• Title/Summary/Keyword: Transgenic soybean

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Analysis of the Glycinin Gy2 Promoter Activity in Soybean Protoplasts and Transgenic Tobacco Plants (대두 원형질체와 형질전환된 담배에서의 대두 glycinin 유전자 Gy2 promoter의 발현조절 기작)

  • Kim, Soo-Jung;Lee, Jee-Young;Kim, Chung-Ho;Choi, Yang-Do
    • Applied Biological Chemistry
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    • v.38 no.5
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    • pp.387-392
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    • 1995
  • To study the regulatory expression mechanism of soybean glycinin gone, Gy2, the 5' upstream region of the gene was searched for the presence of putative regulatory elements by nucleotide sequencing. It revealed various kinds of regulatory sequence elements commonly found in plant storage protein genes. There were canonical promoter sequences, TATA box (TATAAT) and AGGA box (GAAT) which are common in the 5' upstream region of the plant genes. The embryo factor binding sequence, RY repeat, CACA sequences, ${\alpha}$-conglycinin enhancer-like sequences were also found. To delineate the function of these sequences, 5' upstream deletion mutants of Gy2 were prepared and fused to the ${\alpha}$-glucuronidase (GUS) gene. Each chimeric construct was transferred into soybean protoplasts for transient assay, which led to the identification of the sequences between -281 and -223, -170 and -122, of Gy2 promoter as negative regulatory elements, and the sequences between -223 and -170, -122 and -16 as positive regulatory elements. These results are consistent in transformed tobacco plants as well. The serially deleted promoter fragments fused to the GUS were transformed into Nicotiana tabacum by Agrobacterium tumefaciens using the binary vector system. GUS activity of Gy2 promoter deletion constructs was detected only in seeds but not in leaves with different levels of expression as in transient assay. These results suggest that the glycinin Gy2 promoter drives a tissue-specific expression in transgenic tobacco plants.

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Overexpression of GmAKR1, a Stress-Induced Aldo/keto Reductase from Soybean, Retards Nodule Development

  • Hur, Yoon-Sun;Shin, Ki-Hye;Kim, Sunghan;Nam, Kyoung Hee;Lee, Myeong-Sok;Chun, Jong-Yoon;Cheon, Choong-Ill
    • Molecules and Cells
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    • v.27 no.2
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    • pp.217-223
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    • 2009
  • Development of symbiotic root nodules in legumes involves the induction and repression of numerous genes in conjunction with changes in the level of phytohormones. We have isolated several genes that exhibit differential expression patterns during the development of soybean nodules. One of such genes, which were repressed in mature nodules, was identified as a putative aldo/keto reductase and thus named Glycine max aldo/keto reductase 1 (GmAKR1). GmAKR1 appears to be a close relative of a yeast aldo/keto reductase YakC whose in vivo substrate has not been identified yet. The expression of GmAKR1 in soybean showed a root-specific expression pattern and inducibility by a synthetic auxin analogue 2,4-D, which appeared to be corroborated by presence of the root-specific element and the stress-response element in the promoter region. In addition, constitutive overexpression of GmAKR1 in transgenic soybean hairy roots inhibited nodule development, which suggests that it plays a negative role in the regulation of nodule development. One of the Arabidopsis orthologues of GmAKR1 is the ARF-GAP domain 2 protein, which is a potential negative regulator of vesicle trafficking; therefore GmAKR1 may have a similar function in the roots and nodules of legume plants.

Agronomic characteristics and field resistance to bacterial soft rot of transgenic potato overexpressing the soybean calmodulin 4 gene (SCaM4) (SCaM4 과발현 형질전환 감자의 농업적 특성 및 감자 무름병 저항성 평가)

  • Sohn, Hwangbae;Cho, Kwangsoo;Cho, Jihong;Gwon, Ohgeun;Cheon, Chunggi;Choi, Jigyeong;Chung, Woosik;Lee, Shin Woo
    • Journal of Plant Biotechnology
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    • v.39 no.4
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    • pp.295-299
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    • 2012
  • We performed in vitro assay and field trials to assess levels of changes in intrinsic properties and resistance against soft rot of the potato cv. Dejima upon the introduction of a soybean calmodulin 4 gene (SCaM4). Field trials with four lines overexpressing SCaM4 gene were conducted over two seasons, and harvested tubers were evaluated in bioassay for resistance to Pectobacterium carotovorum ssp. carotovorum. The SCaM4 transgenic potato lines inoculated with $10^8$ CFU/ml of P. carotovorum ssp. carotovorum showed enhanced resistance compared to control. Among the SCaM4 transgenic lines, the transgenic line SCaM4-4 exhibited the highest tolerance to soft rot in vitro assays, so did in field trials. In the field trial, the soft rot resistance of SCaM4-4 line was more than 5 times higher compared to that of control cultivar, Dejima. The major agronomic characteristics of the SCaM4 transgenic lines were not different from those of the nontransgenic 'Dejima'. The result demonstrated that the transformation of a calmodulin 4 gene was a successful strategy in development of potato cultivar enhanced to soft rot.

Translational Enhancement by the 5' Leader of Tobacco Mosaic Virus and Soybean Glycinin Gene in Transgenic Tobacco Plants (담배 모자이크 바이러스와 대두 Glycinin 유전자의 5' Leader Sequence를 이용한 외래 유전자의 전이효율 증진)

  • Kang, Hong-Gu;Park, Jee-Won;Kim, Chung-Ho;Lim, Jae-Yun;Choi, Yang-Do
    • Applied Biological Chemistry
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    • v.38 no.3
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    • pp.224-231
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    • 1995
  • To increase the expression of a foreign protein in transgenic plant, the benefits of 5'-untranslated leader sequences of tobacco mosaic virus (TMV) RNA or soybean glycinin gene, Gy2, fused to a protein coding sequence were exploited. pGA643-derived plasmid contains 355 promoter of cauliflower mosaic virus, protein coding sequence of maize 10 kDa zein (10kZ) and Gy2 terminator. The leader from Gy2 or TMV RNA was inserted between the promoter and the coding sequence in each construct. The recombinant DNAs were introduced into tobacco plants by Agrobacterium mediated leaf disc transformation method. Although the transgene without the leader had more transcripts than the others, mRNAs containing the leader were translated more efficiently. It might be due to difference in the length of 5'-untranslated sequence and context surrounding the AUG codon, but could be sequence specific rather. These results suggest that the leader sequences of Gy2 and TMV play important roles as an enhancer in translational control of foreign gene in transgenic tobacco plant.

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Regulation Mechanism of Soybean Storage Protein Gene Expression (대두 저장단백질 유전자의 발현 조절 메카니즘)

  • 최양도;김정호
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.283-307
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    • 1987
  • Glycinin and $\beta$-conglycinin are the most abundant storage protein in soybean. These proteins are known to be synthesized predominantly during germination and cell expansion phase of seed development for short period, and synthesized not in other tissues. Genes encoding these storage proteins are useful system to study the mechanism of development stage and tissue specific gene expression in eukaryotes, especially plants, at the molecular level. The cDNA and genomic clones coding for glycinin have been isolated and regulation mechanism of the gene expression has been studied. Initially, development and tissue-specific expression of the glycinin gene is regulated at the level of transcription. Post-transcriptional processing is also responsible for delayed accumulation of the mRNA. Translational control of the storage protein gene has not been reported. Post-translational modification is another strategic point to regulate the expression of the gene. It is possible to identify positive and/or negative reguratory clements in vivo by producing transgenic plants agter gene manipulation. Elucidation of activation and repression mechanism of soybean storage protein genes will contribute to the understanding of the other plant and eukaryotic genes at molecular level.

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Development of Transgenic Soybean Using Agrobacterium tumefaciens (Agrobacterium tumefaciens을 이용한 대두 형질전환체 개발)

  • Cho, Mi-Ae;Choi, Dong-Woog;Liu, Jang-Ryol;Clemente Tom;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.255-259
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    • 2004
  • Agrobacterium tumefaciens-mediated cotyledonary node transformation was used to produce transgenic soybean. Cotyledonary node explants of three cultivars and one genotype were co-cultivated with strains Agrobacterium (LBA4404, GV3101, EHA101, C58) containing the binary vectors (pCAMBIA3301 and pPTN289) carrying with CaMV 35S promoter-GUS gene as reporter gene and NOS promoter-bar gene conferring resistance to glufosinate (herbicide Basta) as selectable marker. There was a significant difference in the transformation frequency depend on bacteria strain. The EHA101 strain of the bacterial strains employed gave the maximum efficiency (3.6%). One hundred-six lines transformed showed the resistance in glufosinate. Histochemical GUS assay showed that at least 11 plants transformed with the GUS gene were positive response. The soybean transformants were obtained from the Thorne (5 plants), 1049 (5 plants) and Bakun (1 plant), respectively. Southern blot analysis and leaf painting assay revealed that the GUS and bar gene segregated and expressed in their progeny.

Development of Salt-Tolerant Transgenic Rice Using Soybean PR10 Gene (콩의 Pathogenesis-Related 10 유전자를 이용한 내염성 벼 형질전환 계통 개발)

  • Kim, Hyo Jin;Baek, So Hyeon;Shin, Woon Chul;Seo, Chun Sun;Park, Myoung Ryoul;Ko, Jae Kwon;Yun, Song Joong
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.540-546
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    • 2010
  • This study was conducted to understand the role of soybean pathogenesis-related 10 (GmPR10) gene in salt tolerance and to develop salt-tolerant rice using GmPR10 cDNA. GmPR10 transgene was expressed constitutively in the shoot and root of the $T_1$ transgenic rice plants. Interestingly, however, the levels of the transgene expression were increased temporally up to over four- to five-fold in the shoot and root by 125 mM NaCl treatment, peaking at six hours after the treatment and decreasing thereafter. Electrolyte leakage of leaf cells under 125 mM NaCl treatment was lower in all the transgenic lines than in the control variety, Dongjin-byeo. Ability of seedlings to recover from 125 mM NaCl treatment for two weeks was higher in the transgenic plants than in the control plants. These results demonstrated that GmPR10 had function to increase cell integrity and promote growth under the saline stress imposed by NaCl. The transgenic line GmPR10-3 which showed highest ability to recover from the saline stress could be used as a potential source for salt tolerance in rice breeding programs.

Impact of a Glyphosate-Tolerant Soybean Line on the Rhizobacteria, Revealed by Illumina MiSeq

  • Lu, Gui-Hua;Zhu, Yin-Ling;Kong, Ling-Ru;Cheng, Jing;Tang, Cheng-Yi;Hua, Xiao-Mei;Meng, Fan-Fan;Pang, Yan-Jun;Yang, Rong-Wu;Qi, Jin-Liang;Yang, Yong-Hua
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.561-572
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    • 2017
  • The global commercial cultivation of transgenic crops, including glyphosate-tolerant soybean, has increased widely in recent decades with potential impact on the environment. The bulk of previous studies showed different results on the effects of the release of transgenic plants on the soil microbial community, especially rhizosphere bacteria. In this study, comparative analyses of the bacterial communities in the rhizosphere soils and surrounding soils were performed between the glyphosate-tolerant soybean line NZL06-698 (or simply N698), containing a glyphosate-insensitive EPSPS gene, and its control cultivar Mengdou12 (or simply MD12), by a 16S ribosomal RNA gene (16S rDNA) amplicon sequencing-based Illumina MiSeq platform. No statistically significant difference was found in the overall alpha diversity of the rhizosphere bacterial communities, although the species richness and evenness of the bacteria increased in the rhizosphere of N698 compared with that of MD12. Some influence on phylogenetic diversity of the rhizosphere bacterial communities was found between N698 and MD12 by beta diversity analysis based on weighted UniFrac distance. Furthermore, the relative abundances of part rhizosphere bacterial phyla and genera, which included some nitrogen-fixing bacteria, were significantly different between N698 and MD12. Our present results indicate some impact of the glyphosate-tolerant soybean line N698 on the phylogenetic diversity of rhizosphere bacterial communities together with a significant difference in the relative abundances of part rhizosphere bacteria at different classification levels as compared with its control cultivar MD12, when a comparative analysis of surrounding soils between N698 and MD12 was used as a systematic contrast study.