Tetra Null 유전자형을 가진 갈색 종피 콩 육종

리사랏,강재은,이성경,최상우,정종일 경상대학교 농업생명과학연구원 2021 농업생명과학연구 Vol.55 No.1

성숙 콩 종실에는 단백질, 지방, 탄수화물의 3대 영양소와 아이소플라본, 안토시아닌, 레시틴, 사포닌, 루테인, 플라보노이드와 같은 기능성 성분 뿐만 아니라 리폭시지나아제, 쿠니츠트립신인히비터, 렉틴, 7S αʹ-subunit 단백질과 같은 항영양 성분들도 포함하고 있어 콩을 가공하고 섭취하는 데 제약과 어려움이 발생하며 품질과 기능성을 저하시킨다. 본 연구는 성숙 콩 종실에서 항영양 성분인 lipoxygenase, Kunitz Trypsin Inhibitor (KTI), lectin 및 7S αʹ-subunit의 4가지 단백질을 암호하는 유전자들이 tetra null 유전자형(lox1lox2lox3tilecgy1)으로 이들 항영양성 단백질이 축적되지 않고, 갈색 종피인 계통을 육성하기 위하여 진행되었다. 4개의 모본(Jinpumkong#2, Gaechuck#2, PI506876, PI548392)을 이용한 육종집 단이 창성되었다. SDS 전기영동과 western blot 기술로 4가지 단백질(lipoxygenase, KTI, lectin, 7S αʹ-subunit)의 존재 여부를 결정하는 유전자가 모두 열성인 tetra null 유전자형과 갈색 종피를 가진 F2 개체가 선발되었다. 선발개체는 세대진전을 통하여 초형, 성숙기, 탈립, 백립중 및 종자상태와 같은 농업형질을 평가하였고, F5 세대에서 4가지 단백질과 갈색 종피에 대한 유전적 고정을 확인하였다. 선발 계통의 경장은 68cm 정도였으며, 백립중은 29.2g으로 대립에 속하였고, 종피색은 갈색이고 제색은 하얀색이며 성숙 자엽색은 노란색이었다. 본 연구를 통하여 선발된 계통은 성숙 콩 종실에서 품질과 기능성을 저하시키는 lipoxygenase, KTI, lectin 및 7S αʹ-subunit 단백질이 모두 검출되지 않는 유색콩 품종 육성을 위한 중간모본으로 이용될 수 있을 것으로 사료되었다. Soybean [Glycine max (L.) Merr.] seed is one of the major food sources for protein, oil, carbohydrates, isoflavones, and many other nutrients to humans and animals. Also, soybean seeds contain antinutritional factors such as lipoxygenase protein, Kunitz Trypsin Inhibitor (KTI) protein, lectin protein, and 7S αʹ-subunit protein. The genetic elimination of these antinutritional proteins will improve the nutritional value of soybean seed. The objective of this research was to develop new soybean line with brown seed coat and tetra null genotype (lox1lox2lox3tilecgy1) for lipoxygenase, KTI, lectin and 7S αʹ-subunit proteins. Four parents were used to create breeding population. Presence and absence of lipoxygenase and 7S αʹ-subunit proteins were detected by SDS electrophoresis and KTI and lectin proteins were detected by Western Blot technique, respectively. A new tetra null soybean (lox1lox2lox3tilecgy1 genotype) with brown seed coat and lacking of lipoxygenase, KTI, lectin and 7S αʹ-subunit proteins was developed. A new strain has purple flower, brown seed coat, yellow cotyledon and white hilum. Stem height of new strain was 69 cm and 100-seed weight was 29.2g. A new soybean line developed through this research will be used to improve new tetra null cultivar (free of lipoxygenase, KTI, lectin and 7S αʹ-subunit proteins) with brown seed coat and high quality and function.

갈색 종피를 가진 cgy1cgy1titi 유전자형 콩 육종

최상우,채원기,강경영,정종일 경상대학교 농업생명과학연구원 2019 농업생명과학연구 Vol.53 No.4

콩[Glycine max (L.) Merr.] 종자에는 수용성 탄수화물 11%, 단백질 40%, 지방 21% 정도의 3대 영양소가 골고루 함유 되어 있고, 콩 종실 단백질은 양과 질적인 면에서 식물성 단백질 중 가장 우수한 것으로 알려져 있다. 그러나, 성숙 콩 종실에는 Kunitz Trypsin Inhibitor (KTI) 단백질, P34 단백질, 7S 단백질 등 품질과 기능성을 떨어뜨리는 성분이 다수 존재하고 있다. 본 연구는 콩 및 콩 제품의 품 질과 기능성을 떨어뜨리는 성분인 7S α' subunit 단백질과 Kunitz Trypsin Inhibitor (KTI) 단백질이 유전적으로 없으면서 (cgy1cgy1titi 유전자형) 갈색종피 색을 가진 콩 계통을 선발하기 위하여 진행되었 다. 3개의 모본 (장려품종, PI506876, Gaechuck#2)을 이용한 육종집단으로부터 갈색종피, 노란자엽 및 7S α' subunit 단백질과 KTI 단백질이 없는 개체를 선발하였다. 선발개체에 대하여 7S α' subunit 단백질과 KTI 단백질의 유전적 고정과 농업형질 평가를 통하여 갈색 종피를 가지면서 cgy1cgy1titi 유전자형을 가져 7S α' subunit 단백질과 KTI 단백질이 없는 농업형질이 양호한 계통을 선발하였다. 선발된 계통의 초장은 65 cm 정도였으며 백립중은 24.2 g으로 중립이었으며 종피색은 갈색이고 제색은 하얀 색이며 성숙 자엽색은 노란색이었다. 본 연구를 통하여 선발된 계통은 7S α' subunit 단백질 및 Kunitz Trypsine Inhibitor (KTI) 단백질이 동시에 없는 유색콩 품종 육성을 위한 중간모본으로 이용 될 수 있을 것으로 사료되었다. Soybean [Glycine max (L.) Merr.] protein is excellent nutritional factors and is widely used for human and animal feed in the world. Also, antinutritional factors in the raw mature soybean are exist. The presence of antinutritional proteins requires that soybeans to be heat-treated at high temperature. Kunitz trypsin inhibitor (KTI) protein and β-conglycinin (7S α'-subunit) protein 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 new soybean strain (cgy1cgy1titi genotype) with brown seed coat, 7S α' subunit protein free, and KTI protein free. Three parents were used. Presence and absence of 7S α'-subunit and KTI proteins were detected by SDS electrophoresis and Western Blot technique, respectively. One new strain (cgy1cgy1titi genotype) with brown seed coat, lacking of 7S α'-subunit and KTI protein was selected from F4 plant population. New strain has purple flower, brown seed coat, yellow cotyledon, white hilum, 7S α'-subunit protein free, and KTI protein free. Plant height of new strain was 65 cm and 100-seed weight was 24.2 g. New strain selected in this research will be used to improve new cultivar with brown seed coat, 7S α'-subunit and KTI protein free.

갈색종피 및 Penta Null 유전자형을 가진 콩 계통 육종

리사랏,이정환,오현수,김세영,정종일 경상대학교 농업생명과학연구원 2023 농업생명과학연구 Vol.57 No.3

콩 종실에는 레시틴, 아이소플라본, 사포닌, 루테인, 안토시아닌 및 플라보노이드와 같은 생리활성 성분과 약 40%의 단백질이 들어 있어 최근 식물성 대체 단백질의 주요 재료로 이용성이 점차 증가되고 있다. 그러나 lipoxygenase, Kunitz Trypsin Inhibitor (KTI), lectin, 7Sα′subunit 단백질 및 stachyose와 같은 알레르기를 유발시키고 품질 및 기능성을 저하시키는 성분들도 존재한다. 본 연구는 갈색 종피와 penta null 유전자형 (lox1lox2lox3tilecgy1rs2)을 가져 성숙 종실에서 lipoxygenase, KTI, lectin 및 7Sα′subunit의 4가지 단백질이 모두 없으면서 stachyose의 함량이 매우 낮은 콩 계통을 육성하기 위하여 진행되었다. 5개의 자원을 이용하여 창성된 육종집단으로부터 갈색 종피와 penta null 유전자형을 가진 6개의 F2 종자가 선발되었다. 농업형질이 양호한 계통을 선발하여 F7 계통에서 lipoxygenase, KTI, lectin 및 7Sα' subunit의 4가지 단백질에 대한 부재를 확인하였다. 선발계통은 갈색종피와 자주색 꽃 및 유한신육형이며 배꼽색은 흰색이고 성숙 자엽색은 노란색이었다. 선발계통의 경장은 68cm 정도이며 백립중은 29.2g으로 대립이었고 stachyose의 함량은 3.8g/kg으로 매우 낮았으며 수량은 2.75ton/ha 정도였다. 선발된 계통은 성숙 콩 종실에서 콩 가공적성과 품질 및 기능성을 저하시키는 lipoxygenase, KTI, lectin 및 7Sα′subunit의 4가지 단백질이 모두 없으면서 난소화성 당으로 알려진 stachyose의 함량이 매우 낮은 유색콩 품종 및 중간모본으로 이용될 수 있을 것으로 사료되었다. Soybean [Glycine max (L.) Merr.] seed is a major protein source for production of plant-based meat. However, antinutritional factors such as stachyose, lipoxygenase, Kunitz Trypsin Inhibitor (KTI), lectin and 7S α’subunit proteins exist in mature seed. The genetic removal of these antinutritional components is necessary step in soybean breeding. The research was conducted to improve new soybean strain with brown seed coat and penta null genotype (lox1lox2lox3tilecgy1rs2) for lipoxygenase, KTI, lectin, 7S α' subunit and stachyose. Six F2 seeds with brown seed coat and penta null genotype were selected from the breeding population derived from five genotypes. The absence for lipoxygenase, KTI, lectin and 7S α' subunit proteins was confirmed in F7 strain developed. The breeding line has purple flower, determinant growth type, brown seed coat, white hilum color and yellow cotyledon. Stem height of new strain was 68 cm and 100-seed weight was 29.2 g. The content of stachyose was very low at 3.8 g/kg and the yield was about 2.75 ton/ha. A strain developed through this study can be registered as new cultivar and can be used as intermediate parent to improve soybean cultivars with penta null genotype and colored seed coat.

Application of Simple Biological Analyses to Screen Irradiated Brown Rice, Soybean and Sesame Seeds

Ahn, Jae-Jun,Shahbaz, Hafiz Muhammad,Park, Ki-Hwan,Kwon, Joong-Ho The Korean Society for Applied Biological Chemistr 2014 Applied Biological Chemistry (Appl Biol Chem) Vol.57 No.2

The efficacy of biological screening assays such as germination test and direct epiuorescent lter technique (DEFT) and aerobic plate count (APC) was evaluated to detect the irradiation status of different seeds. DEFT/APC help to calculate the difference between dead and living microorganisms in a sample after a possible irradiation treatment. Likewise, the irradiation can significantly affect the physiological and biochemical processes in germinating seeds, which provides the basis for the germination test. In the present study, three different seeds (brown rice, soybean, and sesame) of Korean and Chinese origins were subjected to gamma-irradiation (0.5, 1, 2, and 4 kGy) and the effects on the germination characteristics were evaluated. The results revealed that the growth rate and shoot length decreased with increasing irradiation doses. Particularly, 4 kGy of irradiation had a pronounced effect on all the germination characteristics in all seed samples. The DEFT counts did not change, which were independent of the irradiation dose, whereas the APC counts gradually decreased with dose increment. The results showed the potential of the germination test and DEFT/APC as useful screening methods for irradiated seeds.

Application of Simple Biological Analyses to Screen Irradiated Brown Rice, Soybean and Sesame Seeds

안재준,Hafiz Muhammad Shahbaz,박기환,권중호 한국응용생명화학회 2014 Applied Biological Chemistry (Appl Biol Chem) Vol.57 No.2

The efficacy of biological screening assays such asgermination test and direct epiuorescent lter technique (DEFT)and aerobic plate count (APC) was evaluated to detect theirradiation status of different seeds. DEFT/APC help to calculatethe difference between dead and living microorganisms in asample after a possible irradiation treatment. Likewise, theirradiation can significantly affect the physiological and biochemicalprocesses in germinating seeds, which provides the basis for thegermination test. In the present study, three different seeds (brownrice, soybean, and sesame) of Korean and Chinese origins weresubjected to gamma-irradiation (0.5, 1, 2, and 4 kGy) and theeffects on the germination characteristics were evaluated. Theresults revealed that the growth rate and shoot length decreasedwith increasing irradiation doses. Particularly, 4 kGy of irradiationhad a pronounced effect on all the germination characteristics inall seed samples. The DEFT counts did not change, which wereindependent of the irradiation dose, whereas the APC countsgradually decreased with dose increment. The results showed thepotential of the germination test and DEFT/APC as usefulscreening methods for irradiated seeds.

방사선 조사된 현미, 대두 및 참깨의 저장기간에 따른 방사선 조사여부의 이화학적 판별 특성

이정은 ( Jeong Eun Lee ),안재준 ( Jae Jun Ahn ),권중호 ( Joong Ho Kwon ) 한국식품저장유통학회 ( 구 한국농산물저장유통학회 ) 2012 한국식품저장유통학회지 Vol.19 No.1

The identification characteristics of irradiated (0.5, 1, 2, and 4 kGy) brown rice, soybean, and sesame seeds were investigated using photostimulated luminescence (PSL), thermoluminescence (TL), and hydrocarbon analysis during 12-month storage. PSL-based screening was possible for the irradiated soybean and sesame seed samples up to 6 and 12 months, respectively. The TL glow curve shape, intensity, and ratio enabled the clear dose-dependent discrimination of all the non-irradiated and irradiated samples. The TL intensity decreased during storage, but the TL glow curve did not change qualitatively, which provided enough information to confirm the irradiation treatment of the samples over the storage period. Radiation-induced hydrocarbons were found in all the irradiated samples even at 0.5 kGy, throughout the storage period. 8-Heptadecene (C17:1) and 1,7-hexadecadiene (C16:2) originated from oleic acid, and 6,9-heptadecadiene (C17:2) and 1,7,10-hexadecatriene (C16:3) originated from linoleic acid, can be used as radiation-induced markers in identifying irradiated brown rice, soybean, and sesame seeds.

직박구리(Microscelis amaurotis)가 광나무(Ligustrum japonicum) 발아에 미치는 영향

조숙영(Sook-Young Cho),남현영(Hyun-Young Nam),최창용(Chang-Yong Choi),채희영(Hee-Young Chae) 한국조류학회II 2011 한국조류학회지 Vol.18 No.3

광나무(Ligustrum japonicum)는 우리나라 서·남해안 및 도서지방에 분포하는 상록활엽수로 열매의 결실기는 10월부터 이듬해 1월까지이다. 이 시기에는 조류의 먹이로 이용될 수 있는 과실이 드물어 광나무 열매가 추운 겨울동안 관찰되는 새들에게 중요한 먹이자원이 될 것으로 판단된다. 따라서 전남 홍도에서 겨울철 열매를 맺는 광나무와 이를 이용하는 조류의 상호작용을 파악하기 위해 2009년 12월 하순부터 2010년 1월 초순까지 광나무 열매를 섭식하는 조류에 대하여 조사하였다. 또한 조류에 의한 섭식활동이 광나무 종자의 발아에 어떠한 영향을 미치는지 알아보기 위해 2010년 1월부터 7월까지 180일간 종자의 발아여부 및 시기를 파악하는 실험을 실시하였다. 그 결과 광나무 열매를 섭식하는 조류는 직박구리(Microscelis amaurotis) 1종만이 관찰되었으며 인위적으로 과육을 제거한 종자와 직박구리의 소화기관을 통과한 종자는 자연상태 그대로 파종된 열매와 비교하여 발아율에 차이를 보이지 않았으나 발아시기는 크게 단축되었다. 결국 광나무는 먹이자원이 부족한 겨울철 직박구리에게 먹이를 제공하는 중요한 역할을 하고 직박구리는 광나무 종자의 발아시기를 단축시키는 역할을 하는 것으로 나타났다. 또한 직박구리가 열매를 섭취하고 이동하여 종자를 산포시킴으로써 광나무 군락의 종내 경쟁을 줄이고 새로운 곳에서 정착할 수 있는 기회도 줄 것으로 판단된다. 조류와 식물 간의 이러한 상호작용은 서로에게 이익을 주며 더 나아가 도서지역의 산림보전 및 자연 복원에 활용될 수 있을 것으로 기대된다. The Ligustrum japonicum is a broad-leaved evergreen tree distributed in southern and western parts of Korea. The fruiting season of L. japonicum is a winter between October and January when foods for animals are generally scarce; hence the species may be an importance source of foods for birds wintering in these areas. We aimed to understand interactions between winter-fruiting trees and birds: (1) identification of avian species consuming L. japonicum fruits and (2) the effect of fruit ingestion by birds on its seed germination. We monitored avian species consuming the fruits and carried out germination experiments with seeds in different conditions from December 2009 to July 2010 on Hong-do Island Korea. The Brown-eared Bulbul (Microscelis amaurotis) was the only observed species that consumed the fruits of L. japonicum during the period. Pulp and coat-removed seeds and ingested seeds by bulbuls were germinated significantly earlier than intact seeds. However germination rates of the three groups were not different. This result suggests that the bird ingestion of L. japonicum fruits helps earlier germination or farther dispersal of its seeds so that the tree may be advantageous in intraspecific competition in the community and have the increased opportunity to generate new colonies. Understanding symbiotic interactions between birds and plants on a small remote island may give a clue for future restoration of damaged vegetations on islands in Korea

품종별 자두 종자 추출물의 항갈변 활성

박지원(Ji-Won Park),이태정(Tae-Jeong Lee),이승욱(Syng-Ook Lee) 한국식품영양과학회 2021 한국식품영양과학회지 Vol.50 No.4

본 연구에서는 가공식품 제조 시 생성되는 부산물 중 하나인 자두 종자를 이용하여 식품첨가물 소재로서 활용 가능성에 관한 기초자료를 제공하고자 다양한 항갈변 활성을 측정하였다. 세 품종의 자두 종자로부터 제조한 70% 프레타놀 추출물의 추출 수율은 후무사, 도담, 피자두 모두 유사한 수준이었으며 품종 간 유의적인 차이는 없었다. 감자 상등액에 대한 항갈변 활성은 피자두, 도담, 후무사 순으로 높게 나타났으며 품종 간 유의적인 차이를 보였다. 또한 이들 자두종자 추출물들은 모두 양성대조군인 arbutin에 비해서도 24시간째 우수한 항갈변 활성을 보이는 것으로 확인되었다. PPO 저해 활성은 피자두 종자가 모든 처리 농도에서 도담과 후무사 종자보다 유의적으로 높은 저해 활성을 보였으며, 항산화 활성은 피자두와 후무사 종자 추출물이 각각 ABTS와 DPPH 라디칼에 대해서 상대적으로 높은 저해 활성을 나타내었다. 사과 슬라이스에 대한 항갈변 활성의 경우는 피자두와 도담 종자가 상대적으로 우수하였으나 품종 간 큰 차이는 보이지 않았다. 최종적으로 본 연구에서는 다양한 항갈변 실험을 수행한 결과 피자두 종자 추출물이 후무사와 도담 종자 추출물보다 상대적으로 우수한 항갈변 활성을 나타내는 것을 확인할 수 있었다. 이 결과들은 자두 종자를 이용한 새로운 천연 항갈변제 개발의 기초 자료로 활용될 수 있을 것으로 예상되며, 더 나아가 식품가공 부산물 중 특히 과실 종자들의 새로운 활용방안 구축에도 기여할 수 있을 것으로 기대된다. In the present study, various natural extracts were screened for their anti-browning properties. Plum(Prunus salicina L.) seeds were subsequently selected as a potential material candidate, and the anti-browning activities of 70% ethanol seed extracts from three different plum cultivars (Formosa, Soldam, and Dodam) were analyzed. The average yield of plum seed extract was 3.02∼3.34%, with Formosa seeds showing a relatively high yield. In assays measuring the anti-browning effect on potato supernatant and the inhibitory effect on polyphenol oxidase, the Soldam seed extract showed maximum anti-browning effects. In addition, the Soldam and Formosa seed extracts exerted considerably strong antioxidant activity towards 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) and 2,2-diphenyl-1-picrylhydrazyl radicals, respectively. Taken together, our results indicate the potential of plum seed extracts as sources of natural anti-browning agents.

Screening of Seed Disinfectant for Controlling Brown Leaf Blight in Alisma plantago Double Cropping after Early Rice

Shin, Dong-Young,Kwon, Byung-Sun,Lim, June-Taeg,Hyun, Kyu-Hwan,Lim, Jung-Mook,Seo, Young-Nam The Plant Resources Society of Korea 2006 한국자원식물학회지 Vol.19 No.6

This study was conducted to evaluate the effect of seed disinfectant, in control of brown leaf blight, growth characteristics, and dry root yield in the cultivation of Alisma plantago after early maturing rice cropping. Experimental plot was laid out in split plots design with three replications. The major seed disinfectants were benomyl Wp, 20%, Captan Wp, 50%, Triferine Ec, 17%, Etridia zole Ec, 25%, and Thioplant-mythyl Wp, 50%. Even though seed disinfectant treated had no effect on the growth and flowering date of Alisma plantago, dry root yield was increased largely with benomyl Wp, 20%, in seed disinfectant than in the other seed disinfectants and contorl. All seed disinfectants had no injury with standard dosage. But all seed disinfectants had slight injury in the double dosage level for the Alisma plantago. On the basis of yield, vegetative and disease paramerer, benomyl Wp (20%) ($100g/20{\ell}$) had shown superior performance, however, all the seed disinfectants are effective as compare to without treatment.

Antioxidant and Tyrosinase Inhibitory Activities from Seed Coat of Brown Soybean

Jin Hwan Lee,In-Youl Baek,Jong-Min Ko,Nam Suk Kang,Seong-Hyu Shin,Sea-Gyu Lim,Ki-Won Oh,Sang-Ouk Shin,Keum-Yong Park,Ki Hun Park,Tae Joung Ha 한국식품과학회 2008 Food Science and Biotechnology Vol.17 No.1

Soybeans with brown, black, and yellow seed coats were compared to total phenolic contents and antioxidant activities including 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2"-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radicals. Also, 3 seed coats were examined for inhibitory activities on tyrosinase and lipoxygenase-1 on the basis of spectrophotometric and polarographic methods. Among seed coat extracts, 80% methanol extract of brown soybean seed coat showed the highest total phenolic contents (68.9±3.29 mg GAE/g) as well as exhibited potent scavenging effects on the DPPH (IC₅₀μg/mL) and ABTS (IC₅₀=3.7 μg/mL) radicals. In a polarographic experiment, this extract was potentially inhibited the oxidation of L-tyrosine and L-3,4-dihydroxy-phenylalanin (L-DOPA) catalyzed by mushroom tyrosinase with IC₅₀of 12.4 and 63.7 μg/mL, respectively. It was also detected inhibition of the tyrosinase catalyzed oxidation of L-DOPA with an IC₅₀value of 120.3 mg/mL in UV spectrophotometric experiment. In addition, this extract inhibited the linoleic acid peroxidation catalyzed by lipoxygenase-1 with an IC₅₀ value of 4.0 μg/mL. These results suggest that brown soybean may possess more beneficial effect on human health than black and yellow soybeans.