• Title/Summary/Keyword: IN VITRO SEED GERMINATION

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Screening of Medicinal Plants with Antifungal Activity on Major Seedborne Disease (주요종자전염병 억제를 위한 항균성 약용식물탐색)

  • Paik, Su-Bong;Chung, Il-Min;Doh, Eun-Soo
    • Korean Journal of Medicinal Crop Science
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    • v.6 no.4
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    • pp.277-285
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    • 1998
  • Antifungal activity on major seedborne diasease of crops was screened by the treatment of the extracts from 50 medicinal plants in vitro and in vivo. The extracts of garlic and taxus, Rheum undulatum, Achiranthes japonica, Glycyrrhiza uralensis, Oenothera lamar kiana treated with the blotting filter paper and water agar methods inhibited the growth of Pyricularia oryzae, Alternaria sesamicola, Colletotrichum gloeosporioides, and Alternaria brassicicola among the tested plants. Antifungal activities on infected seeds by soaking methods were shown even at the dilution of the extracts by 10 times. The activity was the highest in soaking seeds at $25^{\circ}C$ for 24 hours. The effect of plant extract on seed germination was not significant as compared with untreated seed. However, early growth of seedling was increased by the treatment of extracts. The extract of taxus slightly inhibited the seed germination of radish and chinese cabbage but those of Achirunthes japonica, Glycyrrhiza uralensis and Oenthfera lamarkiana showed severe damage on the seed germination and early growth of seedling.

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In Vitro Regeneration of Pongamia pinnata Pierre

  • Sujatha, K.;Hazra, Sulekha
    • Journal of Plant Biotechnology
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    • v.33 no.4
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    • pp.263-270
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    • 2006
  • Pongamia pinnata Pierre is a tree legume, having potential in production of raw material for biodiesel. A protocol for in wk propagation of this plant was standardized using seedling explants. Growth regulators (GR) including gibberellic acid $(GA_3),\;N^6-benzylaminopurine(BA)$, thidiazuron (TDZ), and Adenine sulphate (Ads) were tested for optimum germination of seeds. Removal of seed coat prior to germination, controlled fungal growth partially but enhanced bacterial growth. Antibiotic cefotaxime was ineffective in controlling bacterial contamination. Seedling derived nodal explants and cotyledon nodes with attached cotyledons were excised and cultured for induction of shoots. Optimum sprouting and multiplication of shoot buds were obtained in MS medium supplemented with $8.88{\mu}M$ BA. These buds differentiated and rooted on medium devoid of GR. Optimum growth of Pongamia seedling was obtained in cotton plugged culture vessels. Reculturing of the cotyledon node explants produced more shoots from the same site. This process of removing shoots and reculturing of cotyledon node was followed for eight passages yielding 4 to 8 shoots in each cycle. The shoots (75%) rooted on half strength MS basal medium supplemented with 0.22% charcoal. All plants survived on transfer to soil. This is the first report on in vitro regeneration of Pongamia pinnata. This report demonstrates the possibility of coupling more than one parameter in single experiment to hasten the process of standardization. The process of cycling the nodal explant repeatedly for production of large number of shoots from single meristem may find application in genetic transformation experiments wherein meristems are used for transformation.

In vitro Multiplication through Single-Node Culture of Sea-Milkwort (Glaux maritima L.) (갯봄맞이(Glaux maritima L.) 실생의 단마디배양을 통한 기내증식)

  • Bae, Su-Ji;Kang, Beum-Chang;Jeong, Mihye;Kim, Soochong;Kim, Chang Kil;Han, Jeung-Sul
    • Horticultural Science & Technology
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    • v.34 no.3
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    • pp.461-471
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    • 2016
  • This study was conducted to establish an in vitro propagation system for sea-milkwort (Glaux maritima L.), which is an endangered coastal plant species with high horticultural value. Two phenotypes, 'Red type (RT)' and 'Pistachio type (PT)' based on the colors of stem and flower, were obtained from a personal horticulturist in 2009 and used for this study as plant materials. The stock plants showed typical morphologies in flower, capsule, and seed appearances as previously reported. Low temperature treatment at $4^{\circ}C$ for four or more weeks after in vitro sowing maximized seed germination percentage, indicating that imbibition of seed and subsequent low temperature treatment are crucial for its germination. The in vitro seedlings had phenotypic variation, falling into 'RT' and 'PT' classes like the stock plants. Although slight differences depending on genotype and medium were recognized, the fourth or fifth nodes detached from the in vitro seedlings revealed the best multiplication efficacy when estimated on the basis of total number of nodes of newly developed axillary shoots. In addition, the nodes from 'RT' and 'PT' regenerated the most shoots on medium supplemented with $0.5mg{\cdot}L^{-1}$ BA alone and $0.5mg{\cdot}L^{-1}$ BA plus $0.5mg{\cdot}L^{-1}$ IAA, respectively. The node culture-derived plantlets were well acclimatized in a culture room ex vitro and completed the pseudo-annual life cycle coincident with that in the natural salt march habitat with the current cultivation method of applying fresh water-irrigation under an inland environment. This work represents the first report of in vitro propagation of sea-milkwort. Thus, our study will contribute to exo-habitat conservation and natural habitat restoration of this endangered species in addition to development of a horticultural product.

Improvement of Seed Germination in a Spontaneous Autotetraploid of Poncirus and Chlorophyll Fluorescence of Seedlings in Salt Stress (동질 사배체 탱자의 종자 발아 증진과 염류 과잉에 따른 엽록소 형광 반응)

  • Chae, Chi Won;Yun, Su Hyun;Park, Jae Ho;Kim, Min Ju;Han, Seung Gab;Kang, Seok Beom;Koh, Sang Wook;Han, Sang Heon
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1079-1087
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    • 2013
  • Speed germination success and robust vegetative growth of citrus rootstock through improved sowing methods and fertilizer inputs offer the usage of root system for the citrus. The current study evaluated the influence of seed coat removal and different fertilizer concentrations on plant germination and plant growth of spontaneous rootstock siblings. Decoated and coated seeds of diploid and tetraploid plants were sown in tubes. Commercial fertilizer concentrations of 0, 2, 4, 6, 8 and $10g{\cdot}l^{-1}$ were added. The experimental layout followed a randomized block $2{\times}6$ factorial design (seed coat removal ${\times}$ fertilizer concentration) for each rootstock. Fertilizer concentrations were 0, 10, 20 and $30g{\cdot}l^{-1}$ of the fertilizer for the resistance of the strength on the salt level. The germination rate of seeds without testa sown in vitro was improved (67-80%) compared to that of nontreated seeds. The eventual tree height of the seeds without testa in the diploid group was increased due to higher fertilization compared to that in the nontreated group. The removal of seed testa promoted the seed germination of both diploid and tetraploid trifoliate orange and resulted in greater height. Their vegetative development was also increased due to the increased fertilization of the rootstock. The Fv/Fm value for the diploid plants was 0.4 and 0.8 for the tetraploid ones under salt stress after 11 days of treatment. The removal of seed testa may improve the seed germination of trifoliate orange. Tetraploid trifoliate orange appears to possess resistance to salt stress compared to the diploid variety.

Isolation of Cucurbitacin E from Sprouted Pumpkin Seed and Analysis of Its Anti-cancer and Anti-inflammatory Activities (발아 호박씨로부터 Cucurbitacin E의 분리정제 및 항암, 항염증 활성)

  • Sim, Hu-Sung;Jang, Byeong-Chur;Park, Hye-Min;Jeng, Byeong-Yong;Oh, Man-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.7
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    • pp.834-840
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    • 2008
  • In order to improve the use of pumpkin seed, the present study was performed to isolate compositions of the bitter components which were not seen in pumpkin seed itself but newly biosynthesized during germination of the seed. The compositions isolated were then further purified by TLC and preparative HPLC in which a fraction with Rf 0.73 and RT 10.3 was obtained. Cucurbitacin E with molecular weight of 557 from the fraction was finally identified by subsequent structural analysis of LC-MS/MS. The production of cucurbitacin E peaked with 224.7 mg/kg at 4 days of germination at $20^{\circ}C$ with the water supply at ntervals of 48 hrs in the darkness, while that of cucurbitacin E reached 146.7 mg/kg in the brightness. In vitro-cell based assays demonstrated that the isolated and purified cucurbitacin E inhibited proliferation of A549 lung cancer cells and suppressed expression of the IL-$1{\beta}$- or PMA-induced cyclooxygenase-2, an inflammatory protein in A549 cells, suggesting its anti-proliferative and anti-inflammatory activities.

Effect of Basal Medium and Plant Growth Regulator on in vitro Plant Regeneration from Axillary Buds of Walnut New Cultiver "Sinlyeong"

  • Kwon, Young Hee;Lee, Joung Kwan;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong;Huh, Yoon Sun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.15-15
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    • 2019
  • The walnut (Juglans regia L.), a member of the Juglandaceae, is native to the mountain ranges of central Asia. This species of walnut is valued commercially for its nuts and in some areas for its timber. The seeds of walnut are recalcitrant and it has strong integument dormancy and their germination is irregular, making its natural propagation difficult. Low percentage of seed germination and long propagation cycle are the main problems of propagation. This study was conducted medium composition on in vitro plantlet regeneration from axillary buds of walnut. It has proved to be the most generally applicable and reliable method of in vitro propagation. Micropropagation culture that axillary buds are excised aseptically enables faster multiplication of plants. The axillary buds of walnut new cultivar "Sinlyeong" were cultured on two basal media which contained the different plant growth regulators depending on the respective shooting and rooting stage. After 12 weeks, the shoot generation rate was 85.3%, the shoot number and its length were 1.9/explant and 2.7 cm in the most favorable medium composition. The percentage of rooting was 25.4%. From these results, it was found the optimum basal medium and plant growth regulator for in vitro plant regeneration from axillary buds of walnut new cultivar "Sinlyeong". However, we have continued to search the other medium additives to enhance the rate of walnut root.

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Regional Adaptation of the Genus Cypripedium in Korea and Micropropagation with Seed

  • Lee, Joung Kwan;Kwon, Young Hee;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong;Jeong, Mi Jin;Son, Sung Won;Suh, Gang Uk
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.23-23
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    • 2019
  • Cypripedium is an orchid genus of over 50 species widely distributed in the northern temperate zone, with a few taxa extending central America and the Himalayas and the southwesternmost tip of China in Asia. Of the known species approximately two thirds are found in China. In Korea, 4 species were reported to find with rare popularity ; C. macranthos, C. guttatum, C. japonicum, and C. calceolus. The seed has a firm brown fusiform testa with an opening at one end where the micropyle of the ovule was situated. Cypripedium has small, fusiform, wind-dispersed seeds that weigh about $1{\sim}2{\mu}g$ and range from 0.1 to 2 mm long and 0.07 to 0.4 mm across. The germination of Cypripedium was previously thought to be hard to cultivate in one's garden and impossible to propagate artificially, nowadays lab-propagated seedlings are readily available in EU or United States. We had already reported the successful micropropagation of lady's slipper orchids with green pod sowing methods. The collected Cypripedium species and hybrids were planted in Korea National Arboretum (KNA) on Oct. 2018. The 11 species including C. candidum and the 31 hybrids such as C. Ingrid were introduced from Germany with CITES certification. The lady's slipper orchids bloomed on May to June, and the flowers were pollinated in the same species or hybrids to get seeds for in vitro germination on 2~3 days after fully flowering. The green pods were collected after 49~70 days after pollination. The 6 species of Cypripedium seeds were easily germinated on the POM within 70 days after sowing, and the 12 hybrids PLBs were observed with small roots. We had established the scheme of lady's slipper orchids germination in 3 months with green pods, and get the possibility of seedling in 10 months of sowing in vitro.

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Effect of Plant Growth Regulators on Flowering and Micropropagation of Gentiana scabra Bunge In Vitro. (용담의 기내 개화 및 증식에 관한 연구)

  • Son, Beung-Gu;Choi, Young-Whan;Ahn, Chong-Kil;Cho, Dong;Kwon, Oh-Chang;Park, Jung-Ki
    • Journal of Life Science
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    • v.6 no.1
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    • pp.40-47
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    • 1996
  • Experiments were conducted to determine the effects of plant growth regulators on in vitro flowering and micropropagation of Gentiana scabra Bunge which had been used the cut flower, pot flower ornamental and medicinal plants. Flower bud formation was affected by GA$_{3}$ and kinetin. The optimum concentrations for flower bud formation was observed at 0.5 mg/l kinetin and GA$_{3}$ , while kinetin was favorable. More flowerings result from the interaction of GA$_{3}$ and kinetin at in a combination of 0.1 mg/l kinetin + o.1 mg/l GA$_{3}$, but the optimum concentration of GA$_{3}$ and kinetin was decreased. All concentrations of kinetin with 0.1 mg/l GA$_{3}$ or O mg/l GA$_{3}$ + 0.5 mg/l kientin reduced t (weeks needed for 50% plantlets). The plantlet growth was affected by GA$_{3}$ and kinetin during plantlet culture. More lateral shots and better shoot length per plantlet were obtained as GA$_{3}$ and kinetin concentration were increased up to 1.0 mg/l. The number of per plantlet was greater increased in MS medium containing GA$_{3}$ than kinetin. Interaction was exhibited at lower concentration with 0.5mg/l GA$_{3}$ and kinetin, but not in higher concentration with 1.0 mg/l GA$_{3}$ and kinetin. Higher pod diameter increased seed germination, while lower pod diameter was obtained from abnormal plantlet. MA medium containing 0.5 mg/l GA$_{3}$ significantly increased germination without regard to pod diameter.

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Impact of Rhizosphere Competence of Biocontrol Agents upon Diseases Suppression and Plant Growth Promotion

  • Park, Chang-Seuk-
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1994.06a
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    • pp.27-49
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    • 1994
  • Root colonization of biocontrol agents via seed treatment was investigated and a compatible combination, Gliocladium virens G872B and Pseudomonas putida Pf3, in colonizing cucumber rhizosphere was confirmed through the study. Much higher number of fungal and bacterial propagules were detected when two isolates were inoculated together. The presence of Pf3 in root system was greatly helpful to G872B to colonize at root tip. The mechanism of this phenomenon is partially elucidated through the results of in vitro experiments and the observations of scanning electron and fluorescence microscope. Addition of Pf3 cells resulted earlier germination of G872B conidia and increased mycelial growth. And the more number of germinated conidia on seed coat, the more vigorous hypal streching and sporulation on the root surface were observed in coinoculated treatment. The propagules of G872B on the cucumber root when they were challenged against the pathogenic Fusarium oxysporum, were even higher than that of G872B treated alone, and the magnitude of such a difference was getting grater toward the root ip and the population of F. oxysporum on the root was reduced by seed inoculation of G872B. The rhizosphere competence was obviously reflected to disease suppression and plant growth promotion that induced by the given isolate. Green house experiments revealed that the combined treatment provided long-term disease suppression with greater rate and the larger amount of fruit yield than single treatments. Through this study the low temperature growing Pseudomonas fluorescens M45 and MC07 were evaluated to apply them to the winter crops in field or plastic film house. In vitro tests reveal that M45 and MC07 inhibited the mycelial growth of Pythium ultimum, Rhizoctona solani and Phytophthora capsici and enhanced growth of cucumber cotyledon in MS agar. This effect was more pronounced when the bacteria were incubated at 14$^{\circ}C$ than at 27$^{\circ}C$. And disease suppression and plant growth promotion in green house were also superior at low temperature condition. Seed treatment of M45 or soil treatment of MC07 brought successful control of damping-off and enhanced seedling growth of cucumber. The combined treatment of two isolates was more effective than any single treatment.

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Optimal Condition for Pollen Germination of Rare and Endangered Forsythia saxatilis (희귀.멸종위기 산개나리(Forsythia saxatilis) 화분의 최적 발아 조건)

  • Han, Sim-Hee;Kang, Hye Jin;Kim, Gil Nam;Kim, Du Hyun
    • Journal of Korean Society of Forest Science
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    • v.102 no.1
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    • pp.136-142
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    • 2013
  • Optimal condition for pollen germination was suggested as a basic research of flowering physiology in order to identify the characteristics of flowering and seed setting of rare and endangered Forsythia saxatilis Nakai. Pollen samples were collected during flowering time from the end of March to the beginning of April. First, a suitable germination temperature, medium sucrose concentration and germination time were determined for pollen germination and pollen tube elongation in vitro, and then an optimal pH of culture medium. Pollen germination and tube elongation were significantly different among the levels of germination temperature, sucrose concentration and germination time. Interactive effects were observed between germination temperature and time, germination temperature and sucrose concentration, germination time and sucrose concentration. Pollen germination was the highest at $10^{\circ}C$ and increased with the increase of sucrose concentration, whereas it had no relation with germination time. In addition, pollen germination and tube elongation did not increase at more than 15% of sucrose concentration and 24 hours later. Pollen germination was the highest at pH 5 (20.8%) and the lowest at pH 6 (3.8%). In conclusion, $10^{\circ}C$, 15% sucrose and pH 5 were proposed as the optimal condition for pollen germination 24 hours later of pollen culture.