• Title/Summary/Keyword: Transgenic soybean

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Optimization of Regeneration Condition Under Agrobacterium-mediated Transformation in in vitro Cultured Korean Soybean

  • Kantayos, Vipada;Bae, Chang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.36-36
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    • 2018
  • Soybean is the important crop in Asian countries as protein source, oil production and animal feed. Improving soybean using genetic transformation is the principal tool in nowadays. Developing herbicide resistant transgenic soybean plants through Agrobacterium-mediated transformation has been worked in many previous studied. However, the transformation efficiency is still low. Many attempts try to find the optimum media condition for plant regeneration after infection. After transformation, the plant regeneration is very important condition to promote growth of transgenic plant. In this study, we optimized a regeneration condition for two Korean soybean cultivar, Dawonkong and Pungsannamulkong using cotyledon, cotyledonary nodes and hypocotyl as explant. The results showed that shoot regeneration of cotyledonary nodes on B5 medium containing 2 mg/L 6-benzylaminopurine showed the highest percentage of regeneration in Dawonkong (75.8%) while Pungsannamulkong presented high number of shoots 2.12 shoots per explant. For transformation condition, co-cultivation in 7 days showed a high number of GUS positive expression. Most of explants can survived under media including 5 mg/L of glufocinate which refers phosphinotricin for 2-week selection. Washing with 400 mg/L of cefotaxime in several times and selection in plant regeneration media with 400 mg/L of cefotaxime can prevent bacteria growth, effectively.

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Organogenesis and Production of Some Transgenic Legume Plants by Agrobacterium tumefaciens-mediated Herbicide Resistance Gene Transformation

  • Kantayos, Vipada;Lee, Hyo-Yeon;Bae, Chang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.52-52
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    • 2018
  • Development of herbicide resistant transgenic legume plants through Agrobacterium-mediated transformation has been worked in many previous studied. Plant regeneration after infection is the important step to obtain successful transgenic plants. Many attempts try to find the optimum media condition for plant regeneration after infection. However, the transformation efficiency of legume plants is still low. In this study, regeneration of some Korean legume species including two soybean cultivars (Dawon and Pungsan) and pea have been done with organogenesis which is used various kind of explants such as cotyledonary-nodes in soybean and bud-containing tissue in pea. We developed the optimum media condition for plant regeneration regulators under Agrobacterium-mediated transformation using different kind and various concentration of plant growth. As the results, B5 medium containing 2 mg/L of 6-benzylaminopurine was selected in this study for the optimum plant regeneration media. The segments were inoculated with Agrobacterium suspension harbored an IG2 vector containing bar gene which confers resistance to phosphinotricin (PPT) in 3, 5 and 7 days. The transformation efficiency was achieved in Dawon 3.03 % and pea 1.46 % with co-cultivation period of 7 days which is showed a high number of GUS positive expression period.

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Evaluation of the acute toxicity of theoredoxin (TRX) transgenic soybean to Daphnia magna

  • Oh, Sung-Dug;Min, Seok-Ki;Kim, Jae Kwang;Park, Jung-Ho;Kim, Chang-Gi;Park, Soo Yun
    • Korean Journal of Agricultural Science
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    • v.47 no.4
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    • pp.791-802
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    • 2020
  • Theoredoxin (TRX) transgenic soybeans were developed using the human Theoredoxin gene under the control of the ��-conglycinin promoter with a selection marker, the phosphinothricin acetyltransferase (PAT) gene. This study was done to assess the acute toxicity of a genetically modified (GM) soybean using the fresh water planktonic crustacean Daphnia magna. The acute toxicity effect of the TRX soybean and non-GM soybean (Gwangan) on D. magna was investigated at different concentrations (0, 156, 313, 625, 1,250, 2,500, and 5,000 mg·L-1). The TRX soybean used for the test was confirmed to express the TRX/PAT genes by PCR and enzyme-linked immunosorbent assay (ELISA). D. magna feeding tests showed no significant differences in the cumulative immobility or an abnormal response with either the TRX soybean or non-GM soybean. The feeding study showed a similar abnormal response and cumulative immobility of the D. magna between the TRX soybean and Gwangan treatments. Additionally, the 48 h-EC50 values for the TRX and Gwangan soybeans were 755.6 and 778 mg·L-1, respectively. The soybean NOEC (no observed effect concentration) value for D. magna was suggested to be 156 mg·L-1. These results suggest that there is no significant difference in toxicity to Daphnia magna between the TRX soybean and its non-GM counterpart.

Identification of diversified functions of soybean FT homologs in photoperiod-dependent flowering time control

  • Lee, Su Hyeon;Choi, Cheol Woo;Kim, Min Chul
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.100-100
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    • 2017
  • FT is one of the major floral activator in plant flowering. FT determines the changing point of time from vegetative stage to reproductive stage. To understand the role of FT homologs in short-day plant soybean, we identified 10 soybean FT homologous genes and named GmFTs. We figured out that 10 GmFT genes were further categorized into three subclades through phylogenetic analysis. Expression analysis of GmFT genes indicated that they might have different functions in photoperiod-dependent soybean flowering. Most of GmFTs, for example, GmFT2a, GmFT2b, GmFT5a and GmFT6 mainly expressed in soybean leaves at short-day condition. However, interestingly GmFT1a and GmFT4 represented opposite expression pattern to other GmFTs. Arabidopsis transgenic plants overexpressing GmFT2a and GmFT5a exhibited extremely early flowering. In contrast, overexpression of GmFT4 delayed flowering of Arabidopsis transgenic plants. The results suggest that GmFT4 has antagonistic role to other GmFTs in soybean flowering. Interestingly, mRNA level of GmFT2a is higher in early flowering soybean accessions than in late flowering ones. Moreover, the highest point of mRNA level of GmFT2a showed the positive correlation with the timing of flowering of soybean accessions. But that of GmFT4 showed opposite pattern. Here, we report that soybean FT homologs might acquire different functions in photoperiod-dependent flowering through the functional diversification during evolution.

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Influence of β-carotene enhanced transgenic soybean cultivation on the diversity of non-target arthropods in Korea for three years

  • Sung-Dug Oh;Eunji Bae;Soo-Yun Park;Seong-Kon Lee;Doh-Won Yun;Kihun Ha;Minwook Kim;Yeongjin Son;Chang Uk Eun;Young-Kun Kim;Junho Lee;Dongmin Kim;Donguk Kim;Jongwon Kim;Sang Jae Suh
    • Korean Journal of Agricultural Science
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    • v.49 no.4
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    • pp.719-736
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    • 2022
  • Environmental risk assessment of living modified (LM) crops is essential for their cultivation. In this study, we cultivated β-carotene enhanced transgenic soybean (LM soybean) and non-LM soybean (Gwangan) in living modified organism (LMO) isolated fields, and investigated changes in the insect fauna using three types of collection methods for three years. In total, 331,483 individual insects and arachnids, representing 82 families in 14 orders, were captured during the study. Totals of 166,518 and 164,965 individual insects and arachnids were collected from LM soybean and Gwangan, respectively. Throughout the study, although there were differences between the investigation year, region, and methods, there were no significant differences between the population densities of insect pests, natural enemies, and other insects on LM soybean and non-LM soybean. Also, there were no statistically significant differences between varieties in the results of the species diversity analysis. The data on insect species population densities were subjected to multidimensional scaling (MDS), which did not distinguish between the two varieties, LM soybean and the non-LM soybean, in all cultivated fields. However, the results of the MDS analysis were completely divided into six groups based on the yearly survey areas. These results provided the insect diversity for an environmental risk assessment of LM soybean and suggested that the guideline could be useful to detect LM crops.

System for Improvement of Soybean Using Gene Editing Technology

  • Ji Hyun Bae;Gyu Tae Park;Soo-kwon Park;Yu-na Kim;Dool-Yi Kim;Hyeon Jung Kang;Jung Kyung Moon;Mi-Suk Seo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.234-234
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    • 2022
  • Gene-editing is currently one of the most popular technologies in recent years. Development of the new crop using the gene editing have advantage of improved accuracy and efficiency compared with conventional breeding. Soybean (Glycine max L.) is one of the most important crops worldwide used as food and forage. We tried to establish a system for breeding improvement of soybean through gene-editing technology. For the gene-editing system of soybean, i) selection of efficiency gRNA of targeted gene, ii) efficient genetic transformation of the selected gRNA, iii) selection of trans-clean mutant is essential. First of all, we investigated the selection conditions of gRNA with high editing efficiency of targeted gene using isolated protoplast of soybean. Furthermore, we performed the Agrobacterium-mediated genetic transformation of various soybean cultivars. We identified the tissue culture ability in 23 soybean cultivars for genetic transformation of soybean. The six cultivars with high tissue culture ability were selected and confirmed the transgenic plants in four cultivars. Finally, we established a speed-breeding system as a powerful tool for the fast selection of trans-clean mutants from transgenic plants. Our laboratory will provide the valuable system for improvement of soybean by the gene-editing technology.

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Soybean mosaic virus Infection and Helper Component-protease Enhance Accumulation of Bean pod mottle virus-Specific siRNAs

  • Lim, Hyoun-Sub;Jang, Chan-Yong;Bae, Han-Hong;Kim, Joon-Ki;Lee, Cheol-Ho;Hong, Jin-Sung;Ju, Ho-Jong;Kim, Hong-Gi;Domier, Leslie L.
    • The Plant Pathology Journal
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    • v.27 no.4
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    • pp.315-323
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    • 2011
  • Soybean plants infected with Bean pod mottle virus (BPMV) develop acute symptoms that usually decrease in severity over time. In other plant-virus interactions, this type of symptom recovery has been associated with degradation of viral RNAs by RNA silencing, which is accompanied by the accumulation of virus-derived small interfering RNAs (siRNAs). In this study, changes in the accumulation of BPMV siRNAs were investigated in soybean plants infected with BPMV alone, or infected with both BPMV and Soybean mosaic virus (SMV) and in transgenic soybean plants expressing SMV helper component-protease (HC-Pro). In many potyviruses, HC-Pro is a potent suppressor of RNA silencing. In plants infected with BPMV alone, accumulation of siRNAs was positively correlated with symptom severity and accumulation of BPMV genomic RNAs. Plants infected with both BPMV and SMV and BPMV-infected transgenic soybean plants expressing SMV HC-Pro exhibited severe symptoms characteristic of BPMVSMV synergism, and showed enhanced accumulation of BPMV RNAs and siRNAs compared to plants infected with BPMV alone and nontransgenic plants. Likewise, SMV HC-Pro enhanced the accumulation of siRNAs produced from a silenced green fluorescent protein gene in transient expression assays, while the P19 silencing suppressor of Tomato bushy stunt virus did not. Consistent with the modes of action of HC-Pro in other systems, which have shown that HC-Pro suppresses RNA silencing by preventing the unwinding of duplex siRNAs and inhibiting siRNA methylation, these studies showed that SMV HC-Pro interfered with the activities of RNA-induced silencing complexes, but not the activities of Dicer-like enzymes in antiviral defenses.

Transformation of Lettuce (Lactuce sativa L.) Using Iron Storage Protein Ferritin Gene (철 저장단백질 관련 Ferritin 유전자를 이용한 상추의 형질전환)

  • 김성하;노일섭;최장선;강권규
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.147-151
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    • 2001
  • Explants of Lactuce sativa cultivar, chungchima, were co-cultivated with Agrobacterium tumefaciences LBA4404, EHA101 strains containing nptll gene and ferritin gene encoding iron storage protein from soybean for transformation. Through initial selection of regenerated explants by culturing on a kanamycin and carbenicillin containing MS medium, multiple shoots were obtained after 2 months of culture. For a complementary step of selection, putative transgenic shoots were transferred to 1/2 MS basal medium supplemented with 100 mg/L kanamycin and 500 mg/L carbenicillin. The selected shoots were tested with PCR analysis using nptll, ferritin specific primers whether ferritin gene was introduced to genome of the plants. These results confirmed that produced the specific PCR bands in the putative transgenic lines. Additionally the Northern blot showed that transcripts of ferritin gene were detected in mature leaf of the transgenic lines. These results suggest that ferritin gene be successfully integrated and transcribed in the putative transgenic lettuce plants.

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Molecular Breeding for Plant Disease Resistance : Prospects and Problems

  • Park, Hyo-Guen
    • The Plant Pathology Journal
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    • v.17 no.1
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    • pp.1-8
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    • 2001
  • The technique of plant transformation has started to show off its great power in the area of plant breeding by commercially successful introduction of transgenic varieties such as herbicide tolerant soybean and insect resistant corn in USA with an unimaginable speed. However, in contrast with the great success in the commercialization of herbicide tolerance and insect resistance, the transformation works on disease resistance has not yet reached the stage of full commercialization. This review surveys the current status of molecular breeding for plant disease resistance and their limits and problems. Some novel ideas and approaches in molecular breeding for disease resistance are introduced.

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