강남콩 하배체 (下胚體) 절편의 (切片) 부정근형성에 미치는 내재 IAA 의 이종

조덕이(Duck Yee Cho),소웅영(Woong Young Soh) 한국식물학회 1989 Journal of Plant Biology Vol.32 No.4

This work was carried out to elucidate effects of endogenous and exogenous IAA transport on adventitious root formation in Phaseolus vulgaris hypocotyl cuttings. For inverted or normal incubation in distilled water, the adventitious root is always formed at the morphological base, but not at the morphological apex. For inverted incubation, in both distilled water and certain chemical solution, the root formation is retarded more at the first stage (0-24 hr) than at the second stage (24-48 hr). When p-chlorophenoxyisobutyric acid (PCIB) was applied to the cuttings at the first stage, the root formation was inhibited more than at the second stage. Treatment of 2,3,5-triiodobenzoic acid (TIBA) markedly inhibited the adventitious root formation in Phaseolus vulgaris hypocotyl cuttings. This inhibition influenced the root according to the applied stage and period. Therefore, the root formation is more related to the stage of root primordium formation than to the stage of root elongation from the primordium. Inhibition of auxin transport or action by TIBA or PCIB could also be reversed when hypocotyl cuttings are incubated in exogenously applied IAA solution.

구기자나무(Lycium chinense Mill.) 접합자 배로부터 체세포배 및 부정아 발생

이재동,조덕이,소웅영,Lee, Jae-Dong,Cho, Duck-Yee,Soh, Woong-Young 한국식물생명공학회 2002 식물생명공학회지 Vol.29 No.4

구기자나무의 접합배의 자엽절편을 MS 배지에 오옥신 (2,4-D, NAA, IAA)와 싸이토키닌 (zeatin, kinetin, BAP)를 단독 또는 조합처리하여 체세포배와 부정아를 유도하였다. 0.5 mg/L 2,4-D를 첨가한 배지에서 배발생능 캘러스를 유도되었고 이로부터 체세포배가 발생되었다. 반면에 부정아는 0.01 mg/L 2,4-D+0.5 mg/L zeatin을 조합한 배지에서 발생하였다. 접합자배의 여러 부위별로 배양한 결과 자엽 절편이 가장 높은 형태형성을 보였으며, 기관형성은 자엽>유근>배축>배 전체 순이었다. 해부학적으로 관찰한 결과 심장형배를 관찰할 수 있었고 정단분열조직과 부정아를 확인할 수 있었다. Somatic embryos or adventitious buds were formed from the segments of zygotic embryo of Lycium chinense Mill. on Murashige and Skoog (MS) medium supplemented with auxins (2,4-D, NAA, IAA) and / or cytokinins (zeatin, kinetin, BAP). Embryogenic callus formed on MS medium containing 0.5 mg/L 2,4-D and then differentiated into somatic embryos without transfer to hormone free medium. On the other hand, adventitious buds were formed on medium with 0.01 mg/L 2,4-D and 0.5 mg/L zeatin. Among various parts of zygotic embryos, the morphogenic potential was higher in the cotyledonary region, and the most organogenic potential was found in cotyledon followed by radicle, hypocotyl, and whole embryo. Histologically, bipolar structure of the heart-shaped embryos were confirmed and in adventitious buds only shoot apical meristems were found to exist.

카사바의 미세증식에서 기내 발생 부정근의 절단이 순화에 미치는 영향

윤실,조덕이,소웅영,Yoon, Sil,Cho, Duck-Yee,Soh, Woong Young 한국식물생명공학회 2001 식물생명공학회지 Vol.28 No.2

카사바 (Manihot esculenta Crantz cv. MColl 22)의 마디절편을 제아틴 0.01%가 첨가된 MS 기본배지에서 2주간 현탁배양하여 경엽부 길이가 1.5∼2.5 cm로 생장된 유식물의 부정근을(1) 1∼l.5 cm길이만 남기고 짧게 전정, (2) 1∼2 mm 이내로 제거, (3)뿌리 발생부 전체를 절단한 후 마사토를 담은 유리병에 이식하여 비무균조건에서 순화생장시켰다. 그 결과 이식 7∼10일경부터 각 조건에서 새 뿌리가 발생되기 시작하여 6주 후에는 조건에 따라 73∼93%의 생존율을 나타냈다. 기내 발생 뿌리 가운데 손상 없이 토양에 이식된 것은 그대로 생존을 계속하여 유식물 생장에 도움을 주었으며, 제거된 부정근은 이식 후 신장이 중단되고 새로 뿌리가 발생되었다. 그러나 간혹 전정되고 남은 부분에 측근이 발생된 것이 관찰되었다. 각 조건에서 새로 발생된 기외 뿌리의 수와 길이 및 경엽부 길이와 마디 수에서는 서로 유의성 있는 차이를 나타내지 않았으나, 생체중에서는 전정>제거>절단의 순으로 약간의 차이를 보였다. 미세증식된 카사바의 기내발생 부정근을 적절히 전정 또는 제거한 후 이식하여 순화시키는 것은, 작업을 간편하게 할 뿐만 아니라 유식물의 생존율이 높고 생장에도 지장이 거의 없으므로 경제적인 미세증식 방법으로 유용하게 이용될 수 있다. The in vitro plantlets of cassava (Manihot esculenta Crantz cv. MColl 22) could be regenerated from nodal explant cultures in a liquid MS basal medium containing 0.01 mg/L zeatin for 2 weeks. The plantlets of 1.5∼2.5 cm in shoot length were transplanted to a glass bottle containing fine sand and acclimated under non-sterile conditions after excising their intact roots by: 1) prune leaving roots base of 1∼1.5 cm; 2) complete removal of roots; and 3) cutting off the rooting zone. The majority of in vitro-formed intact roots continued growth after transferred to soil, and all of the damaged roots stopped further growth. The plantlets with excised roots began to develop new roots within 7∼10 days after being transferred to a glass bottle, and a few of the pruned roots developed lateral roots from the remaining portion. Pruning and removal of in vitro roots resulted in a high survival rate (>87%), and did not significantly affect ex vitro root regeneration and acclimation, but the plantlets in which the rooting zone had been cut-off showed 73% survival rate. Pruning or removal of in vitro roots before transfer of plantlets is recommended for useful method of commercial micropropagation because of easier handling and high survival rate of plantlets.

카사바의 절간절편 배양에서 부정근 발생이 결정되는 시기의 판별

윤실,조덕이,소웅영,Yoon, Sil,Cho, Duck-Yee,Soh, Woong Young 한국식물생명공학회 2001 식물생명공학회지 Vol.28 No.1

카사바 (Manihot esculenta Crantz, cv. MCol 22)의 마디 및 절간절편으로부터 부정근 발생이 분화결정되는 시기를 조사하였다. 1개의 액아를 가진 10mm 길이의 마디절편을 MS 기본배지에서 8일간 배양하면 직접 부정근이 발생되었으나, 액아를 갖지 아니한 절간절편은 동일한 배지상에서 부정근이 형성되지 않았다. 반면에 마디절편을 MS 기본배지에서 5일간 배양한 뒤, 마디절편의 중간부로부터 절단된 하부의 절간절편으로부터는 부정근이 발생되었다. 이에 따라 마디절편의 중간 부를 일정시간 단위로 절단하는 방법으로 조사된 분화결정 시기는 3∼5일이었다. 한편 절간절편을 여러가지 농도의 IBA가 첨가된 배지에서 일정 기간 배양한 후 MS 기본배지로 이식했을 때 부정근이 발생되는 시기는 옥신 농도와 배양기간 에 따라 달랐다. 0.5 mg/L IBA 첨가에서는 5일 이상 배양되어야 90% 이상의 절편에서 부정근이 발생되고, 1 mg/L IBA 첨가에서는 2.5일 이상 처리에서, 그리고 2mg/L IBA에서는 1.5일의 처리에서 90% 이상의 부정근이 발생되었다. 따라서 부정근 발생을 분화결정시킨 외래 옥신의 처리기간은 분화결정 시기이라기보다는 근원기 유도에 필요한 적정량의 옥신을 흡수축적한 기간으로 사료된다. The timing for the determination in root formation from nodal and internodal explants of cassava (Manihot esculenta Crantz, cv. MCol 22) was investigated. Nodal explants about 10 mm with an axillary bud formed adventitious roots directly on MS basal medium for 8 days of cultures. But internodal segments without an axillary bud did not develop the adventitious roots on the same medium, and most internodal segments excised from nodal explants after cultures of 5 days on MS basal medium developed adventitious roots. On the other hand, the internodal segments rooted at 90% after cultures on medium with 0.5 mg/L IBA for 5 days, with 1 mg/L IBA for 2.5 days, and with 2 mg/L IBA for 1.5 days respectively. Thus the period of culture on medium with IBA and its IBA concentration affected the rooting rate. Therefore, it is suggested that the determination for root formation occurred before the differentiation of root primordia on medium with IBA, and root inducing factors in medium were absorbed and accumulated during the period of determination for root primordium differentiation in internodal segment of cassava.

마디 절편의 현탁배양에 의한 카사바의 미세증식

윤실,조덕이,소웅영,Yoon, Sil,Cho, Duck-Yee,Soh, Woong Young 한국식물생명공학회 2000 식물생명공학회지 Vol.27 No.3

For the micropropagation, node explants of cassava were cultured in liquid MS medium with various concentrations of cytokinins on a rotary shaker (100 rpm) for 2 weeks. The adventitious roots and shoots from the explants were differentiated more efficiently in liquid medium than in solid. But root formation was not inhibited in medium with BAP and kinetin at low concentration (>0.05 mg 1/sup -1/), while in medium added with BAP and zeatin at high level (<0.25 mg 1/sup -1/), it was inhibited by callus forming on cut end of the cuttings. However, all of plantlets grown in liquid medium for more than 2 weeks showed symptoms of hyperhydricity. The plantlets grown in liquid medium were transferred into culture bottles filled with fine sand or artificial soil (pitmoss:perlite:vermiculite, 1:1:1 v/v) wetted with half strength of Knop's solution. After transplanted to culture bottles, some of vitriscent leaves were defoliated and new leaves were normally formed from shoot apex. Most of plantlets (>95%) were hardened-off successfully only in culture bottles with fine sand, and grew into 3-5 cm seedlings possessing 4-6 nodes after 4 weeks. Thus, the mass propagation of cassava on medium containing cytokinin could be established based on the suspension culture using node explants. 카사바 (Manihot esculenta Crantz, cv. MCol 22)의 마디 절편을 MS 기본배지에 3종류의 사이토키닌을 여러 농도로 첨가한 조건에서 2주동안 현탁배양한 결과, 고체배지에서 3주 배양된 것보다 훨씬 생장이 효과적이었다. 현탁배양된 유식물에서는 과수화현상이 나타났으나, 이들을 세사 (마사토 + 1/2농도의 크놉씨액)를 담은 유리배양병에 이식하여 생육시킨 결과, 유경에서 새 잎이 신장되면서 과수화를 벗어나 4주후에는 4~6개의 마디를 가진 유경길이 3~5 cm의 정상 유묘로 생장됨으로서, 목본식물인 카사바의 경우 마디절편의 현탁배양에 의한 대량증식의 가능성을 확인했다. 현탁배양시 저농도의 제아틴 (0.01-0.05 mg 1$^{-1}$) 처리는 BAP 최적조건에서 보다 2배나 길게 부정아 길이를 신장시켰다. 사이토키닌 처리로 부정아가 잘 발생된 유식물은 토양이식 작업을 간편하게 했으며, 토양 이식 후 부정근을 활발히 발생시켜 one step culture시스템 배양이 가능하게 했다. 과수화된 유식물은 기외에서 순화가 어려웠으나 유기물이 혼입되지 않은 세사를 담은 유리배양병에서는 비무균조건일지라도 95% 이상 순화될 수 있었다.

한국산 고등균류에 관한 연구(제 1보) -능이버섯의 단백분해효소 활성-

은재순,양재헌,조덕이,이태규,Eun, Jae-Soon,Yang, Jae-Heon,Cho, Duck-Yee,Lee, Tae-Kyu 한국약제학회 1988 Journal of Pharmaceutical Investigation Vol.18 No.3

This study was undertaken to investigate the proteolytic enzyme from Neungee mushroom [Sarcodon aspratus (Berk) S. Ito]. The proteolytic activity of Neungee was higher than other several edible mushrooms under various pHs. The potency of proteolytic enzyme of Neungee was same as the digestive drugs containing protease. So the proteolytic activity of the enzyme was increased in neutral or weak alkaline pH, whose characteristics would be alkaline protease. The specific activity of the purified enzyme obtained by using Tris acryl CM-cellulose ion exchange increased 20 times as compared with that of the crude extract. The proteolytic enzyme was stable at room temperature, but decomposition was fast when incubated at higher temperature more than $40^{\circ}C$. The half life of the enzyme was longest in neutral pH and rate constant was increased in acidic or alkaline solution.

더덕의 체세포배발생에서 전형층분화와 자엽수의 관계

최필선,소웅영,조덕이,유장렬,Choi Pil-Son,Soh Woong-Young,Cho Duck-Yee,Liu Jang-Ryol 한국식물생명공학회 2005 식물생명공학회지 Vol.32 No.2

Embryogenic callus was obtained from cotyledonary explants of Codonopsis lanceloata on Murashige & Skoog's medium supplemented with 1 mg/L 2,4-D. Suspension cultures of the embryogenic calli were grown on a shaker at 100 strokes/min, and then the calli were subcultured for 2 weeks in 2,4-D-free medium to produce somatic embryo. In somatic embryos at the globular stage, cotyledon initials began to differentiate themselves in the near distal end of the procambial strand. Dicotyledons, tricotyledon, tetracotyledon and fused cotyledon were differentiated from the distal ends of two, three, four and circular procambial strands, respectively. Nearly circular procambial strand in lower hypocotyls were independently differentiated into two, three, four procambial tissues at cotyledonary node and cotyledons to form somatic embryos with dicotyledon, tricotyledon, tetracotyledon. If the distal subepidermal cells of globular embryo exclusively became cotyledon initials, the torpedo or cotyledonary embryo was characterized by somatic embryos with fused cotyledon. 더덕 자엽절편을 1 mg/L 2,4-D가 첨가된 MS기본배지에 배양하여 배발생캘러스를 얻었고, 호르몬이 첨가되지 않은 MS액체배지에서 구형기의 체세포배를 얻었다. 구형기에서 초기 심장형기로 발달할 때 자엽의 시원세포는 전형성층조직으로부터 분화되기 시작 하였으며, 하배축에서 관찰되는 원통형 전형성층대는 2개의 자엽을 형성할 경우 2개의 전형성층대가, 3개의 자엽은 3개의 전형성층대가, 4개의 자엽은 4개의 전형층대가 각각 독립적으로 분화되어 자엽절과 자엽부위까지 연결되어 있었다. 만약 구형기 체세포배에서 자엽시원세포가 원통형으로 분화될 경우 합생자엽을 갖는 체세포배가 형성되었다.

당근의 배양세포로부터 형성된 체세포배의 다자엽 구조

소웅영(Woong Young Soh),조덕이(Duck Yee Cho),이은경(Eun Kyong Lee) 한국식물학회 1996 Journal of Plant Biology Vol.39 No.1

Embryogenic callus was selected from callus induced from hypocotyl segment cultures of Daucus carota seedlings on MS basal medium supplemented with 1.0 ㎎/L 2,4-D. Cell clumps prepared from the embryogenic callus were transferred to MS medium without 2,4-D for somatic embryo development. Cotyledonary abnormalities were frequently observed on somatic embryos developed after the incubation of cell clumps in MS basal medium with 1.0 ㎎/L 2,4-D for one week and then subculture in the same medium but without 2,4-D for two weeks. The percentage of abnormalities was as follows: 5% one cotyledon, 21% three cotyledons, 6% four cotyledons, 5% five cotyledons, 0.2% six cotyledons and 1% trumpet-like cotyledons. On the other hand, the normal somatic embryo with two cotyledons appeared at 63%. The germination rate of somatic embryos was higher in two cotyledon somatic embryos than in multicotyledonary embryos. Trumpet-like somatic embryos did not germinate normally showing limited elongation and enlargement of roots and cotyledons without shoot development. From anatomical examination circula procambium in the root of somatic embryo began to branch around the middle regions of the hypocotyl which extended into the cotyledons through the cotyledonary nodes and the number of branched procambial strands in hypocotyl was equal to the number of cotyledons. Monocotyledonous somatic embryo always had larger cotyledon than that of somatic embryos with multicotyledons and had horseshoe-like cotyledons where the procambium was of the same structure.

Agrobacterium rhizogenes 에 의한 Hairy Root 형성에 대한 생리학적 연구 - 1. A. rhizogenes 에 감염된 당근 조직에서의 IAA 함량과 형태적인 특징과의 관계

황백(Baik Hwang),조덕이(Duck Yee Cho),홍성식(Sung Sik Hong) 한국식물학회 1986 Journal of Plant Biology Vol.29 No.4

Inoculation of carrot discs with Agrobacterium rhizogenes harboring Ri plasmid resulted in transformation of cells, as revealed by the tumors and hairy roots arising from them. Measurements of IAA content using HPLC indicate that it is higher in tumors and inoculated tissues than in uninoculated tissue. A lot of meristemoids and vessel elements formed in tumor tissue and the hiary roots differentiated from meristemoids. IAA content in tumor tissues is decreased with hairy root and vessel elements formation from them. Formation of wound callus in uninoculated tissues resulted on wound healing but no formation of vessel elements and hairy roots. Tumor tissues show continuous growing on hormone free medium.

은단풍 ( Acer saccharinum L. ) 유식물의 뿌리에서 유관속 형성층의 초기발생

소웅영(Woong Young Soh),홍성식(Sung Sik Hong),조덕이(Duck Yee Cho) 한국식물학회 1991 Journal of Plant Biology Vol.34 No.4

The origin of cambial initials from procambium was studied in the seedling root of Acer saccharinum. In transverse view, the first periclinal divisions of procambial cells occurred just outside of each early metaxylem and resulted in meristematic strips. As root development progressed, the division activities appeared subsequently outside of each late metaxylem and then in pericycle cells opposite the four protoxylem poles. Eventually, such meristematic strips were connected completely each other. Thus, a nearly rectangular shaped meristematic layer in outline was formed outside the xylem in a whole root transectioned. In tangential section, early procambium showed a homogeneous structure consisted of uniform short cells with transverse end walls. However, some of the procambial cells did elongate, whereas others divided transversely. The former became more elongate, tapered, and vacuolated. Finally, they differentiated into fusiform initials. Short cells consisting axial strands divided continuousely in transverse plane and became ray initials, while some short cells elongated and transformed into long cells. The early ontogeny of vascular cambium in Acer saccharinum root was interpreted to be established by a gradual process.