• Title/Summary/Keyword: Soybean mutant

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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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Evaluation of Nitrogen Fixation between Supernodulating Soybean Mutants and their Wild-Types Using $^{15}N$ in Field Conditions

  • Youn, Jong-Tag;Van, Kyu-Jung;Ryu, Yong-Hwan;Shimada, Shinji;Lee, Choon-Ki;Kim, Sun-Lim;Seo, Sea-Jung;Lee, Suk-Ha
    • Journal of Crop Science and Biotechnology
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    • v.11 no.1
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    • pp.31-38
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    • 2008
  • Improvement of $N_2$ fixation by symbiotic rhizobia is considered an effective means for enhancing its productivity without high input of nitrogen(N) fertilizer. Several methods to improve $N_2$ fixation have been proposed including the use of supernodulating mutants. The objective of this research was to identify the varietal difference in N and $N_2$-fixation ability among the soybean supernodulating mutants, SS2-2 and Sakukei 4, with different nodulation abilities using $^{15}N$ in field conditions in both Korea and Japan. The dry weight(DW) was higher in wild-type soybeans. The distribution rate of DW in each plant part was high in seeds of supernoduating and wild-type soybeans but high in stems and leaves of non-nodulating mutants. Although the supernodulating mutants had a low DW rate at maturity, they showed a similar $N_2$ fixation ability compared with wild-type. Supernodulating mutant plants mainly obtained N from $N_2$ fixation, while soil N was the main resource for obtaining N in non-nodulating mutants. The percentage of N derived from atmospheric dinitrogen(Ndfa) was higher in supernodulating mutants than in wild-type and relatively high in seeds between plant parts at maturity. In particular, supernodulating mutants showed higher N content in roots than those of wild-type and non-nodulating mutants. It was considered that supernodulating mutants have the advantage of saving nitrate in soil and being beneficial for N absorption of subsequent crops due to their conserving more N in the field and releasing considerable amounts of N from roots and leaves fallen to the soil.

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Development of Seed Culture Using Soybean for Mass Production of Lovastatin with Aspergillus terreus ATCC 20542 Mutant (대두를 이용한 Lovastatin 대량생산용 Seed Culture의 제조기술)

  • Kim, Soo-Jung;Ko, Hee-Sun;Kim, Hyun-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.5
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    • pp.666-670
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    • 2008
  • Lovastatin (Mevinolin, Monacolin K) is a well-known drug for the therapy of hypercholesterolemia. It is an important fungal secondary metabolite as it inhibits 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase, EC 1.1.1.34) which catalyzes a major rate-limiting step in the biosynthesis of cholesterol. Both soybeans and black soybeans with Aspergillus terreus ATCC 20542 mutant were used as the seed culture for the mass production of lovastatin. The production of lovastatin in soybean seed culture of Asp. terreus was twofold compared to that of black soybean seed culture. The effect of two different vessels (petri dish and Erlenmeyer flask) on lovastatin production was also studied. The production of lovastatin on petri dish was tenfold to that of Erlenmeyer flask. Furthermore, the most lovastatin production on rice bran was achieved when the soybean seed culture was treated by heat shock at $30^{\circ}C$ for 1 hour, representing 82% of HMG-CoA reductase inhibition in the koji extract. We estimated that the heat treated soybean seed culture could be a new method for the mass production of lovastatin.

Mutagenesis of Nodulation and Other Growth Characters with Ethyl Methanesulfonate in Soybean (EMS 처리에 관한 콩 근류착생 및 기타 형질의 돌연변이 출현 양상)

  • 이홍석;구자환;이석하;김석동
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.38 no.5
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    • pp.442-448
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    • 1993
  • The aims of the work were to examme the variability induced by EMS (ethyl methanesulfonate) mutagenesis of soybean plants, and to isolate mutants altered in nodulation and other growth characters. Seeds of two soybean cultivars, ‘Hwanggeumkong’ and ‘Baegunkong’ were treated with 30 and 50mM EMS(pH 7.0) for 6 hours and were planted directly in the field. Field emergency of$M_1 seed was averaged to be 61.0%, and frequency of plants with chlorophyll-deficient sectors of the first trifoliolate is about 0.7%. Regardless of varieties and does of EMS, $M_1 plant injury at harvest was present in plant height, pod and seed number per plant when compared to those of original-type soybean plants. The $M_2 variability of nodulation process induced by EMS treatment was found to be narrower than that of shoot dry weight. On the basis of the occurrence of chlorophyll-deficient plants, mutated cell frequency within $M_1 seed ranged from 5.3% to 84.2%, suggesting that mutation frequency on the $M_1 seed induced by EMS occurred partly and randomly regardless of varieties and doses of EMS. The putative mutant, which had more nodulation than original-type plant, was short in plant height. Sparse-nodulating soybean mutant was lower in leaf chlorophyll content and showed reduced growth.

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Improved Production, and Purification of Aclacinomycin A from Streptomyces lavendofoliae DKRS

  • Kim, Wan-Seop;Youn, Deok-Joong;Cho, Won-Tae;Kim, Myung-Kuk;Kim, Hak-Ryul;Rhee, Sang-Ki;Choi, Eui-Sung
    • Journal of Microbiology and Biotechnology
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    • v.5 no.5
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    • pp.297-301
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    • 1995
  • An anthracycline antibiotic, aclacinomycin A (aclarubicin), was produced from a mutant strain of Streptomyces lavendofoliae. The mutant strain which showed a 4-fold higher productivity of aclacinomycin A compared with the parent strain was also found to produce a significantly higher amount of aclacinomycin A than the reported production strain, Streptomyces galilaeus. The aclacinomycin A was produced up to 125 mg/l using potato starch and soybean meal as carbon and nitrogen sources, respectively, on a 3 liter scale fermentation in a 5 liter jar fermentor. The mutant strain also produced significant amount of aclacinomycins Band Y. Aclacinomycin A was isolated from the culture broth by solvent extractions and further purified by silica gel column chromatography. The yield of aclacinomycin A with over 99$%$ purity was found to be over 60$%$ starting from 3 liters of culture broth.

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Gamma Radiation Induced Mutagenesis of Lysobacter enzymogenes for Enhanced Chitinolytic Activity

  • Lee, Young-Keun;Kim, Kyoung Youl;Senthilkumar, M.
    • Journal of Radiation Industry
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    • v.4 no.1
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    • pp.65-71
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    • 2010
  • Two chitinase producing strains CHI2 and CHI4 were isolated from soybean rhizosphere soil. Both the strains belonged to Lysobacter enzymogenes as indicated by 16S rDNA sequence analysis. Though strain CHI2 and CHI4 produced extracellular chitinase, they differ in their chitinolytic activity. CHI4 produced approximately three times the higher amounts of enzyme than that of CHI2 under specified conditions. CHI2 produced $535.67U\;l^{-1}$ of chitinase after 48 h incubation with a specific activity of $3.91U\;mg^{-1}$ of protein while strain CHI4 produced $1584.13U\;l^{-1}$ of chitinase with a specific activity of $10.88U\;mg^{-1}$ protein. SDS-PAGE analysis indicated that the molecular weight of chitinase enzyme was approximately 45 kDa. A faint band with a molecular weight of 55 kDa reveals the possibility for the presence of another kind of chitin binding protein. Mutant library was developed by exposing the isolates to gamma rays at their $LD_{99}$ value (0.23 kGy). Totally, 11 mutants of CHI2 and CHI4 are reported to have enhanced chitinase activity. Several leaky mutant clones with decreased enzyme activity and a defective mutant (CHI2-M16) with complete loss of chitinase activity were also identified. CHI4-M18, CHI4-M8 and CHI4-M29 showed 78.8, 41.5, and 31.9% increased chitinase activity over wild type CHI4.

Transfer of foreign Genes into the Bradyrhizobium japonicum and their Inoculation Effects on Soybean Plants (Bradyrhizobium japonicum에 외부유전자(外部遺傳子)의 도입(導入)과 대두(大豆)에 대한 접종효과)

  • Kim, Yong-Woong;Kim, Kil-Yong;Rhee, Young-Hwan;Kim, Kwang-Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.25 no.4
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    • pp.387-393
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    • 1992
  • The fate of inoculum strain of Bradyrhizobium japonicum was studied by using genetically marked strain. RJB6 $str^rnal^rneo^r$. A spontaneous mutant of B. japonicum isolated from nodules was made to have antibiotic resistance against streptomycin and nalidixic acid. In order to make genetically marked strain, neomycine resistant gene(Tn5) was introduced into this spontaneous mutant by conjugation with E. coli containing pSUP2021. The southern hybridization was carried out to confirm the plasmid insertion. Hybridization of chromosome DNA using pSUP2021(Tn5) as a probe showed that Tn5 was located on the 4.9kb fragment of chromosome. Soybean seeds were planted into a soil previously cultivated with soybean and inoculated with different cell densities of marked strain. Fourty days after planting, the inoculation effects on nodule number, nodule fresh weight, plant height and nitrogen content in the plot inoculated with heavy cell suspension was a little better than those in the plot with low inoculation. The recovery percentage of the marked strains was about 12% in the plot inoculated with heavy density cell suspension, while 5% in the plot inoculated with low cell suspension.

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Media Optimization and Fed-Batch Fermentation for Riboflavin Overproduction by Ashbya gossypii (Ashbya gossypii로부터 riboflavin 대량생산을 위한 배지 최적화와 유가식 배양)

  • 남수완;장형욱;반재구;민태익;김익환
    • Microbiology and Biotechnology Letters
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    • v.21 no.6
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    • pp.615-621
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    • 1993
  • In order to maximize the riboflavin production by a mutant strain Ashbya gosspyii, the optimization of medium and fed-batch fermentation were performed. As carbon sources, glucose and soybean oil were necessary for the riboflavin overproduction. Optimal concentrations of glucose and soybean oil in the flask cultures were found to be 3.0% and 0.5%, respectively, in a complex medium containing corn steep liquor(CLS) 1%. Among the various organic nitrogen sources tested, CSL was the most effective one both for the cell growth and riboflavin overproduction.

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ANTIMUTAGENIC EFFECT OF KOCHUJANG (KOREAN RED PEPPER SOYBEAN PASTE) AND KOCHUJANG INGREDIENTS IN THE AMES TEST

  • Jung, Keun-Ok;Kim, So-Ja;Yoon, Suk-Kwon;Park, Kun-Young
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.10a
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    • pp.97-97
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    • 2001
  • The antimutagenicities of methanol extracts from traditional and commercial kochujang (Korean Red Pepper Soybean Paste) and their ingredients were evaluated in Salmonella/mammalian microsome assay system. The traditional kochujang showed higher antimutagenic effects than the commercial one against Ν-methyl-Ν'-nitro-Ν-nitrosoguanidine (MNNG) in the Salmonella typhimurium TA100, a base-pair substituted mutant strain.(omitted)

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