• Journal of Forest and Environmental Science
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• 제39권1호
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• pp.49-54
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• 2023

For unknown reasons, a few trees in a private chestnut orchard in Icheon si, Gyunggi-do suffered leaf chlorosis and growth decline. Based on symptoms, phytoplasma was a probable cause. Leaf samples were collected from two symptomatic and non-symptomatic trees in the orchard for phytoplasma detection. An amplicon of about 1.2 bp size was obtained from both symptomatic trees by PCR with the universal 16S rDNA primers. Sequences of these amplicons were found to have 99% nucleotide sequence identity to the corresponding genomic region of 16SrIII (X-disease group). More than 100 phytoplasma isolates, such as Candidatus phytoplasma pruni, Milkweed yellows phytoplasma, Goldenrod yellows phytoplasma, Tsuwabuki witches'-broom phytoplasma, Candidatus Phytoplasma trifolii, etc. were involved in the list. Phylogenetic analysis revealed that the sequence obtained in this study closely clustered with Candidatus phytoplasma groups. While one of the amplicons shared 91% identity with the Candidatus phytoplasma castaneae, the other shared only 47%. It needs further analysis and investigation to determine the exact taxonomy. Meanwhile, based on the analysis of the sequences, chlorosis, and small leaves were associated with phytoplasma.

• The Plant Pathology Journal
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• 제23권2호
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• pp.106-108
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• 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
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• 제96권6호
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• pp.737-741
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• 2007

전형적인 파이토플라스마성 빗자루증상을 보이는 아까시나무가 전북지방에서 발견되었으며, 일반적으로 작은 잎들이 총생하였고, 마디와 마디 사이가 좁고, 가지가 위축되어 있는 증상을 보였다. 파이토플라스마 검출 primer인 P1/P7(약 1.8 kb)과 R16F2n/R2(약 1.2 kb)를 이용하여 증폭한 결과 빗지루증상을 보인 아까시나무에서 파이토플라스마 검출되었으며, 건전한 아카시에서는 PCR산물이 증폭되지 않았다. 염기서열 분석에 의한 유의성 검정결과 아까시나무 빗자루병 파이토플라스마는 aster yellow, 뽕나무오갈병, maize bushy stunt, 물푸레나무 빗자루병, 붉나무 빗자루병 파이토플라스마와 99.2% 이상의 유의성을 보였으며, 아까시나무 빗자루병 (GeneBank Accession No. AF 244363) 및 대추나무 빗자루병 파이토플라스마와는 각 각 88.6% 및 87.7%의 유의성을 보여 본 연구에서 분석한 아까시나무 빗자루병 파이토플라스마는 Candidatus phytoplasma asteris(16Sr I) group에 속하였으며, 16Sr III(peach X-disease phytoplasma group) group에 속하는 아카시나무 빗자루병 파이토플라스마와는 다른 계통으로 분류되었다.

• The Plant Pathology Journal
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• 제38권2호
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• pp.146-158
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• 2022

Changes in physiological and biochemical patterns in lucerne plants caused by the presence of 'Candidatus Phytoplasma australasia', which is one of the significant pathogens causing yield losses in lucerne plants, were investigated. Significant differences were evident in total chlorophyll, chlorophyll a, chlorophyll b, and protein amounts between 'Ca. Phytoplasma australasia'-positive and negative lucerne plants. Stress-related metabolites such as phenol, malondialdehyde, and proline accumulations in 'Ca. Phytoplasma australasia'-positive plants were remarkably higher than those of phytoplasma-negative plants. As a response to disease attack, phytoplasma-positive plants exhibited higher antioxidant enzymes (peroxidase and catalase) and nonenzymatic metabolite responses such as jasmonic and salicylic acids. We state that partial disease responses were revealed for the first time to breed resistant lucerne lines infected by 'Ca. Phytoplasma australasia'.

• The Plant Pathology Journal
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• 제28권3호
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• pp.239-247
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• 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.

• The Plant Pathology Journal
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• 제27권4호
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• pp.367-371
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• 2011

Multiplex PCR assays were developed for the simultaneous detection of ten important Korean quarantine phytoplasmas. The species-specific primers were designed based on ribosomal protein, putative preprotein translocase Y, immunodominant protein, elongation factor TU, chaperonin protein and the 16S rRNA genes of 'Candidatus (Ca.) Phytoplasma' species. Three main primer sets were prepared from ten designed primer pairs to limit nonspecific amplification as much as possible. The multiplex PCR assay using the three primer sets successfully amplified the correct conserved genes for each 'Ca. Phytoplasma' species. In addition, ten important 'Ca. Phytoplasma' species could be easily determined by recognizing band patterns specific for each phytoplasma species from three primer sets. Moreover, a high sensitivity of multiplex PCR for each primer set was observed for samples containing a low DNA concentration (10 ng/${\mu}l$). This study provides the useful multiplex PCR assay as a convenient method to detect the presence of ten important quarantine phytoplasmas in Korea.

• The Plant Pathology Journal
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• 제35권3호
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• pp.274-279
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• 2019

Artemisia sieberi showing symptoms resembling those caused by phytoplasma were observed in Geno Mountain, Hormozgan Province, Iran, and were examined for phytoplasma presence by PCR assays. In addition, the essential oils hydrodistilled from the aerial parts of phytoplasma-infected and healthy plants have been analyzed and compared by GC and GC/MS. Phylogenetic and virtual RFLP analysis of the 16S rRNA gene sequences revealed that the phytoplasma associated with A. sieberi witches' broom (AsWB) was a strain of 'Candidatus Phytoplasma aurantifolia'. The presence of the disease, however, induced a further enrichment (from 4.9 to 45.2%, a relative increase of 90%) of the entire monoterpene class as compared to the abundance in healthy samples. Conversely, a matching decrease in monoterpenoid (from 48.7 to 2%, a relative decrease of 90.2%) was observed in the infected plants. Besides the first report of phytoplasma infection of A. sieberi, the changes of its essential oils are reported.

• 생명과학회지
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• 제27권6호
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• pp.694-701
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• 2017

알려진 단독배양이 가능한 원핵생물 중 최소게놈을 가지고 있는 Mycoplasma genitalium보다 보존적 유전자 수가 적은 14개 원핵생물의 유전자를 보존적 유전자 관점의 COG (Clusters of Orthologous Group of proteins)로 검토하였다. 분석대상은 M. genitalium, 초고온성 고세균으로 세포외공생을 하는 Nanoarchaeum equitans, 진정세균으로 식물의 세포내에 기생하는 병원균인 Candidatus Phytoplasma 속 4개와 식물의 수액을 섭취하는 곤충의 세포내에 공생하는 9종이었다. M. genitalium이 가진 367개의 보존적 유전자 중에서, 284개가 비교대상 다른 원핵생물과 공통이었다. M. genitalium 등 분석대상 원핵생물 모두에 보존적 유전자는 29개로, 이들은 리보솜 구성단백질 22개 등 번역관련 25개, RNA 중합효소의 소단위체 3개, 단백질 접힘관련 1개 등으로 단백질의 중요성을 알 수 있었다. 분석대상 15개 원핵생물 중 Candidatus Phytoplasma속 4개 균주 모두에만 존재하는 COG는 40개 였다. 속(genus)이 서로 다른 나머지 9개의 Candidatus는 곤충에 공생한다는 공통점이 있지만 COG0539 (Ribosomal protein S1) 하나만 공통적이었고, 이는 곤충 세포내 공생체들 사이에 보존적 유전자가 다양함을 나타내는 것으로 판단되었다. 본 연구의 결과는 배양이 불가능한 세균의 보존적 유전자 이해에 대한 단서와 함께 아미노산, 항생제, 의약품, 유기합성 전구체 등을 효율적으로 합성하는 원핵생물의 조작에 필요한 기초자료로 활용이 가능할 것이다.

• Journal of Microbiology and Biotechnology
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• 제21권1호
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• pp.81-88
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• 2011

The anti-phytoplasma activities of surfactin (derived from Iranian native Bacillus subtilis isolates) and tetracycline towards Candidatus "Phytoplasma aurantifolia", the agent of lime Witches' broom disease, were investigated. HPLC was used to quantify the surfactin production in four previously characterized native surfactin-producing strains, and the one producing the highest amount of surfactin (about 1,500 mg/l) was selected and cultivated following optimized production and extraction protocols. Different combinations of purified surfactin and commercial tetracycline were injected into artificially phytoplasmainfected Mexican lime seedlings using a syringe injection system. An absolute quantitative real-time PCR system was developed to monitor the phytoplasma population shifts in the lime phloem during 3 months following the injections. The results revealed that the injections of surfactin or tetracycline had a significant inhibitory effect on Candidatus "P. aurantifolia". However, the combined treatment with both surfactin and tetracycline (1:1) resulted in the highest inhibition due to a synergic effect, which suppressed the phytoplasma population from about $2{\times}10^5$ to less than 10 phytoplasma units/g plant tissue.

• The Plant Pathology Journal
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• 제36권4호
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• pp.364-377
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• 2020

Sugarcane is an important sugar crop contributes more than 80% of world sugar production. Mosaic, leaf fleck, and yellow leaf (YL) are the major viral diseases affecting sugarcane, amongst YL occurrence is widely reported in all the sugarcane growing countries. It is caused by Sugarcane yellow leaf virus (SCYLV) and detailed works were done on complete genome characterization, transmission, and management. However, in countries like Egypt, South Africa, Cuba, Mauritius and Hawaii, the disease was reported to the cause of sugarcane yellow leaf phytoplasma (SCYP) and/or SCYLV as single/combined infections. Hence, we have investigated in detail to identify the exact Candidatus phytoplasma taxon associated in Indian cultivars affected with YL. The sequencing results and the restriction fragment length polymorphism pattern of the PCR products using the universal phytoplasma primers confirmed presence of sugarcane grassy shoot (SCGS) phytoplasma (16SrXI group) in the YL-affected plants. Mixed infection of SCYLV and SCGS phytoplasma was estimated as 32.8% in YL affected plants. Evolutionary genetic relationship between SCYP and SCGS phytoplasma representatively taken from different countries showed that SCYP from South Africa and Cuba were diverged from others and had a highest similarity with SCGS phytoplasma. Although we wanted to identify SCYP from YL affected Indian sugarcane cultivars, the study clearly indicated a clear absence of SCYP in YL affected plants and we found SCYLV as the primary cause for the disease.