Korean J. Plant Res. 30(1):50-57(2017) https://doi.org/10.7732/kjpr.2016.30.1.050 Print ISSN 1226-3591 Online ISSN 2287-8203 Original Research Article 신소림, 임윤경, 권혁준, 김유리, 김수영 * 국립생물자원관 Morphological Characteristics and Germination Conditions of Seeds in Arabis pendula L. So Lim Shin, Yun Kyung Lim, Hyuk Joon Kwon, Yu Ri Kim and Soo-Young Kim* National Institute of Biological Resources, Incheon 22689, Korea Abstract - This study aimed to investigate the morphological characteristics and optimal germination conditions in seeds of Arabis pendula L., a traditionally edible and medicinal plant. The external seed shape was circular-obovate with narrow wings and dark brown. The seed length and width were 1.25 mm and 1.47 mm, respectively. The seeds were exalbuminous and the embryo was a bent type. Seed germination was the highest (49.7%) at 20 C under dark conditions among the various temperature and light conditions applied. However, under the dark condition, the seedling was weak, overgrown, and the cotyledons were small and folded. To improve the germination and growth of seedlings, the seeds were pre-treated with GA 3 solutions of varying concentrations (0, 200, 500, and 1,000 mg /L). The seed germination and seedling growth were effectively improved by GA 3 pre-treatment. The germination rate was the highest (97.3%), mean germination time was the shortest (8.1 days), and a vigorous growth of seedlings was observed upon pre-soaking the seeds in 500 mg /L GA 3 solution. In conclusion, the best method for germination was pre-soaking in 500 mg /L GA 3 (4, dark, 24 h) and incubating the seeds at 20 for 15 days. Key words - Brassicaceae, Dark germination, Gibberellic acid, Tetrazolium test 서언 늘어진장대 (Arabis pendula L.) 는십자화과장대나물속의다년생초본으로, 우리나라중부이북의산과들의습한곳에서자라며, 국외에서는러시아, 몽골, 일본, 중국, 유럽에분포한다 (Lee, 1996). 국내자생하는장대나물속식물은갯장대, 늘어진장대, 묏장대, 바위장대, 산장대, 섬장대, 자주장대나물, 장대나물, 주걱장대, 참장대나물, 털장대및흰장대나물등총 12종이있으며 (NIBR, 2011), 이중식용식물은 3종 ( 늘어진장대, 장대나물및털장대 )(MFDS, 2016) 이고, 약용식물은늘어진장대뿐이다 (Moon, 1999). 늘어진장대의어린잎은식용으로먹을수있으며, 민간에서전초를달여위염, 여성질환, 두통, 관절염, 해독및해열에사용해왔다 (Moon, 1999). 중국에서는늘어 * 교신저자 : sy7540@korea.kr Tel. +82-32-590-7111 진장대미숙과를수과남개 ( 垂果南芥 ) 라하여청열 ( 清热 ), 해독 ( 解毒 ) 및소종 ( 消肿 ) 에효과가있고특히부기를가라앉히고피부궤양, 부스럼, 종기의치료에사용한다 (NBTCMEB, 1999). 또한수과남개를달여결절종의치료약으로사용하거나, 수과남개와형개, 만형자, 익모초, 둥굴레, 쑥을함께곱게분쇄한후달여질염 ( 특히 Trichomonas 에의한질염 ) 의치료에사용하고있다 (NUTCM, 2006). 최근천연물유래약리물질탐색연구가증가하면서야생식물의유용생리활성에관한연구및이를이용한천연물신약개발등산업화가증가하고있는데 (Yoo et al., 2016), 식물소재를산업화하기위해서는반드시대량증식방법이확립되어야한다. 초본식물은주로종자로증식하는데, 늘어진장대와같은야생종자는재배작물에비해강한휴면상태인경우가많으므로발아율이낮으며, 상대적으로복잡한휴면체계를가지고있어발아효율이낮기때문에효율적증식방법에어려움이많다 c 본학회지의저작권은 ( 사 ) 한국자원식물학회지에있으며, 이의무단전재나복제를금합니다. This is an Open-Access article distributed under the terms of the Creative Commons -50- Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
(Koornneef et al., 2002). 따라서야생식물을산업적으로재배하기위해서는발아율을높이고, 짧은기간안에발아될수있는발아조건을확립하여입묘과정에서빠르게적정입묘율을확보할수있는방법이개발되어야한다 (Kim et al., 2014). 늘어진장대는민간에서식 의약소재로활용해왔지만, 발아및증식에대한체계적인연구는보고된바없다. 우리나라에서는장대나물속식물의발아및재배에대한연구가보고된바없지만, 해외에서는일부 Arabis 속종자의휴면유형과발아에대해보고되었다. Arabis laevigata 의종자는저온층적처리에의하여휴면타파가가능한생리적휴면 (Physiological dormancy, PD) 이며, 5 에서 10주저온층적처리하여 20/10 의변온조건에파종했을때발아가가장왕성하였다 (Bloom et al., 1990). 한편 A. hirsuta, A. scabra 및 A. glabra 종자는전처리없이 10 20 에서 80% 이상발아하였고, A. petraea는 23/9 의변온조건에서 90% 발아하는것으로보고되었다 (MSB, 2016). 따라서 Arabis 속종자는비교적저온에서발아가왕성하지만종에따라발아적온이다르며, 휴면의유무도다르므로각종에적합한발아및증식조건을확립할필요가있다. 본연구는우리나라에서자생하는늘어진장대의종자를이용한효율적증식방법확립을위해종자의형태적특성관찰과활력검정을수행하였으며, 발아율과발아효율을증가시킬수있는적정발아온도와지베렐린처리농도등최적발아조건을규명하고자수행되었다. 재료및방법실험재료연구재료로사용된늘어진장대종자는 2013 년 9월에강원도태백시에서채집되었고, 파쇄립, 저질, 병충해립, 감염립, 이물질등을제거하여건조실 (15, 10%) 에서수분함량 5 10% 미만으로건조후밀봉하여국가야생식물종자은행중기수장고 (4, 상대습도 40%) 에보존후연구에사용하였다. 종자의형태학적특성및활력검증종자형태는해부현미경 (SMZ1500, Nikon, Tokyo, Japan) 으로관찰하였다. 종자의외부형태를디지털카메라 (DS-RiI, Nikon, Tokyo, Japan) 로촬영하였고, 이미지분석프로그램 (NIS-Elements BR 3.10 software, Nikon GmbH, Vienna, Austria) 을이용하여무작위선발한종자 20립의길이와너비를측정하여평균과표준오차를구하였다. 종자의천립중은종자 계수기 (Contador, Pfeuffer GmbH, Waldenburg, Germany) 로계수한후정밀전자저울 (MS1003S, Mettler Toledo GmbH, Greifensee, Switzerland) 로무게를 3회반복측정하여평균과표준오차를구하였다. 종자의활력은국제종자검정협회 (International Seed Test Associate) 의 Tetrazolium Test ( 이하, TZ 테스트 ) 방법을응용하여검증하였다 (ISTA, 2014). 늘어진장대종자상단에단면도 (Single edge blades DN-52, Dorco, Korea) 로얇은칼집을내고멸균수에침지한후 (20, 18시간 ), 1% 의 2, 3, 5-Triphenyl tetrazolium chloride (T8877, Sigma Chemical Co., St. Louis, MO., USA) 용액에넣어 30 에서 18시간동안침지후종단면을잘라해부현미경 (SMZ1500, Nikon, Tokyo, Japan) 으로종자내부배의염색정도와상태를관찰하여활력을판단하였다. 종자발아특성분석종자는파종전 70% 에탄올을넣고 30초흔들어준다음에탄올과이물질을제거하고 2% 의 NaOCl 을넣어 10분동안소독후멸균수로 5회수세 (1회당 1분이내 ) 하고멸균수를제거하여침지처리에사용하였다. 침지처리는연구목적에따라침지용액을멸균수또는 GA 3 희석액으로달리하였으며, 저온저장고 (NRD-450RF, Lassele, Ansan, Korea) 에서침지처리 (4, 암조건, 24시간 ) 하였다. 침지처리후침지용액을모두제거하고종자를멸균수로 1회수세하여파종하였다. 파종은종자 50립을무작위선발하여여과지 (Whatman No. 2, Whatman Ltd., Kent, England) 를넣은페트리디쉬 (10090, SPL Life Science Co., Pocheon, Korea) 내에치상하였으며, 이를 3반복으로수행하였다. 종자발아는 4실배양기 (WIM-R4, Daihan Labtech, Namyangju, Korea) 에서수행하였으며, 배양기는광주기 24시간으로명조건 (100 μmol m -2 s -1, Fluorescent) 또는배양기에조명을하지않은암조건으로설정하였다. 늘어진장대종자의발아적온및온도별발아특성을규명하고자종자를멸균수에침지처리후파종하여각기다른온도조건 (15, 20, 25, 30 ) 의명조건에서 30일동안발아연구를수행하였다. 종자발아에영향을미치는광조건을규명하기위하여멸균수에침지처리한종자를파종하여 20 의명조건또는암조건에서 30일동안발아연구를수행하였다. 식물생장조절물질중하나인 GA 3 의발아촉진효과를연구하기위하여소독된종자를각기다른농도의 GA 3 희석액 (0, 200, 500 및 1,000 mg /L) 에침지처리후 20 의광조건에서발아연구 -51-
韓資植誌 Korean J. Plant Res. 30(1) : 50~57(2017) 를수행하였다. 종자파종후 2 3 일간격으로발아상태를조사하였으며, 발아율 (Germination percentage, GP), 평균발아소요일수 (Mean germination time, MGT), 발아속도지수 (Germination speed index, GSI), 발아균일도 (Germination uniformity, GU) 와 T 50 ( 파종후최종발아율에대한 50% 발아까지도달하는데소요되는기간 ) 을아래와같은공식을이용하여구한후발아특성을분석하였다 (Coolbear et al., 1984; Gavassi et al., 2014). GP = ( n/n) 100 MGT = (Tx Nx)/ n GSI = (Nx/Tx) GU = [MGT-Tx) 2 Nx]/( n-1) T 50 = Ti+[(N/2)-ni](Tj-Ti)/(nj-ni) N: 총치상한종자수, n: 발아수, Tx: 치상후조사일수, Nx: 조사당일발아수, Ti: ni까지소요된기간, Tj: nj까지소요된기간, ni: 최종발아율대비 50% 미만까지의누적발아수, nj: 최종발아율대비 50% 초과된누적발아수통계처리연구에사용된소독된종자는 50립씩무작위선발하여치상하였으며, 3반복으로발아실험을수행하였다. Microsoft Office Excel 2007 (Microsoft Corporation, Redmond, WA, USA) 을사용하여평균과표준오차를구하고, 통계프로그램 (Statistical Analysis System version 9.2, SAS Institute, Cary, NC., USA) 을이용하여 ANOVA (Analysis of variance) 및 DMRT (Duncan's multiple range test) 를수행하였으며, 5% 유의수준에서각처리간의유의성을비교하였다. 결과및고찰종자의형태학적특성및활력검증늘어진장대종자는원형 계란형 (circular obovate) 에날개가붙어있는형태였으며, 종피는진갈색이다 (Fig. 1A). 크기는길이 1.25±0.020 mm, 너비 1.47±0.011 mm이고, 천립중은 320.96±8.151 mg이었다. 종자의단면을확인한결과, 배는굴곡형 (bent type) 이었으며, 무배유종자로 TZ 테스트에서모두배가전체적으로붉은색으로염색되었고배의성숙과발달상태가매우양호하여활력이있는것으로판단되었다 (Fig. 1B). 발아적온및온도조건에의한발아특성늘어진장대종자는종피가열개된후유근이먼저돌출하는유형으로발아하였다 (Fig. 2). 명조건 (24시간) 에서온도조건을달리하여실험한결과, 20 에서가장먼저발아가시작되었다. 발아는파종 5일후부터시작되었으며, 발아율은 3.9% 였으며, 최종발아율또한 20 에서가장높았으며, 파종 16일후 27.4% 발아되었다 (Fig. 3). 최종발아율은 20 와 25 에서통계적으로유의차가없었으나, 20 에서발아소요기간이가장짧고, 발아속도가가장빠르며, 발아균일도가낮았다. 발아가빨라져전체적인발아일수가짧아지고발아균일도가낮아지는경우에는파종포지에서효율적인생산관리가가능하므로 (Jeon et al., 2013), 발아효율을고려하여늘어진장대의발아적온을 20 로판단하였다 (Table 1). 온도는종자발아율및발아속도에영향을미치는중요한요인으로 (Washitani, 1985; Phartyal et al., 2003), 종자는일반적으로 15 30 의범위에서발아하는것으로알려져있고야생식물은재배종보다발아적온이낮은경향을보이는것으로보고되어있다 (Copeland and McDonald, 2001). 본연구에서 A B C Fig. 1. Seed morphology and viability by tetrazolium test of Arabis pendula. (A) Seed morphology, Bar, 0.5 mm, (B) Median longitudinal section through the seed after TZ test, (C) Tetrazolium-stained embryo. All bars = 0.5 mm. -52-
A B Fig. 2. Seed germination of A. pendula. (A) Sowing after 7 days at 20 with water pre-treatment, (B) Sowing after 7 days at 20 with GA3 500 mg /L pre-treatment. All bars = 0.5 mm. Table 1. Effects of temperature on the seed germination percentage, time, speed and uniformity in A. pendula Temperature ( ) GP z MGT y v GSI x GU w T 50 (%) 15 N.G u N.G N.G N.G N.G 20 27.3 a t) 9.8 b 3.2 a 12.5 b 9.3 b 25 26.7 a 12.1 a 2.7 b 20.7 a 12.0 a 30 16.0 b 10.3 b 1.8 c 13.2 b 9.1 b z GP: Germination percentage of 20 days after sowing. y MGT: Mean germination time. x GSI: Germination speed index. w GU: Germination uniformity. v T 50 : The time to 50% of final germination. u NG: Not germinated. t Data represent the means ± S.D (n=3). Different letters are significantly different according to Duncan s Multiple Range Test (p < 0.05). 늘어진장대는저온 (15 ) 에서는발아하지않았으며, 20 25 에서발아율이높았으나발아속도는 20 에서가장빠르고온도가상승할수록발아속도는낮아지는것으로나타났다. Fig. 3. Effect of temperature on the germination of A. pendula under continuous fluorescent light for 30 day. Bars represent means±sd, n=3 replicates, each with 50 seeds. 광조건에따른발아율및발아특성광조건에의한늘어진장대종자의발아특성을확인하고자 20 에서파종후 24시간광조건또는암조건에서발아특성을분석하였다. 늘어진장대종자는암조건에서발아율이높았으며, 특히파종 5일에서 10일사이에다수의종자가발아되었다 (Fig. 4). 암조건의최종발아율은 49.7% 로명조건보다발아율이 1.8배높았다. 특히암조건에서는발아속도가높고종자가 -53-
韓資植誌 Korean J. Plant Res. 30(1) : 50~57(2017) Table 2. Effects of light on the seed germination percentage, time, speed and uniformity in A. pendula GP z MGT y v GSI x GU w T 50 (%) Light 27.0 a u 9.8 a 3.2 b 12.5 a 8.5 a Dark 49.7 b 8.0 b 6.9 a 6.6 b 6.5 b z GP: Germination percentage of 20 days after sowing. y MGT: Mean germination time. x GSI: Germination speed index. w GU: Germination uniformity. v T 50 : The time to 50% of final germination. u Data represent the means ± S.D (n=3). Different letters are significantly different according to Duncan s Multiple Range Test (p < 0.05). Fig. 4. Effect of light on the germination of A. pendula. Bars represent means ± SD, n=3 replicates, each with 50 seeds. 짧은기간에동시에발아하여발아효율이우수한것으로확인되었다 (Table 2). 그러나발아된유묘의줄기가길고가늘게웃자랐으며, 자엽크기가명조건에서자란종자보다작아유묘의생육이비정상적인것으로판단되었다 (Fig. 5). 일반적으로야생종자는호광성으로알려져있으나 (Park and Chung, 1996), 식물종에따라발아에필요한광요구도가다르며일부야생종자및다수의재배종은광조건에노출될경우발아가억제되는것으로보고되어있다 (Rollin, 1972). 광에의해발아가억제되는혐광성종자는 GA 3 처리를통해광조건에서발아율이회복되기도하는데 (Chen and Chang, 1972; Macchia et al., 2001; Yamaguchi and Kamiya, 2002), 본연구에서는늘어진장대종자의 GA 3 처리를통한발아율및발아효율향상을확인하고자 GA 3 처리농도를달리하여발아시험을수행하였다. Fig. 5. The seedling of A. pendula grown under light (left) and dark conditions (right) at 20 sowing after 10 days. In dark conditions, seedling showed long and emaciated stems with tiny yellow cotyledon. Bar, 5 mm. GA 3 처리에의한발아촉진및발아특성 Gibberellic acid (GA 3 ) 는종자휴면을제어하고발아를유도하는주요호르몬으로 (Iglesias and Babiano, 1997), 종자내가수분해효소인 α-amylase 활성을증진시켜배의발달을촉진하고발아를유도하는것으로알려져있다 (Abeles, 1986; Kim et al., 2009; Kim and Lee, 2013). GA 3 에의해발아가촉진되는종자의유형으로는광발아종자, 암발아종자, 층적처리가필요한종자및후숙이필요한종자등다양한유형의종자가 GA 3 에의해발아가촉진되며 (Chen and Chang, 1972), GA 3 의발아촉진효과는예냉처리를동시에처리했을때상승하는것으로알려져있다 (Bretzloff and Pellett, 1979). 늘어진장대종자를 24시간동안다양한농도의 GA 3 희석액 (0, 200, 500, 1,000 mg /L) 에침지후 (4, 암조건 ) 파종한결과, -54-
Table 3. Effects of gibberellic acid (GA 3 ) treatment on the seed germination percentage, time, speed and uniformity in A. pendula GA 3 GP z MGT y v GSI x GU w T 50 ( mg /L) (%) 0 27.4 d u 9.8 a 3.3 d 12.5 a 8.3 a 200 61.7 c 8.5 b 6.8 c 8.2 b 8.8 a 500 97.3 a 8.1 b 13.6 a 7.8 b 6.6 b 1,000 85.0 b 8.4 b 12.0 b 11.7 a 7.6 ab z GP: Germination percentage of 20 days after sowing. y MGT: Mean germination time. x GSI: Germination speed index. w GU: Germination uniformity. v T 50: The time to 50% of final germination. u Data represent the means ± S.D (n=3). Different letters are significantly different according to Duncan s Multiple Range Test (p < 0.05). 물에적합한처리조건과방법에따라적용할필요가있다. 본연구에서늘어진장대종자는종피에날개가있는구형또는심방형의무배유종자이며, 4 에서 2년이상저장하여도활력이유지되었다 (Fig. 1B, Tables 1-3). 발아적온은 20 였으며, GA 3 희석액 (500 mg /L) 에침지 ( 암조건, 4, 24시간 ) 후 2 0 광조건에서파종했을때 97.3% 발아하여발아율이가장높았으며, 발아속도또한향상되어효율적인생산관리가가능할것으로판단되었다. Fig. 6. Effect of GA 3 on the germination of A. pendula. Bars represent means ± SD (n=3), each with 50 seeds. GA 3 처리에의하여발아세와발아율이증가되었으며, GA 3 500 mg /L 에서최종발아율이 97.3% 로가장높았다 (Fig. 6). GA 3 500 mg /L 처리는발아속도와발아동시성도크게향상시켰으며, 평균발아소요기간이 8일내외로암조건에파종했을때와같은기간동안 2배이상의유묘를생산할수있었다 (Table 3). 또한 GA 3 는유묘의자엽분화및뿌리생육을촉진하여유묘의생육에도효과적이었다. GA 3 는대체로종자발아를향상시키지만식물종과처리농도에따라발아에미치는영향이크게다르다. 숙은노루오줌 (Astilbe koreana) 종자와눈잣나무 (Pinus pumila) 종자는 GA 3 침지처리에의하여발아가촉진되어본연구와유사한경향을보였다 (Lim et al., 2015; Jang et al., 2016). 반면배풍등 (Solanum lyratum) 종자는저농도 (10 mg /L) 에서발아가촉진되었으나, 처리농도가증가할수록발아가억제되었다 (Lee et al., 2014). 따라서종자발아율및발아속도증진을위해서는각식 적요본연구는식의약소재로활용가치가높은늘어진장대 (Arabis pendula L.) 의대량증식을위한기초연구로종자의형태적특성과발아특성을규명하고, 최적발아조건을규명하고자수행되었다. 늘어진장대종자는종피에날개가있는원형으로색은진갈색이며, 너비 1.47 mm, 길이 1.25 mm이고, 배는굴곡형이며무배유종자이다. 온도와광조건실험에서, 종자발아는 20, 암조건에서가장우수하였다 (49.7%). 그러나암조건에서는유묘가약하고웃자랐으며, 자엽이작고말려있는특징을보였다. 종자발아와유묘의생육을촉진하기위하여종자를다양한농도의 GA 3 희석액 (0, 200, 500, 1,000 mg /L) 에침지하여전처리한후파종하였다. 연구결과, GA 3 희석액 500 mg /L 처리구에서종자발아율이가장높고 (97.3%), 발아소요기간이짧았으며 (8.1일), 유묘의생육또한왕성하였다. 연구의결과, 늘어진장대종자의최적발아조건은 GA 3 500 mg /L희석액에침지처리 (4, dark, 24시간 ) 하여 20 에파종하 -55-
韓資植誌 Korean J. Plant Res. 30(1) : 50~57(2017) 여 15일동안발아시키는것으로확인되었다. 사사본연구는국립생물자원관 국가야생식물종자확보및은행운영 (NIBR201619101) 사업의지원을받아수행되었습니다. References Abeles, F.B. 1986. Role of ethylene in Lactuca sativa cv Grand Rapid seed germination. Plant Physiol. 81:780-787. Bloom, C.T., C.C. Baskin and J.M. Baskin 1990. Germination ecology of the facultative biennial Arabis laevigata variety laevigata. American Midl. Nat. 124:214-230. Bretzloff, L.V. and N.W. Pellett. 1979. Effect of stratification and gibberllic acid on the germination of Carpinus caroliniana Walt (angiosperm trees). HortScience 14:621-622. Chen, S.S.C. and J.L.L Chang. 1972. Does gibberellic acid stimulate seed germination via amylase synthesis?. Plant Physiol. 49:441-442. Coolbear, P., A. Francis and D. Grierson. 1984. The effect of low temperature pre-sowing treatment on the germination performance and membrane integrity of artificially aged tomato seeds. J. Experim. Bot. 35:1609-1617. Copeland, L.O. and M. McDonald. 2001. Principles of Seed Science and Technology. Springer Science Business Media LLC, New York, USA. p. 78. Gavassi, M.A., G.C. Fernandes, C.C. Monteiro1, L.E.P. Peres and R.F. Carvalho 2014. Seed germination in tomato: A focus on interaction between phytochromes and gibberellins or abscisic acid. American J. Plant Sci. 5:2163-2169. Iglesias, R.G. and M.J. Babiano. 1997. Endogenous abscisic acid during the germination of chickpea seed. Physiol. Plant 100:500-504. International Seed Testing Association (ISTA). 2014. International Rules for Seed Testing. Chapter 6: The tetrazolium test. International Seed Testing Association. Bassersdorf, Switzerland. pp. 1-25. Jang, B.K., J.S. Cho and C.H. Lee. 2016. Effect of environmental conditions and chemical treatments on seed germination of Astilbe koreana (Kom.) Nakai. Korean J. Plant Res. 29:235-240 (in Korean). Jeon, K.S., K.S. Song, C.H. Kim, J.H. Yoon and J.J. Kim. 2013. Effects of seed pre-treatment and germination environments on germination characteristics of Ligularia fischeri seeds. Protected Hort. Plant Fact. 22:262-269 (in Korean). Kim, D.H., B.J. Ahn, H.J. Ahn, Y.S. Ahn, Y.G. Kim, C.G. Park, C.B. Park, S.W. Cha and B.H. Song. 2014. Studies on seed germination characteristics and patterns of protein expression of Lithospermum erythrorhizon by plant growth regulators and seed primings. Korean J. Med. Crop Sci. 22:435-441 (in Korean). Kim, Y.H. and I.J. Lee. 2013. Influence of plant growth regulator application on seed germination of dandelion (Taraxacum officinale). Weed & Turfgrass Science 2:152-158 (in Korean). Kim, Y.H., M. Hamayun, A.L..Khan, C.I. Na, S.M. Kang, H.H, Han and I. Lee. 2009. Exogenous application of plant growth regulators increased the total flavonoid content in Taraxacum officinale Wigg. African J. Biotech. 8:5727-5732. Koornneef, M., L. Bentsink and H. Hilhorst. 2002. Seed dormancy and germination. Curr. Opin. Plant Biol. 5:33-36. Lee, S.G., H.Y. Kim and J.J. Ku. 2014. Effects of seed storage methods and GA 3 application on seed germination and seedling growth of Solanum lyratum Thunb. Korean J. Plant Res. 27:365-370 (in Korean). Lee, W.C. 1996. Coloured Standard Illustrations of Korean Plants. Academy Publishing Co., Seoul, Korea. p. 350 (in Korean). Lim, H.I., G.N. Kim, K.H. Jang and W.G. Park. 2015. Effect of wet cold and gibberellin treatments on germination of dwarf stone pine seeds. Korean J. Plant Res. 28:253-258 (in Korean). Macchia, M., L.G. Angelini and L. Ceccarini. 2001. Study on germination characteristics of Phacelia tanacetifolia Bentham seeds obtained from different growing seasons. Italian J. Agron. 4:61-66. Millennium Seed Bank (MSB). 2016. UK germination toolbox. Royal Botanic Gardens, Royal botanic gardens, Kew. http://www.kew.org/science-conservation/research-data/res ources/uk-germination-toolbox (cited by November 07, 2016). Ministry of Food and Drug Safety (MFDS). 2016. Standards and Specifications for Food Contact Materials: Attached Table 1. Osong, Korea. pp. 25-96 (in Korean). Moon, G.S. 1999. Constituents and Uses of Medicinal Herbs. Ilweolseogak. Seoul, Korea. p. 322 (in Korean). Nanjing University of Traditional Chinese Medicine (NUTCM). 2006. Dictionary of Traditional Chinese Medicine ( 中药大辞 -56-
典 ). Shanghai Science and Technology Press, Shanhai, China (in Chinese). National Bureau of Traditional Chinese Medicine Editorial Board (NBTCMEB). 1999. Chinese Materia Medica ( 中华本草 ). Shanghai Science and Technology Publishers. Shanhai, China (in Chinese). National Institute of Biological Resources (NIBR). 2011. National List of Species of Korea. National Institute of Biological Resources. Incheon, Korea. pp. 168-169 (in Korean). Park, Y.J. and Y.O. Chung. 1996. Studies on the characteristics of seed germination of Lycoris genera. Korean J. Med. Crop Sci. 4:163-171 (in Korean). Phartyal, S.S., R.C. Thapliyal, J.S. Nayal, M.M.S. Raeat and G. Joshi. 2003. The influences of temperatures on seed germination rate in Himalayan elm (Ulmus wallichiana). Seed Sci. Technol. 31:83-93. Rollin, P. 1972. Phytochrome Control of Seed Germination. In Mitrakos, K. and W. Shropshire Jr. (eds.), Phytochromes, Academic Press, New York, USA. pp. 229-254. Washitani, I. 1985. Germination-rate dependency on temperature of Geranium carilinianum seeds. J. Exp. Bot. 36:330-337. Yamaguchi, S. and Y. Kamiya. 2002. Gibberellins and light-stimulated seed germination. J. Plant Growth Regul. 20:369-376. Yoo, B.K., K. Kwon, Y.H. Ko, H.G. Kim, S. Lee, K.H. Park and O.Y. Kwon. 2016. Screening of natural product libraries for the extension of cell life-span through immune system. J. Life Sci. 26:359-363 (in Korean). (Received 16 August 2016 ; Revised 9 November 2016 ; Accepted 7 December 2016) -57-