• BMB Reports
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• 제41권2호
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• pp.112-118
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• 2008

Camptothecin is an anti-cancer monoterpene indole alkaloid. The gene encoding 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (designated as CaHDR), the last catalytic enzyme of the MEP pathway for terpenoid biosynthesis, was isolated from camptothecin-producing Camptotheca acuminata. The full-length cDNA of CaHDR was 1686 bp encoding 459 amino acids. Comparison of the cDNA and genomic DNA of CaHDR revealed that there was no intron in genomic CaHDR. Southern blot analysis indicated that CaHDR belonged to a low-copy gene family. RT-PCR analysis revealed that CaHDR expressed constitutively in all tested plant organs with the highest expression level in flowers, and the expression of CaHDR could be induced by 100 ${\mu}M$ methyl-jasmonate (MeJA), but not by 100 mg/L salicylic acid (SA) in the callus of C. acuminata. The complementation of CaHDR in Escherichia coli ispH mutant MG1655 demonstrated its function.

• Plant Biotechnology Reports
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• 제2권2호
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• pp.163-169
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• 2008

Roots of Ophiorrhiza prostrata D. Don serve as a rich source of camptothecin (CPT), an anticancer drug. Because of the large-scale collection of its roots, the plant has become a threatened species. The present study accomplishes the induction of adventitious roots as a means for the production of CPT as well as for the large-scale propagation of this anticancer drug plant using leaf and internode explants. The biomass yield and CPT content of adventitious roots induced from different explants were compared to roots developed on ex vitro rooted stem cuttings. Adventitious roots were produced on half-strength Murashige and Skoog (MS) medium supplemented with $10.74{\mu}M$ ${\alpha}-naphthaleneacetic$ acid and $2.32{\mu}M$ kinetin at mean fresh weights of 0.753, 0.739 and 0.748 g roots from leaf, internode and shoot, respectively. CPT yield from in vitro derived roots after 50, 80 and 120 days of incubation (0.028, 0.06 and 0.1% dry weight, respectively) was not significantly different from those harvested at the same age from ex vitro rooted (0.03, 0.06 and 0.13%, respectively) stem cuttings. CPT from subcultured roots derived from solid (0.08%) medium was lower than from suspension culture medium (0.12%). Subsequent cultures of the adventitious roots showed a stable production of CPT (0.16%). The yield of CPT from 360-day-old plant-derived roots was 0.19%. Elicitation using methyl jasmonate and acetyl salicylic acid exhibited no enhancement in CPT yield. In vitro propagation through direct shoot regeneration was achieved from the adventitious roots upon transfer to MS medium with $8.87{\mu}M$ $N^6-benzyladenine$ (BA) and $2.46{\mu}M$ indole-3-butyric acid (IBA) with a mean of 21.2 shoots per culture in 50 days. The shoots upon subculture on medium having the same level of BA and IBA underwent rapid proliferation. The shoots transferred to field conditions after in vitro rooting exhibited 95% survival. Adventitious root induction, from leaf and internode explants, enables the feasible production of CPT as well as the large-scale rapid propagation of this species which can safeguard it from extinction.

• 한국잡초학회지
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• 제18권2호
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• pp.136-145
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• 1998

잔디밭에서 문제잡초로 알려져 있는 향부자의 효과적인 방제를 위한 기초 자료를 제공하고자 향부자의 생육특성, 생장조절제 처리가 맹아 및 생육에 미치는 영향, 제초제의 선발실험, 향부자의 일반성분을 조사하였다. 향부자의 개화는 이식 시기에 관계없이 이식 30일부터 시작되었고 괴경은 기온이 $10^{\circ}C$이하로 내려가면서 형성되기 시작하였다. 생장조절제 BA, ABA, TS-303, PDJ 등의 4종을 향부자 괴경에 처리한 결과 맹아에는 영향을 미치지 않았으며, 이식 60일째부터 경엽에 4회 처리시 4종 공히 신초 수와 지하부 건물중을 다소 증가시켰으나 신초 건물중과 괴경 수에는 영향을 미치지 않았다. Imazaquin 30g ai/10a 처리는 괴경으로부터 다량(multi-shooting)의 신초를 발생시켰으나 pyrazosulfuron-ethyl과 bensulfuron-methyl 처리는 multi-shooting 현상을 보이지 않았다. Pyrazosulfuron-ethyl 10g ai/10a 처리는 향부자의 생육을 90% 가까이 억제하여 bensulfuron-methyl 10g ai/10a 보다 높은 방제율을 보였다. 향부자 성숙한 괴경의 구성성분은 수분 61.83%, 탄수화물 31.60%, 조단백질 4.03%, 조지방 1.57%, 조회분 0.97% 이었다.