• Title/Summary/Keyword: In vitro germination

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Development of Peptides from the Germinated Black Rice and Applications as Cosmetics Ingredients (발아 흑미 유래 펩타이드의 개발과 화장품 응용에 대한 연구)

  • Dong-hwan, Lee;Jin-hwa , Kim;Jun-tae, Bae;Sung-min, Park;Hyeong-bae, Pyo;Tae-boo, Choe;Bum-chun, Lee
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.241-246
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    • 2004
  • To develop novel anti-aging peptides from the germinated black rice, we treated with bromelain, papain and Pronase E. And we investigated the effects of the germinated black rice peptide (GBRP) as anti-aging cosmetic ingredients, and compared with the non-germinated black rice protein (NBRP). We investigated the effects on in vitro inhibition of matrix-metalloprotease (MMP), proliferation of human skin fibroblasts, stimulation of collagen synthesis and expression of UVA-induced MMPs in human skin fibroblasts, UVA induced MMP-1 expression and collagen contents in human skin fibroblasts were analyzed by enzyme-linked immunosorbent assay (ELISA). As a result, the molecular weight distributions of GBRP and NBRP were determined by gel permeation chromatography to be approximately 900 and 10,000 daltons. GBRP increased skin cell proliferation about 40% and reduced UVA-induced MMP-1 expression about 50%. Also the collagen protein level of cells, which were cultured with GBRP, was increased about 25%. These results suggest that the geminated plant seed peptides can be novel anti-aging ingredients for cosmetics.

Plant Regeneration via in Vitro Culture of Ovule Obtain by Intergeneric Crossing Between Citrus junos Sieb. et Tanaka and Poncirus trifoliata Raf. (유자와 탱자의 속간교잡후 배주배양에 의한 식물체 유기)

  • 이만상;남궁승박
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.6
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    • pp.317-322
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    • 1995
  • As a basic research for breeding new varieties, reciprocal -intergeneric crosses between Citrus junos and P.trifoliata were made. F$_1$ hybrid production using in vitro ovule culture, gametogenesis, and fertilization phenomena were investigated. Frequency of fruit set resulting from crossing of Citrus junos and Poncirus Trifoliata was 16.6% while that of Poncirus Trifoliata and Citrus junos was 11.7%. Callus formation occurred well when ovules at the 6th week after pollination were cultured on MT (Murashige and Tucker) medium supplemented with zeatin 0.5 mg/L and NAA 1.0 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4-D 0.1 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4 D 0.1 or 0.5 mg/L. The invitro germination rates of 20-week-old ovules set C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 54.5% and 48.6%, respectively. The emergence ratios of trifoliate hybrids obtained by C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 56.7% and 100%, respectively. The chromosome number of C. junos and P. Trifoliata was n = 9 or 2n = 18, and the sizes of their pollen grain were 33.75 $\mu$ and 25.0 $\mu$. The length and width of embryo sac in C. junos and P. Trifoliata were 69.38~79.23 $\mu$ and 27.50~38.56 $\mu$, and those of egg cells were 17.50~41.50 $\mu$ and 6.25~8.12$\mu$. Fertilization of C. junos and P. trifoliata terminated 72 h after pollination.

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Evaluation of Streptomyces padanus IA70-5 Strain to Control Hot Pepper Anthracnose (Colletotrichum acutatum) (고추 탄저병 (Colletotrichum acutatum) 방제를 위한 Streptomyces padanus IA70-5의 평가)

  • Chi, Tran Thi Phuong;Choi, Okhee;Kwak, Youn-Sig;Son, Daeyoung;Lee, Jeung Joo;Kim, Jinwoo
    • Journal of agriculture & life science
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    • v.46 no.3
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    • pp.37-45
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    • 2012
  • To select bacterial strains with antifungal activity against an anthracnose fungal disease causing damage severely on hot pepper, previous isolates obtained from plant root samples were screened. Among 457 isolates, IA70-5 isolate was finally selected and identified as Streptomyces padanus based on 16S rDNA sequence analysis. Strain IA70-5 is non-pigmenteous, non-mobile, and filamentous. S. padanus IA70-5 inhibited effectively the mycelium growth, spore germination, and appressorium formation of Colletotrichum acutatum in vitro. The results of this study demonstrated that IA70-5 strain, especially applied on fruit of hot pepper, decreased disease incidence 90% for pre-inoculation before pathogen treatment. Taken together, S. padanus IA70-5 strain is a promising biological control agent to control of a major fungal pepper disease, anthracnose.

Protective Effect of Iminoctadine tris(albesilate) and Kresoxim-methyl Fungicides to Citrus Postharvest Diseases caused by Penicillium spp. (저장 감귤의 부패에 관여하는 Penicillium spp.에 대한 Iminoctadine tris(albesilate)와 Kresoxym-methyl의 방제 효과)

  • Hyun, Jae-Wook;Lee, Seong-Chan;Ihm, Yang-Bin;Kim, Dong-Hwan;Ko, Sang-Wook;Kim, Kwang-Sik
    • The Korean Journal of Pesticide Science
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    • v.5 no.2
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    • pp.37-44
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    • 2001
  • The biological effects of the iminoctadine tris (albesilate) and kresoxim-methyl for the protection of citrus postharvest diseases caused by penicillium spp. were assayed. In vitro tests, $EC_{50}$ values of iminoctadine tris(albesilate) were $0.01{\sim}0.02\;and\;0.01{\mu}g$ a.i./mL against mycelial growth of P. italicum and P. digitatum, respectively, but iminoctadine tris(albesilate) at $0.64{\mu}g$ a.i. /mL inhibited a little mycelial growth of unknown Penicillium sp. which produced another symptom different to blue and green mold caused by P. italicum and P. digitatum, respectively. And against germination and growth of germ tube of P. italicum and P. digitatum, $EC_{50}$ value of iminoctadine tris(albesilate) was $0.0013{\sim}0.0025{\mu}g$ a.i./mL. But spore germination of unknown Penicillium spp. was not nearly inhibited at $0.2{\mu}g$ a.i./mL. $EC_{50}$ values of kresoxim-methyl were $0.08{\sim}0.16$, 0.04 and $0.16{\mu}g$ a.i./mL against mycelial growth of P. italicum, P. digitatum and unknown Penicillium sp., respectively, and $0.04{\sim}0.08{\mu}g$ a.i./mL and $0.01{\sim}0.02{\mu}g$ a.i./mL against germination and growth of germ tube of P. italicum and unknown Penicillium sp., and P. digitatum, respectively. Iminoctadine tris(albesilate) and kresoxim-methyl were markedly effective to control the postharvest disease by 7 days spray prior to harvest. When the fruits were sprayed with iminoctadine-tris(albesilate) ($200{\mu}g$ a.i./mL) and kresoxim-methyl ($155{\mu}g$ a.i./mL) 7 days prior to harvest and subsequently stored for 90 days, the percentage of diseased fruit by Penicillium spp. was $3.6{\pm}1.8%$ in treatment of kresoxim-methyl and $5.9{\pm}1.8%$ in iminoctadine-tris(albesilate), respectively. On the other hand, tile percentage of diseased fruit was relatively high, $20.3{\pm}10.0%\;and\;19.5{\pm}9.6%$ in thiophanate-methyl ($700{\mu}g$ a.i./mL) and non-treatment, respectively. Maximum residue amount (ppm) among fruits (flesh and peel) assayed 0, 30, 60 and 90 days after storage was 0.45 and 0.10 ppm in treatment of kresoxim-methyl and iminoctadine, respectively.

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Bioactivity of the Extract of Coptis chinensis: In-vitro Antifungal Activity against Phytophthora capsici and Growth-promotion Effect in Red-pepper (황련 추출물의 고추역병에 대한 In-vitro 항진균 활성 및 고추 생육촉진 효과)

  • Ahn, Seon-Mi;Lee, Dong-Sin;Kim, Mi-Sun;Choi, Su-Ji;Choi, Chung-Sik;Lee, Jung-Bok;Jang, Han-Su;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.37 no.3
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    • pp.280-286
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    • 2009
  • To investigate anti-phytopathogenic fungal activity of Coptis chinensis, the methanol extract and its organic solvent fractions were prepared. Using the extract and the fractions, in-vitro spore-germination inhibition and mycelial-growth inhibition activities were evaluated against Colletotrichum gloeosporioides, Phytohpthora capsici, Pyricularia grisea, Rhizoctonia solani, Botryosphaeri dothidea, Glomerella cingulata, respectively. Treatment of the methanol extract (500 mg/mL) into the spore of phytopathogenic fungi completely inhibited germinations for 5 days, except B. dothidea, and showed strong antifungal activities against P. grisea and B. cinerea, and antioomycetes activity against P. capsici. The minimal growth inhibition concentrations of the methanol extract against P. grisea, B. cinerea and P. capsici were 300, 300, and 500 mg/mL, respectively. For practical application of C. chinensis in red-pepper field, the hot-water extract (1,000 mg/mL) was prepared in commercial facility, after evaluation of heat stability and solvent-extraction yields of antifungal substances. The 3-times leaf-spray of the extract from June to August, 2008 did not show any deleterious effect to red-pepper. In fact, the leaf-spray promoted plant growth including leaf, root and fruit. The average weight and rind of each fruit were increased to 119% and 117% comparison to those of without treatments. Our results suggest that C. chinensis is a useful source for control of red-pepper diseases and plant growth.

Practical Propagation Methods for Production of Prothalli and Sporophytes in Deparia pycnosora (Christ) M. Kato

  • Jang, Bo Kook;Park, Kyungtae;Cho, Ju Sung;Lee, Cheol Hee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.43-43
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    • 2019
  • Deparia pycnosora (Christ) M. Kato is a fern used as ornamental plant. In addition, it is called "Teol-go-sa-ri" in Korean name. The aim of this study was to develop a practical propagation method of D. pycnosora using tissue culture technique. Prothallus obtained from spore germination was the used as experiment materials. The prothalli (300 mg) used in all experiments were sub-cultured for 8-week intervals. The most suitable media for prothallus propagation were identified by culturing 300 mg of prothalli in $1/4{\times}$, $1/2{\times}$, $1{\times}$, $2{\times}$ MS medium and in Knop medium for 8 weeks. Also, the prothalli were cultured by chopping with a scalpel. In addition, sucrose, activated charcoal, and total nitrogen source were added in different concentrations based on the culture medium selected. Cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $30{\times}1.0{\mu}mol-m-2{\cdot}s-1$, and a photoperiod of 16/8 h (light/dark) in in vitro. The results showed that optimum was achieved prothallus fresh weight and development in $1{\times}$ MS medium. When other components were added to the basic $1{\times}$ MS medium, prothallus propagation was maximized in $1{\times}$ MS medium supplemented with 2% sucrose, 0.2% activated charcoal, and 60 mM total nitrogen. To select a suitable soil mixture for sporophyte formation, 1.0 g of prothallus was blended with distilled water, spread on five combinations of different soil substrates (decomposed granite, horticultural substrates, peat moss, and perlite), and cultivated for 12 weeks. The sporophyte cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $43{\pm}2.0{\mu}mol-m-2{\cdot}s-1$, humidity of $84{\pm}1.4%$, and a photoperiod of 16/8 h (light/dark). As a results, horticultural substrate alone, 2:1 (v:v) mixtures of horticultural substrate and perlite, and 2:1 mixtures of horticultural substrate and decomposed granite induced 208.0, 201.3 and 248.8 sporophytes per pot, respectively. Therefore, this result could provide a practical mass propagation method of D. pycnosora

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Biological Control of Sclerotinia sclerotiorum in Lettuce Using Antagonistic Bacteria (길항세균을 이용한 상추 균핵병의 생물학적 방제)

  • Chon, Bong-Goan;Park, Suji;Kim, Jin-Won
    • Research in Plant Disease
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    • v.19 no.1
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    • pp.12-20
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    • 2013
  • To isolate antagonistic bacteria against sclerotinia rot of lettuce, caused by Sclerotinia sclerotiorum, soil samples were collected from the diseased greenhouse field in Namyangju city, Gyeong-gi province from 2007 to 2008. A total of 196 bacterial isolates were isolated using serial dilution method. In dual culture assay in vitro, 26 isolates showed more than 80% of inhibition rates of mycelial growth of S. sclerotiorum. Based on 16S rDNA sequence analysis, the 26 isolates were identified as Bacillus megaterium, B. cereus, B. subtilis, Arthrobacter nicotianae, A. ramosus, Pseudomonas filiscindens, Stenotrophomonas maltophilia, Brevibacterium frigoritolerans and Sphingobacterium faecium. The 26 isolates inhibited the mycelial growth of S. sclerotiorum up to 80% and the sclerotial germination 0-100%. In the greenhouse pot test of ten isolates conducted in summer, 2 isolates B. megaterium (DK6) and B. cereus (C210) showed control efficacy on sclerotia viability of S. sclerotiorum, 20% and 35%, respectively. In the greenhouse pot test in winter, the disease incidence of the control group was 80%, whereas those of 9 isolates among 26 were approximately 20%. From the result, the 9 isolates are expected as potentially antagonistic bacteria for biological control of sclerotinia rot of lettuce caused by S. sclerotiorum.

Screening for Effective Organic Farming Materials for the Control of Cucumber Scab Caused by Cladosporium cucumerinum (오이 검은별무늬병 방제에 효과적인 유기농업자재 선발)

  • Park, So-Hyang;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Park, Jong-Ho;Han, Eun-Jung;Kim, Yong-Ki
    • Research in Plant Disease
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    • v.23 no.2
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    • pp.159-167
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    • 2017
  • This study was conducted to evaluate the control efficacy of the organic farming materials on cucumber scab caused by Cladosporium cucumerinum PT1 (KACC 48094). The antifungal activities in vitro as well as the suppressive effect of 43 organic farming materials on the spore germination and germ tube growth by inoculating spore suspension on cucumber seedlings in vivo were investigated. Thirteen organic farming materials inhibited the mycelial growth of C. cucumerinum and nine of these were microbial agents. In the screening using cucumber seedlings, six organic farming materials were very effective with control efficacy value of 90%. Among them, Bacillus amyloliquefaciens M27 provided suppressive effect on both mycelial growth and spore germination against cucumber scab. Finally, nine organic farming materials were selected to test the protective and curative effects, and all chosen organic farming materials significantly suppressed disease incidence when applied in the preventive action, in comparison with the curative action. Especially, Bordeaux mixture I and III gave excellent protective control efficacy with control values of 96.7% and 73.3%, respectively, whereas its curative control effect was significant low. Among these, only Thymus quinquecostatus+Sophora extract showed curative activity, although the control value was as low as 50%. This study suggests that cucumber scab can be controlled by some organic farming materials in the farmhouses under comparatively cold and wet condition and protective treatment is more important and efficient.

In vitro Variant Induction and Its Content of Gentiopicroside of Gentiana scabra BUNGE (용담(龍膽)의 기내변이주(器內變異株) 유도(誘導)와 변이주(變異株)의 Gentiopicroside 함량(含量))

  • Seong, Nak-Sul;Park, Chung-Heon;Kim, Kwan-Su;Lee, Seoung-Tack;Chang, Yeong- Hee
    • Korean Journal of Medicinal Crop Science
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    • v.3 no.1
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    • pp.40-44
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    • 1995
  • This study was aimed to mutant induction in Gentiana scabra BUNGE through the tissue culture techniques and comparison of gentiopicroside content in each mutant plants derived from tissue culture. In order to induce the mutants of Gentiam scabra, seed, apical and lateral bud were soaked in $NaN_3$, EMS and MNU solutions and cultured on MS basal medium containing 0.5mg/l NAA, BA $NaN_3$, MNU-treated buds survived about 50% but not survived in EMS treatment. Seed germination rate was extremely low in EMS and MNU treatments but various types of mutant plantlets were obtained by those treatments. 63% of elongated type and 37% of dwarf type were obtained from tissue culture treated by various mutagen. Gentiopicroside contents of regenerated plantlets was analyzed. Root of dwarf type contained more gentiopicroside(1.383%) than that of elongation type (0.083%).

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Arabidopsis MAP3K16 and Other Salt-Inducible MAP3Ks Regulate ABA Response Redundantly

  • Choi, Seo-wha;Lee, Seul-bee;Na, Yeon-ju;Jeung, Sun-geum;Kim, Soo Young
    • Molecules and Cells
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    • v.40 no.3
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    • pp.230-242
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    • 2017
  • In the Arabidopsis genome, approximately 80 MAP3Ks (mitogen-activated protein kinase kinase kinases) have been identified. However, only a few of them have been characterized, and the functions of most MAP3Ks are largely unknown. In this paper, we report the function of MAP3K16 and several other MAP3Ks, MAP3K14/15/17/18, whose expression is salt-inducible. We prepared MAP3K16 overexpression (OX) lines and analyzed their phenotypes. The result showed that the transgenic plants were ABA-insensitive during seed germination and cotyledon greening stage but their root growth was ABA-hypersensitive. The OX lines were more susceptible to water-deficit condition at later growth stage in soil. A MAP3K16 knockout (KO) line, on the other hand, exhibited opposite phenotypes. In similar transgenic analyses, we found that MAP3K14/15/17/18 OX and KO lines displayed similar phenotypes to those of MA3K16, suggesting the functional redundancy among them. MAP3K16 possesses in vitro kinase activity, and we carried out two-hybrid analyses to identify MAP3K16 substrates. Our results indicate that MAP3K16 interacts with MKK3 and the negative regulator of ABA response, ABR1, in yeast. Furthermore, MAP3K16 recombinant protein could phosphorylate MKK3 and ABR1, suggesting that they might be MAP3K16 substrates. Collectively, our results demonstrate that MAP3K16 and MAP3K14/15/17/18 are involved in ABA response, playing negative or positive roles depending on developmental stage and that MAP3K16 may function via MKK3 and ABR1.