• Title/Summary/Keyword: Kunitz trypsin inhibitor (KTI)

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Identification of Molecular Markers Linked to Ti Locus in Soybean

  • Kim Myung Sik;Park Min Jung;Hwang Jung Gyu;Jo Soo Ho;Ko Mi Suk;Chung Jong Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.5
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    • pp.419-422
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    • 2004
  • Soybean is a major source of protein meal in the world. Kunitz trypsin inhibitor (KTI) protein is responsible for the inferior nutritional quality of unheated or incompletely heated soybean meal. The objective of this research was to identify RAPD markers linked to KTI protein allele using bulked segregant analysis. Cultivar Jinpumkong2 (TiTi) was crossed with C242 (titi, absence of KTI protein) and F. seeds were planted. The $F_1$. plants were grown in the greenhouse to produce $F_2$ seeds. Each $F_2$ seed from $F_1$. plants was analysed electrophoretically to determine the presence of the KTI protein band. The present and absent bulks contained twenty individuals each, which were selected on the basis of the KTI protein electrophoresis, respectively. Total 94 $F_2$ individuals were constructed and 1,000 Operon random primers were used to identify RAPD primers linked to the Ti locus. The presence of KTI protein is dominant to the lack of a KTI protein and Kunitz trypsin inhibit protein band is controlled by a single locus. Four RAPD primers (OPAC12, OPAR15, OPO12, and OPC08) were linked to the Ti locus. RAPD primer OPO12 was linked to Ti locus, controlling kunitz trypsin inhibitor protein at a distance of 16.0 cM. This results may assist in study of developing fine map including Ti locus in soybean.

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.

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.

A Stack of Recessive Alleles of Kunitz Trypsin Inhibitor, Lectin, and Stachyose in Soybean (콩에서 쿠니츠트립인히비터, 렉틴 및 스타키오스에 대한 열성 유전자의 집적)

  • Choi, Sang Woo;Chae, Won Gi;Kang, Gyung Young;Chung, Jong Il
    • Journal of Life Science
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    • v.29 no.7
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    • pp.774-778
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    • 2019
  • Soybean [Glycine max (L.) Merr.] is one of the major food sources of protein, oil, carbohydrates, isoflavones, and other nutrients for both humans and animals. However, soybean seeds contain antinutritional factors, such as lectin protein, Kunitz Trypsin Inhibitor (KTI) protein, and stachyose. The objective of this research was to stack recessive alleles for development a triple recessive genotype, titilelers2rs2, with low KTI protein, lectin protein, and stachyose contents. Three parents (Gaechuck#2, PI200508, and 14G20) were used to develop the breeding population. The presence or absence of the lectin and KTI proteins was detected by western blotting. The stachyose content in mature seeds was determined by HPLC. Agronomic traits, such as plant type, plant height, maturity date, lodging, seed quality, and 100-seed weight, were evaluated for the four $F_3$ plant strains. One $F_4$ plant strain with the desired agronomical traits was selected. One new strain with the triple recessive titilelers2rs2 genotype was developed. The plant height of the new strain was 51 cm and the 100-seed weight was 31.0 g. The new strain had a yellow seed coat and yellow hilum. The stachyose content of the new strain was 3.8 g/kg. One strain developed in this research will be used to produce improved yellow soybean cultivars that are free of lectin and KTI proteins and low in stachyose content.

Selection of rs2rs2titi Soybean Genotype with Yellow Seed Coat (rs2rs2titi 유전자형을 가진 노란 콩 계통 선발)

  • Choi, Sang Woo;Park, Jun Hyun;Chung, Jong Il
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1285-1289
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    • 2018
  • Soybean [Glycine max (L.) Merr.] seed is an important dietary source of protein, oil, carbohydrates, isoflavones, and other nutrients for humans and animals. But, antinutritional factors in the raw mature soybean are exist. Kunitz trypsin inhibitor (KTI) protein and stachyose are main antinutritional factors in soybean seed. The genetic removal of the antinutritional factors will improve the nutritional value of soybean seed. The objective of this research was to breed a new yellow soybean strains (rs2rs2titi genotype) with the traits of lacking of KTI protein and low content of stachyose. Breeding population was developed from the cross of "Jinyangkong" and 15G1 parents. Presence or absence of KTI protein was detected based on Western Blot technique. Content of stachyose in mature seed was detected by HPLC. Total four new strains (603-1, 603-2, 625, and 694) with KTI protein free and low content of stachyose were selected. Four strains (603-1, 603-2, 625, and 694) have yellow seed coat and hilum. Plant height of 603-1 strain was 65 cm and 100-seed weight was 29.2 g. Plant height of 603-2 strain was 66 cm and 100-seed weight was 26.2 g. Plant height of 625 strain was 64 cm and 100-seed weight was 27.1 g. Content of stachyose for four new strains was 3.0~3.50 g/kg. Four strains selected in this research will be used to improve new yellow soybean cultivar with KTI protein free, and low content of stachyose.

Physicochemical Characteristics and Antioxidant Activity of Kanjang made from Soybean Cultivars Lacking Lipoxygenase and Kunitz Trypsin Inhibitor Protein (Lipoxygenase와 Kunitz Trypsin Inhibitor 단백질 결핍콩으로 제조한 간장의 이화학적 특성 및 항산화 활성)

  • Hwang, Cho-Rong;Lee, Soo-Jung;Kang, Jae-Ran;Kwon, Min-Hye;Kwon, Hyo-Jin;Chung, Jong-Il;Sung, Nak-Ju
    • Journal of agriculture & life science
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    • v.46 no.5
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    • pp.111-125
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    • 2012
  • In order to evaluate suitability by processing for non-GM soybean cultivars such as Gaechuck#2, Jinyangkong and CJ#1 lacking lipoxygenase (LOX) and kunitz trypsin inhibitor (KTI) protein, physicochemical characteristics and antioxidant activity of Kanjang made from above soybean were compared to Kanjang made from a conventional cultivar (Taekwangkong). Proximate compositions of soybeans were similar for the 4 kinds cultivars. Total phenol and flavonoid contents were significantly higher in cultivars lacking LOX and KTI protein than the Taekwangkong. In Kanjang, contents of total and reducing sugar were higher in Taekwangkong Kanjang than Kanjang from made cultivars lacking LOX and KTI protein. Contents of total and amino type nitrogen were the highest in the Jinyangkong Kanjang. Mineral contents were higher in the Jinyangkong and CJ#1 Kanjangs, amino acid contents were higher in the Kanjang made from 3 cultivars lacking LOX and KTI protein than the Taekwangkong. Taste of the Jinyangkong Kanjang with higher sweety and savory was also found to be superior to that of others in overall acceptability evaluation. Total phenol and flavonoid contents in Kanjang were significantly higher in the Kanjang made from cultivars lacking LOX and KTI protein than the Taekwangkong. Radical scavenging activity of Kanjang was increased in the total phenol contents dependent on. Reducing power by ferric-reducing antioxidant potential (FRAP) was significantly higher the Kanjang made from Gaechuck#2 and CJ#1 than the Taekwangkong. $Fe^{2+}$ chelating activity was higher in Taekwangkong Kanjang than the other cultivars, but its activity was similar to Jinyangkong Kanjang. Therefore, higher nutritional composition, total phenol and flavonoids contents and antioxidant activity in the Kanjang made from soybean cultivars lacking LOX and KTI protein might be provide better benefit for manufacture of another their products.

Independent Inheritance between df2 gene and ti gene in Soybean

  • Han, Eun-Hui;Sung, Mi-Kyung;Kim, Kyung-Roc;Park, Jung-Soo;Nam, Jin-Woo;Chung, Jong-Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.1
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    • pp.14-17
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    • 2011
  • Dwarfuess and Kunitz trypsin inhibitor (KTI) protein in soybean is useful traits for basic studies. df2 and ti gene control dwarfness and the expression of Kunitz trypsin inhibitor (KTI) protein in soybean, respectively. The objective of this research was to verify genetic linkage or independent inheritance of df2 and ti loci in soybean. The $F_2$ population was made by cross combination between "Gaechuck#2" (Df2Df2titi genotype, KTI protein absence and a normal growth type) and T210 (df2df2TiTi genotype, a dwarf growth type and KTI protein present). A total of 258 $F_2$ seeds were analyzed for the segregation of KTI protein using SDS-PAGE. And so, 198 $F_2$ plants were recorded for the segregation of dwarfness. The segregation ratio of 3 : 1 for Ti locus (201 Ti_ : 57 titi) and Df2 locus (143 Df2_ : 55 df2df2) was observed. Segregation ratio of 9 : 3 : 3 : 1 (116 Ti_Df2_: 44 Ti_df2df2: 27 titiDf2_: 11 titidf2df2) between df2 gene and ti gene was observed ($x^2$=3.53, P = 0.223). These results showed that df2 gene was inherited independently with the ti gene in soybean.

Inheritance of Kunitz Trypsin Inhibitor and P34 Protein in Soybean Seed (콩 종자에서 쿠니츠트립신인히비터와 P34 단백질의 유전)

  • Han, Eun-Hui;Sung, Mi-Kyung;Baek, Woon-Jang;Shim, Sang-In;Kim, Min-Chul;Chung, Jong-Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.1
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    • pp.78-82
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    • 2012
  • Soybean [$Glycine$ $max$ (L.) Merr.] protein is a high quality source for food and feed. But, antinutritional factors in the raw mature soybean are exist. Kunitz trypsin inhibitor (KTI) protein is a main antinutritional factor in soybean seed. Also, P34 protein, referred as $Gly$ $m$ Bd 30K, has been identified as a predominant immunodominant allergen. Genetic relationship between KTI protein and P34 protein could be useful in soybean breeding program for the genetic elimination or reduction of these factors. The objective of this study was to determine the independent inheritance or linkage between KTI protein and P34 protein in soybean seed. A total of 479 $F_2$ seeds were obtained from the cross of 07B1 and PI567476 parents. KTI protein and relative amount of P34 protein were analysed from $F_2$ seeds harvested from the F1 plants by using SDS-PAGE and Western blot analysis. The segregation ratios of 3 : 1 for KTI protein (353 KTI protein present : 126 KTI protein absent) and relative amount of P34 protein (363 normal amount of P34 protein : 116 low amount of P34 protein). The segregation ratio of 3 : 1 suggested that KTI protein and relative amount of P34 protein in mature soybean seed were controlled by a single major gene. The segregation ratios of 9 : 3 : 3 : 1 (266 KTI protein present, normal amount of P34 protein: 88 KTI protein present, low amount of P34 protein: 102 KTI protein absent, normal amount of P34 protein: 23 KTI protein absent, low amount of P34 protein) and Chi-square value (${\chi}^2$=3.31, P=0.346) were observed in $F_2$ seeds. This data showed that KTI protein was inherited independently with relative amount of P34 protein in soybean. These results will be helpful in breeding program for selecting the line with lacking KTI protein and reduced amount of P34 protein in soybean.

Breeding of a Recessive Soybean Genotype (titirs2rs2) with Green Cotyledons and Black Seed Coats (titirs2rs2 열성 유전자형을 가진 속푸른 검정콩 계통 육성)

  • Choi, Sang Woo;Kim, Jin A;Shim, Sang In;Kim, Min Chul;Chung, Jong Il
    • Journal of Life Science
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    • v.29 no.2
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    • pp.147-151
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    • 2019
  • Soybean [Glycine max (L.) Merr.] is grown worldwide for its high protein and oil content. Anthocyanins from black soybean seed coats are known to have many pharmaceutical effects. Soybean cultivars with large seed sizes and black seed coats are needed by soybean farmers. However, antinutritional factors, like protein, stachyose, and Kunitz trypsin inhibitor (KTI) exist in raw mature soybeans. Genetic elimination or reduction of these components is needed in soybean breeding. The objective of this research was to develop new a soybean strain with black seed coats and green cotyledons that was KTI protein free and low in stachyose. Six parents were used. The presence or absence of KTI protein was detected using the Western blot technique. The content of stachyose in mature seeds was detected using HPLC. One new strain was selected from 11 $F_2$ plants with black seed coats and green cotyledons that lacked KTI protein. The new strain had black seed coats and green cotyledons and was KTI protein free and low in stachyose. The plant height of the new strain was 66 cm, and its 100-seed weight was 28.4 g. The stachyose content of the new strain was 2.59 g/kg. The new strain developed in this research will be used to develop new cultivars that are KTI protein free and low in stachyose.

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.