• Title/Summary/Keyword: In vitro germination

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Development of assay method for the activities of new compounds, and the effect of several fungicides against spore germination, adhesion, and myceial growth of Colletotrichum sp. causing red pepper anthracnose (고추 탄저병균의 포자 발아와 부착, 균사 생장에 미치는 화합물의 활성 검정법 확립 및 살균제의 효과)

  • Kim, Jae-Jeung;Kim, Joon-Tae;Park, Sung-Woo;Park, Eun-Suk;Kim, Heung-Tae
    • The Korean Journal of Pesticide Science
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    • v.7 no.3
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    • pp.159-168
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    • 2003
  • With microtiter plate, the assay method was developed for detecting the fungicidal activity of new compounds against spore germination, spore adhesion and mycelial growth of Colletotrichum sp. JC24 cal1Sing red pepper anthracnose. Also, the effects of some commercialized fungicides on fungal development like above mentioned were investigated by measuring the optical density of mycelia grown into wells of microtiter plate. For the standardization of assay method, some factors, such as the treatment of MTT and/or propanol, inodulum density and incubation period, affecting on mycelial optical density were investigated. For obtaining precise and consistent mycelial optical density, it was necessary the treatment of MTT for 12 hrs and propanol for 1 hr. inoculum density adjusted to $1\times10^5$ spores/mL and incubation period for 36 hrs at $25^{\circ}C$. For fungicidal activities, 6 protective fungicides, 6 ones inhibiting sterol biosynthesis, and one inhibiting respiration were used in this study. While mancozeb, chlorothalonil and dithianon among 6 protective fungicides inhibited strongly spore germination, adhesion, and mycelial growth at $6.25{\mu}g/mL$, propineb, iminoctadine and fluazinam inhibited intermediately spore germination and mycelial growth at $100{\mu}g/mL$. Washing above 3 fungicides with new PD broth, their activity against spore adhesion decreased. With hexaconazole, tebuconazole and myclobutanil, the tendency of the activity against fungal differentiation of the early infection stage was similar to the latter group of protective fungicides, showing the decrease of the inhibitory activity against spore adhesion by washing 2 hrs after incubation. However, kresoxim-methyl inhibited spore adhesion distinctly, depending on the applied concentrations. Based on these results, it might be able to assess the fungicidal activity of many compounds against spore germination, adhesion and mycelial growth by the use of microtiter plate in vitro. Using the assay developed in this report, it was possible to investigate the inhibitory activity of some commercialized fungicides, too.

Conservation of Swertia chirata through direct shoot multiplication from leaf explants

  • Chaudhuri, Rituparna Kundu;Pal, Amita;Jha, Timir Baran
    • Plant Biotechnology Reports
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    • v.2 no.3
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    • pp.213-218
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    • 2008
  • Swertia chirata is an endangered gentian species that prefers to grow at higher altitudes. This ethnomedicinal herb is known primarily for its bitter taste caused by the presence of important phytochemicals that are directly associated with human health benefits. Due to a continuous loss of habitat and inherent problems of seed viability and seed germination, alternative strategies for propagation and conservation are urgently required to prevent the possible extinction of this species. We have formulated a reproducible protocol for the rapid propagation and conservation of this plant using leaves taken from in vitro shoot cultures. Direct induction of more than seven shoot buds per explant was achieved for the first time when the explants were placed on MS medium supplemented with $2.22{\mu}M$ N-6-benzyladenine, $11.6{\mu}M$ kinetin, and $0.5{\mu}M$ ${\alpha}-naphthalene$ acetic acid. Direct organogenesis was noted exclusively from the adaxial surface of the basal segments of leaves. Leaves closer to the apical meristem were more responsive than those farther away from the meristem. Plants raised through direct organogenesis were evaluated for their clonal fidelity by chromosomal analysis and DNA fingerprinting. Complete plants were successfully transferred to the field condition and produced viable seeds. Given the enormous potential of this age-old medicinal plant in terms of potential health-benefitting drugs, this protocol can be used for commercial propagation purposes and to initiate future genetic improvement studies.

Ex-situ conservation and cytotoxic activity assessment of native medicinal orchid: Coelogyne stricta

  • Thapa, Bir Bahadur;Thakuri, Laxmi Sen;Joshi, Pusp Raj;Chand, Krishna;Rajbahak, Sabari;Sah, Anil Kumar;Shrestha, Resha;Paudel, Mukti Ram;Park, So Young;Pant, Bijaya
    • Journal of Plant Biotechnology
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    • v.47 no.4
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    • pp.330-336
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    • 2020
  • Ex-situ conservation of the ornamental and medicinal orchid, Coelogyne stricta, was performed by mass propagation using seed culture. Propagation stages were optimized using full- and half-strength solidified MS medium with different phytohormones. Maximum seed germination (88 ± 0.5% over 6 weeks of culture) was achieved on half-strength MS medium supplemented with 15% coconut water. Maximum shoot numbers were found on full-strength MS medium supplemented with 1 mg/L BAP, 2 mg/L Kinetin, and 10% coconut water, while the longest root was developed on full-strength MS medium with 1.5 mg/L IBA. A 2:1:1 combination of coco-peat, pine bark, and sphagnum moss was found to be a suitable potting mixture resulting in 80% seedling survivability. The cytotoxic activity of extracts of both wild plants and in vitro-developed protocorms was determined using an MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay on a cervical cancer cell line. The wild plant extract inhibited the growth of 41.99% of cells, showing that this extract has moderate cytotoxic activity toward cervical cancer cells.

Propagation of Acorus gramineus from Seeds and In vitro Culture (종자 및 기내배양을 이용한 석창포 증식)

  • Park, Young-Chul;Kim, Jeong-Seon;Yang, Seok-Chul;Cho, Youn-Dong;Kim, Yong-Duk;Park, Jae-Kweon
    • Korean Journal of Plant Resources
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    • v.21 no.5
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    • pp.347-351
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    • 2008
  • Rhizomes of Acorus gramineus Soland have been used as sedatives, analgesics, and stomachics in Korean medicine. Even though A. gramineus produced in Korea is known as having better efficacy of a remedy than that in China, its cultivation area has not increased because of the lack of seedlings. To solve this problem, seed propagation method was tested. Seeds were harvested and sowed three times in June, 2005. The best results showed at greenish yellow seed stage harvested in mid June, and its germination ratio was 90.7%. Seeds were well stored at $5^{\circ}C$ for 30 days. As for in vitro culture, multiple shoots were induced first in MS basal medium supplemented with $2.0mg{\cdot}L^{-1}$ NAA plus $0.1mg{\cdot}L^{-1}$ BA, and then roots were induced in MS basal medium containing $0.1mg{\cdot}L^{-1}$ BA or $0.5mg{\cdot}L^{-1}$ NAA plus $1.0mg{\cdot}L^{-1}$ BA. Growth of A. gramineus seedlings in Wonyesangto, potting material sold in markets for horticultural plants, was superior to vermiculite.

Useful Applications Decursinol Derivatives as Biochemical Fungicides (데커시놀 파생물질들의 항징균제로서의 활용)

  • Yun-Jeong Shin;Jiyoung Shin;Hyewon Jang;Hokyoung Son;Yonghoon Kwon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.30-30
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    • 2022
  • Natural products decursin and decursinol angelate were recently reported as benign fungicides for controlling rice blast. Inspired by the structural similarity of the cumarin compounds and gained hint from the skeletal motifs, we designed and prepared synthetic compounds to increase the natural product efficacy and evaluated their antifungal activities against various plant disease pathogens in vitro. Synthetically prepared compound 4 and 5 indeed suppressed the mycelial growth of B. cinerea, F. oxysporum, P. italicum, and R. quercus-mongolicae. Additionally, compound 5 effectively prevents the growth of C. coccodes and C. parasitica. Furthermore, both 4 and 5 possess better inhibitory activities on spore germination of F. oxysporum and M. oryzae than the natural product decursin. These results suggest that the effect of the lead compound for plant disease protection can be improved by tuning the structure of the original natural product and decursinol chloroacrylates 4 and 5 are candidates for the control of F. oxysporum and M. oryzae.

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Decursinol Derivatives Useful as Fungicides (데커시놀 파생물질들의 항징균제로서의 활용)

  • Yun-Jeong Shin;Jiyoung Shin;Hyewon Jang;Hokyoung Son;Yonghoon Kwon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.98-98
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    • 2022
  • Natural products decursin and decursinol angelate were recently reported as benign fungicides for controlling rice blast. Inspired by the structural similarity of the cumarin compounds and gained hint from the skeletal motifs, we designed and prepared synthetic compounds to increase the natural product efficacy and evaluated their antifungal activities against various plant disease pathogens in vitro. Synthetically prepared compound 4 and 5 indeed suppressed the mycelial growth of B. cinerea, F. oxysporum, P. italicum, and R. quercus-mongolicae. Additionally, compound 5 effectively prevents the growth of C. coccodes and C. parasitica. Furthermore, both 4 and 5 possess better inhibitory activities on spore germination of F. oxysporum and M. oryzae than the natural product decursin. These results suggest that the effect of the lead compound for plant disease protection can be improved by tuning the structure of the original natural product and decursinol chloroacrylates 4 and 5 are candidates for the control of F. oxysporum and M. oryzae.

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Herbicidal Activity and KAPAS Inhibition of Juglone with Potential as Natural Herbicide (천연 Naphthoquinone계 Juglone의 KAPAS 저해 및 제초활성 특성)

  • Choi, Jung-Sup;Lim, Hee-Kyung;Seo, Bo-Ram;Kim, Jin-Seog;Choi, Chun-Whan;Kim, Young-Sup;Ryu, Shi-Yong
    • Korean Journal of Weed Science
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    • v.31 no.3
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    • pp.240-249
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    • 2011
  • The potential of juglone a plant naphthoquinone as a natural herbicide on new target, 7-keto-8-amino pelargonic acid synthetase (KAPAS) in the early step of biotin biosynthesis pathway, was performed in vitro and in vivo. Juglone effectively inhibited KAPAS activities in vitro and the $IC_{50}$ was $9.5{\mu}M$. Foliar application of juglone showed very good herbicidal activity to the eight-tested weed species. Among them, Solanum nigrum was completely controlled at a concentration of $250{\mu}g\;mL^{-1}$ with main symptoms of desiccation or burndown. Digitaria sanguinalis and Aeschynomene indica were also sensitive to juglone treatment. All eight weed species were controlled by 90~100% at a concentration of $500{\mu}g\;mL^{-1}$. However, soil application of juglone to Digitaria sanguinalis did not show any herbicidal symptoms. Cellular leakage from cucumber leaf squares treated with juglone increased depending on the concentrations increased from 6.25 to $100{\mu}M$ after 24 hours incubation with or without light. However, chlorophyll loss in cucumber leaf squares was negligible. Biotin supplements significantly rescued the inhibition of germination rate of Arabidopsis thaliana seeds previously inhibited by the juglone. Our results suggest that the juglone is a possible environmental friendly herbicide candidate with a new target KAPAS inhibiting activity.

Plant Regeneration from the Segments of Petioles of Cacalia firma (병풍쌈의 엽병 조직 절편으로부터 식물체 재분화)

  • Choi, Soo-Wan;Lim, Soon;Park, Wan-Geun;Choi, Yong-Eui
    • Korean Journal of Plant Resources
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    • v.24 no.5
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    • pp.483-488
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    • 2011
  • Cacalia firma recently has been used increasingly as leaf vegetables but endangered in natural forest. In this work, we established the plant regeneration via adventitious shoot formation from petiole segments of seedling and in vitro plantlets. Wounding of seed coats and $GA_3$ treatments were effective to induce in vitro germination of seeds, whereas, seed did not germinate at all without these treatment. When cotyledon, leaf, petiole, and root segments of seedling were cultured on medium with 2 $mg{\cdot}L^{-1}$ benzyl adenine (BA) and 0.5 $mg{\cdot}L^{-1}$ naphthaleneacetic acid (NAA), petiole segments showed highest number of shoots per explant among the other segments. Among the various kinds of cytokinins, BA, isopentyl adenine (2-ip), kinetin, zeatin, thidiazuron (TDZ), TDZ and BA treatments were effective to induce high frequency of adventitious shoot formation from petiole segments of in vitro propagated plants. NAA stimulated the frequency of adventitious shoot formation but not for number of adventitious shoots per explants compared to TDZ or BA treatment alone. Most of adventitious shoots were developed directly from surfaces of explants. Adventitious shoots were transferred on medium with IBA for root formation, thereafter the plantlets were successfully transferred to soil.

Effect of Plant Growth Regulators, Media and Celling Agents on In Vitro Microtuber Production of Pinellia ternata Breit (1식물생장조절제, 배지와 고형지지물이 반하의 기내 소괴경 생산에 미치는 영향)

  • Kim, Yong-Kyung;Lee, Ji-Yeon;Kim, Eung-Hwi;Cho, Dong-Ha;Park, Sang-Un
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.3
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    • pp.210-214
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    • 2007
  • The study was carried out to establish in vitro microtuber production from leaf and petiole explant cultures of Pinellia ternata. Culture conditions were optimized by investigating the effect of plant growth regulators, different media, and gelling agents on the efficiency of microtuber indurtion. Among the different combinations of plant growth regulators tested, the combination of 0.1 mg/l 2,4-Dichlorophenoxyacetic (2,4-D) and 0.5 mg/l 6-benzylaminopurine (BAP) showed the highest yield fer microtuber production from leaf (3.9) and petiole (4.7) explant culture on MS medium far 6 weeks. SH(Schenk and Hildebrandt) medium was the most effective medium for microtuber induction and the half strength SH medium was better than SH medium for microtuber production from both leaf and petiole culture. Gelrite was better than agar in the formation of microtubers and 4% Gelrite showed the highest number of microtubers per explant frome leaf (5.9) and petiole (7.8) culture. Germination rate of microtubers after cold storaged for one months long was 86% from in vitro culture and 43% from autoclaved soil.

A Tetraploid Induction in Hypericum patulum Thunberg by Colchicine Soaking Treatment (콜히친 침지처리에 의한 '망종화'의 4배체 식물유도)

  • Kwon, Soo Jeong;Cho, Kab Yeon;Kim, Hag Hyun
    • Korean Journal of Plant Resources
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    • v.26 no.2
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    • pp.284-288
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    • 2013
  • This study aimed to get the basic data on the breeding of good varieties in Hypericum patulum Thunberg. The optimum materials, concentration and soaking time were examined to identify the effective approach to induce tetraploid plant by colchicine treatment to cultivate the varieties. For the seed germination rate of seed by colchicine treatment, the higher colchicine concentration was and the longer soaking time was, the more the germination rate decreased. While individuals were germinated in 16 test groups except control group (no treatment group), all the plants were diploid and no tetraploid was induced. For the plant regeneration rate by colchicine treatment on the explant of Hypericum patulum Thunberg that was under in vitro culture, the higher the colchicine concentration increased, the ress the regeneration rate. While total 147 individuals were regenerated in all treatment, when the explant was soaking treatment in more than 0.05% for over 6 hours, tetraploid could be obtained. In the soaking treatment of 0.05% for over 6 hours, tetraploid could be obtained. In particular, for the soaking treatment in 0.05% for 12 hours, 8 tetraploids were induced, which was about 47.1% of the number of plant regenerated. In accordance with the observation on doubling of DNA contents in leaf in order to identify polyploidy, the peak DNA content of G1 phase was 94.5 for diploid and 192.5 for tetraploid. It confirmed doubling of DNA content. Furthermore, the number of chloroplasts per guard cell depending on polyploid was around 10 in diploid and 17 to 19 in tetraploid, which were around 1.7 to 1.9 times as much as diploid.