Journal of Radiation Industry 3 (3) : 227~232 (2009) 감마선조사에의한다양한스프레이국화품종의돌연변이육종 김상훈 정성진 이긍주 1 김동섭 김진백 강시용 * 한국원자력연구원방사선과학연구소방사선식품육종연구부 1 목포대학교자연과학대학생명과학부 Mutation reeding of Various Spray Chrysanthemum Cultivars by Gamma-ray Irradiation Sang Hoon Kim, Sung Jin Chung, Geung-Joo Lee 1, Dong Sub Kim, Jin-aek Kim and Si-Yong Kang* Division of Food Irradiation and Radiation reeding, dvanced Radiation Technology Institute, Korea tomic Energy Research Institute, Jeongeup 580-185, Korea 1 College of Natural Science, Mokpo National University, Muan 534-830, Korea bstract - This study was carried out to analyze mutation rate and spectrum of chrysanthemum by gamma-ray irradiation. Five flower types, 16 cultivars including Geumsu, Hwiparam, Ilwol, Magic, Moonlight, Noblewine, Pinky, Progy, Sangtte, Waterfog, Whitneypangpang, Yelloweye, Yellowpangpang, Yesmiso, Yesmorning, and Yestogether were irradiated as 30, 50, and 70 Gy dose during 24 hours. s a result, mutation rate was identified as the highest in single type among five flower types, but there was a little difference according to cultivars. Mutation rate was increased in proportion to irradiation dose in anemone, pompon, and spider type cultivars, but there wasn t clear in single and semi-double type cultivars. Mutation spectrum was identified as the highest in the cultivar Noblewine. The most sensitive cultivars to radiation were revealed as Noblewine and Yesmorning and the least were Moonlight, Waterfog, and Yellowpangpang. Consequently, there was much difference in radio-sensitivity according to cultivars of chrysanthemum and flower type was correlated a little with mutation rate. Key words : Chrysanthemum, Flower type, Gamma ray, Mutation breeding, Radio-sensitivity 서 론 국화 (Chrysanthemum; Dendranthema grandiflorum) 는 The first two authors contributed equally to this work. * Corresponding authors: Si-Yong Kang, Tel. +82-63-570-3310, Fax. +82-63-570-3319, E-mail. sykang@kaeri.re.kr 전세계3대절화류에속하며, 국내에서도장미다음으로생산량이높은중요한화훼작물로최근에는분화로서의이용도증가되고있는추세이다 (MIFFF 2005). 또한다양한꽃의색깔과형태등으로인하여국내절화류시장의 1/4 가량을차지하고있으나재배및국내에서유통되고있는대부분이네덜란드나일본등에서육성된외국품종으로막대한로열티의증가는화훼재배농가에 227
228 김상훈 정성진 이긍주 김동섭 김진백 강시용 큰부담으로작용하고있다. 하지만최근들어로열티부담의경감과고부가가치국산신품종의개발및보급의필요성에따라다양한국내육성품종들의품종보호출원이증가되고있는추세이다 (Lim et al. 2008). 그러나국내에서수행되는국화의신품종개발은주로교배육종에의해이뤄졌으며, 교배육종의보조수단혹은교배육종으로불가능한품종에한해돌연변이육종을통한신품종개발이이뤄지고있다 (Schum and Preil 1998; Goo et al. 2003). 돌연변이육종은 colchicine이나 ethyl methanesulphonate (EMS) 등의화학적돌연변이원또는감마선이나 X선등의방사선을처리하여돌연변이를유기한다. 이러한돌연변이육종은열성유전자를이용하거나단백질함량등특수성분을개량할경우또는교배육종의한계성을해결하려고할경우에이용되며, 화훼류에서는다양한화색을통한신품종의시리즈화를위해주로이용되고있다 (Schum and Preil 1998). FO/IE 돌연변이품종데이터베이스 (http://www-mvd.iaea.org/mvd/ Default.htm) 에의하면국화를포함한화훼류가약 570 여품종이등록되어있으며, 종자번식작물다음으로 2 위를차지하고있다. 화훼류중에서도국화가차지하는비중이전체품종의 40% 이상을차지하고있으며, 네덜란드의경우돌연변이체나이를교배모본으로활용하여육성한품종이화훼시장의 30~40% 를차지할정도로화훼류의신품종을육종하는데돌연변이육종방법이중요한역할을하고있다고할수있다 (Park and Song 2005; Park et al. 2007). 따라서본연구에서는다양한스프레이국화품종에방사선처리를통한돌연변이체발생빈도를조사하여품종별적정선량을탐색하고, 유용한돌연변이체는선발, 증식및세대진전을거쳐신품종으로개발하는것을목적으로하고있다. 재료및방법실험에사용한재료로는 5가지화형의스프레이국화 Geumsu, Hwiparam, Ilwol, Magic, Moonlight, Noblewine, Pinky, Progy, Sangtte, Waterfog, Whitneypangpang, Yelloweye, Yellowpangpang, Yesmiso, Yesmorning, Yestogether 의 16개품종을대상으로 2008년 7월과 8월에각각방사선 ( 감마선 ) 조사를실시하였다. 방사선조사는한국원자력연구원방사선과학연구소내저준위감마선조사시설 ( 60 Co) 을이용하여국화삽수체에 30, 50, 70 Gy 선량으로 24시간동안실시하 였다. 선량별방사선조사개체수는품종에따라 100 ~300개정도의삽수체를대상으로실시하였다 (Table 1). 조사된삽수체는한국원자력연구원방사선과학연구소방사선육종시험장포장에정식하였으며, 방사선감수성조사는돌연변이발생률그리고화색및화형돌연변이스펙트럼등을대상으로실시하였다. 돌연변이체선발은방사선조사후개화시기에실시하였다. 결과및논의기존연구결과에서국화의돌연변이유기를위한적정조사선량인반치사선량 (LD 50) 이국화5개품종을이용하여실험한결과스프레이계통의경우 30 Gy 정도, 스탠다드계통의경우 40 Gy 정도로밝혀졌으며, 품종에따라약간의차이가나는것으로보고되었다 (Goo et al. 2003). 스프레이국화 rgus 의경우돌연변이유기를위하여배양체에 30~50 Gy를조사하였으며그결과조사선량에관계없이 97% 이상의높은생존율이확인되었고 50 Gy 조사구에서가장높은변이율이확인되었다 (Park et al. 2007). 그리고국화 Taihei 를이용하여총조사선량과단위시간당조사선량이돌연변이유기와 DN 함량에미치는영향을보기위하여 15~60 Gy 선량으로조사가되었으며돌연변이율은총조사선량에영향을받는것으로확인되었다 (Yamaguchi et al. 2008). 따라서본연구에서는기존연구를바탕으로서로다른 5 가지화형의 16개국화품종을이용하여 30, 50, 70 Gy 를조사선량으로선정하여조사를하였고개화후품종, 꽃의형태, 조사선량별로방사선감수성과돌연변이율, 변이스펙트럼을조사하였다. 그결과품종, 꽃의형태, 조사선량별로다양한돌연변이율과변이스펙트럼이확인되었다 (Table 1). 꽃의형태에따른돌연변이율은홑꽃형이반겹꽃형, 아네모네형, 폼폰형, 스파이더형에비하여상대적으로높은것으로보이며품종에따라다소차이는있는것으로판단된다. 홑꽃형, 반겹꽃형, 폼폰형에서는주로화색변이의돌연변이가관찰되었으나, 아네모네형과스파이더형에서는화색변이와화형변이가같이관찰되었다. 품종별로는 Yesmorning 이 70 Gy 조사선량에서약 30%, Noblewine 이 30 Gy 조사선량에서약 22% 의돌연변이체가확인되었다 (Table 1). 반면 Moonlight, Waterfog, Yellowpangpang 의경우모든조사구에서변이체가전혀발견되지않아방사선감수성이가장낮은것으로관찰되었으며, 돌연변이유기를위해서는보다높은선량이조사되어야할것으로판단된다. 조
감마선이용국화돌연변이육종 229 Table 1. Mutation frequency and type on chrysanthemum according to cultivar and irradiation dose Frequency of flower mutation (%) Flower type Cultivar 30 Gy 50 Gy 70 Gy Type of flower mutation Single Noblewine 21.9 (28/128) x - y NT z Color Single Yesmorning 13.0 (19/146) 13.8 (42/304) 29.9 (43/144) Color Single Hwiparam 11.4 (17/149) 6.7 (21/312) 16.5 (18/109) Color Single Moonlight 0.0 (0/149) 0.0 (0/309) 0.0 (0/155) - Semi-double Magic 7.8 (10/128) 1.6 (2/128) NT Color Semi-double Sangtte 4.2 (6/144) 0.0 (0/142) NT Color Semi-double Pinky 2.3 (3/128) 6.0 (15/251) 4.7 (6/128) Color Semi-double Yelloweye 1.6 (2/128) 1.6 (2/128) NT Color Semi-double Waterfog 0.0 (0/162) 0.0 (0/323) 0.0 (0/161) - nemone Ilwol 0.7 (1/152) 1.3 (4/316) 3.5 (5/144) Color, shape nemone Geumsu 0.0 (0/144) - NT - Pompon Progy 0.0 (0/152) 1.0 (3/309) 4.4 (7/160) Color Pompon Whitneypangpang 0.0 (0/129) 0.0 (0/295) 8.1 (10/124) Color Pompon Yesmiso 0.0 (0/144) 0.0 (0/296) 1.9 (3/160) Color Pompon Yellowpangpang 0.0 (0/123) 0.0 (0/241) 0.0 (0/101) - Spider Yestogether 0.7 (1/149) 3.9 (12/312) 5.8 (9/155) Color, shape x Number of flower mutants/number of investigated plants are shown in parentheses. y Gamma-ray is irradiated, but not scored because of growing deterioration. z Not tested Table 2. Mutation frequency on chrysanthemum according to flower type and irradiation dose Dose (Gy) Flower type 30 50 70 Single 11.6±9.0 x 6.8±6.9 15.5±15.0 Semi-double 3.2±3.0 1.8±2.5 2.4±3.3 nemone 0.4±0.5 1.3 3.5 Pompon 0.0±0.0 0.3±0.5 3.6±3.5 Spider 0.7 3.9 5.8 x Mean±standard deviation 사선량에따라서는아네모네형, 폼폰형, 스파이더형이조사선량이증가함에따라돌연변이율이증가하는경향이있으나홑꽃형과반겹꽃형의경우뚜렷한경향치가확인되지않았고품종별로다소차이가있는것으로확인되었다 (Table 2). 이는조사선량이증가할수록돌연변이율이반드시비례적으로높아지는것은아니라는것을의미하며이러한결과는기존의연구결과와도일치한다 (Goo et al. 2003; Yamaguchi et al. 2008). 품종별변이스펙트럼비율의경우 Noblewine 이 5.46% 로가장높은변이스펙트럼이확인되었으며, 이는변이의폭이넓은것으로육종과정에서다양한변이체의선발효율이가장높다는것을의미한다 (Table 3). 주요품종별변이의종류는다음과같다. 홑꽃화형인 Noblewine 의경우대조구의설상화가연핑크바탕에자주줄무늬가있는것이특징인데반해변이체의경우노랑바탕에자주줄무늬, 빨강등의설상화화색변이가확인되었다 (Fig. Table 3. Mutation spectrum of chrysanthemum cultivars by gammaray irradiation Cultivar Flower type Mutation spectrum (%) Noblewine Single 5.46 x Yesmorning Single 0.84 Magic Semi-double 0.78 Pinky Semi-double 0.78 Sangtte Semi-double 0.69 Progy Pompon 0.64 Yelloweye Semi-double 0.39 Hwiparam Single 0.35 Yestogether Spider 0.32 Ilwol nemone 0.32 Whitneypangpang Pompon 0.18 Yesmiso Pompon 0.16 Moonlight Single 0.00 Waterfog Semi-double 0.00 Geumsu nemone 0.00 Yellowpangpang Pompon 0.00 x It is calculated using number of different kind of mutants and several same color or shape mutants are considered as only one. 1). 복색반겹꽃화형인 Magic 의경우대조구의설상화가자주색, 흰색혼합인것이특징인데반해변이체의경우연자주, 노랑혼합, 다홍등의설상화화색변이가확인되었다 (Fig. 2). 아네모네화형인 Ilwol 의경우대조구의설상화가노랑색, 관상화가노랑색바탕에빨강색중심이특징이지만변이체의경우빨강색, 노랑색혼합의관상화변이체와다홍색, 빨강색의설상화변이체가확인되었다 (Fig. 3). 그리고폼폰화형인 Progy 의경우대조구의설상화가연두색인것이특징인데반해변
230 김상훈 정성진 이긍주 김동섭 김진백 강시용 C D E F G H Fig. 1. Mutation spectrum of flower color induced by gamma-ray irradiation in single type chrysanthemum Noblewine. Left. Original (wild) type. ~H, mutants in flower color:, color (stripe);, color (not stripe); C, color (purple); D and F, color (yellow); E, color (red); G and H, color (chimera). C D E F Fig. 2. Mutation spectrum of flower color induced by gamma-ray irradiation in semi-double type chrysanthemum Magic. Left. Original(wild) type. ~F, mutants in flower color:, color (white and yellow);, color (dark red); C, color (yellow); D, color (dark purple and yellow); E, color (dark purple); F, color (yellow and purple).
감마선이용국화돌연변이육종 231 이체의경우노랑, 연두혼합, 흰색등의설상화화색변이가확인되었다 (Fig. 4). 스파이더화형인 Yestogether 의경우대조구의설상화가연분홍, 관상화가아이보리, 연두혼합이특징인데반해변이체의경우설상화와관상화에서화색변이와화형변이가같이확인되었다 (Fig. 5). 이러한결과들은기존의 rgus 의돌연변이연구에서관상화의화색변이가설상화보다다양하게확인된결과와상충되는결과이다 (Park et al. 2007). 하지만감마선조사시화형변이보다는화색변이가많이발생한것은기존연구결과와일치한다 (roertjes and van Horten 1998; Goo et al. 2003; Park et al. 2007). 본연구결과에서는국화품종별돌연변이를유기하기위한적정선량을일부품종에서확인할수있었으며, 품종별방사선감수성의차이를확인할수있었다. 또한다양한화색변이체를선발함으로써국화신품종의시리즈화가가능할것으로판단된다. 결 론 Fig. 3. Mutation spectrum of flower color and shape induced by gamma-ray irradiation in anemone type chrysanthemum Ilwol. Left. Original (wild) type. ~C, mutants in flower color and shape:, color and shape (yellow);, color (red); C, color (chimera). C 본연구는다양한국화품종에서감마선조사선량에따른돌연변이율과변이스펙트럼을조사하고품종별적정조사선량을확립하기위하여수행되었다. 그결과품종, 꽃의형태, 조사선량에따라다양한돌연변이율과변이스펙트럼이확인되었다. 꽃의형태에따른돌연변이율은홑꽃형이반겹꽃형, 아네모네형, 폼폰형, 스파이더형에비하여상대적으로높은것으로확인되었으나품종별로다소차이를보이는것이확인되었다. 조사선량에따라서는아네모네형, 폼폰형, 스파이더형의경우조사선량이증가함에따라돌연변이율이증가하는경향이있으나홑꽃형과반겹꽃형의경우조사선량과돌연변이율과의상관관계가불명확한것으로보인다. 품종별변이스펙트럼의경우꽃의형태에따른경향치는확인되지않았으며, 16개의국화품종중 Noblewine 의변이스펙트럼 C D Fig. 4. Mutation spectrum of flower color induced by gamma-ray irradiation in pompon type chrysanthemum Progy. Left. Original (wild) type. ~D, mutants in flower color:, color (yellow);, color (white); C and D, color (chimera).
232 김상훈 정성진 이긍주 김동섭 김진백 강시용 참고문헌 Fig. 5. Mutation spectrum of flower color and shape induced by gamma-ray irradiation in spider type chrysanthemum Yestogether. Left. Original (wild) type. ~, mutants in flower color and shape:, color (pale pink);, color and shape (yellow). 폭이가장넓은것으로확인되었다. 방사선감수성이가장높은품종은 Noblewine 과 Yesmorning 으로확인되었으며, 가장낮은품종은 Moonlight, Waterfog, Yellowpangpang 으로모든조사구에서변이체가확인되지않았다. 결과적으로국화품종에따라방사선감수성의차이가많이나는것으로판단되며, 꽃의형태또한돌연변이율과상관관계가있는것으로판단된다. 사 본연구는한국원자력연구원기관고유사업및농촌진흥청바이오그린 21사업 (Code 20070301034033) 의연구비지원에의해수행되었으며, 이에감사드립니다. 사 농림수산식품부 (MIFFF). 2009. 2005 년화훼류재배현황보고. roertjes E and van Horten M. 1998. pplied mutation breeding for vegetatively propagated crops. Elsevier, rmsterdam, The Netherlands. Goo DH, Yae W, Song HS, Park IS, Han H and Yu HJ. 2003. Color change in chrysanthemum flower by gamma ray irradiation. J. Kor. Soc. Hort. Sci. 44(6):1006-1009. Hwang JC, Chin YD, Chung YM, Kim SG and Jeong R. 2009. new spray chrysanthemum cultivar, Noble Wine with good color, single type and bi-color petals for cut flower. Flower Res. J. 17(2):137-140. The FO-IE Mutant Varieties Database. http://www-mvd. iaea.org/mvd/default.htm. 2004. Lim JH, Shin HK, Park SK, Cho HR, Rhee HK, Kim MS and Joung HY. 2008. new spray chrysanthemum cultivar, Forest roma, an anemone flower type with vigorous and large, green center and white petals for cut flower. Kor. J. Hort. Sci. Technol. 26(Suppl. II):108-111. Park IS, Lee GJ, Kim DS, Chung SJ, Kim J, Song HS, Goo DH and Kang SY. 2007. Mutation breeding of a spray chrysanthemum rgus by gamma-ray irradiation and tissue culture. Flower Res. J. 15(1):52-57. Park IS and Song HS. 2005. Current trends of mutation breeding by radiation technology at domestic and foreign ornamentals. J. Kor. Flower Res. Soc. 13(3):184-200. Schum and Preil W. 1998. Induced mutations in ornamental plants. Somaclonal variation and induced mutations in crop improvement. p. 332-366. Kluwer cademic Publ., Dordrecht, The Netherlands. Yamaguchi H, Shimizu, Degi K and Morishita T. 2008. Effects of dose and dose rate of gamma ray irradiation on mutation induction and nuclear DN content in chrysanthemum. reeding Science 58:331-335. Manuscript Received: September 25, 2009 Revision ccepted: September 30, 2009