine Oocytes

  • Jeon, Ye-Eun;Hwangbo, Yong;Cheong, Hee-Tae;Park, Choon-Keun
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.2
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    • pp.86-92
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    • 2019
  • The aim of this study was to investigate effects of hyaluronidase during IVM on oocyte maturation, oxidative stress status, expression of cumulus expansion-related (PTX, pentraxin; GJA1, gap junction protein alpha 1; PTGS2, prostaglandin-endoperoxide synthase 2) and fatty acid metabolism-related (FADS1, delta-6 desaturase; FADS2, delta-5 desaturase; PPARα, peroxisome proliferator-activated receptor-alpha) mRNA, and embryonic development of porcine oocytes. The cumulus-oocyte complexes (COCs) were incubated with 0.1 mg/mL hyaluronidase for 44 h. Cumulus expansion was measured at 22 h after maturation. At 44 h after maturation, nuclear maturation, intracellular glutathione (GSH) and reactive oxygen species (ROS) levels were measured. Gene expression in cumulus cells was analyzed using real time PCR. The cleavage rate and blastocyst formation were evaluated at Day 2 and 7 after insemination. In results, expansion of cumulus cells was suppressed by treatment of hyaluronidase at 22 h after maturation. Intracellular GSH level was reduced by hyaluronidase treatment (p < 0.05). On the other hand, hyaluronidase increased ROS levels in oocytes (p < 0.05). Only PTGS2 mRNA was enhanced in COCs by hyaluronidase (p < 0.05). Population of oocytes reached at metaphase II stage was higher in control group than hyaluronidase treated group (p < 0.05). Both of cleavage rate and blastocyst formation were higher in control group than hyaluronidase group (p < 0.05). Our present results showed that developmental competence of porcine oocytes could be reduce by hyaluronidase via inducing oxidative stress during maturation process and it might be associated with prostaglandin synthesis. Therefore, we suggest that suppression of cumulus expansion of COCs could induce oxidative stress and decrease nuclear maturation via reduction of GSH synthesis and it caused to decrease developmental competence of mammalian oocytes.

Production of transgenic cucumber expressing phytoene synthase-2A carotene desaturase gene

  • Jang, Hyun A;Utomo, Setyo Dwi;Kwon, Suk Yoon;Ha, Sun-Hwa;Xing-guo, Ye;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.341-346
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    • 2016
  • The objectives of this study were to 1) evaluate the efficiency of the protocol of Agrobacterium-mediated transformation of cucumber to introduce phytoene synthase-2a carotene desaturase (PAC genes); 2) demonstrate the integration of PAC genes into the genome of putative transgenic cucumber based on growth on selection medium, PCR and Southern analysis; 3) evaluate the expression of PAC genes in transgenic cucumber based on the analysis of RT-PCR and Northern blot hybridization. Out of 5,945 cotyledonary-node explants inoculated with Agrobacterium, 65 (1.1%) explants produced 238 shoots. Integration of PAC genes into the genome of the cucumber was demonstrated based on the analysis of gDNA-PCR, 21 out of the 238 plants regenerated; while 6 plants proved positive for Southern blot hybridization. Transgene expression was demonstrated based on analysis of RT-PCR, 6 plants proved positive out of the 6 plants analyzed; while 4 plants out of 6 proved positive during Northern blot hybridization. This study successfully demonstrated the production of transgenic cucumber, integration, and expression of the PAC gene in cucumber.