Isolation of Protoplasts and Subprotoplasts from Callus of Nicotiana glauca

여읍동,소웅영 ( Up Dong Yeo,Woong Young Soh ) 한국식물생명공학회 ( 구 한국식물조직배양학회 ) 1988 한국식물생명공학회 춘계학술연구발표회 Vol.1988 No.-

담배 ( Nicotiana glauca Graham ) Callus 배양시 Nicotine 생성에 미치는 2 , 4-D 및 NAA 의 영향

여읍동(Up Dong Yeo),김경호(Kyong Ho Kim) 한국식물학회 1991 Journal of Plant Biology Vol.34 No.2

Effects of two auxins, 2,4-dichlorophenoxyacetic acid (2,4-D) and α-napthaleneacetic acid (NAA) on nicotine production during callus culture of a wild tobacco (Nicotiana glauca) were investigated using a high performance liquid chromatography (HPLC). The high concentration (11.5 μM) of 2,4-D and NAA had peaks of nicotine contents at 4th and 2nd week, respectively. Thereafter, the concents decreased and the nicotine was metabolized to other alkaloids, The low concentration (1.5 μM) of 2,4-D on the medium supplemented with 0.1 mM of L-aspartic acid of L-arginine inhibited nicotine production. However, the low NAA promoted it only when the medium was supplemented with L-aspartic acid. From these results, it could be concluded that both auxins exhibit different action mechanisms on nicotine production pathway and the low NAA promotes the activities for the pathway with L-aspartic acid as a precursor.

담배 CMS line의 원형질체로부터 cytoplast의 유도 및 이와 타품종 원형질체와의 융합에 관한 연구

소상섭(Sang-Sup So),여읍동(Up-Dong Yeo) 한국생물공학회 1993 KSBB Journal Vol.8 No.2

담배(Nicotiana glauca) 카루스로 부터 Protoplast 및 Subprotoplast의 分離

呂邑東,蘇雄永 전북대학교 유전공학연구소 1988 遺傳工學硏究所報 Vol.1 No.1

As tools for the genetic modification of plants, the protoplasts were isolated from the green callus of Nicotiana glauca and were partitioned to subprotoplasts by gentle press operation. The isolation of protoplasts was taken for 5 hours in the enzyme solution(cellulase Onozuka R-10, 2%, macerozyme R-10, 1%, D-mannitol, 13% pH, 5.8)on the shaking bath(30℃, 70 strokes/min). The mean diameter was 45. 47±23.28μm and the mean volume was 4.92×10^-8 ㎤. The vacuoles was more easily observed by means of the deletion of cytoplasm and the destrution of cytoplasmic strands. After treated for 5 hours with the same enzyme solution for isolation of protoplasts, the callus clusters were pressed for 5 minutes by pincette through cover glass on them. The protoplasts were partitioned to about five subprotoplasts containing miniprotoplasts and cytoplasts. The ranges of diameter of miniprotoplasts and cytoplasts were 8.53-46.18μm and 8,58-52.18μm, respectively. One protoplast was partitioned to miniprotoplasts and cytoplasts at the ratio of 1:3 by the mean volume of protoplasts plsamolysed. But the observed ratio of miniprotoplast to cytoplast was 1:0.44. This result indicates that cytoplasts were more easily destructed than miniprotoplasts containing nucleus.

홍당무우(Beta vulgaris) 잎切片에 由來 Callus의 Anthocyanin 生成 : Callus 生長 및 Anthocyanin 生成에 미치는 Auxin과 Cytokinin의 組合效果 Effects of the Combinations of Auxins and Cytokinins on Callus Growth and Anthocyanin Production

呂邑東,黃世炫 전북대학교 유전공학연구소 1989 遺傳工學硏究所報 Vol.2 No.1

The effects of the combinations of auxins and cytokinins on growth and anthocyanin production was investigated in callus induced from red radish (Beta vulgaris) leaf explants. The callus induction was arrived at early stationary phase for 20 days on MS medium supplemented with 1.0 mg/l of 2, 4-D. And the anthocyanin contents of them was ca, four-fold higher than in the leaves of the donor planr. The combinations of low concentrations(0.5-1.0 mg/l) of 2, 4-D and kinetin (0.5-2.0 mg/l) were effective on callus growth and anthocyanin production, and the most effective combination was 1.0 mg/l of 2, 4-D and 0.5 mg/l of kinetin. In the other hand, the combinations of IBA as a rooting auxin and cytokinins (kinetin, BAP) were more effective on anthocyanin and adventitious root formation than those of combinations of 2, 4-D and kinetin. And the combinations of IBA, 1.0 mg/l and cytokinins were more effective than those of IBA, 0.5 mg/l, and the combinations of IBA with BAP were also better than those with kinetin. The most effective combination was 1.0 mg/l of IBA and 1.0 mg/l of BAP. There was some correlation between the anthocyanin contents and the numbers of adventitious root (r=0.62^*).

강남콩子葉 있어서 카이네친과 카드뮴이 葉錄素形成에 미치는 影響

呂邑東 全北大學校 基礎科學硏究所 1982 基礎科學 Vol.5 No.1

In order to investigate the influences of kinetin and Cadmium upon chlorophyll formation, the exciesed cotyledons of kidney bean (Phaseolus vulgaris) grown in darkness were illuminated with fluorescent light after pretreatment of different concentrations of kinetin (0, 1, 2, 4, 6, 8, 10mg/ℓ), Cadmium (0, 1, 2, 5, 5, 10, 20, 40mg/ℓ) and kinetin 4mg/ℓ above Cadmium concentrations in darkness. Then, the chlorophyll contents of the cotyledons were measured. The main results are as follows: 1. Kinetin 4mg/ℓ was most effective on chlorophyll formation, especially chlorophyll a. 2. The low concentrations of Cadmium(0-10mg/ℓ) promoted the chlorophyll formation, especially chlorophyll a, whereas the high concentrations(20-40mg/ℓ) inhibited it. 3. The low concentrations of Cadmium(0-5mg/ℓ) promoted kinetin activity. And the high concentrations(10-40mg/ℓ) inhibited it.

光合成系의 遺傳情報

呂邑東 전북대학교 유전공학연구소 1988 遺傳工學硏究所報 Vol.1 No.1

칼라코에 줄기切片의 器官分化에 미치는 生長調節物質의 影響

李康燮,呂邑東,蘇雄永 全北大學校 基礎科學硏究所 1991 基礎科學 Vol.14 No.1

In order to elucidate the effects of growth regulators on organogenesis in the stem explants of Kalanchoe daigremontiana, we have cultured the explants for 10 weeks on the MS medium with or without ammonium nitrate, supplemented with simultaneous combinations of auxin(IAA), gibberellin(GA_3) and cytokinin(BAP)The cocentrations of these growth regulators were 0.001-10.0 ㎎/l, respectively. The callus formation and organogenesis in the explants are mainly achieved under the simultaneous actions of the auxin and cytokinin. The optimal combination of growth regulators for callus formation was IAA 10.0 ㎎/l, GA_3 0.001 ㎎/l and BAP 0.1 ㎎/l. The optimal combinations for the adventitious root and bud formation are IAA 10.0 ㎎/l, GA_3 0.01 ㎎/l, BAP 0.01 ㎎/l and IAA 0.1 ㎎/l, GA_3 0.01 ㎎/l, BAP 1.0 ㎎/l, respectively. On the MS medium without ammonium nitrate, the callus and adventitious bud formation in the explants are better than those on the MS medium with ammonium nitrate, but the adventitious root formation is worse. The correlation between callus formation and organogenesis is negative on the both media.

嶺東地方에 있어서 PTC 味盲의 頻度 및 血液型과의 關係

李金永,呂邑東 全北大學校 1981 論文集 Vol.23 No.-

韓國人類集團의 PTC (phenylthiocarbamide) 味覺에 대한 頻度의 平均値와 味盲과 血液型과의 關係를 糾明하기 위하여 그 一環으로 嶺東地方에 있어서 男女 中高等學校 學生을 對象으로 調査하여 다음과 같은 結果를 얻었다. 1. PTC 味覺은 크게 taster와 non-taster(味盲)로 區分된다. 그 中 taster는 쓴맛, 신맛, 단맛, 기타의 맛 頻度順으로 나타났다. PTC 濃度 C_1(1,300.00 ㎎/ℓ)에서 味盲의 頻度는 9.9%이었고 期待와는 달리 C_10에서는 훨씬 낮은 頻度를 나타내었다(61.2%). 쓴맛과 味盲의 頻度曲綠은 C_6과 C_7 사이에서 交叉되었다. C_13 以上의 低濃度에서의 味盲의 頻度는 거의 一定한 頻度의 傾向을 나타내었다. 2. 신맛, 단맛, 기타의 맛의 頻度는 PTC 濃度에 關係없이 男子가 女子보다 더 높았다. 특히, C_1에서의 味盲의 頻度는 男子가 9.6%, 女子가 10.6%를 나타내었고 高濃度(C_1~C_5)에서는 女子가 男子보다 PTC 味覺에 더 銳敏함을 나타내는 반면에 低濃度(C_6~C_17)에서는 그 反對現象이였다. 3. C_1에서의 血液型 A型의 味盲頻度는 11.8%, O型이 9.8%, AB型이 9.6%, B型이 8.5%였다. 이와 같이 이러한 頻度는 PTC 濃度가 낮아짐에 따라 거의 一定한 頻度의 傾向을 나타내었다. To investigate the frequency of PTC (phenylthiocarbamide) non-taster and relationship between non-taster and blood type, middle and high school students in Yeongdong population of Gangwondo province, Korea were studied. The results are as follows : 1. Most of the tasters tasted PTC as bitter. However some tasted as sour, sweet and other in the order of frequency. Frequency of non-taster of C_1 was 9.9%. That of C_10 was unexpectedly low (61.2%). The curves of bitter and non-taster was crossed between C_6 and C_7. the In lower concentrations than C_13, the frequencies of non-taster were inconsistent. 2. The frequencies of sour, sweet and other taster in male were higher than in female. Especially, the frequency of non-taster of C_1 was 9.6% in male and 10.6% in female. Whereas male was more sensitive to the high concentrations(C_1~C_5) of PTC, female was more sensitive to the lower concentrations(C_6~C17). 3. Among the non-taster of C_1, the frequency of blood type A was 11.8%, O 9.8%, AB 9.6%, and B 8.5%. This frequencies invariably changed as the PTC concentration become lower.

담배(N. glauca Graham) 잎 切片 由來 callus의 生長 및 nicotine 生成에 미치는 두가지 auxin과 光照射의 同時效果

金敬鎬,呂邑東 전북대학교 유전공학연구소 1990 遺傳工學硏究所報 Vol.3 No.1

Simultaneous effects of two auxins(2,4-D and NAA) and irradiation on growth and nicotine production of callus derived from Nicotiana glauca Graham leaf explants on the modified MS medium cintaining 4 organic acids were investigated. For growth of callus, NAA(115μm) which had promoted adventitous root formation after 2nd weeks of culture, was more effective than 2,4-D(11.5μm). And irradiation(16/8 hrs) was more effective than in dark. However, for nicotine profuction, 2, 4-D was more effective than NAA. The both 2, 4-D and NAA had a peak of incotine contents on the 4th and 3rd week of callus growth, respectively. Thereafter, the nicotine contents was decreased. It means that nicotine has been transformed to other alkaloids and excreted into the medium. And irradiation(16/8 hrs)was also more effective than in dark. This results show that the biosynthesis of nicotine is mediated by phytochromes.