• Journal of Korean Society of Forest Science
• /
• v.80 no.2
• /
• pp.232-236
• /
• 1991

The efficacy of berberine sulphate, a fluorochrome having binding properties with both DNA and RNA, was investigated for the detection of mycoplasma-like organisms(MLOs) in jujube(Zizyphus jujuba), paulownia(Paulownia tomentosa), mulberry(Morus alba) trees and periwinkle (Catharanthus roseus) plant. When examined under fluorescence microscope, berberine sulphate-stained sections of diseased samples showed distinct MLO-specific fluorescent particles in the phloem area, while such fluorescence was absent in the healthy ones. This staining technique was proved to be a very accurate method for the diagnosis of MLO infections in woody and herbaceous plants. Furthermore, the cheap and easy procedure could be used to test a great number of samples on MLO infections with reliability and rapidity.

• The Korean Journal of Mycology
• /
• v.13 no.1
• /
• pp.49-51
• /
• 1985

Mycoplasma-like organisms (MLO) were identified from three plants. The disease symptoms show stunting, yellowing and witche's broom on Cnidium officinale Makino collected from Ulreung Island, Bupleurum falcatum L. collected from Jangsu, and Plantago asiatica L. collected from Daegwanreung. The particles of MLO were observed in the phloem tissues by a Hitachi Hu-11E electron microscope. C. officinale infected with MLO was frequently observed in Ulreung Island.

• Korean Journal of Veterinary Research
• /
• v.26 no.2
• /
• pp.283-292
• /
• 1986

Isolation and identification of Mycoplasma were performed to clarify Mycoplasma infection of mice fed by conventional feeding at two ($K_1$, $K_2$) institutes in Korea. The twenty mice to be tested were randomly sampled from each of 10 breeding colonies in respective institute. Identification of the Mycoplasma strains isolated from the nasal cavity, lung and synovia of mice was made according to the morphology of colonies, biological and biochemical properties with special reference to M. pulmonis, M. arthrotodis and M. neurolyticum. In addition, growth inhibition test was performed using hyperimmune rabbit antisera to the strain PG-22 of M. pulmonis, the strain PG-6 of M, arthritidis and the strain PG-28 of M. neurolyticum and also differentiation of isolates from L-form bacteria was dont by Dieses staining and culture method with passage of the isolates on liquid media eliminated antibacterial drug. On the other hand, a total of 13 strains out of the 44 isolated M. pulmonis from mice was investigated for their susceptibility against 16 antibiotics in vitro. The antibiotic sensitivity test was made using $3{\times}10^4$ organisms/0.3ml on each plate(90mm diameter) with antibiotic mono-or tri-disk. The results obtained are summarized as follows: 1. Out of 20 mice from 10 breeding colonies in Kl institute, mycoplasma-like strains from the nasal cavity of 16 mice(80%) and from the lung of 8 mice(40%) were isolated, while out of 20 mice in K2 institute, M-like strains were isolated from the nasal cavity of 14 mice(70%) and from the lung of 6 mice(30%). However, no mycoplasma-like organisms were isolated from the synovia of the 40 mice examined. All the 44 strains isolated were identified as the organisms of M. pulmonis. 2. Out of the 16 antibiotics tested, penicillin, oleandomycin and bacitracin showed no activity against all the 13 M. pulmonis strains. On the contrary, lincomycin, clindamycin, chloramphenicol, tetracycline, minocycline, kanamycin, gentamycin and tobramycin showed high activity with three different antibiotic concentration of tridisk, but amikasin and spiramycin showed intermediate activity. Other antibiotics such as polymyxin B and colistin showed low activity, while erythromycin showed lower activity than others.

• Korean Journal Plant Pathology
• /
• v.1 no.1
• /
• pp.12-16
• /
• 1985

Attempts were made to evaluate the efficacy of three fluorochromes, i.e., DAPI (4'-6-diamidino-2-phenylin-dole-2HCl), aniline blue and quinacrine(quinacrine mustard dihydrochloride) for the detection of mycoplasma infections in jujube (Zizyphus jujuba), mulberry (Mows alba) trees and periwinckle (Catharanthus roseus) plant by fluorescence microscopy. Stem sections from these plants infected with mycoplasma-like organisms (MLO) produced distinct fluorescence in the phloem when stained with DAPI, aniline blue or quinacrine, while fluorescence was absent in the healthy plants. The use of these fluorochromes provided simple and efficient techniques for the diagnosis of MLO infections. IOf the three fluorochromes tested, DAPI was found to be most efficient.

• Journal of Korean Society of Forest Science
• /
• v.67 no.1
• /
• pp.28-30
• /
• 1984

Distribution of mycoplasma-like organisms (MLO) in the phloem tissue of witches'-broom infected jujube trees was investigated by fluorescence microscopy applying new fluorochrome DAPI (4'-6-diamidino-2-phenylindole, 2HCl). MLO were detected from the phloem of leaf and stem sections of diseased branches exhibiting typical witches'-broom symptom but not detected from those of symptomless branches of an infected tree. MLO were also present in the healthy looking leaves frequently found in the diseased shoots. Fluorescence microscopy revealed the presence of MLO in the phloem of root sections of infected trees.

• Korean Journal Plant Pathology
• /
• v.1 no.3
• /
• pp.184-189
• /
• 1985

Transmission of mulberry dwarf mycoplasma (MDM) from diseased mulberry to 5 herbaceous plants (periwinkle, white clover, Ladino clover, red clover, and Chinese milk vetch) through insect vector, Hishimonus sellatus, was confirmed by symptom expression and microscopic evidences. The earliest symptom appearance was noticed on periwinkle in which incubation period was 25-30 days, while it ranged 35-40 days in the other plant species. The common symptoms of MDM infected plants were characterized by poor plant growth with accompanying discolorations of leaves (chlorosis with vein clearing in periwinkle, reddish in red clover, brownish in white and Ladino clovers, and yellowish in Chinese milk vetch). Mycoplasmal infections were diagnosed light microscopically by Dienes' and toluidine blue staining of hand-cut and Epon-embedded sections, respectively. In Dienes' stain, all the plants infected with MDM showed specific staining reaction in phloems. In toluidine blue stain, mycoplasmal existence was noted by granular appearance in sieve tubes which were confirmed to be mycoplasma-like organisms under an electron microscope.

• Journal of Korean Society of Forest Science
• /
• v.83 no.1
• /
• pp.1-5
• /
• 1994

Sixteen kinds of piercing-sucking mouth type insects were observed at witches'-broom diseased paulownia (Paulownia tomentosa) trees. Among ten insect species tested, a leafhopper, Empoasca sp., transmitted paulownia witches'-broom mycoplasma-like organism(PWB-MLO) to periwinkle(Catharanthus roseus) plant. Adults of this leafhopper were collected from PWB-diseased paulownia trees. The leafhpoppers were allowed to feed on the PWB-infected paulownia trees for three weeks to acquire PWB-MLO. These insects were then transferred to healthy seedlings of paulownia and periwinkle plants for inoculation-feeding. About 30-40days after the inoculation, "little-leaf" symptoms developed on periwinkle plants, but no symptoms developed on paulownia seedlings. MLO infection in the diseased periwinkle tissue was diagnosed by fluorescence microscopy. MLOs were observed in the sieve tubes by electron microscopy, proving the insect transmission of PWB-MLO.

• Applied Microscopy
• /
• v.12 no.2
• /
• pp.11-22
• /
• 1982

The morphological development of Mycoplasma pneumoniae attached to solid surfaces was examined by light and electron microscopies. Critical point drying and carbon replication techniques revealed that during the growth cycle of developing microcolonies, the morphological form coincided with the pH of the culture. M. pneumoniae appeared to have a well defined morphology associated with age of the culture. The organisms were dimorphic, with round cells capable of reproduction and segments consisting of a spindle shaped body with one pointed and one knob-like end. Starting with single cells, there were the following stages in the development of a culture: replication stage through binary fission and segmentation, stage of confluency, and a degeneration stage into rough spherical forms. The round cells appearrd to replicate by binary fission during the lag and early log phases of growth, while spindle segments replicated by segmentation during most of the logarithmic growth. The growth of the filaments and replication of the segments occured at the knob-like ends, showing a type of polarity, and formed a meshwork across the surface. This development could be cycled under favorable growth conditions, but the culture aged and when the conditions became adverse(e.g. pH 6.8 or lower), filamentous cells converted to spherical forms, losing their reproductive capability.

• Journal of Korean Society of Forest Science
• /
• v.78 no.2
• /
• pp.103-118
• /
• 1989

The occurrence of witches' broom in Ligustrum ovalifolium was first noticed in Korea by author in 1984. The present study was carried out with particular emphasis on the symptomatology, etiology, transmission of the disease and antibiotic treatments. The infected tissue was observed by the fluorescence and electron microscopy and its biochemical characteristics were compared with healthy one by electrophoresis. The results are summarized as follows : 1. symptoms of the infected trees were characterized by the dwarfing of the organs, yellowing and brooming of the foliage. 2. The observation by the trans electron microscopy on the witches' broom of L. ovalifolium revealed the occurrence of numerous mycoplasma-like organisms(MLOs) in the phloem tissue cells of the midribs of infected leaves. 3. The MLOs were surrounded by a single unit membrane, and they appeared to be multiplied by binary fission. 4. The presence of crystals unidentified in the phloem parenchyma cells was noticed by electron rnicroscopy, 5. The disease was able to be transmitted by budding, crown, and greenwood graftings to L. ovalifolium, L. obtusifolium, L, japonicum and also transmitted, even when the stocks and scions were not completely grafted. 6. Insect transmission on L. ovalifolium and L, obtzrsifolium was carried by Hishimonus sellatus. 7. The infected roots dipped in the 1,000 ppm of teracyclin solution was only temporarily effective in controlling the disease. 8. Infected plant with MLOs showed specific fluorescent reactions in phloems with DAPI stain. 9. The protein and peroxidase separated by electrophoresis showed strikingly distinctive difference between the healthy and diseased leaves.

• Journal of Korean Society of Forest Science
• /
• v.47 no.1
• /
• pp.1-15
• /
• 1980

The occurrence of witches' broom of Rhus javanica was first noticed in Korea by the author in 1979. Subsequently, studies were made on the symptomatology, etiology, and transmission of the disease, as well as the effect of some antibiotics on the disease development. The results of these studies are summarized as follows: 1. Symptoms of the infected plant were characterized by dwarfing of the tree accompanied by yellowing and brooming of the foliage. 2. Electron microscopy of witches' broom diseased Rhus javanica plant revealed the occurrence of numerous mycoplasma-like organisms (MLO's) in the phloem tissue cells (sieve tube elements and phloem parenchyma cells) of the rachis and midribs of infected leaves. 3. The MLO's were bounded by a single unit membrane and contained ribosome-like granules and strands presumed to be DNA. It also appears that the MLO multiply possibly by budding as well as binary and plurinary fission. 4. In the midrib of healthy leaves, vascular bundles were collaterally discontinuous. In the diseased leaves, however, xylems were connected to each other and phloem cells showed an atrophy. Granules, which were prominent in the normal abaxial epidermis, were not observed in the peidermis of diseased leaves. 5. Electron microscopy revealed crystals or osmopholic granules in the phloem parenchyma cells, and that normal stacks of grana were not developed in the chloroplasts of infected levels. 6. The disease was experimentally transmitted by grafting. Budding was more effective than crown grafting for transmitting the disease. The disease has been transmitted by grafting even when complete union of stocks and scions has not taken place. The disease agent was not transmitted by sap inoculation. Insect transmission has not been confirmed. 7. Dipping the roots of infected plants into the 500 ppm and 1,000 solutions of either tetracycline HCI or oxytetracycline, HCI was more effective on temporary remision of the symptoms than spraying the 100 ppm and 200 ppm solutions of the same antibiotics. A greater effect was achieved through dipping into 1,000 ppm than into 500 ppm.