보이젠베리(Rubus ursinus)의 휴면아 절편체로부터 다경유도를 통한 식물체 재분화

조란,김효진,조한직,김이엽,오승철 한국원예학회 2016 한국원예학회 학술발표요지 Vol.2016 No.5

섬시호(Bupleurum latissimum Nakai)의 조직배양을 통한 대량생산

김효진,조한직,김이엽,김무열,박학봉 한국자원식물학회 2007 한국자원식물학회지 Vol.20 No.4

- This study was carried out to establish the micropropagation system of Bupleurum latissimumNakai that is aKorean native endangered species. Callus were induced from the leaf, petiole and floral bud and the percentage of callusformation was highest in the floral bud on the MS medium containing 2.0 mg· L-1 2,4-D. Especially, callus inducedfrom floral bud was formed 77.8% and the percentage of shoot formation was 42.6% on the MS medium containing 2.0mg· L-12,4-D plus 1.0 mg· L-1TDZ. For simultaneously callus formation and shoot regeneration, 1/2 MS medium wasmore effective than MS medium. The percentage callus formation, shoot regeneration and rooting were 46.3%, 13.0%,13.0% in 1/2 MS medium, respectively. Soot regeneration from callus was good in 1/2 MS medium supplemented with2.0 mg· L-12,4-D plus 1.0 mg· L-1BA where percentage of shoot regeneration was 74.1%, and the number of shoot perexplant was 2.4. The percentage of rooting was lowest (57.8%) in control while it was highest (97.8%) in 1.5 mg· L-1NAA. In acclimatization of regenerated plantlets, the percentage of survived plantlets was highest (86.1%), and plantheight, root length and fresh weight were good in the soil for horticulture.Key words - Bupleurum latissimum, Callus, Shoot, Regeneration, Propagation韓資植誌 Korean J. Plant Res. 20(4):367∼374(2007)한 거의 이루어진 적이 없다.따라서 본 연구는 멸종 위기에 처한 섬시호의 대량 증식을 위해 조직배양을 통한 번식체계를 확립하고자 적합한 배양 절편체를 선정하고 선정된 절편체로부터 캘러스 유도, 식물체 재분화,발근에 효과적인 식물생장조절물질의 종류와 농도를 구명하며재분화 식물체의 생존율을 향상시킬 수 있는 순화용토를 선발하고자 하였다.재료 및 방법식물재료본 실험에서 사용한 섬시호는 종자를 2005년 8월 환경부 서식지외 보전기관인 기청산식물원에서 채취하여 전북대학교 온실에서 발아시킨 후 실험재료로 사용하였다.치상 절편체 부위 및 생장조절물질에 따른 캘러스 유도캘러스 형성률이 높은 치상 절편체를 찾기 위하여 식물체가약 30cm로 자랐을 때 5∼7번째의 잎과 엽병을 채취하여 잎은 5

가시없는 블랙베리(Rubus fruticosus L. × R. parvifolius L.)의 근맹아를 이용한 다경유도와 절간배양을 통한 식물체증식 조건확립

김효진(HyoJin Kim),조란(Lan Cho),조한직(HanJig Cho),김이엽(EeYoup Kim),오승철(SeungCheol Oh) 한국원예학회 2016 한국원예학회 학술발표요지 Vol.2016 No.5

가시없는 블랙베리(Rubus fruticosus L. cv. BB21)의 근맹아를 이용한 다경유도와 절간배양을 통한 식물체 증식조건의 확립

이강섭,김효진,박대현,오승철,조한직,김이엽,Lee, Kang Seop,Kim, Hyo Jin,Park, Dae Hyun,Oh, Seung Cheol,Cho, Han Jig,Kim, Ee Youb 한국식물생명공학회 2018 식물생명공학회지 Vol.45 No.2

본 연구에 사용한 식물체는 Rubus fruticosus L. cv. BB21은 Rubus fruticosus L. ${\times}$ R. parvifolius L. 교잡 블랙베리 V3품종에서 방사선을 처리하여 돌연변이로 선발 육종된 V9개체 중 다시 방사선을 처리하여 선발육종 된 품종이다. 이러한 우량계통의 신속한 증식을 이루기 위하여 조직 배양을 통한 효과적인 방법을 구명하고자 배지 및 생장조절물질 등의 처리에 따른 안정적인 생산을 위해 본 연구가 수행되었다. 땅속에서 채집된 근맹아 절편체를 0.1, 0.5, 1.0, 2.0 mg/L BAP가 첨가된 MS와 1/2MS 배지에 치상하여 6주간 배양한 후, 다경의 형성률을 조사하였다. 형성된 다경으로부터 산업적으로 이용하기 위한 안정된 생산을 위하여, 신초의 잎을 제거한 후 절간배양을 수행하여 다수의 소식물체를 증식하고자 여러종류의 배지 즉, AD (Anderson 1980), B5 (Gamborg 1968), MS (Murashige and Skoog 1962), SH (Schenk and Hildebrandt 1972), QL (Quoirin and Lepoivre 1977), WPM (Lloyd and McCown 1981)배지와 상기의 각각의 동일배지의 무기염류의 수준을 절반으로 한 1/2배지에 배양하였다. 초기의 생육(30일간 배양 시)에 있어서 생장은 배지 간에 차이를 나타내어 1/2AD, 1/2QL 배지에서 가장 양호한 초장을 나타내었으며, 절간수도 약 3.8, 4.4개를 나타내었으며, 뿌리, 생중량 다경유도 모두 양호하였다. 90일간 배양한 경우에서는 MS배지가 초장이 가장 양호하였으며 AD, SH 순으로 양호한 결과를 나타내었다. 식물체의 토양 순화는 신초와 뿌리가 잘 발달된 정상적인 식물체를 모래, 토양, 그리고 버뮤큘라이트가 혼합된 토양(1:1:1, v/v)에 각각 직접 이식하여 실시한 결과, 식물체의 발근율과 생존률(약98%)을 나타내었다. 이상의 결과를 통하여 블랙베리 묘목의 대량생산이 가능하여 농가에 원활한 공급이 가능할 것으로 예상된다. This study was conducted to develop a simple, rapid, and reliable method for in vitro propagation of disease-free and true-to-type clones from sucker explants of thornless blackberry (Rubus fruticosus L. ${\times}$ R. parvifolius L.). To induce multiple shoots, the sucker explants were sterilized in 1% NaOCl solution, and then were aseptically cultured on the full and 1/2 MS solid medium supplemented with BAP (0.1, 0.5, 1.0, 2.0 mg/L). After six weeks of culture, the highest frequency (85.4%) of shoot formation from sucker explants was obtained on the full-strength MS medium with 1.0 mg/L BAP. Node explants obtained from multiple shoots were cultured on the various media of full- or half-strength of AD, B5, MS, SH, QL, WPM media, respectively. After 30 days of culture, plant growth was good on the half-AD, half-QL medium. After 90 days of culture, plant growth was good on the full MS and full SH medium. The survival rate of the plantlets after transfer to plastic pots containing soil mixture (sand: soil: vermiculite was 1:1:1, vol.) in the greenhouse was 98%. The results indicate that a multiple-shoot procedure can be applied for an efficient mass propagation of Rubus fruticosus L. ${\times}$ R. parvifolius L.

가시없는 블랙베리(Rubus fruticosus L. cv. BB21)의 근맹아를 이용한 다경유도와 절간배양을 통한 식물체 증식조건의 확립

이강섭,김효진,박대현,오승철,조한직,김이엽 한국식물생명공학회 2018 JOURNAL OF PLANT BIOTECHNOLOGY Vol.45 No.2

This study was conducted to develop a simple, rapid, and reliable method for in vitro propagation of disease-free and true-to-type clones from sucker explants of thornless blackberry (Rubus fruticosus L. × R. parvifolius L.). To induce multiple shoots, the sucker explants were sterilized in 1% NaOCl solution, and then were aseptically cultured on the full and 1/2 MS solid medium supplemented with BAP (0.1, 0.5, 1.0, 2.0 mg/L). After six weeks of culture, the highest frequency (85.4%) of shoot formation from sucker explants was obtained on the full-strength MS medium with 1.0 mg/L BAP. Node explants obtained from multiple shoots were cultured on the various media of full- or half-strength of AD, B5, MS, SH, QL, WPM media, respectively. After 30 days of culture, plant growth was good on the half-AD, half-QL medium. After 90 days of culture, plant growth was good on the full MS and full SH medium. The survival rate of the plantlets after transfer to plastic pots containing soil mixture (sand: soil: vermiculite was 1:1:1, vol.) in the greenhouse was 98%. The results indicate that a multiple-shoot procedure can be applied for an efficient mass propagation of Rubus fruticosus L. × R. parvifolius L.

블랙베리(Rubus fructicosus L.) 돌연변이 유전자원의생육특성과 형태학적 변이 분석

류재혁,김동섭,하보근,김진백,김상훈,정일윤,조한직,강시용 한국방사선산업학회 2012 방사선산업학회지 Vol.6 No.3

This study was carried out to evaluate growth, morphological characteristics, andsugar content among fifty six blackberry (Rubus fructicosus) genotypes derived from gamma-raytreatment (fifty two lines), MNU treatments (three lines), and cross breed R. fructicosus×R. parvifolius (one line). While 56 genotypes had similar stem diameter with origin variety, the mostof mutants showed reduced leaf size. Also, they showed a wide range of variation in fruit size andone hundred fruit weight compared with origin variety. In fruit size and sugar content, twomutant lines showed large fruit sizes and ten mutant lines showed 20% higher sugar content thanthe original variety. In the correlation analysis, positive relationships were determined betweensugar content and stem diameter, fruit length, leaf wide and fruit length/fruit wide ratio. However, leaf length/leaf wide ratio showed a negative correlation with sugar content. The resultswill be used as fundamental data for Rubus fructicosus breeding program.

감마선 및 화학적 돌연변이원 처리가 스테비아(Stevia rebaudiana Bert.)의 종자 발아 및 초기 생장에 미치는 영향

윤태영,김이엽,김영호,최진수,현경섭,성윤희,조한직,김동섭,강시용,고정애 한국방사선산업학회 2012 방사선산업학회지 Vol.6 No.2

This study was carried out to develop the improved useful mutants for yield orcomposition of stevia plants using the gamma ray or chemical mutagens treatments. The seeds ofstevia ‘Suwon No. 11’ were irradiated up to 400 Gy of gamma ray. Chemical mutagens weretreated on the seeds of the ‘Suwon No. 11’ using 0.07% colchicine, 10 mM sodium azide, or 10mM NMU for various durations. The germination rate, and shoot and root growth of seedlingwere estimated at 30 days after gamma ray irradiation or chemical mutagen treatment, and theplant height, the number of branches, and leaf length and width were examined at 3 months aftermutagenesis treatments. In the case of gamma ray treatments, the germination rate and earlystagegrowth were decreased as the increase of radiation dose, and the 50% lethal dose was foundto be 200 Gy. the plant height was decreased as the increase of radiation dose, while the numberof branches per plant and leaf length were increased. Leaf shape was modified to the relativelylonger one compared to the control, which was identified more apparently at the treatments ofhigher than 150 Gy. In the treatment of chemical mutagens, the rate of germination and survivalwere decreased as the increase of incubation time. The 50% lethal dose for germination rate wereidentified as the conditions of the 15 hours incubation in 0.07% colchicine, the 4 hrs in 10 mMsodium azide, and the 2 hrs in 10 mM NMU, in the three chemical mutagens treatments. Chemicalmutagens had no influence on shoot growth, while root growth was increased, especially as theincubation time was extended. The highest root growth occurred in the NMU treatment at 6 hrsincubation time. The plant height was decreased as the increase of incubation time in the chemicalmutagens treatments. Among the chemical mutagens, NMU was the most effective to induce themutants with long-shaped or the least lobed leaves.

국내 육성 블랙베리 품종의 생육 및 과실 특성

김은주(Eun Ju Kim),최소라(So Ra Choi),조한직(Han Jik Cho),최창학(Chang Hak Choi),조종현(Chong Hyeon Cho),허병수(Byong Soo Heo),김정만(Jeong Man Kim) 한국원예학회 2016 한국원예학회 학술발표요지 Vol.2016 No.10

블랙베리(Rubus fruticosus L.)의 맹아절편체로부터 다경유도를 통한 식물체 재분화

신정순(Jeong-Sun Shin),심옥경(Ock-Kyeong Sim),이종천(Jong-Chon Lee),조한직(Han-Jik Cho),김이엽(Ee-Yup Kim),이강섭(Kang-Seop Lee) 한국자원식물학회 2005 한국자원식물학회지 Vol.18 No.3

블랙베리의 다경유도를 통한 식물체 재생을 위하여, 노지에서 생육되고 있는 맹아를 채취하여 1.2% NaOCl 용액에 침지하여 표면 살균한 후, kinetin과 BA가 각각 포함된 MS 배지에서 4주간 배양하여 절편당 형성된 신초의 수를 조사하여 BA가 효과적임을 밝혔다. 동일방식으로 맹아를 IBA와 BA를 첨가한 MS 고체배지에서 4주간 배양하여 신초를 유도하였는데 1.0 mg/L BA가 첨가된 배지에서 신초유도는 100% 이루어졌다. 기내배양한 신초의 절편체를 1.0 mg/L BA가 첨가된 MS 배지에 치상하여 형성된 신초의 수는 절편체당 5.3개로 최고를 나타내었다. 약 10주 정도 배양을 더 오래하였을 때, 다경 형성율이 더 높아질 뿐 아니라 절편체당 신초 수도 많아졌다. 형성된 신초로부터 뿌리 발생을 위하여 glycine의 농도를 달리한 (0~2.0mg/L) MS 기본배지에서 4주간 배양하였을 때, 0.5 mg/L glycine을 첨가한 배지에서 고빈도 (85%의 뿌리발생을 관찰 할 수 있었다. 재생된 식물체는 모래 : 토양 : 버뮤큘라이트 (1:1:1, vol.) 혼합토양에서 순화시켜, 토양에 이식하였을 때 95%의 식물체 생존율을 나타내었다. This study was carried out to induce plant regeneration via shoot formation from sucker explants of Rubus fruticosus L. To induce adventitious shoots, sucker explants were sterilized in 1.2% NaOCl solution, and cultured on the MS solid medium supplemented with kinetin (0.5, 1.0, 3.0 mg/L) and BA (0.5, 1.0, 3.0 mg/L), respectively. As above, to induce adventitious shoots, sucker explants were cultured on the MS solid medium supplemented with IBA (0, 0.1, 1.0 mg/L) and BA (0, 0.1, 1.0, 2.0 mg/L). After 4 weeks of culture, the highest frquency (100% of shoot formation from sucker explants was obtained from the medium with 1.0 mg/L BA. The highest shoot number per explant from in vitro shoot explants was 5.3. After 10 weeks of culture, the number of shoot per explant was increased. The highest frequency (85% of root formation was obtained at 0.5 mg/L glycine medium, when the explant with shoot were cultured on the MS medium containing glycine at various concentrations from 0 to 2.0 mg/L. The survival rate of the plantlets after transfer to plastic pots containing sand, soil, and vermiculite (1:1:1, vol.) was 95%. The results indicate that micropropagation procedure can be applied for an efficient mass propagation of Rubus fruticosus.

감마선 조사 유래 블랙베리(Rubus fructicosus L.) 돌연변이 계통의 농업부산물 사료가치 분석

류재혁,김동섭,하보근,김진백,김상훈,안준우,정일윤,강시용,조한직,김이엽 한국방사선산업학회 2013 방사선산업학회지 Vol.7 No.1

We examined the forage quality to provide the basic information for the utilization ofblackberry by-products (leaf and stem) among thirty-five mutant lines derived from gamma-ray. The leaves had higher crude protein (CP) contents than stems. In addition, the leaves showed broadranges of variation in crude fat, crude fiber and crude ash. The overall forage quality of the stemswere similar to those of the original variety. The mean of neutral detergent fiber (NDF) content ofleaves and stems in mutant lines were 25.88% and 58.98%, respectively. Also, the mean of aciddetergent fiber (ADF) content of leaves and stems in mutant lines were 15.43% and 49.27%, respectively. Forage quality grades of all blackberry leaf were special quality class. However, the blackberrystem ranked 1st to 5th in quality class. The stem diameter was negatively (P‹0.05) correlatedwith the moisture and crude protein. Also, the fruit length was highly negatively (P‹0.01) correlatedwith the RFV (relative feed value) in blackberry leaf. The one hundred fruit weight was highly negatively(P‹0.01) correlated with the crude ash. Based on these results, the by-products of blackberrycould provide high quality forage for feeding of live stock.