• 한국육종학회지
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• 제49권3호
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• pp.170-179
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• 2017

분자농업은 식물을 일종의 공장개념으로 확대 적용하여 산업적으로 가치가 높은 유용물질들을 대량생산하는 분야로 최근 많은 각광을 받고 있다. 그 중에서 벼 캘러스를 이용한 단백질 발현 시스템은 대량 배양이 가능하고 목적 단백질의 높은 발현율로 인한 산업화가 가능한 기술이다. 본 연구는 이러한 벼(Oryza sativa L.) 캘러스의 활용도를 높이기 위한 효율적인 형질전환 시스템을 구축하기 위해 수행되었다. 형질전환에 이용된 종자의 종피 제거 시 손을 이용함으로써 캘러스 유도율을 높여주었고, 6년 정도의 오래된 종자에서도 원활한 캘러스 유도가 가능하였다. 목적 유전자가 도입된 캘러스의 선발은 최소한 3주 이상의 배양기간을 필요로 하였고 가장 효율적인 것은 한번의 계대를 포함한 6주 배양 후 선발하는 것이었다. 이러한 선발은 유전자가 식물세포의 genomic DNA 안에 안정적으로 삽입되어 이루어졌음을 서던 블롯 분석 및 후대검정 등을 통하여 확인할 수 있었다. 이러한 장기적이고 대량의 배양이 가능한 벼 캘러스의 효율적인 선발 시스템은 벼를 이용한 유용 재조합 단백질의 산업화에 실속있는 기술로 기여할 수 있을 것이다.

• 한국자원식물학회지
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• 제11권1호
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• pp.15-21
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• 1998

This study was carried out to determine the effects of plant growth regulators on cell culture and organogenesis from Sicyos angulatus L. using explants of leaves, stems and cotyledons. Optimal callus induction for S. angulatus was obtained on MS medium with 0.1mg/$\ell$ BA and 2.0mg/$\ell$ 2,4 -D from cotyledons, 0.1mg/$\ell$ BA and 5.0mg/$\ell$ NAA from leaves explants, Optimal media for subculture and growth of S. angulatus callus were 1/2 MS medium with 0.1mg/$\ell$ BA and 1.0mg/$\ell$ 2,4 -D for solid culture, and 0.1mg/$\ell$ 2,4-D for suspension culture. Many adventitious roots with some shoots were formed were formed from leaf and cotyledon explants of S. angulatus during callus induction with optimal combinations of plants growth regulators.

• Journal of Plant Biotechnology
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• 제30권2호
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• pp.151-154
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• 2003

This study was conducted to determine the optimal concentrations of plant hormones (2,4-D, picloram, dicamba, NAA, kinetin) and the suitable explants among seeds, hypocotyl, and cotyledons on calls formation and plant regeneration of stevia(Stevia rebaudiana Bertoni). The frequency of cellus formation was higher in the young leaf-explants then the older ones, and in the seeds then the hypocotyls and cotyledons on MS medium with 1mg/L 2,4-D. After transfer of seed-derived stevia callus producing embryogenic callus on plant-regeneration medium, the frequency of plant regeneration from callus was 23.8% in MS medium with 1mg/L NAA and 3mg/L kinetin.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2023년도 임시총회 및 춘계학술대회
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• pp.53-53
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• 2023

Abeliophyllum distichum, one of the Korean endemic plant, is a significant pharmaceutical plant resource. A. distichum with phenylethanoid glycoside can use to regulate the development of cancer, DNA damage with radicals, and the generation of inflammatory mediators. In this study, we investigated whether the biomass, content of phenylethanoid glycoside, and growth rate of callus derived from A. distichum (cultivar Okhwang1, CAD) change in the absence or presence of plant hormones (2,4-Dichlorophenoxyacetic acid; 2, 4-D and 1-Naphthaleneacetic acid; NAA). The results showed that the best biomass, the growth rate of callus, and the contents of phenylethanoid glycoside were cultivated on Murashige and Skoog (MS) growth medium fortified with 1 ppm 2,4-D + 2 ppm NAA after 4 weeks. In a further study, CAD was cultivated on MS growth medium fortified with an elicitor (Methyl Jasmonate, MeJA). The results showed that CAD turned to brown color and fragile form with the elicitor. HPLC-PDA analysis revealed that the contents of phenylethanoid glycoside in the elicitor-treated group were higher than in the elicitor-non-treated group. These results are consistent with the findings of Arano-Varela H et al.,'s study which is that acteoside production can increase after the treatment of MeJA. Therefore, this study can be used to develop an effective and sustainable production of useful substances as an alternative to plant cultivation.

• Journal of Plant Biotechnology
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• 제30권2호
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• pp.115-121
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• 2003

In this study, plant regeneration and Agrobacterium tumefaciens-mediated transformation Kentucky bluegrass(Poa pratensis L.) were evaluated. Three different types of calli were produced depending on the combinations of growth regulators. They were non-friable brown or gray-colored callus (type I), compact, friable and yellow or white-colored callus (typeII), and soft, watery translucent callus with differentiated structure (typeIII). The highest regenerable organogenic callus (typeII) was obtained on the medium containing 1mg/L, 2,4-D and 0.1mg/L BA. Additionally, the production of typeII calli increased significantly when AgNO$_3$ was added to the callus induction and growth medium. The highest frequency of multiple shout formation from typeII callus was obtained on MS medium containing 1mg/L BA and 1mg/L Thidiazuron(TDZ). The organogenic calli(typeII) were inoculated with Agrobacterium tumefaciens strain EHA101 harboring the binary vector pIG121Hm with $\beta$-glucuronidase gene, and various factors were found to influence the transfer-DNA delivery efficiency. The highest transient GUS activity was observed on typeIIcallus. In the present work, we reported the first transient GUS activity of Kentucky bluegrass mediated by Agrobacterium tumefaciens. Our system may contribute to genetic improvement for breed-recalcirtrant grass species, Kentucky bluegrass.

• Journal of Plant Biotechnology
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• 제4권1호
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• pp.29-32
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• 2002

The ability to form embryiod from callus in satsuma mandarin is low and unstable. In this study, the conditions of embryogenic calli induced from nucellar tissue for promotion of plant regeneration in satsuma mandarin were investigated. The calli of I, II and III line were divided into two sizes of 0.5 mm and 1.0 mm in diameter and two weight gradients of percoll at 40% and 50% though the filter mesh. The frequency of embryo formation from $\phi$ 1.0mm-40% was slightly higher than callus that from others. Adventitious embryoids developed to a globular stage were transferred to regeneration medium. In 'Miyagawa Wase', the embryos from I and II line developed into a heart stage from most of $\phi$ 0.5 mm- 40% and $\phi$1.0 mm-40% calli, but it failed in 'Sugiyama Unshu'. In the cultivar of 'Miyagawa Wase', 63% of adventitious embryos transferred to the regeneration medium developed into the heart stage from the most $\phi$ 1.0 mm-40% calli of I line, but of 'Sugiyama Unshu' failed in some calli condition. The embryoids from two callus lines developed further to shoots and plantlets, while the embriods from III line abnormal failed to regenerate in the cultivar. From these results, it is suggested that the plant regeneration from embryogenic callus in satsuma mandarin could be affected by callus conditions.

• 한국작물학회지
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• 제39권6호
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• pp.537-541
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• 1994

식용 또는 약용으로 그 이용가치를 충분히 가지고 있는 강활의 미숙화서, 줄기, 그리고 엽병으로부터 캘러스 유도와 식물체 발생에 관하여 실험한 결과를 요약하면 다음과 같다. 1. 캘러스 유도와 증식정도는 미숙화서를 2,4-D 2mg/l 첨가배지에 치상했을 때 가장 양호하였 다. 2. 2,4-D 1mg/l와 2mg/l 첨가배지에서 미숙화서로부터 유도된 담황색의 유연한 캘러스 표면에 희고 단단한 embryogenic 캘러스가 발생하였고, 그외 배지에서는 발생하지 않았다. 3. 줄기 발생은 2,4-D 0.1mg/l와 Kinetin 1mg/l 그리고 2,4-D 0.5mg/l와 Kinetin 2mg/l를 각각 조합한 배지에서 가장 효율적이었다. 4. 캘러스 유도와 식물체 재생에 가장 적합한 재료는 미숙화서라 생각된다.

• 식물조직배양학회지
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• 제27권3호
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• pp.233-238
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• 2000

야생 흰제비꽃의 엽병 유래 callus를 장기 계대배양하는 과정 중에 발생하는 순화된 friable callus와 분화능이 높은 compact callus를 비교 관찰한 바, friable callus는 연초록색으로 부서지기 쉬운 부드러운 callus이고, compact callus는 진녹색으로 단단한 callus였다. 동결처리 한 시료를 주사전자 현미경에서 동일하게 200배로 관찰하여 보면, friable callus 는 작은 세포집단으로 이루어진 세포군의 주변부에 고도로 액포화된 세포가 광범위하게 분포되어 있는 반면, compact callus는 거의 균일한 세포들로 구성되어 세포구성이 치밀한 것으로 관찰되었다. 또한 friable callus와 compact callus로 부터의 체세포배형성은 배양세포에서 배가 발생하여 배양기간이 지남에 따라 식물체로 분화하였다. 이와 같은 과정은 배양세포의 세포질이 보다 충만한 부위에서 배유사체 (embryo like body)의 발생이 이루어지는 것으로 관찰되었다.

• 한국자원식물학회지
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• 제7권2호
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• pp.171-175
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• 1994

The effects of medium, hormones, and PFP on the proliferation of red callus in leaf tissue cultures of Garden orach(Atriplex hortensis L.) was investigated. As a result,88% of leaf tissues formed eallus on MS nledium containing 2mg/$\ell$ 2,4-D. Fresh weight of callus was higher on MS medium than on Bsand NN media. It was also found that 2, 4-D was more effective than Dicamba and Picloram. The op-timum concentrations of hormones for callus proliferation depended on culture media. Isolated red cal-lus grew markedly both on MS medium supplemented with 1-2mg/$\ell$ 2, 4-D and Bs medium contain-ing 2-4111g/$\ell$ 2,4D. Callus proliferated on B5 and NN media containing Dicabma Img/$\ell$ as well as onthe same media containing 2mg/$\ell$ Picioram. The addition of PFP concentrations of 2, 5, and 40mg/ $\ell$rcspectiely to culture medium caused increase of callus fresh weight, especially under light condition.

• Journal of Plant Biology
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• 제33권4호
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• pp.253-257
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• 1990

Callus-inducing ability of Alnus hirsuta was examined by culturing various tissues (leaf, hypocotyl, cotyledon and seed) on NT (Nagata & Takebe) medium, supplemented with 2.5$\mu$M 2,4-D. Leaf-originated callus was cultured on media varying in auxin (IBA and NAA) and cytokinin (BAP) concentrations to examine the effects of auxin and cytokinin on callus growth. Maximum growth was obtained at 10 $\mu$M IBA+10$\mu$M BAP and 10$\mu$M NAA without cytokinin. Cell suspensions established from cotyledon-originated callus yielded viable protoplasts after incubation for 16-18 hours in an enzyme mixture (1% (w/v) Onozuka R-10 0.5% (w/v) Macerozyme, CPW salts and 13% (w/v) mannitol, pH 5.8). Protoplasts were cultured on NT medium, supplemented with glucose, hormones and coconut milk. After 6 weeks of culture, protoplasts sustained cell divisions to form microcallus, which showed various colors from red to white.