• Korean journal of applied entomology
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• v.5_6
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• pp.1-7
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• 1968

The present investigation was initiated to find out the possible insect vector of the paulownia witches'-broom virus disease. 1. Among a number of piercing-sucking mouth type insects that were feeding on the witches'-broom infected Paulownia leaves, the tobacco leaf bug, Cyrtopeltis tenuis REUTER, was most abundant. Eggs, nymphs and adults of Cyrtopeltis tenuis were observed from the mid-August through the end of October. 2. It was found that Paulownia witches'-broom virus is transmitted by Cyrotopeltis tenuis. 3. On young Pnulownia plants, first symptom appeared twenty days after exposure to virus-bearing colonies of Cyrtopeltis tenuis. Forty days after exposure to these viruliferous insects, the characteristic symptom of Paulownia witches'-broom was apparent. 4. Paulownia witches' -broom virus was also transmitted by Cyrtopeltis tenuis to calendula and morning glory. No app"rent symptom was observed from tomato, carrot. aster, zinnia, cosmos, soybean and plantago at least until sixty days after exposure to the viruliferous colonies of Cyrtopeltis tenuis. 5. Calendula and morning glory appear to be highly suitable test plants for Paulownia witches'-broom vims because of their relatively short incubation period and distinct symptom appearance.

• The Microorganisms and Industry
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• v.25 no.2
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• pp.11-18
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• 1999

The sweet potatoes infected with witches'-broom disease were collected in the growing fields in Jeon-bug area, Korea. As a possible control plant, Ipomoea batatas L.var. Suwon 147 was selected. The pathogen was identified by various methods ; such as mechanical transmission, antibiotic reactions and electron microscopy. In the results attained the author believed the pathogen of the sweet potato infected with witches' broom to be a mycoplasma-like organism. the results are as follows : 1. Mycoplasma-like bodies were occurred in the phloem region of the sweet potatoes infected with witches'-broom and its particles were sized in the range of about 200-2,500m.mu.. The membrane of the pathogen was observed to be made of an unit. 2. Responsibilities to the antibiotices were sensitive in case of tetracycline and terramycin, and root dipping method showed remarkable than foliage spray. 3. The infection was developed by the grafting transmission but by the insects, Myzus persicae and Cicadella viridis. 4. rosette, witches'-broom, stunt, yellowish, mosaic and necrosis were observed as the symptomps of the disease.

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

• The Plant Pathology Journal
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• v.21 no.4
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• pp.383-386
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• 2005

In 2003 typical phytoplasma symptoms of witches' broom and flower malformation were observed on statice (Limonium sinuatum) plants grown at commercial greenhouses in Busan, South Korea. The DNA extracted from the infected leaves was amplified using universal primer pair of Pl/P6 derived from conserved 16S rRNA gene of Mollicutes giving the expected Polymerase chain reaction (PCR) product of 1.5 kb. In the nested PCR assays, the expected DNA fragment of 1.1 kb was amplified with the specific primer pair 16Fl/Rl that was designed on the basis of aster yellows (AY) phytoplasma 16S rDNA sequences. The 1.1 kb PCR products were cloned and nucleotide sequences were determined. The sequences were identical to that of Onion yellows OY phytoplasma (GenBank accession no. D12569) isolated from Onion in Japan. Electron microscopy of thin sections of leaf veins showed phytoplasma bodies in the phloem. Statice witches' broom symptom occurred on statice in commercial greenhouses in Korea was confirmed as infection of AY phytoplasma by transmission electron microscopy observation, and by determination of 16S rRNA gene sequences of phytoplasma.

• Korean Journal Plant Pathology
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• v.14 no.5
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• pp.382-385
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• 1998

Molecular identification and genetic relationships between a phytoplasma associated with chestnut little leaf (CLL) and phytoplasma isolates of other trees in Korea were amplified by polymerase chain reaction (PCR). These 16S rDNA sequences amplified from the various phytoplasmas were used in DNA heteroduplex mobility assays (HMA). In DNA HMA combined with PCR, the mobility shift was observed for a heteroduoplex formed in combined with CLL and jujube witches broom, but not for those formed in combined with CLL and each of sumac witches broom, paulownia witches broom, and mulberry dwarf. HMA combined with PCR has been shown to be a very useful method for detection and differentiation of phytoplasmas.

• Korean journal of applied entomology
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• v.15 no.3 s.28
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• pp.107-110
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• 1976

A Witches'-broom disease of jujube (Zizyphus jujuba) with which mycoplasma-like bodies are associated is wide spread in South Korea. Jujube trees with witches'-broom symptoms were selected from 15-20 year old planting and treated with solution of oxytetracycline hydrochloride (Terramycin). Treatments were 1) one injection on April; 2) one postharvest injection on October; 3) two injections (April and July); 4) three injections (April, June and August) and 5) an untreated control. Each injection consisted of 500ml of 1,000 p.p.m. solution of oxytetracycline HCl transfused into affected trees from plastic reservoir through plastic tubes connected to 3 small holes drilled in the basal part of the tree trunks. Complete remission of witches'-broom symptoms was accomplished within one growing season by one spring (April) injection. One postharvest (Oct.) injection also prevented the symptoms in the following growing season. Two and three injection treatments prevented the symptoms for at least two growing seasons and restored previously severely diseased trees to normal or near normal conditions. Remission of symptoms was found only above the injection site while current season witches'-brooms developed from areas below the injection site and from untreated main scaffolds. The results of this experiment demonstrates that transfusion treatment with oxytetracyclin HCl is feasible for the practical control of witches'-broom of jujube.

• Journal of Microbiology and Biotechnology
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• v.23 no.8
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• pp.1047-1054
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• 2013

Witches' broom of lime is a disease caused by Candidatus Phytoplasma aurantifolia, which represents the most significant global threat to the production of lime trees (Citrus aurantifolia). Conventional disease management strategies have shown little success, and new approaches based on genetic engineering need to be considered. The expression of recombinant antibodies and fragments thereof in plant cells is a powerful approach that can be used to suppress plant pathogens. We have developed a single-chain variable fragment antibody (scFvIMP6) against the immunodominant membrane protein (IMP) of witches' broom phytoplasma and expressed it in different plant cell compartments. We isolated scFvIMP6 from a naïve scFv phage display library and expressed it in bacteria to demonstrate its binding activity against both recombinant IMP and intact phytoplasma cells. The expression of scFvIMP6 in plants was evaluated by transferring the scFvIMP6 cDNA to plant expression vectors featuring constitutive or phloem specific promoters in cassettes with or without secretion signals, therefore causing the protein to accumulate either in the cytosol or apoplast. All constructs were transiently expressed in Nicotiana benthamiana by agroinfiltration, and antibodies of the anticipated size were detected by immunoblotting. Plant-derived scFvIMP6 was purified by affinity chromatography, and specific binding to recombinant IMP was demonstrated by enzyme-linked immunosorbent assay. Our results indicate that scFvIMP6 binds with high activity and can be used for the detection of Ca. Phytoplasma aurantifolia and is also a suitable candidate for stable expression in lime trees to suppress witches' broom of lime.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.136.2-137
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• 2003

In order to diagnose and differentiate jujube witches' broom (JWB) phytoplasma rapidly, oligonucleotide primer pair, 16Sr(V) F/R, for polymerase chain reactions (PCRs) was designed on the basis of 165 rRNA sequences of JWB phytoplasma. The PCR employing phytoplasma universal primer pair P1/P7 consistently amplified DNA in all tested phytoplasma isolates. But no phytoplasma DNA was detected in healthy jujube seedlings. The nested PCR, the primer pair 16S(V) F/R, about 460 bp fragment, amplified DNA in all tested JWB and related phytoplasmas including LiWB phytoplasma of the 165 rRNA group V, but no DNA amplification was detected from other phytoplasma strains such as group 16SrI (Aster yellows) and group 16SrⅩII (Stolbur group) phytoplasmas in which mulberry dwarf phytoplasma and chrysanthemum witches broom phytoplasma are belonged to, respectively The same results were obtained from both Korean- and Chinese-isolates of JWB. Nested-PCR using phytoplasma universal primer pair P1/P7 and 16S rRNA group V specific primer pair 16S(V) F/R could detect group V phytoplasma rapidly and easily, in particular JWB phytoplasma.

• The Plant Pathology Journal
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• v.17 no.5
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• pp.295-299
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• 2001

The relative density of phytoplasmas in witches'broom (WB)-infected jujube trees was investigated using compatitive polymerase chain reaction (PCR). During dormant and defoliating seasons, the densities of phytoplasmas were about the same in roots and twigs. In early growing season, the density showed the highest rates in roots, then in twigs and in petioles. however, the density was highest in petioles and the lowest in roots during actively growing season. Throughout the year, root samples did not show any serious fluctuation compared with that of t2wigs and petioles. Density was lowest during actively growing season in root samples. In contrast, petiole sample densities varied to a great extent depending on the season, very high during actively growing season, but very low during the early growing season, In twig samples, the densities were very high and almost the same in both defoliating and dormant seasons. Among the parts of the trees, phytoplsma density was the most stable in root samples throughout the year. The highest densities of phytoplasmas were about the same in all tree parts. These results suggest that the phytoplasmas may overwinter not only in roots but also in twigs, and that multiplication rate of phytoplsma becomes very high right after the early growing season.

• The Plant Pathology Journal
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• v.25 no.1
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• pp.21-25
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• 2009

Jujube trees infected with phytoplasma exhibit symptoms of typical witches' broom, such as yellowing, abnormally small leaves, short internodes and proliferation of shoots. A 1.2 kb fragment of the 16S rDNA from jujube phytoplasma was generated by R16F2n/R16R2 primer pair from earlier amplified P1/P7 PCR products of cloned jujube witches' broom phytoplasmas. Enzymatic restriction fragment length polymorphism (RFLP) and sequence analysis of 16S rDNA revealed that the jujube tree was infected with 16S rDNA I and V groups of phytoplasmas. Extensive comparative analyses of restriction enzyme profiles from Alu I, Hha I, Msp I, and Rsa I clearly classified the two into different phytoplasma groups. The phylogenie analyses based on 16S rDNA showed that the similarity of the two different clones was 87.5%. This is the first report of a mixed phytoplasmal infection in a single jujube tree.