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1 Korean Journal of Breeding Science Korean J. Breed. Sci. 50(3): ( ) Online ISSN: Print ISSN: 종피가터지지않고탈립에강한녹자엽검정콩 청자 4 호 김현태 한원영 서정현 * 강범규 신상욱 김홍식 이영훈 이병원 윤홍태 박장환 곽도연 농촌진흥청국립식량과학원 Pod Shattering Tolerant Black Soybean Cheongja4 with Green Cotyledon and without Seed Coat Cracking Hyun Tae Kim, Won Young Han, Jeong Hyun Seo *, Beom Kyu Kang, Sang Ouk Shin, Hong Sik Kim, Yeong Hoon Lee, Byoung Won Lee, Hong Tai Yun, Chang Hwan Park, and Do Yeon Kwak National Institute of Crop Science (NICS), RDA, Jeonju 55365, Korea Abstract Cheongja4 is a black soybean cultivar developed via a cross between Milyang148 and Daemang in F 1 and F 2 populations were grown for 2 years, and promising lines were selected using the pedigree method from F 3 to F 5. The preliminary yield trial (PYT) and advanced yield trials (AYT) were conducted from 2011 to 2012, and regional yield trial (RYT) in 7 regions was conducted from 2013 to Cheongja4 is determinate and has purple flowers, a green cotyledon, and black spherical seeds. Flowering date and maturing date were Aug. 5 and Oct. 19, respectively. With respect to its quantitative characteristics, Cheongja4 has more pods (50) and smaller seed size (30.1g/100-seed weight) than Cheongja3 (38 and 35.2g/100-seed weight). Cheongja4 was revealed to be more tolerant to pod shattering than Cheongja3 in RYT field and indoor test. Cheongja4 had high levels of resistance to bacterial pustule and soybean mosaic virus in field and inoculation test. Although its seed quality-related characteristics were similar to those of Cheongja3, it contained more anthocyanin in the seed coat; this is the main functional component of black soybean. The mean yield of Cheongja4 in RYT was 314kg/10a, 123% of the yield of Cheongja3. Cheongja4 is expected to be widely used for variable functional purposes, such as cooking with rice and the production of processed food. Keywords Soybean,, Cheongja4, Green cotyledon, Black seed coat Received on May 10, Accepted on July 20, * Corresponding Author ( next0501@korea.kr, Tel: , Fax: ) 서언콩 [Glycine max (L.) Merr.] 은한반도북부가원산지인작물로서약 5,000년전부터재배가시작되었다. 콩종자에는양질의단백질과지방이다량함유되어있으며이러한성분조성의특이성으로인하여콩은다양한용도의원료곡으로사용되어왔다. 우리나라에서는특히전통장류, 두부, 콩나물등각종가공식품의원료곡으로서주식인쌀의부족한영양분을채워주는역할을해왔다. 이중에서도검정콩은콩의한종류로서종피색이검은색인콩을말하며일반콩과달리예로부터혼반용, 자반용, 떡소용등종실자체를소비하는형태로발전해왔다. 따라서종실의외관이일반콩에비해매우중요한구매요인으로작용하고있으며특히소비자들은종피안의자엽색이녹색인콩을선호한다 (Kim et al. 2006). 이처럼자엽의색이녹색이면서종실이크고숙기가늦은재래종검정콩을 서리태 라고부르며무름성이좋고당도가높아주로혼반용으로선호도가높다 (Kim et al. 2006). 국내에서검정콩은주로이러한재래종서리태를개량하는것을목표로꾸준히연구되어왔으며특히서리태가가지고있는농업적불량형질인쓰러짐, 탈립등의형질을개선하는것을주된목표로육종연구사업을진행해왔다. 국내에서는최초의녹자엽검정콩품종인 청자콩 (Baek et al. 2001) 을시작으로녹자엽검정콩개발을시작하였으며 (Han et al. 2017), 이후 청자2호 (Baek et al. 2004), 청자3호 (Yun et al. 2005) 등이개발되었다. 우리나라 2017년콩의재배면적은약 45,000 ha로 2012년이후꾸준한감소추세를보이고있으며콩가공품의국산원료비중은 2015년기준 5.9% 에불과하다 (KREI 2018). 이러한현상의주요인은콩의수익성감소로인한재배기피와저렴한수입산 Copyright c 2018 by the Korean Society of Breeding Science This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

2 韓育誌 (Korean J. Breed. Sci.) 50(3), 2018 원료사용의증가때문이다. 따라서국산콩의수익성을향상시키고수입산콩에대한경쟁력을갖게하기위해서는콩의용도다변화를통한새로운수요를창출하는것이필요하다. 검정콩은종피에함유되어있는각종기능성성분과그효능들이연구되면서건강기능성식품으로서인식되고있으며 (Kim et al. 2008, Chung et al. 2011, Jeon et al. 2011, Jeong et al. 2014), 일반콩에비하여높은시장가격을형성하고있다. 최근에는기존의혼반용, 반찬용으로만활용되었던검정콩의용도를넘어서두부, 두유, 생청국장등각종콩가공식품으로보다높은가격을받으며판매되고있고이에대한연구가활발히진행되고있다 (Im & Kim 2003, Kim & Jeon 2005, Bang & Jeong 2007, Kim et al. 2011, Song et al. 2011). 이러한검정콩의소비잠재력에대응하기위해서는안정적인원료곡을공급하는것이중요하다. 기존의재래종서리태와개발된검정콩품종들은탈립, 도복등재해에취약하거나숙기가늦어안정적인수확이힘든경우가많았다. 이러한서리태를개량한검정콩품종중 가장널리보급되어있는것은 2004년개발된 청자3호 이지만환경에따른종피의열피현상, 성숙기탈립등불량형질이발견되어재배및유통과정에어려움을겪고있다. 이에농촌진흥청국립식량과학원에서는 청자3호 의불량형질인종피의열피현상이개선되고성숙기꼬투리탈립에강하여재배특성이우수한품종인 청자4 호 를개발하여그육성과정과특성에대해소개하고자한다. 재료및방법육성과정 청자4호 는녹자엽대립검정콩품종육성을목표로하여 100 립중이 22.0 g으로중립인녹자엽검정콩 밀양148호 를모본으로, 100립중이 31.4 g으로대립인두부 장류용콩 대망 (Yun et al. 2005) 을부본으로 2005년에인공교배하였다 (Fig. 1). '06-'07 년도에 F 1, F 2 세대를양성하고, F 3 부터계통육종법으로선발한 Fig. 1. Genealogy of Cheongja4. Fig. 2. Pedigree diagram of Cheongja4, z: Preliminary yield trial, y: Advanced yield trial, x: Regional yield trial. 290

3 종피가터지지않고탈립에강한녹자엽검정콩 청자 4 호 YS1758-2B 계통이다. '11-'12년도에실시한생산력검정시험에서불마름병등에강하고대립인녹자엽검정콩으로유망시되어 밀양263호 의계통명을부여한후 '13-'15년 3년동안지역적응시험을실시한결과, 녹자엽대립인검정콩으로서탈립에매우강한특성을나타내어 2015년 12월농작물직무육성신품종선정위원회에서품종으로서가치가인정되어 청자4호 로명명하였다 (Fig. 2). 생산력검정시험 2011년생산력검정예비시험 (PYT, Preliminary Yield Trial), 2012년생산력검정본시험 (AYT, Advanced Yield Trial) 을수행하였다. 생산력검정예비시험은시험구면적 8.4 m2, 재식거리 cm, 1주2 본으로재배되고난괴법 2반복으로배치하였다. 생산력검정본시험은시험구면적 11.2 m2, 재식거리 cm, 1주2본으로재배되고난괴법 3반복으로배치하였다. 생육특성과수량성은농업과학기술연구조사분석기준에따라조사하였다 (RDA 2012). 지역적응시험지역적응시험 (RYT, Regional Yield Trial) 은수원, 춘천, 연천, 나주, 달성, 예산, 칠곡등 7개지역에서 2013년부터 2015년까지 3년간수행되었다. 재배방법은시험구면적 11.2 m2, 재식거리 cm, 1주2본으로재배되고달성과수원지역은난괴법 4반복으로배치하였으며이외지역은난괴법 3반복으로배치하였다. 파종기는 6월중순이며농업과학기술연구조사분석기준에따라생육특성및수량성을조사하고, 대비품종인 청자3호 와비교하였다. 내병성검정병해검정을위해 5월상순에검정포장에파종하여콩모자이크바이러스와불마름병의자연이병률을조사하였다. 콩모자이크바이러스병저항성검정을위해각시험전년도에접종및증식한후저온보관해둔콩모자이크바이러스 G5 (Lim et al. 2003), G6H (Seo et al. 2009), G7H (Kim et al. 2003) 균계를지닌잎을마쇄하여 2 mole 농도의인산나트륨버퍼에녹인후즙액을만들고, 온실에서초생엽전개시 Carborandom을이용하여잎표면에상처를낸후접종하였다. 접종 2주경과후접종한하위엽을, 10일뒤접종한상위엽을병반없음 (-), 갈색병반 (L), 엽맥병반 (V), 모자이크증상 (M), 고사 (N) 등 5 가지병징으로구분하고조사하였다. 불마름병저항성검정을위해 8ra를증식하여증류수에희석하여 OD값 으로농도를맞추어제2본엽기에충분히분무접종하고습식처리하였다. 균주접종후 10일이경과하고 2회에걸쳐잎에나타난병징발현정도를조사하였다. 내도복성및내탈립성검정내도복성검정을위해 2 수준의 cm( 표준 ) 와 cm(2배밀식 ) 를설정하고난괴법 2반복으로배치하여도복정도를조사하였다. 도복정도는성숙기에 45 이상기울어진개체의비율로등급화 (1: 5% 이하, 3: 6~10%, 5: 11~50%, 7: 51~75%, 9: 76% 이상 ) 하였다. 반복및연차간도복정도등급을평균하여반올림후 1~9로최종등급을표시하였다. 협개열성정도는내도복성검정시험구에서각계통별꼬투리 30개를성숙기에채취하여 40 건조기에 48시간건조한후꼬투리의개열정도를조사하였다 (Kang et al. 2005). 품질분석단백질함량은질소분석기 (Elementar Analysen system, US/RapidN111, Germany) 를이용하여분석하였으며지방함량은자동유지추출장치 (Soxhlet System: BUCHI Labotechnik, B-811, AG, Switzerland) 에넣고 n-hexane으로 3시간열수추출한후지방함량을구하였다. 당성분분석은분쇄한시료 1 g을정밀히칭량하여튜브에넣은후 70% 에탄올 10 ml를첨가하여실온에서 shaking 하여전처리물을추출하였다. 추출된시료를 24시간후여과하여증류수와여과된시료각 0.7 ml를마이크로튜브에혼합하여이중 0.7 ml를액체크로마토그래피 (Dionex, Ultimate 3000, USA) 에넣고분석하였다. 이소플라본성분함량분석을위해분쇄한시료 1.0 g을 50% methanol 용액 20 ml에 12시간실온에서교반한뒤추출액을여과지 (Whatman NO. 2) 를이용하여여과하였다. 여과된시료는다시 HPLC 분석을위해 0.2 μm필터를하였다. 이소플라본분석은액체크로마토그래피 (Agilent, 1100, USA) 로분석하였다. 안토시아닌분석을위하여콩껍질 0.1 g을정밀히칭량하여튜브에넣고 40% 메탄올용매 30 ml를첨가하여 rpm에서 48시간 shaking 하여추출하였다. 추출된시료용액을 No2 여과지로여과한후여액 1.0 ml 을취하여 0.45 μm필터로여과하였고안토시아닌은액체크로마토그래피 (Dionex, Ultimate 3000, USA) 로분석하였다. 수분흡수율은건조종자 50립의무게를잰후상온의물에 291

4 韓育誌 (Korean J. Breed. Sci.) 50(3), 2018 침종하여 2, 6, 12시간후무게를각각측정하여구하였으며 (( 침종후종자무게 -침종전종자무게 )/ 침종전종자무게 100%), 12시간후부풀지않은종자수를세어경실종자비율을구하였다. 결과및고찰고유특성및생육특성 청자4호 는유한신육형초형으로꽃색은자색, 엽형은난형, 모용색은갈색, 협색은갈색, 종실은구형이며종피색은검정이다 (Table 1). 표준품종인 청자3 호 와달리종피의피분이없는특징이있으며열피현상이나타나지않는다 (Fig. 3), 자엽색은녹색으로소비자의기호에알맞은특징을가지고있다. 개화기는 8월 5일, 성숙기는 10월 19일인중만생종이다. 경장, 마디수, 협수, 협고는각각 72 cm, 16개, 50개, 19 cm로표준품종인 청자3 호 에비해 협수가많고협고가높은특징이있으며, 100립중이 30.1 g으로 청자3호 에비해약 5 g 작은중대립종이다 (Table 2). 내도복 내탈립및내병성 청자4호 는시험포장과밀식검정포장에서각각 2, 4로평가되어 청자3호 (2,4) 와같이도복에강한것으로나타났다. 협개열성정도는포장에서 2로평가되고실내검정에서 7.0% 로측정되어 청자3호 (75.2%) 보다탈립에매우강하였다 (Table 2). 청자4호 의불마름병저항성은시험포장및인공접종시에 청자3 호 보다강하였고, 콩모자이크바이러스접종시에모자이크증상이나타났으나검정포장에서는나타나지않았다. 검은뿌리썩음병에는다소취약한특성을나타내었다 (Table 3). Fig. 3. Pictures of Cheongja4 in the comparison of seed coat cracking (left) and at the maturing stage in the field (right). Table 1. Qualitative characteristics of Cheongja4. Growth habit Leaflet shape Flower color Pubescence color Seed coat color Cotyledon color Seed shape Cheongja4 Determinate Oval Purple Brown Black Green Spherical Cheongja3 Determinate Oval Purple Brown Black Green Spherical Table 2. Mean values of quantitative characteristics of Cheongj4 in the regional yield trial from 2013 to Flowering Date (Month. Day) Maturity Date (Month. Day) Height (cm) No. of nodes No. of branches No. of pods First pod height (cm) 100-seed weight (g) Lodging at field (1-9) z Shattering Ratio by Normal Dense Field culture culture (1-9) z Oven test y Cheongja a x 16a 2.9b 50a 19a 30.1b Cheongja a 16a 3.4a 38b 13b 35.2a z (1) Tolerant ~ (9) Susceptible. y the ratio of shattered pod by dry oven test at 40 for 48hr. x same letters were not significantly different between Cheongja4 and Cheongja3 by student t-test at

5 종피가터지지않고탈립에강한녹자엽검정콩 청자 4 호 Table 3. The resistance of Cheongja4 to the diseases of soybean evaluated from 2013 to Root rot Inoculation Field Inoculation Field(0-9) z x G5 G6H G7H Cheongja4 0 3 M / M - / M - / M Cheongja / M - / M - / M z (0) Tolerant~(9) Susceptible. y Reaction of inoculated leaf/upper leaves, (-) No symptom, (L) Local reaction, (M) Mosaic. x Root rot and damaged seed that were naturally infected in the field were estimated. Table 4. Seed quality-related characteristics of Cheongja4 evaluated from 2013 to Crude Protein Crude Oil Hard seed ratio Ratio of water absorption Sugar content (mg/g) Anothocyanin content ( μg /g) 2hr 6hr 12hr Sucrose Total Cy3glc Total Cheongja4 40.2a z 17.9a 1.7a 119a 156a 184a 56.9a 101.5a 18,513a 25,366a Cheongja3 41.4a 17.2a 0.0a 139a 182a 194a 60.0a 106.5a 15,978a 23,972a z same letters were not significantly different between Cheongja4 and Cheongja3 by student t-test at Table 5. Yield of Cheongja4 in the preliminary and advanced yield trials in Daegu. Yield (Ton/ha) PYT( 11) z AYT('12) y Mean Index x Cheongja Cheongja t-value w ** -3.77* - - z (PYT) preliminary yield trial. (AYT) advanced yield trial. x (Index) mean yield of Cheongja4 /mean yield of Cheongja w ** and * denote significantly different between Cheongja4 and Cheongja3 at 0.01 and 0.05 by student t-test, respectively. Table 6. Yield of Cheongja4 in the regional yield trials at 7 locations. Location Cheongja4 Cheongja Mean Index z Mean Suwon Chuncheon y Yeoncheon Dalseong Yesan Naju Chilgok Mean t-value x ** z (Index) mean yield of Cheongja4 / mean yield of Cheongja y deletion due to the growth disorder caused by poor decomposed compost. x ** denotes significantly different between Cheongja4 and Cheongja3 at 0.01 by student t-test. 293

6 韓育誌 (Korean J. Breed. Sci.) 50(3), 2018 종실품질및가공적성 청자4호 의조단백질함량은 40.2% 로 청자3호 보다 0.8% 낮았고, 조지방함량은 17.9% 로 0.7% 높았지만이에대한통계적유의성은없었다. 수분흡수율은 12시간 184% 로 청자3 호 (194%) 와비슷하였고, 경실종자는없었다. 종실의총당함량은 mg/g, sucrose 함량은 56.9 mg/g으로 청자3 호 에비해다소낮았으나, 종피의총안토시아닌함량은 25,366 μg /g으로 청자3호 (23,972 μg /g) 에비해높았다 (Table 4). 수량성 청자4호 는 '11-'12 년 2년간생산력검정시험에서 284 kg/10a 의수량성을나타내어표준품종인청자 3호에비해 25% 증수하였다 (Table 5). '13-'15년 3년간지역적응시험결과전국 7개소평균 314 kg/10a로 청자3호 (257 kg/10a) 보다 23% 증수된수량성을보였고통계적으로유의한차이를나타내었다 (Table 6). 재배상의유의점종실의크기가다소작은단점이있으므로 6월상 ~ 중순경파종하고극만파를지양하여종실무게가감소되지않도록해야하며검은뿌리썩음병에약하므로배수가불량한포장에서의재배를삼가고배수관리를철저히해야한다. 적요 청자4 호 는대립인녹자엽검정콩품종육성을목표로 2005년에 밀양148호 를모본으로 대망 을부본으로교배하여계통육종법에의해선발되었다. '11-'12 년도에실시한생산력검정시험에서는대립인녹자엽검정콩으로탈립및도복에강하여 밀양 263호 의계통명을부여하였고, '13~'15년 3년간 7개소에서수행한지역적응시험결과대립인녹자엽검정콩으로탈립에매우강한특성을나타내어 청자4호 로명명되었다. 청자4 호 는유한신육형, 화색이자색, 종피는검은색, 자엽색은녹색이면서구형인검정콩품종이다. 개화기는 8월 5일, 성숙기는 10월 19일로중만생종이며, 협수가 청자3 호 에비해많고협고가높은특성이있다. 100립중은 청자3호 에비하여 5 g 작았다. 도복정도는시험포장과밀식검정포장에서 청자3 호 와비슷하여강하였고, 협개열성정도는실내검정에서 청자3 호 에비해매우낮았다. 불마름병, 콩모자이크바이러스등병해에 청자3 호 와대등하거나더강하였으나, 검은뿌리썩음병에다소 취약한특성을나타내었다. 청자4 호 는 12시간침종후경실종자가없었으며, 당함량은다소낮았으나검정콩의기능성성분인안토시아닌함량이 청자3호 보다높았다. 수량성은전국 7개지역평균 314 kg/10a로청자3호보다 23% 증수되었다. 사사본연구는농촌진흥청연구사업 ( 과제명 : 콩전통식품가공적성신품종육성 (2단계), 과제번호 : PJ ) 의지원에의해수행되었다. REFERENCES 1. Baek IY, Kang ST, Shin DC, Choung MG, Han WY, Kwack YH, Moon HP A new black soybean variety with green cotyledon, early maturity and large seed size Cheongjakong. Korean J Breed Sci 33: Baek IY, Han WY, Kang ST, Shin DC, Choung MG, Oh SK, Shin SO, Suh DY, Kim SC A new black soybean cultivar, Cheongja2 with green cotyledon, early maturity and high anthocyanin. Korean J Breed 36: Bang BH, Jeong EJ A study on manufacturing black soybean yogurt. Korean J Food & Nutr 20: Chung MJ, Ha TJ, Choi HN, Lee JS, Park YI Inhibitory effects of anthocyanins isolated from black soybean (Glycine max L.) seed coat on degranulation and cytokine generation in RBL-2H3 cells. J Korean Soc Food Sci Nutr 40: Han WY, Seo JH, Kang BK, Kim HT, Shin SO, Kim HS, Lee YH, Lee BW, Ha TJ, Seo MJ, Baek IY, Ko JM, Kwak DY Green seed-coat colored soybean variety Jungmo3005 with tolerance to lodging and shattering. Korean J Breed Sci 49: Im JG, Kim YH Quality characteristics of bread prepared by the addition of black soybean powder. J East Asian Soc Dietary Life 13: Jeon AJ, Lim TG, Jung SK, Lee EJ, Yeom MH, Park JS, Choung MG, Lee HJ, Lim Y, Lee KW Black Soybean (Glycine max cv. Heugmi) seed coat extract suppresses TPA or UVB-induced COX-2 expression by blocking mitogen activated protein kinases pathway in mouse skin epithelial cells. Food Sci Biotechnol 20:

7 종피가터지지않고탈립에강한녹자엽검정콩 청자 4 호 8. Jeong JH, Kim HJ, Park SK, Jin DE, Kwon OJ, Kim HJ, Heo HJ An investigation into the ameliorating effect of black soybean extract on learning and memory impairment with assessment of neuroprotective effects. BMC Complementary and Alternative Medicine 2014, 14: Kang ST, Kim HK, Baek IY, Chung MG, Han WY, Shin DC, Lee SH Genetic analysis of pod dehiscence in soybean. Korean J Crop Sci 50: Kim YH, Kim OS, Lee BC, Moon JK, Lee SC, Lee JY G7H, a new soybean mosaic virus strain: its virulence and nucleotide sequence of ci gene. Plant Dis 87: Kim J, Jeon JR Quality characteristics of tofu added with black soybean hull powder. Korean J Food Culture 20: Kim YH, Lee JH, Lee YS, Yun HT Antioxidant activity and extraction efficiency of anthocyanin pigments in black soybean. Korean Soybean Digest 23: Kim YH, Kim DS, Woo SS, Kim HH, Lee YS, Kim HS, Ko KO, Lee SK Antioxidant activity and cytotoxicity on human cancer cells of anthocyanin extracted from black soybean. Korean J Crop Sci 53: Kim HE, Han SY, Jung JB, Ko JM, Kim YS Quality Characteristics of Doenjang (Soybean Paste) Prepared with germinated regular soybean and black soybean. Korean J Food Sci Technol 43: KREI (Korea Rural Economic Institute) Agricultural Outlook 2018 Korea; Lim WS, Kim YH, Kim KH Complete genome sequences of the genomic RNA of soybean mosaic virus strains G7H and G5. Plant Pathol. J 19: RDA (Rural Development Administration) Agricultural science technology standards for investigation of research. RDA, Jeonju, Korea. 18. Seo JK, Ohshima K, Lee HG, Son M, Choi HS, Lee SH, Sohn SH, Kim KH Molecular variability and genetic structure of the population of soybean mosaic virus based on the analysis of complete genome sequence. Virology 393: Song NE, Song YR, Kim JH, Kim YE, Han AR, Jeong DY, Baik SH Development of sugar-soaked black soybean snack and its quality change on functional components. J Korean Soc Food Sci Nutr 40: Yun HT, Park KY, Moon JK, Kim YH, Kim SL, Ku JH, Lee YH, Ryu YH, Baek IY, Han WY, Kim HT, Cho JH, Ko JM, Shin DC, Suh DY, Kim HY, Kim SK, Ha KS, Lee SS, Son CK, Kim YD A new black soybean cultivar, Cheongja3 with green cotyledon, medium-late maturity and high anthocyanin. Korean J Breed 37: Yun HT, Park KY, Moon JK, Seo MJ, Lee YH, Kim SL, Ryu YH, Kim YH, Lee ES, Ha KS, Son CK, Kim ES, Kim SD, Lee MH A new soybean cultivar for fermented soyfood and tofu, Daemang with large seed and high yield. Korean J Breed 37:

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<31332E28C7B0C1BE29B9E8BCAEBAB92E687770> 韓育誌 (Korean J. Breed. Sci.) 40(2) : **~** (2008) 단경 내병 다수성 땅콩 품종 풍산 배석복 * 정영근 2 박장환 이명희 황정동 심강보 정찬식 강철환 박금룡 박충범 최규환 4 이재철 5 김인재 6 김제규 영남농업연구소, 2 호남농업연구소, 작물과학원, 4 전라북도노업기술원, 5 충청남도농업기술원, 6 충청북도 농업기술원,