• 한국자원식물학회지
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• 제35권4호
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• pp.508-514
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• 2022

Bulbophyllum auricomum Lindl. is very popular among orchid growers due to the attractive fragrance of its flowers and has become an endangered orchid in Myanmar. In this study, we carried out an aseptic technique that can be used to conserve this endangered orchid species. The seeds of B. auricomum Lindl. were obtained from artificial pollination and cultured in MS basal medium for seed germination. The effect of coconut water and BAP in MS basal medium on callus induction was investigated. The highest callus induction was found at 2.0 mg/L BAP. The maximum growth of protocorm-like bodies (PLBs) was evaluated, and the best response was observed on MS medium supplemented with 150 mL/L coconut water at pH 5.6. MS basal medium supplemented with 150 mL/L coconut water along with 2.0 mg/L BAP and 1.0 mg/L NAA (MCBN) showed the highest number of plantlets at 15℃ at the second week of culture. At the second and third week of culture, MS medium supplemented with 2.0 mg/L BAP and 1.0 mg/L NAA (MBN) showed the best result in terms of the number of leaves and the longest leaves at 15℃ and 25℃, respectively. The present study showed evidence of successful in vitro propagation of B. auricomum Lindl.

• Journal of Plant Biotechnology
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• 제46권1호
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• pp.22-31
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• 2019

에케베리아 라우이(Echeveria laui)와 엘레강스(Echeveria elegans)의 기내 식물체 대량생산을 위하여 캘러스와 신초 유도, 신초생장과 발근에 영향을 미치는 생장조정제 종류 및 농도를 구명하고 형성된 식물체의 기외 순화조건을 구명하고자 본 연구를 수행하였다. 캘러스 유도에 적합한 부위를 구명하기 위해 잎을 상부, 중부, 하부로 나누어 NAA와 BA의 농도별로 치상한 결과, E. laui는 엽상중부를 이용 하였을 때 $NAA\;1\;mgL^{-1}+BA\;1\;mgL^{-1}$의 혼용처리에서 캘러스가 100% 형성되었으며, E. elegans는 엽중부에서 $NAA\;2\;mgL^{-1}+BA\;4\;mgL^{-1}$ 혼용배지가 83.3%로 가장 높은 캘러스 형성을 보여 캘러스 형성에 효율적일 것으로 생각된다. 캘러스로부터 신초 형성율은 E. laui에서는 $NAA\;0.1\;mgL^{-1}+BA\;3\;mgL^{-1}$ 혼용배지, E. elegans는 $NAA\;0.3\;mgL^{-1}$ 단용배지에서 가장 높았다. 또한 신초의 증식에 적합한 배지는 E. laui는 $NAA\;1\;mgL^{-1}+BA\;1\;mgL^{-1}$ 혼용배지에서 신초가 10.4개, E. elegans는 $NAA\;1\;mgL^{-1}+BA\;0.1\;mgL^{-1}$ 혼용배지에서 12.0개로 신초가 가장 많이 형성되었으며, 형성된 재분화체는 정상적인 식물체로 생장하였다. 기외 순화조건의 경우, 용토에 따라 생육은 큰 차이를 보이지 않았으며, 관상가치를 고려하여 차광막 35 ~ 55%에서 순화 처리하는 것이 적합할 것으로 판단된다. 이러한 결과는 최적의 기내 배양 조건 및 재분화 식물체 순화조건 체계확립을 통해 고부가 다육식물인 E. laui와 E. elegans의 대량생산이 가능할 것으로 기대된다.

• 한국육종학회지
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• 제40권1호
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• pp.31-38
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• 2008

본 실험은 보리 약배양에서 모식물체의 생육환경과 전처리 방법에 따른 배양효율을 검토코자 수행되었다. 캘러스 유기정도는 품종에 따라 0~5.6%의 범위를 보였고 식물체 분화율은 캘러스 대비 0~30.0%이었으나 그중 녹색체는 0~4.4%로 낮았다. 동절기 온실과 춘계포장에서 각각 재배한 모식물체의 약배양 캘러스 유기율은 춘계포장 모식물체의 약에서 많았지만 식물체 분화율은 각각 30.4%와 21.3%로 동절기 온실시료의 배양에서 높게 나타났다. 또한 정상상태에서 자란 모식물체와 신장기에 한발 스트레스를 받은 모식물체의 캘러스 유기율은 각각 19.2%와 7.2%로서 약유래 모식물체의 생육환경에 따라 큰 차이를 보였다. 약 채취 시기는 외형적인 식물체의 엽이간장이 5~10 cm정도로 지엽 다음 엽의 엽이와 엽초 내 이삭의 가장 불룩한 중앙부위가 일치하는 시기(출수전 3~4일)를 기준으로 약을 선별하여 치상 하는 것이 유기효율이 높았다 전처리는 직경 10cm의 롤(roll) 비닐봉지에 5~10개체의 이삭을 밀봉하여 $4^{\circ}C$에서 4 3주간 실시하는 것이 효과적이었으며 관행 페트리디쉬 보관에 비하여 약 6배 정도의 노력절감효과를 보였다 전처리 기간 및 온도별 시료의 상태는 $4^{\circ}C$에서 4주까지, $7^{\circ}C$는 2주까지, $10^{\circ}C$는 1주까지 신선도 유지가 가능하였고, 전처리 기간은 $4^{\circ}C$, 3주에서 두원찹쌀보리(2조)와 새쌀보리(6조)조가 각각 4.8%, 1.7%의 캘러스가 유기되어 가장 높았다.

• 한국자원식물학회지
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• 제12권1호
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• pp.1-9
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• 1999

인삼 뿌리 절편으로 부터 모상근 유기를 위한 최적 조건을 확립하고자 Agrobacterium rhirogenes와 인삼 뿌리 절편의 항생제 내성 조사 및 최적의 모상근 유도 배지를 조사하기 위하여 수행하였다. NaOCl로 인삼 뿌리를 멸균하였을 때, 오염 정도가 감소하면서 조직의 손상이 일어나지 않는 NaOCl의 농도는 7% NaOCl에서 15-20분, 9% NaOCl에서 5분으로 나타났다. 인삼근은 년수가 증가할수록 오염 정도가 심하였으며, 특히 6년근중 표피가 있는 처리구는 오염 정도가 매우 높았다. Agrobacterium의 성장억제를 위한 항생제는 tetracycline이 가장 효과적이었으며, 30mg/L 이상의 농도에서 균의 성장이 억제되었다. 하지만 30mg/L tetracycline에서 인삼 조직이 고사하였으며, cefotaxime(500mg/L), carbenicillin(500mg/L)에서 균의 성장을 완전히 억제하였으며, 조직의 손상이 일어나지 않았다. 3년근 인삼에서 모상근 유도을 위한 배지로는 1/2MS 배지에 500mg/L의 cefotaxime이 첨가된 배지가 가장 좋았으며, 인삼 뿌리 절편에 Agrobacterium을 발라주는 것 보다는 균과 공동배양할때가 절편이 좋았다. Agrobacterium접종 2주 후부터 callus가 유기되기 시작한 후, 다시 2주 후에 모상근이 유도되었다. 유도된 hairy roots는 PCR에 의하여 rol C유전자를 조사함으로서 형질전환체임을 확인하였다.

• KSBB Journal
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• 제6권3호
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• pp.289-297
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• 1991

The insecticidal protein gene in the pKL-20-1 clone derived from Bacillus thuringiensis serovar. kurstaki plasmid was subcloned in the plant shuttle vector, pGA643. The 7.3 kb fragment was cloned in the BglII and Hpal sites of pGA643 vector and expressed in E. coli S17-1, which produced insecticidal proteins killing Bombyx mori larvae. The clone was named pHL-20. The protoplast formation, calli induction and plantlet regeneration of Nicotiana tabacum was carried out. A tremendous number of mesophyll protoplasts of N. tabacum were formed, up to 7$\times$105 protoplast per ml, for 20 hours in darkness in the enzyme solution of 0.5% cellulase and 0.1% macerosin, pH 5.8. The viabilities of the protoplasts were maintained above 80% for 6 days in the media containing 2mg/1 of NAA and 1mg/1 of kinetin. Calli were induced from the protoplasts and leaves of the N. tabacum on MS medium containing 0.5mg/1 BAP. Under the culture conditions the protoplasts underwent repeated cell division into calli. Plantlets were regenerated from callus cultures derived from protoplast and leaves. Shoots were induced in a medium containing 1mg/1 of BAP.

• Journal of Life Science
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• 제12권1호
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• pp.19-21
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• 2002

The pathway of embryos formed anther culture in herbaceous peony was influenced by addition of 2,4-D. MS medium with 2,4-Dichlorophenoxy acetic acid (2,4-D) alone did not arise direct embryogenesis, but was proliferate callus. Embryos through calli were produced on medium containing 0.2 mg/1 zeatin or without growth regulators. Direct embryogenesis was obtained from MS basal medium. However, after the anthers were cultured on medium with 0.1 mg/1 2,4-D, 3 g/1 AC, 30 g/1 sucrose, 2 g/1 gelrite for 40 days. Its efficiency (32.3 %) was markedly improved when anthers cultured on medium without 2,4-D. Embryo morphology was also affected by the 2,4-D used in medium. The induction of normal embryos with two cotyledons was higher in the embryos formed through direct embryogenesis than those formed callus. The embryos formed from calli were mainly showed abnormal embryo with one, three, four cotyledons or hors and bowling pin type.

• BMB Reports
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• 제30권1호
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• pp.21-25
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• 1997

The growth. freezing resistance and electrophoretic protein patterns of carrot callus cultures were investigated following treatment with NaCl for various' intervals at 20$^{\circ}C$. Following 7 day exposure to 250 mM NaCl. freezing tolerance increased, which was measured by 2.3.5-triphenyl tetrazolium chloride (TTC) assay and fresh weight was reduced compared to control cells. Changes of electrophoretic patterns of total and boiling stable proteins were investigated using one or two dimensional gel system. Several proteins with molecular weight of 43 and 21 kDa increased by NaCl treatment. The most prominent change was detected in 21 kDa protein. The steady state level of this protein increased in NaCl treated cells, but decreased in control cells. Twenty one kDa protein was detected only in the NaCl treated cell when boiling stable protein was analyzed. The isoelectric point of 21 kDa protein was identified as 5.7. The timing of increase of 21 kDa protein was correlated to freezing resistance which implied the role of this protein in the induction of freezing resistance of the cell.

• Advances in Traditional Medicine
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• 제7권2호
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• pp.197-204
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• 2007

The present study focused on the establishment of micropropagation protocol for the high value Pluchea (P.) indica (L.) Less., genotype, an important medicinal plant and evaluation of the diuretic activity of the leaf extract of the tissue cultured plant. Leaf explants, nodal segments and shoot tips were cultured in MS medium supplemented with auxin and cytokinin and their combinations. With the objective of inducing callus giving rise to new adult plants, naphthalene acetic acid was found to be most effective for (80%) for callus induction. The methanolic extract of leaves of the micropropagated P. indica was investigated for its diuretic activity in Wistar albino rats. Urinary excretion parameters were studied for evaluation of diuretic activity using Frusemide (20 mg/kg, p.o.) as standard. The extract showed significant diuretic activity at the doses of 100, 200 and 300 mg/kg. p.o. An oral acute toxicity study for the extract was carried out and the $LD_{50}$ value was found to be 2,825 mg/kg body weight.

• Journal of Plant Biotechnology
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• 제6권2호
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• pp.113-118
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• 2004

Somatic embryos were formed from calli obtained from axillary shoots (raised from nodal segments of glasshouse-grown plants under aseptic conditions), internodal segments (from in vitro-raised plants), and root and coty-ledonary leaf segments (from in vitro-raised seedlings) after 8 weeks of initial culture. Embryo formation was the highest (97.33%) from cotyledonary leaf callus on Mura-shige and Skoog's (MS) medium containing kinetin (KN) (3 mg/L). Somatic embryo induction was lesser with different combinations of auxins while it increased to 100% in internodal segment and cotyledonary leaf calli with 6-benzyladenine (BA) (2mg/L) along with 2,3,5-triiodobenzoic acid (TIBA) (2mg/L). The shoots were induced from somatic embryos raised from root, coty-ledonary leaf and internodal segment calli grown on MS medium containing BA in combination with indole-3-acetic acid (IAA). Maximum of 66.67% cultures formed shoots on MS medium containing BA (1mg/L) in combination with IAA (2mg/L). The shoots raised from somatic embryos were rooted on MS medium supplemented with indole-3-butyric acid (IBA) (2mg/L). The plantlets transferred to the field showed 70% survival rate after one year.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권2호
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• pp.154-159
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• 2006

Interleukin-18 (IL-18), otherwise known as interferon-gamma-inducing factor (IGIF), is one of several well characterized and important cytokines that contribute to host defenses. The complementary DNA (cDNA) of mature human interleukin-18 gene (hIL-18) was fused with the signal peptide of the rice amylase 1A gene (Ramy1A) and introduced into the plant expression vector under the control of a duplicated CaMV 35S promoter. The recombinant plasmid was transformed into tobacco (Nicotiana tabacum L. cv Havana) using the Agrobacterium-mediated transformation method. The integration of the hlL-18 gene into the genome of transgenic tobacco plants was confirmed by polymerase chain reaction (PCR) amplification and its expression was observed in the suspension cells that were derived from the transgenic plant callus by using Northern blot analysis. The hlL-18 protein was detected in the extracts of the transgenic callus and in the medium of the transgenic tobacco suspension culture by using immunoblot analysis. Based upon enzyme-linked immunosorbant assay (ELISA) results, the expression level of the hlL-18 protein approximated $166{\mu}g/L$ in the suspension culture medium. Bioassay results from the induction of $interferon-{\gamma}$ from a KG-1 cell line indicated that the hlL-18 secreted into the suspension culture medium was bioactive.