• Title/Summary/Keyword: single cell plant

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Biological Control of Tea Anthracnose Using an Antagonistic Bacterium of Bacillus subtilis Isolated from Tea Leaves

  • Kim, Gyoung-Hee;Lim, Myoung-Taek;Hur, Jae-Seoun;Yum, Kyu-Jin;Koh, Young-Jin
    • The Plant Pathology Journal
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    • v.25 no.1
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    • pp.99-102
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    • 2009
  • An antagonistic bacterium of Bacillus subtilis BD0310 against Colletotrichum theae-sinensis was isolated from the phylloplane of tea trees at a tea plantation in Korea. SC (suspension concentrate)-type biofungicide was formulated with the antagonist. Cell viability and antifungal activity of B. subtilis were maintained in the formulation more than 12 months at room temperature. The antagonist was sensitive only to copper sulfate among the chemical pesticides currently registered for tea trees in Korea. Greenhouse application demonstrated that the biofungicide controlled more effectively the disease in a protective mode than in a curative mode. Field trial showed that alternate applications of the biofungicide and chemical fungicide were more effective in controlling tea anthracnose than single application of the biofungicide or chemical fungicide with less use of chemicals. This study suggests that the biofungicide of B. subtilis 8D0310 is an effective method for biological control of anthracnose in tea plantations.

Establishment of reverse transcription polymerase chain reaction for detection of Getah virus infection in livestock

  • Lee, Seung Heon;Yang, Dong-Kun;Kim, Ha-Hyun;Choi, Sung-Suk;Cho, In-Soo
    • Korean Journal of Veterinary Research
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    • v.57 no.1
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    • pp.37-42
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    • 2017
  • Getah virus (GETV) infection causes sporadic outbreaks of mild febrile illness in horses and reproductive failure in pigs. In this study, we established a reverse transcription polymerase chain reaction (RT-PCR) method to detect GETV from suspected virus-infected samples. The reaction conditions were optimized and validated by using RNA extracted from GETV propagated in cell culture. A GETV-specific GED4 primer set was designed and used to amplify a 177 bp DNA fragment from a highly conserved region of the E1 glycoprotein gene in the GETV genome. RT-PCR performed with this primer set revealed high sensitivity and specificity. In the sensitivity test, the GED4 primer set detected GETV RNA at the level of $10^{2.0}\;TCID_{50}/mL$. In the specificity test, the GED4 primer set amplified only a single band of PCR product on the GETV RNA template, without non-specific amplification, and exhibited no cross-reactivity with other viral RNAs. These results suggest that this newly established RT-PCR method is useful for accurate identification of GETV infection in animals.

Effects of Abscisic Acid on the Epidermal Structure and Ontogeny of Stomata in Orostachys malacophyllus Leaves (둥근바위솔(Orostachys malacophyllus) 잎의 표피구조와 기공발생에 미치는 Abscisic Acid의 영향)

  • 김진성
    • Journal of Plant Biology
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    • v.30 no.1
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    • pp.21-30
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    • 1987
  • The effects of abscisic acid(ABA) spraying for 12 weeks on the stomatal types and frequencies of O. malacophyllus leaves were summarized as follows. ABA inhibited the growth of O. malacophyllus. The prominent effect of ABA on the epidermal structure was the promotion of senescence such as thickness of cell walls, smooth sinuosity of cell walls, and large size of epidermal cells. The stomatal frequency was decreased to 23% by 10$\mu\textrm{g}$ ml-1 ABA and to 48% by 100$\mu\textrm{g}$ml-1, and also the stomatal size was more or less smaller than that of control. The developing secondary stomatal mother cell was not found in both 10 and 100$\mu\textrm{g}$ml-1ABA, but the arrested secondary stomatal mother cell was rarely found in 10$\mu\textrm{g}$ ml-1 ABA. The formation of normal stomatal types such as helico-eumesogenous and aniso-eumesogenous was found in both 10 and 100 $\mu\textrm{g}$ ml-1 ABA asin well as control. Also nine abnormal stomatal types were found, and the frequencies were promoted to 6% by 10 $\mu\textrm{g}$ ml-1 ABA and to 17% by 100 $\mu\textrm{g}$ ml-1 ABA. Among these abnomal stomata, four types such as aborted stomata, single-aborted guard cells, arrested stomata, and modified stomatal complexes were found in control as well as in 10 and 100 $\mu\textrm{g}$ ml-1 ABA, but five types such as wrenched stomata, unequal stomata, wavy guard cells, guard cells overlapped by guard cells, and dissolved cell wall stomata were found in both 10 and 100 $\mu\textrm{g}$ ml-1ABA. The modified stomata complexes were abnormal stomatal types which were newly found and also were varied in types.

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Analysis of temperature-dependent abnormal bursting patterns of neurons in Aplysia

  • Hyun, Nam Gyu;Hyun, Kwangho;Oh, Saecheol;Lee, Kyungmin
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.4
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    • pp.349-362
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    • 2020
  • Temperature affects the firing pattern and electrical activity of neurons in animals, eliciting diverse responses depending on neuronal cell type. However, the mechanisms underlying such diverse responses are not well understood. In the present study, we performed in vitro recording of abdominal ganglia cells of Aplysia juliana, and analyzed their burst firing patterns. We identified atypical bursting patterns dependent on temperature that were totally different from classical bursting patterns observed in R15 neurons of A. juliana. We classified these abnormal bursting patterns into type 1 and type 2; type 1 abnormal single bursts are composed of two kinds of spikes with a long interspike interval (ISI) followed by short ISI regular firing, while type 2 abnormal single bursts are composed of complex multiplets. To investigate the mechanism underlying the temperature dependence of abnormal bursting, we employed simulations using a modified Plant model and determined that the temperature dependence of type 2 abnormal bursting is related to temperature-dependent scaling factors and activation or inactivation of potassium or sodium channels.

Antifungal and Plant Growth Promotion Activities of Recombinant Defensin Proteins from the Seed of Korean Radish (Raphanus sativus L.)

  • Hwang, Cher-Won
    • Korean Journal of Environmental Agriculture
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    • v.28 no.4
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    • pp.435-441
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    • 2009
  • In the present study, we analyzed the defensin protein deduced from Korean radish (Raphanus sativus L.) seeds.To express the genes in E. coli, we constructed a recombinant expression vector with a defensin gene, named rKRs-AFP gene isolated from Korean radish seeds. Over expressed rKRs-AFP proteins was separated by SDS-PAGE to determine the purity, and protein concentration was determined by the Bradford method. Antifungal activity was assessed by disk assay method against the tested fungi. As a result, when 500 mL of cell culture were disrupted by sonicator, 32.5 mg total proteins were obtained. The purified protein showed a single band on SDS-PAGE with estimated molecular weight about 6 KDa, consistent with the molecular mass calculated from the deduced amino acid sequence. The purified rKRs-AFP protein showed remarkable antifungal activities against several fungi including Aspergillus niger, Botrytis cinerea causing the gray mold disease, and Candida albicans. In field tests using the purified rKRs-AFP protein, the protein showed the reducing activity of disease spot and the mitigating effect of spreading of disease like agrichemicals. The immuno-assay of rKRs-AFP protein showed that the purified protein entirely accumulated at B. cinerea cytoplasm through the hyphal septa shown by fluorescence imaging. There was no fluorescence inside the cell, when the hypha was incubated without the protein. These all results indicate that the recombinant rKRs-AFP proteins can be utilized as a potential antifungal drug to control harmful plant fungal pathogens.

A Study on Structure and Differentiation of Seed Coat of Panax ginseng C.A. Meyer (인삼(Panax ginseng C.A. Meyer) 종피의 구조 및 분화에 관한 연구)

  • 김우갑
    • Journal of Plant Biology
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    • v.29 no.4
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    • pp.295-315
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    • 1986
  • Structure and differentiation mechanism of the seed coat of Panax ginseng are studied with light and electron microscopes to clarify the developmental processes of seed coat and the structural changes during the differentiation of the seed. The seed coat of ginseng is differentiated from the inner cell layers of ovary wall, which can be compared with the seed coat differentiated from integument(s) in other plants. The single integument is differentiated into endothelium, which is degenerated to one layer of 4${\mu}{\textrm}{m}$ in thickness, composed of remants of cell wall components in fully ripened seed. The ripened seed coat is composed of three layers; fringe layer, inner layer and palisade layer, and all of the them are crossed at right angles with one another. This may be the cause of protection of the kernel from other mechanical injuries. The thickness of fully ripened seed coat is about 300~600 ${\mu}{\textrm}{m}$, and arrangements of sclereids are irregular. However, the raphe region of seed coat is thin about 200 ${\mu}{\textrm}{m}$ in thickness and sclereids in that region are arranged regularly. This is the important cause for the cleavage of the seed coat during post-maturation process. The vascular bundles on the raphe are still remaining after sarcocarps are removed, and one of the branches of vascular bundles entered into the seed coat through the hilum and extended to chalazal region. During post-maturation process, the supply of water being necessary for growth of embryo may be accompolished by the vascular bundles entered into the seed coat through the opened hilum.

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Ultrasound-Assisted Liquid-Liquid Extraction for Recovery of Paclitaxel from Plant Cell Cultures (식물세포배양으로부터 파클리탁셀 회수를 위한 초음파를 이용한 액-액 추출)

  • Ha, Geon-Soo;Kim, Jin-Hyun
    • Korean Chemical Engineering Research
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    • v.54 no.2
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    • pp.229-233
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    • 2016
  • In this study, an efficient ultrasound-assisted liquid-liquid extraction process was developed for recovering of paclitaxel from plant cell cultures. The optimal ultrasonic power and operating time were 250 W and 15 min at fixed ratio of bottom phase, methylene chloride to top phase, MeOH (25%, v/v). Under the optimal conditions developed in the present method, most of the paclitaxel (~92%) was recovered from crude extract by a single extraction step. Due to the synergistic effect of ultrasound by the addition of inorganic salt, an appropriate inorganic salt concentration and the ultrasonic power were found to be required for the effective recovery of paclitaxel using ultrasound-assisted liquid-liquid extraction.

Plant genome analysis using flow cytometry

  • Lee Jai-Heon;Kim Kee-Young;Chung Dae-Soo;Chung Won Bok;Kwon Oh-Chang
    • Proceedings of the Korean Society of Crop Science Conference
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    • 1999.05a
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    • pp.162-163
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    • 1999
  • The goal of this research was (1) to describe the conditions and parameters required for the cell cycle synchronization and the accumulation of large number of metaphase cells in maize and other cereal root tips, (2) to isolate intact metaphase chromosomes from root tips suitable for characterization by flow cytometry, and (3) to construct chromosome-specific libraries from maize. Plant metaphase chromosomes have been successfully synchronized and isolated from many cereal root-tips. DNA synthesis inhibitor (hydroxyurea) was used to synchronize cell cycle, follwed by treatement with trifluralin to accumulate metaphase chromosomes. Maize flow karyotypes show substantial variation among inbred lines. thish variation should be sueful in isolating individual chromosome types. In addition, flow cytometry is a useful method to measure DNA content of individual chromosomes in a genotyps, and to detect chromosomal variations. Individual chromosome peaks have been sorted from the maize hybrid B73/Mol7. Libraries were generated form the DOP-PCR amplification product from each peak. To date, we have analyzed clones from a library constructed from the maize chromosome 1 peak. Hybridization of labeled genomic DNA to clone inserts indicated that $24\%,\;18\%,\;and\;58\%$ of the clones were highly repetitive, medium repetitive, and low copy, respectively. Fifty percent of putative low cpoy clones showed single bands on inbred screening, blots, and the remaining $50\%$ were low copy repeats. Single copy clones showing polymorphism will be mapped using recombinant inbred mapping populations. Repetitive clones are being characterized by Southern blot analysis, and will be screened by in situ hybridization for their potential utility as chromosome specific markers.

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Ultrastructural Aspects of the Mixed Infections with Turnip mosaic virus and Ribgrass mosaic virus in Oriental Cabbage

  • Kim, Jeong-Soo;Cho, Jeom-Deog;Park, Hong-Soo;Kim, Kyung-Soo
    • The Plant Pathology Journal
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    • v.17 no.4
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    • pp.201-204
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    • 2001
  • Ultrastructural observation was conducted for the cells of oriental cabbage, Brassica campestris ssp. pekinensis 'Chungawang', inoculated simultaneously with Turnip mosaic virus (TuMV-ACT2-4vq) and Ribgrass mosaic virus (RMV-Ca1dn2) which were known as major destructive viruses of oriental cabbage in Korea. In cells infected with RMV alone, the virus particles were located as bundle or scattering in cytosols and vacuoles, which were typical ultrastructures of tobamovirus. Vessels of xylem were compacted with RMV particles. The cells infected only with TuMV had the cluster of virus particles scarcely and the typical potyvirus inclusions of scrolls, pinwheels, tubes and laminated aggregates in cytosols. The TuMV particles were jammed lineally between tonoplasts. In double infection, the two unrelated viruses of TuMV-ACT2-4vq and RMV-CA1dn2 were located together in a cell, and typical properties of each virus were also observed. The potyvirus inclusions and the tobamovirus particles were mixed entirely in cytoplasm. The virus particles of RMV wre presented strikingly near and in the center of potyvirus inclusions. In vascular cells, the tobamovirus particles were located abundantly than those in single infection. The potyvirus inclusions were embedded in the cluster of RMV particles in phloem parenchyma cells and the vascular elements were degenerated severely.

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First Report of Gray Mold Disease on Endangered Species Cypripedium japonicum

  • Jeon, Chang-Wook;Kim, Da-Ran;Gang, Geun Hye;Kim, Byung-Bu;Kim, Nam Ho;Nam, Seong-Yeol;Kwak, Youn-Sig
    • Mycobiology
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    • v.48 no.5
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    • pp.423-426
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    • 2020
  • Cypripedium japonicum is known to be the indigenous plant to Korea, Japan, and China. However, C. japonicum represents the most critically endangered plant species in South Korea. The plant is esthetically pleasing due to its flower, which is larger than any other orchidaceous species. Disease symptoms relating to gray mold were observed on C. japonicum in May 2019. The suspected pathogen was successfully isolated from the symptomatic leaf tissue and conducted a pure culture of the fungi. The conidia formed consisted of a colorless or light brown single cell, which was either egg or oval-shaped with a size of 7.1 to 13.4 × 5.2 to 8.6 ㎛. Molecular phylogenetic relationship analysis was also confirmed that the pathogen concerned belonging to the family of Botrytis cinerea. Therefore, the findings confirmed that the pathogen isolated from C. japonicum was consistent with the unique properties of B. cinerea.