• Title/Summary/Keyword: ${\alpha}^{\prime}$-subunit

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Inheritance of 7S α' - subunit Protein in Soybean Seed (콩의 7S α' - subunit 단백질의 유전)

  • Sung, Mi-Kyung;Kim, Kyung-Roc;Park, Jung-Soo;Hwang, Kyo-Jin;Chung, Jong-Il
    • Journal of agriculture & life science
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    • v.43 no.5
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    • pp.39-42
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    • 2009
  • Soybean is an important sources of plant proteins for human and animal nutrition. The use of soybean proteins has been expanded in the food industry due to their excellent nutritional benefits. But, Soybeans contain allergenic proteins that cause allergies to sensitive individuals. ${\beta}$-conglycinin(7S globulin) and glycinin(11S globulin) are the major components of storage protein in soybean. ${\beta}$-conglycinin consists of three subunits, ${\alpha}^{\prime}$, ${\alpha}$, ${\beta}$ and exhibits poorer nutritional and food processing properties than glycinin. There is a great deal of interest in the development of soybean lines with reduced amounts of ${\beta}$-conglycinin. The objective of this study was to determine the inheritance of ${\alpha}^{\prime}$-subunit protein in 7S globulin. F2 population was developed from the cross of "Jinpumkong2ho"(${\alpha}^{\prime}$-subunit presence) and PI506876(${\alpha}^{\prime}$-subunit absence) parent. Total 98 of F2 seeds were obtained and analyzed for the segregation of ${\alpha}^{\prime}$-subunit protein by SDS-PAGE. Among 98 F2 seeds, 70 F2 seeds showed ${\alpha}^{\prime}$-subunit protein and 28 F2 seeds did not show ${\alpha}^{\prime}$-subunit protein. The segregation ratios of 3 : 1 for presence and absence of ${\alpha}^{\prime}$-subunit protein were observed(${\chi}^2=0.667$, P=0.414). These data indicate that presence and absence of ${\alpha}^{\prime}$-subunit protein is controlled by a single major gene and might be useful for strain selection of 7S protein reduced soybean.

The development of new soybean strain with ti and cgy1 recessive allele

  • Choi, Sang Woo;Park, Jun Hyun;Chung, Jong Il
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.328-332
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    • 2018
  • Soybean [Glycine max (L.) Merr.] seed is an important dietary source of protein, oil, carbohydrate, isoflavone and other various nutrients for humans and animals. However, there are anti-nutritional factors in the raw mature soybeans. Kunitz trypsin inhibitor (KTI) protein and stachyose are the main anti-nutritional factors in soybean seed. The ${\alpha}^{\prime}$-subunit of ${\beta}$-conglycinin protein exhibit poor nutritional and food processing properties. The genetic removal of the KTI and ${\alpha}^{\prime}$-subunit proteins will improve the nutritional value of the soybean seed. The objective of this research was to develop a new soybean strain with KTI and ${\alpha}^{\prime}$-subunit protein free ($titicgy_1cgy_1$ genotype) and proper agronomic traits. A breeding population was developed from the cross of the Bl-1 and 15G1 parents. A total of 168 $F_2$ seeds from the cross of the BL-1 and 15G1 parents were obtained. The segregation ratios of 9: 3: 3: 1 ($104Ti\_Cgy_{1\_}:\;30Ti\_cgy_1cgy_1:\;21cgy_1cgy_1Ti\_:\;13titicgy_1cgy_1$) between the Ti and $Cgy_1$ genes in the $F_2$ seeds were observed (${\chi}^2=5.12$, P=0.5-0.10). Two $F_4$ plant strains with proper agronomical traits and $titicgy_1cgy_1$ genotype (free of both KTI and ${\alpha}^{\prime}$-subunit protein) were selected and harvested. 2 strains (S1 and S2) had yellow seed coats and hilum. The plant height of the S1 strain was 65 centimeters. The 100-seed weight was 29.2 g. The plant height of the S2 strain was 66 centimeters and 100-seed weight was 26.2 g. The two strains selected in this research will be used to improve the new cultivar that will be free of the KTI and ${\alpha}^{\prime}$-subunit proteins.

Selection of Lipoxygenase, Kunitz Trypsin inhibitor and 7Sα′-subunit Protein Free Soybean Strain (Lipoxygenase, Kunitz Trypsin inhibitor, 7Sα′-subunit 단백질이 결핍된 콩 계통의 선발)

  • Sung, Mi-Kyung;Kim, Kyung-Roc;Park, Jung-Soo;Han, Eun-Hui;Nam, Jin-Woo;Chung, Jong-Il
    • Journal of agriculture & life science
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    • v.44 no.5
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    • pp.29-33
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    • 2010
  • Soybean (Glycine max (L.) Merr.) seed is the main source of protein and oil for human and animal. The use of soybean protein has been expanded in the food industry due to their excellent nutritional benefits. But, antinutritional and allergenic factors are exist in the raw mature soybean. Lipoxygenase, Kunitz trypsin inhibitor (KTI) protein, and ${\alpha}^{\prime}-subunit$ of 7S globulin are main antinutritional factors in soybean seed. Breeding of a new soybean strain with lacking these components is needed. The objective of this research was to select new soybean line with lipoxygenase-free, KTI-free, and ${\alpha}^{\prime}-subunit$ free (lx1lx1lx2lx2lx3lx3titicgy1cgy1 genotype). Total 434 $F_2$seeds were obtained from the cross of cultivar, "Gaechuck#2" and PI506876. Presence and absence of lipoxygenase, KTI protein, and ${\alpha}^{\prime}-subunit$ of 7S globulin was tested by SDS electrophoresis using a partial seed of each $F_2$seed. Only one $F_2$seed with lacking these three components was selected and was planted to $F_2$plant. Absence of lipoxygenase, KTI, and ${\alpha}^{\prime}-subunit$ protein was confirmed on the $F_3$seeds harvested. Selected line with lx1lx1lx2lx2lx3lx3titicgy1cgy1 genotype might be useful for soybean breeding.

Inheritance of Cgy1 gene and Ti gene in Mature Soybean Seed

  • Sung, Mi-Kyung;Han, Eun-Hui;Kim, Kyung-Roc;Park, Jung-Soo;Hwang, Kyo-Jin;Nam, Jin-Woo;Chung, Jong-Il
    • Korean Journal of Breeding Science
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    • v.42 no.1
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    • pp.35-39
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    • 2010
  • Soybean proteins are widely used for human and animal feed in the world. ${\beta}$-conglycinin protein exhibits poor nutritional and food processing properties and Kunitz trypsin inhibitor (KTI) protein is a main anti-nutritional factor in soybean seed. The objective of this research was to identify the inheritance of $cgy_1$ gene and ti gene for the improvement of soybean cultivar with no KTI proteins and low amount of ${\beta}$-conglycinin. $F_2$ population was made by crossing between "Gaechuck2ho" (${\alpha}^{\prime}$-subunit present $Cgy_1Cgy_1$, KTI protein absent titi) and PI506876 (${\alpha}^{\prime}$-subunit absent $cgy_1cgy_1$, KTI protein present TiTi) parent. A total of 434 $F_2$ seeds were obtained and analyzed for the segregation of ${\alpha}^{\prime}$-subunit protein and KTI protein using SDS-PAGE. The segregation ratio of 3 : 1 for $Cgy_1$ locus (310 $Cgy_1$_ : 124 $cgy_1cgy_1$) and Ti locus (339 Ti_ : 95 titi) were observed. Segregation ratios of 9 : 3 : 3 : 1 (241 $Cgy_1$_Ti_: 69 $Cgy_1$_titi: 98 $cgy_1cgy_1$Ti_: 26 $cgy_1cgy_1titi$) between $Cgy_1$ gene and Ti gene in $F_2$ seeds were also observed (${\chi}^2= 5.367$, P = 0.10 - 0.20). This data showed that $Cgy_1$ gene was inherited independently with the Ti gene in soybean. These results will be useful in breeding program for selecting the line that does not exhibit or lacks both ${\alpha}^{\prime}$-subunit protein and KTI protein in soybean.

Physiological Functions of the COPI Complex in Higher Plants

  • Ahn, Hee-Kyung;Kang, Yong Won;Lim, Hye Min;Hwang, Inhwan;Pai, Hyun-Sook
    • Molecules and Cells
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    • v.38 no.10
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    • pp.866-875
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    • 2015
  • COPI vesicles are essential to the retrograde transport of proteins in the early secretory pathway. The COPI coatomer complex consists of seven subunits, termed ${\alpha}-$, ${\beta}-$, ${\beta}^{\prime}-$, ${\gamma}-$, ${\delta}-$, ${\varepsilon}-$, and ${\zeta}$-COP, in yeast and mammals. Plant genomes have homologs of these subunits, but the essentiality of their cellular functions has hampered the functional characterization of the subunit genes in plants. Here we have employed virus-induced gene silencing (VIGS) and dexamethasone (DEX)-inducible RNAi of the COPI subunit genes to study the in vivo functions of the COPI coatomer complex in plants. The ${\beta}^{\prime}-$, ${\gamma}-$, and ${\delta}$-COP subunits localized to the Golgi as GFP-fusion proteins and interacted with each other in the Golgi. Silencing of ${\beta}^{\prime}-$, ${\gamma}-$, and ${\delta}$-COP by VIGS resulted in growth arrest and acute plant death in Nicotiana benthamiana, with the affected leaf cells exhibiting morphological markers of programmed cell death. Depletion of the COPI subunits resulted in disruption of the Golgi structure and accumulation of autolysosome-like structures in earlier stages of gene silencing. In tobacco BY-2 cells, DEX-inducible RNAi of ${\beta}^{\prime}$-COP caused aberrant cell plate formation during cytokinesis. Collectively, these results suggest that COPI vesicles are essential to plant growth and survival by maintaining the Golgi apparatus and modulating cell plate formation.

In Vitro Characterization of Protein Kinase CKII β Mutants Defective in β-β Dimerization

  • Kim, Tae-Hyun;Lee, Jae-Yong;Kang, Beom Sik;Bae, Young-Seuk
    • Molecules and Cells
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    • v.19 no.1
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    • pp.124-130
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    • 2005
  • Protein kinase CKII is composed of two catalytic (${\alpha}$ or ${\alpha}^{\prime}$) subunits and two regulatory (${\beta}$) subunits. The ${\beta}$ subunit mediates tetramer formation through ${\beta}-{\beta}$ homodimerization and ${\alpha}-{\beta}$ heterodimerization. In a previous study R26 and R75, point mutants of $CKII{\beta}$ defective in ${\beta}-{\beta}$ dimerization, were isolated. In the present work we characterized these $CKII{\beta}$ mutants in vitro. Purified R26 and R75 bound to $CKII{\alpha}$ but were defective in binding to $CKII{\beta}$. R75 stimulated the catalytic activity of CKII whereas R26 gave little stimulation, and poly-L-lysine increased the stimulation of catalytic activity by R26 or R75. Circular dichroism and intrinsic fluorescence data pointed to different conformational changes in R26 and R75. Molecular modeling of these mutants provides an explanation of the difference in their ability to interact with $CKII{\beta}$ and to activate $CKII{\alpha}$.

Upregulation of miR-760 and miR-186 Is Associated with Replicative Senescence in Human Lung Fibroblast Cells

  • Lee, Young-Hoon;Kim, Soo Young;Bae, Young-Seuk
    • Molecules and Cells
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    • v.37 no.8
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    • pp.620-627
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    • 2014
  • We have previously shown that microRNAs (miRNAs) miR-760, miR-186, miR-337-3p, and miR-216b stimulate premature senescence through protein kinase CK2 (CK2) downregulation in human colon cancer cells. Here, we examined whether these four miRNAs are involved in the replicative senescence of human lung fibroblast IMR-90 cells. miR-760 and miR-186 were significantly upregulated in replicatively senescent IMR-90 cells, and their joint action with both miR-337-3p and miR-216b was necessary for efficient downregulation of the ${\alpha}$ subunit of CK2 ($CK2{\alpha}$) in IMR-90 cells. A mutation in any of the four miRNA-binding sequences within the $CK2{\alpha}3^{\prime}$-untranslated region (UTR) indicated that all four miRNAs should simultaneously bind to the target sites for $CK2{\alpha}$ downregulation. The four miRNAs increased senescence-associated ${\beta}$-galactosidase (SA-${\beta}$-gal) staining, p53 and $p21^{Cip1/WAF1}$ expression, and reactive oxygen species (ROS) production in proliferating IMR-90 cells. $CK2{\alpha}$ overexpression almost abolished this event. Taken together, the present results suggest that the upregulation of miR-760 and miR-186 is associated with replicative senescence in human lung fibroblast cells, and their cooperative action with miR-337-3p and miR-216b may induce replicative senescence through $CK2{\alpha}$ downregulation-dependent ROS generation.

The Development of a New Soybean Strain Without Kunitz Trypsin Inhibitor, Lectin, and 7S α' Subunit Protein (쿠니츠트립인히비터, 렉틴 및 7S α' 서버유닛 3가지 단백질이 없는 콩 계통의 개발)

  • Chae, Won Gi;Choi, Sang Woo;Kang, Gyung Young;Chung, Jong Il
    • Journal of Life Science
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    • v.30 no.7
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    • pp.592-597
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    • 2020
  • Soybean [Glycine max (L.) Merr.] seeds contain an average of 40% protein on a dry weight basis, but they also contain antinutritional elements such as lectin, Kunitz trypsin inhibitor (KTI), and 7S α'- subunit protein. The objective of this research was to develop a new soybean genotype with triple recessive alleles for these elements. Three parents (Gaechuck#2, PI506876, and Le-16) were used to develop the genetic population, and the presence of lectin and KTI protein was detected using Western blot while 7S α' subunit protein was detected using SDS-PAGE. One F3 plant strain with proper agronomical traits such as type, height, seed quality, and 100-seed weight was selected. The genotype of the developed strain is titilelecgy1cgy1, that is KTI, lectin, and 7S α' subunit protein free. The new strain has a purple flower, determinate growth habit, and light yellow pods at maturity. The seed has a buffer hilum and is yellow in color. The new strain's height was 58 cm compared to the Daewonkong cultivar at 46 cm, and its 100-seed weight was 27.1 g, smaller than the Daewonkong at 29.0 g. This is the first new soybean strain with the titilelecgy1cgy1 genotype, and it can be used to improve yellow soybean cultivars of high quality and function.

Selection of a Soybean Line with Brown Seed Coat, Green Cotyledon, and Tetra-Null Genotype (갈색종피와 녹색자엽 및 Tetra Null 유전자형을 가진 콩 계통 선발)

  • Sarath Ly;Hyeon Su Oh;Se Yeong Kim;Jeong Hwan Lee;Jong Il Chung
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.68 no.3
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    • pp.114-120
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    • 2023
  • Soybean is the one of the most important crops for providing quality vegetable protein to umans and livestock. Soybean cultivars with a brown seed coat have a wide range of antioxidant benefits because of the flavonoid components. However, they also contain lectin, 7S α′ subunit, lipoxygenase, and Kunitz trypsin inhibitor (KTI) proteins that can be allergenic and digestive inhibitors and reduce processing aptitude. Genetic removal of these four proteins is necessary in soybean breeding. Therefore, this study was conducted to select a new line with brown seed coat, green cotyledon, and tetra-null genotype (lecgy1lox1lox2lox3ti) for lectin, 7S α′ subunit, lipoxygenase, and KTI proteins in the mature seed. Five germplasms were used to create breeding population. From a total of 58 F2 plants, F2 plants with lele genotype were selected using a DNA marker, and F3 seeds with a brown seed coat, green cotyledon, and the absence of 7S α′ subunit protein were selected. Three lines (S1, S2, and S3) were developed. Genetic absence of lectin, 7S α′ subunit, lipoxygenase, and KTI proteins was confirmed in F6 seeds of the three lines, which had a brown seed coat, green cotyledons, and a white hilum. The 100 seed weights of the three lines were 26.4-30.9 g, which were lower than 36 g of the check cultivar - 'Chungja#3'. The new S2 line with 30.9 g hundred seed weight can be used as a parent to improve colored soybean cultivars without antinutritional factors such as lectin, 7S α′ subunit, lipoxygenase, and KTI proteins.