• 한국산림과학회지
• /
• 제101권4호
• /
• pp.635-639
• /
• 2012

뽕나무 오갈병 파이토플라스마 매개곤충을 알아보기 위하여 뽕나무 재배지에서 채집된 곤충 중 흡즙성 곤충 3 종(Hishimonas sellatus, Bothrogonia japonica and Balclutha punctata)를 선발하였다. 이들 흡즙성 곤충을 뽕나무 이병주에서 흡즙시킨 다음 건전 뽕나무와 일일초로 전반시킨 결과, 마름무늬매미충(H. sellatus)과 B. punctata이 파이토플라스마를 전반하였다. 뽕나무에서는 전형적인 오갈 증상을 보였지만 일일초에서는 가지가 가늘게 늘어지며, 잎은 총생하고 작아지는 빗자루증상이 나타났다. 매개충과 매개전염시킨 뽕나무와 일일초에 대하여 R16F2n/R2 프라이머로 PCR 검정 결과 끝검은말매미충을 제외한 모든 시료에서 파이토플라스마가 검출되었다.

• Plant Resources
• /
• 제3권3호
• /
• pp.173-178
• /
• 2000

This test checked jujube witches'-broom disease, sumac witches'-broom disease, paulonia witches'- broom disease, and mulberry dwarf disease whether or not they were infected by phytoplasma, using universal and specific primers. Upon treatment of DNA amplified by PCR of phytoplasma with Alu I , Hpa II and Sat I restricted enzymes, distinction of phytoplasmas was possible. Particularly, phytoplasma of each host was distinguishable by treatment of Hpa II restricted enzyme. Meanwhile, analysis of restricted enzymes of jujube witches'-broom disease showed a higher infectivity of phytoplasmas of two origins. There were a lot of relations between jujube witches'-broom disease and sumac witches'-broom disease, and between paulonia witches'-broom disease and mulberry dwarf disease.

• The Plant Pathology Journal
• /
• 제23권2호
• /
• pp.106-108
• /
• 2007

The Symptoms of witches' broom disease caused by phytoplasma including general stunting and yellowing, were observed in leafy lespedeza (Lespedeza cyrtobotrya M.) on Doam-myeon, Pyeongchang-gun, in 2006. Based on the sequence analysis of PCR-amplified 16S ribosomal DNA and 16S-23S spacer region DNA products using universal phytoplasma primers, the phytoplasma associated with leafy lespedeza witches' broom (LLWB) disease was identified as a member of Candidatus Pytoplasma trifolii. It was most closely related to alsike clover proliferation phytoplasma (99.8% similarity, accession no. AY390261), Candidatus Pytoplasma trifolii strain. RFLP patterns generated with AluI, HpaII clearly differentiated LLWB phytoplasma from the referenced phytoplasma strains, water dropwort witches' broom, mulberry dwarf, glehni aster yellow dwarf and jujube witches' broom. This paper is the first report on Candidatus Phytoplasma trifolii in leafy lespedeza identified at a molecular level.

• 한국산림과학회지
• /
• 제100권4호
• /
• pp.623-629
• /
• 2011

파이토플라스마에 감염된 뽕나무의 각 부위를 nested-PCR법과 경쟁 PCR법을 이용하여 파이토플라스마의 검출 범위 및 밀도 변화를 조사하였다. 파이토플라스마에 부분 감염된 뽕나무를 총 5부위(A. 빗자루병이나 오갈병 증상을 보인 잎의 엽병, B. 병징을 보인 잎이 있는 가지의 아래쪽 건전해 보이는 잎의 엽병, C. 병징을 보인 잎의 가지 부위, D. 병징을 보이지 않은 가지 쪽의 영병이나 가지, E. 잔뿌리)로 구분하였다. AS-1/AS-2 프라이머를 이용한 nested-PCR 결과, 파이토플라스마는 모든 시료에서 검출되었으며, 파이토플라스마의 검출범위는 P1/P7 프라이머를 이용한 direct-PCR 산물에서는 희석액이 $10^4$배까지 검출되었고, AS-1/AS-2 프라이머를 이용하여 nested-PCR를 수행한 결과, PCR 산물을 $10^{13}$배 희석한 시료까지 파이토플라스마가 검출되었다. 경쟁 PCR 결과, 뽕나무 파이토플라스마의 밀도는 $7.94{\times}10^{18}-10^{12}copies/{\mu}L$의 범위내에 존재하였으며, 년 중 파이토플라스마가 모든 시료에서 존재하였다. 초기생장시기에는 "B"부위와 "C"부위에서 파이토플라스마 밀도가 높게 나타났으나 "A"부위와 "D"부위에서는 이보다 낮게 나타났다. 휴면기에는 "C"부위와 "E"부위가 "D"부위보다 밀도가 높았다. 모든 시료 중 "C"부위와 "A"부위에서 년 중 파이토플라스마 밀도가 안전적으로 존재하였다. 본 연구 결과, nested-PCR 법과 경쟁 PCR법은 식물체내의 파이토플라스마의 정확한 검출과 밀도변화를 알아볼 수 있는 유용한 방법이다.

• The Plant Pathology Journal
• /
• 제18권2호
• /
• pp.98-101
• /
• 2002

Using phytoplasma universal primer pair Pl and P7, a fragment of about 1.8 kb nucleotide sequences of 16S rRNA gene and 16S-23S rRNA intergenic spacer region, and a portion of 23S rRNA gene of jujube witches'broom (JWB) and mulberry dwarf(MD) phytoplasmas were determined. The nucleotide sequences of JWB and MD were 1,850 bp and 1,831 bp long, respectively. The JWB phytoplasma sequence was aligned with the homologous sequence of MD phytoplasma. Twenty-eight base insertions and nine base deletions were found in the JWB phytoplasma sequence compared with that of MD phytoplasma. The similarity of the aligned sequences of JWB and MD was 84.8%. The near-complete 16S rRNA gene DNA sequences of JWB and MD were 1,529 bp and 1,530 bp in length, respectively, and revealed 89.0% homology. The 16S-23S rRNA intergenic spacer region DNA sequences were 263 bp and 243 bp in lengths respectively, while homology was only 70% and the conserved tRNA-lle gene of JWB and MD was located into the intergenic space region between 16S-23S rRNA gene. The nucleotide sequences were 77 bp long in both JWB and MD, and showed 97.4% sequence homology. Based on the phylogenetic analysis of the two phytoplasmas, the JWB phytoplasma belongs to the Elm yellow phytoplasma group (16S rV), whereas, the MD phytoplasma belongs to the Aster yellow group (16S rI).

• 한국식물병리학회:학술대회논문집
• /
• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
• /
• pp.73.2-73
• /
• 2003

Typical whiches broom symptoms caused by phytoplasma were observed in Ash (Fraxinus rhynchophylla Hence) in Korea. The symptoms were showing abnormally small leaves, short internodes, and proliferation of shoots. Fluorescence and electron microscopy of leaf midribs revealed phytoplasma positive DAPI fluorescence and numerous phytoplasma bodies localized in the phloem sieve tubes. Phytoplasma DNA of 1.8 Kb was detected consistently from all symptomatic samples by the amplification of phytoplasma DNA with the phytoplasma specific primer pair Pl/P7. But no phytoplasma DNA was detected in healthy ash seedlings. Based on sequence analyses of an amplified region, this phytoplasma is closely related to Eqilodium phyllody, Mulberry dwarf, and Aster yellows phytoplasmas with the homology of 99.95 ％, 99.79 ％ and 99.78 ％, respectively, This phylogenetic analyses indicate that ash witches broom phytoplasma but is evidently distinct from the ash yellows group 16SrⅦ and should be classified into the Aster yellows group 16SrⅥ.

• 한국산림과학회지
• /
• 제94권2호통권159호
• /
• pp.103-107
• /
• 2005

Typical phytoplasma whiches' broom symptoms were observed in Ash (Fraxinus rhynchophylla Hence) in Korea. The symptoms of the disease were showing abnormally small leaves, shorted internodes and proliferation of shoots. Examination of fluorescent and electron microscopy of leaf midribs revealed numerous phytoplasma bodies localized in the phloem tube cells. The phytoplasmas were detected in all the symptomatic samples by the amplification with phytoplasma specific primer pair P1/P7 consistently, and the expected size was 1.8 kb. However, the phytoplasma DNA was not detected in healthy seedlings. Based on sequence analysis of amplified region, this phytoplasma has close homologies with eqilodium phyllody, mulberry dwarf, and aster yellow phytoplasmas, 99.95%, 99.79% and 99.78%, respectively, This phylogetic analysis indicates that ash witches' broom phytoplasma should be classified in the aster yellow group 16SrVI and clearly distinct from the ash yellow group 16SrVII.

• The Plant Pathology Journal
• /
• 제21권1호
• /
• pp.55-58
• /
• 2005

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 16S 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 from 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 ligustrum witches' broom phytoplasma of the 16S rRNA group V, but no DNA amplification was detected from other phytoplasma strains such as groups 16SrI (Aster yellows) and 16SrXII (Stolbur group) in which mulberry dwarf phytoplasma and chrysanthemum witches' broom phytoplasma belong to, respectively. The same results were obtained from both Korean and Chinese isolates of JWB phytoplasma. Nested-PCR using phytoplasma universal primer pair P1/P7 and 16SrV group-specific primer pair 16S(V) F/R could detect group V phytoplasmas rapidly and easily, in particular JWB phytoplasma.

• 한국식물병리학회:학술대회논문집
• /
• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
• /
• pp.136.2-137
• /
• 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
• /
• 제28권3호
• /
• pp.239-247
• /
• 2012

Phytoplasmas have been associated with more than 46 plant species in Korea. Several vegetables, ornamentals, fruit trees and other crop species are affected by phytoplasma diseases. Six 16Sr groups of phytoplasmas have been identified and these phytoplasmas are associated with 63 phytoplasma diseases. Aster yellows phytoplasmas are the most prevalent group and has been associated with more than 25 diseases in Korea. Jujube witches' broom, paulownia witches' broom and mulberry dwarf diseases cause economic losses to host trees throughout the country. So far, Korean phytoplasmas belong to six species of 'Candidatus Phytoplasma'; 'Ca. P. asteris', 'Ca. P. pruni$^*$', 'Ca. P. ziziphi', 'Ca. P. trifolii', 'Ca. P. solani$^*$' and 'Ca. P. castaneae'. The diseases are distributed throughout the country and most of them were observed in Gyeongbuk and Chonbuk provinces. At least four insect vectors; Cyrtopeltis tenuis, Hishimonus sellatus, Macrosteles striifrons and Ophiola flavopicta have been identified for phytoplasma transmission.