• Journal of Plant Biotechnology
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• 제2권1호
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• pp.29-33
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• 2000

To develop an efficient propagation method for yew tree, zygotic embryos were cultured under various conditions. When dissected embryos were cultured on GA$_3$ containing media, the highest germination frequency was observed on WPM medium contaning 1.0 mg/L GA$_3$. For germination of the embryos, two different conditions were compared; culturing embryos with endosperm (Method I), and 2) culturing embryos only (Method II). Maximum germination was achieved in 0.5 mg/L GA$_3$ when embryos with endosperm were cultured on the media. Of the media tested, White and WPM medium were the most suitable on germination of embryos. The abnormality of yew embryos found was observed when it cultured on GA$_3$ or culture media. About 40% of the precociously germinated embryos could be developed into full seedlings. Seedlings contained taxol in high quantity (535 $\mu\textrm{g}$/g dry weight). In vitro techniques will be sewed as a useful tool for the development of transformed root cultures and biosynthesis studies.

• The Plant Pathology Journal
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• 제33권6호
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• pp.582-588
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• 2017

The objective of this study was to determine inhibitory activities of four volatile plant essential oils (cinnamon oil, fennel oil, origanum oil and thyme oil) on in vitro growth of Fusarium oxysporum f. sp. fragariae causing Fusarium wilt of strawberry plants. Results showed that these essential oils inhibited in vitro conidial germination and mycelial growth of F. oxysporum f. sp. fragariae in a dose-dependent manner. Cinnamon oil was found to be most effective one in suppressing conidial germination while fennel oil, origanum oil and thyme oil showed moderate inhibition of conidial germination at similar levels. Cinnamon oil, origanum oil and thyme oil showed moderate antifungal activities against mycelial growth at similar levels while fennel oil had relatively lower antifungal activity against mycelial growth. Antifungal effects of these four plant essential oils in different combinations on in vitro fungal growth were also evaluated. These essential oils demonstrated synergistic antifungal activities against conidial germination and mycelial growth of F. oxysporum f. sp. fragariae in vitro. Simultaneous application of origanum oil and thyme oil enhanced their antimicrobial activities against conidial germination and fungal mycelial growth. These results underpin that volatile plant essential oils could be used in eco-friendly integrated disease management of Fusarium wilt in strawberry fields.

• Journal of Plant Biotechnology
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• 제45권4호
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• pp.347-356
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• 2018

Chionanthus retusus is a small deciduous tree that is widely used in landscaping due to its beautiful white spring flowers and ornamental value. Conventional propagation through seeds requires one to two years of breaking dormancy. The objective of this study was to determine the conditions of in vitro germination in C. retusus. In vitro embryo culture was carried out to investigate the effects of six factors: basal media (McCown Woody Plant Medium (WPM) and Murashige and Skoog (MS)); plant growth regulators (different combinations and concentrations of naphthaleneacetic acid (NAA), 6-Benzylaminopurine (BA), and gibberellic acid ($GA_3$)); embryo age (collected weekly beginning 36 days after fruit setting); low temperature pretreatment (storing $4^{\circ}C$ for 1, 2, 3, and 4 weeks); coconut additives (100, 200, and $300ml{\cdot}L^{-1}$); and genotype (grouping plants depending on their flowering nature). The basal medium used in this study was WPM with $2mg{\cdot}L^{-1-1}\;GA_3$, $20g{\cdot}L^{-1}$ sucrose, and $6g{\cdot}L^{-1}$ Agar. WPM medium mixed with $GA_3$, resulted in higher germination rate as compared to when using a combination of auxin and cytokinin. $GA_3$ at $2mg{\cdot}L^{-1}$ was the most effective of all combinations and concentrations of PGRs. WPM medium with $2mg{\cdot}L^{-1}GA_3$ resulted in better and faster germination (75.93%). Embryos collected at 57 days after fruit setting had the highest percent of germinated seeds (87.04%) while low-temperature pretreatment of fruits at $4^{\circ}C$ for two weeks produced the highest germination (95.37%). These results of this study could be an open ground for development of an efficient protocol for commercial production of the ornamental tree.

• Journal of Plant Biotechnology
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• 제34권4호
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• pp.339-346
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• 2007

식물화분발아는 수분수정을 통한 식물의 종자형성과 식물개체의 지속적 세대유지를 위한 중요한 요소이다. 따라서, 본 연구에서는 boric acid를 이용하여 형질전환 하지 않은 (NT) 식물과 항암활성 (anti-colorectal cancer mAb CO17-1A, anti-breast cancer mAb BR55) 혹은 항바이러스 활성을 갖는 항체 (anti-rabies virus mAb57)의 유전인자를 발현하는 형질전환식물 간의 in vitro pollen germination과 Pollen tube growth의 차이를 알아보았다. Boric acid의 화분발아에 미치는 영향을 보기 위하여 0, 5, 10, 15, 20, $40{\mu}g/mL$의 농도를 갖는 발아용액 (germination buffer)을 담배식물 화분에 처리하여 pollen germination rate을 본 결과 $20{\mu}g/mL$에서 가장 높게 발아율 (49.5%)을 보였다. Boric acid ($20{\mu}g/mL$)에 대해 형질전환식물과 NT 식물에서 화분발아율을 관찰한 결과, boric acid를 처리하지 않은 화분에 비해 형질전환여부에 관계없이 발아율이 3-10배 증가하였다. 또한, boric acid $20{\mu}g/mL$농도에서 형질전환식물과 NT 식물의 pollen tube length의 성장률이 가장 높게 나타났다. 시간 별로 형질전환 식물들의 Pollen tube length를 확인 한 결과, germination buffer ($20{\mu}g/mL$)를 처리하고 난 후 3시간까지 가장 빠른 성장률을 보였으며, 3시간 후에는 그 성장률이 둔화됨을 확인하였다. Western blot를 이용하여 식물의 화분과 잎에서 황체유전자 단백질 발현을 확인하였다. 화분과 잎에서의 발현을 비교해 본 결과, 잎에서 항체의 안정한 발현을 확인하였다. 본 연구결과를 통해 $20{\mu}g/mL$의 boric acid가 항체발현 형질전환담배식물의 in vitro pollen germination을 보기 위해 가장 적절한 농도라는 것을 제시하였다. 궁극적으로 의료용 항체유전자를 발현하는 형질전환식물의 in vitro pollen germination viability를 확인함으로써 형질전환식물의 화분의 안정적인 수분수정이 이루어질 수 있음을 간접적으로 제시하였다.

• 생물환경조절학회지
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• 제17권4호
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• pp.297-305
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• 2008

치커리와 쑥갓에 있어 기내 파종 시 발아 및 무균묘 생산을 증진시키기 위한 적절한 방법을 제안하기 위해 일련의 실험들이 수행되었다. NaOCl의 적정 처리농도와 시간 검정을 통해 치커리 'Precole'과 'Chiavari' 및 쑥갓 'Okiku 3' 모두 Naocl 5%로 15분간 처리하는 것이 발아 및 유묘형성을 촉진시키고 오염발생을 억제하는 것으로 관찰되었다. 배지조성에 있어 MS와 1/2 MS처리구들의 발아반응성은 뚜렷이 차이가 나타나지 않았으며, sucrose 농도가 높아짐에 따라 발아 및 유묘형성이 지연되었다. 배양용기에 따른 반응성의 차이는 치커리 'Precole'의 경우 petri-dish에서 양호한 발아율 및 유묘형성율을 보여주었지만, 치커리 'Chiavari'와 쑥갓 'Okiku 3'에서는 용기에 큰 영향을 받지 않았다. 초음파 처리는 치커리의 발아율 및 유묘형성율을 증가시켰지만, 쑥갓 'Okiku 3'에서는 초음파 처리효과가 나타나지 않았다. 따라서 치커리와 쑥갓 모두에서 NaOCl 5%, 15분간 종자소독하고, sucrose가 무첨가된 1/2 MS 배지로 petri-dish를 이용하며, 초음파 처리는 NaOCl 5% 용매에서 120분간 하는 것이 기내 발아향상에 적당한 조건으로 선정되었다.

• Journal of Plant Biotechnology
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• 제41권1호
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• pp.44-49
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• 2014

In this study, we investigated the effects of sodium hypochlorite (NaOCl) and red or blue light-emitting diode (LED) light on embryo swelling and germination of Oreorchis patens (Lindl.) Lindl. A method for determining the swelling and protocorm formation in O. patens seeds through in vitro examination of immature seeds was established. NaOCl treatment of immature seeds was found to significantly enhance the extent of embryo swelling and protocorm formation in immature zygote embryos compared to those in the untreated controls. Additionally, the effects of white fluorescent light, and red and blue LED lights on embryo swelling and protocorm formation in in vitro cultured seeds were examined and compared to the conditions with or without NaOCl treatment. The most suitable light for embryo swelling and protocorm formation was the red LED light.

• Journal of Plant Biotechnology
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• 제43권1호
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• pp.132-137
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• 2016

The genus Cypripedium, known as lady's slipper orchid, has been considered to have a considerable marketability as a potted and garden plant with beautiful flowers; however, this species is becoming endangered and, in some places extinct, due to habitat destruction and illegal collections. As such, an optimized artificial propagation system is necessary for its conservation and horticultural cultivation. For the establishment of the in vitro proliferation of the endangered Cypripedium macranthos Sw., native to Korea, the effects of medium strength and sucrose concentrations on germination, protocorm formation and seedling growth were investigated through asymbiotic seed culture. The highest germination rate and protocorm formation rate were obtained with a 1/4 MS medium; higher MS medium strengths did not generate the favorable conditions required for germination and protocorm growth. The overall growth characteristics of roots and buds developed from protocorms were best in cultures of 1/4 MS medium. On this medium containing $10g{\cdot}L^{-1}$ sucrose, both the germination rate and protocorm formation rate increased significantly. The general growth properties of seedlings developed on the medium with $10g{\cdot}L^{-1}$ sucrose were best, showing the highest bud formation rate and root number. Our results demonstrate that the 1/4 MS medium, supplemented with $10g{\cdot}L^{-1}$ sucrose, could improve in vitro germination, and facilitate the growth of seedlings developed from the protocorms of C. macranthos Sw.

• Journal of Forest and Environmental Science
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• 제34권5호
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• pp.388-394
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• 2018

An efficient and reproducible protocol for in vitro germination of seeds of Couroupita guianensis has been developed in the present study. Couroupita guianensis is a medicinally important tree and categorized as threatened plant species by the IUCN. The seeds were surface sterilized using 0.1% $HgCl_2$ and inoculated on MS (Murashige and Skoog) and Woody Plants (Llyod and McCown) medium. The seeds with partially removed seed coat responded better (100%) on half strength MS medium as compared to the full (51%), one-fourth strength MS (79%) and the WP medium. Half strength MS medium conjunct with $2.0mg{\cdot}L^{-1}$ indole-3 butyric acid (IBA) was found best among the different types of auxins and cytokinins tested for seed germination, as all the seeds germinated on this medium combination. The seedlings were successfully hardened in the greenhouse and restored in the field with 83% survival rate. The protocol can be used as conservation measure for large scale multiplication of this threatened forest tree species.

• Plant Biotechnology Reports
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• 제2권2호
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• pp.153-161
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• 2008

The factors influencing somatic embryo maturation, high frequency somatic embryo germination, and plantlet formation were studied in Terminalia chebula Retz. Maturation of somatic embryo were influenced by a number of factors such as in vitro culture passage, concentrations of sucrose, levels of abscisic acid (ABA), basal media and media additive combinations. Maximum frequency of somatic embryo maturation ($57.22{\pm}2.02$), was obtained on MS medium supplemented with 50 g/l sucrose. Different factors such as strengths of MS nutrients, plant growth regulators, media additives and their combinations controlling somatic embryo germination and plantlet formation were studied. High frequency of germination and plantlet formation ($58.80{\pm}1.47$) were achieved by subsequent subculture of mature somatic embryos on MS medium containing 30 g/l sucrose and 0.5 mg/l benzyl-adenine (BA). However, although duration of in vitro passage of the callus tissue was critical, contribution of the combinations of plant growth regulators and media additives showed nugatory effect on somatic embryo maturation and germination as evident from variable responses.

• 식물병연구
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• 제24권4호
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• pp.292-301
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• 2018

Alternaria leaf spot disease has been a concern during a tomato production in greenhouse. In vitro antifungal activities of vapours of four plant essential oils, cinnamon oil, fennel oil, origanum oil and thyme oil, were investigated during in vitro conidial germination and mycelial growth of Alternaria alternata causing the tomato leaf spots to find eco-friendly alternatives for chemical fungicides. The four plant essential oils showed different antifungal activities against in vitro conidial germination of A. alternata in dose-dependent manners, and cinnamon oil vapour was most effective to suppress the conidial germination. The four plant essential oils showed similar antifungal activities against the in vitro mycelial growth of A. alternata in dose-dependent manners, but low doses of thyme oil vapour slightly increased in vitro mycelial growth of A. alternata. Necrotic lesions on the A. alternata-inoculated tomato leaves were reduced differently depending on kinds and concentrations of plant essential oils. Delayed conidial germination and germ-tube elongation of A. alternata were found on the tomato leaves treated with cinnamon oil and origanum oil vapours at 6 hpi. These results suggest that volatiles from cinnamon oil and origanum oil can be provided as alternatives to manage Alternaria leaf spot during the tomato production eco-friendly.