• Title/Summary/Keyword: United States isolate

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Generation of an Infectious Clone of a New Korean Isolate of Apple chlorotic leaf spot virus Driven by Dual 35S and T7 Promoters in a Versatile Binary Vector

  • Kim, Ik-Hyun;Han, Jae-Yeong;Cho, In-Sook;Ju, HyeKyoung;Moon, Jae Sun;Seo, Eun-Young;Kim, Hong Gi;Hammond, John;Lim, Hyoun-Sub
    • The Plant Pathology Journal
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    • v.33 no.6
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    • pp.608-613
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    • 2017
  • The full-length sequence of a new isolate of Apple chlorotic leaf spot virus (ACLSV) from Korea was divergent, but most closely related to the Japanese isolate A4, at 84% nucleotide identity. The full-length cDNA of the Korean isolate of ACLSV was cloned into a binary vector downstream of the bacteriophage T7 RNA promoter and the Cauliflower mosaic virus 35S promoter. Chenopodium quinoa was successfully infected using in vitro transcripts synthesized using the T7 promoter, detected at 20 days post inoculation (dpi), but did not produce obvious symptoms. Nicotiana occidentalis and C. quinoa were inoculated through agroinfiltration. At 32 dpi the infection rate was evaluated; no C. quinoa plants were infected by agroinfiltration, but infection of N. occidentalis was obtained.

Characterization of Host Specificity and Reproductive Compatibility Among Different Isolates of Belonolaimus longicaudatus in Southeastern United States (미국 동남부지역에 분포하는 Belonolaimus longicaudatus 개체군들간의 기주특이성과 생식교합능력 비교)

  • Han, Hye-Rim;Donald, W. Dickson
    • The Korean Journal of Soil Zoology
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    • v.9 no.1_2
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    • pp.12-15
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    • 2004
  • Five isolates of Belonolaimus longicaudatus collected from different geographical locations and hosts in southeastern United Stated were tested to determine their host specificity and reproductive compatibility. Both the Georgia and Lake Alfred isolates reproduced well on cotton, bermudagrass, and citrus. On the other hand, the North Carolina and Gainesville isolates reproduced poorly on all crops. The Hastings isolate reproduced well on cotton and potato. The Lake Alfred and Hastings isolates showed greater host specificity to their original host of citrus and potato, respectively. Except for the Hastings isolate, all isolates reproduced poorly on potato, which clearly differentiated the Hastings isolate from others (P>0.05). Hybridization test presented reproductive compatibility among five different isolates of B. longicaudatus by generating F$_1$ offspring. The Gainesville males showed comparatively poor ability to fertilize other females and generated a lower number of offspring. In contrast, the North Carolina males were attracted by all the other isolates of sting nematode, and produced the highest number of juveniles.

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Differences in isolates of Tomato yellow leaf curl virus in tomato fields located in Daejeon and Chungcheongnam-do between 2017 and 2018

  • Oh, June-Pyo;Choi, Go-Woon;Kim, Jungkyu;Oh, Min-Hee;Kim, Kang-Hee;Park, Jongseok;Domier, Leslie L.;Hammond, John;Lim, Hyoun-Sub
    • Korean Journal of Agricultural Science
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    • v.46 no.3
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    • pp.507-517
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    • 2019
  • To follow up on a 2017 survey of tomato virus diseases, samples with virus-like symptoms were collected from the same areas (Buyeo-gun, Chungchungnam-Do and Daejeon, Korea) in 2018. While in 2017 mixed infections of Tomato mosaic virus with either Tomato yellow leaf curl virus (TYLCV) or Tomato chlorosis virus were detected, only TYLCV was detected in symptomatic samples in 2018. TYLCV amplicons of c.777 bp representing the coat protein (CP) coding region were cloned from the TYLCV positive samples, and the sequence data showed a 97.17% to 98.84% nucleotide and 98.45% to 99.22% amino acid identity with the 2017 Buyeo-gun isolate (MG787542), which had the highest amino acid (aa) sequence identity of up to 99.2% with four 2018 Buyeo-gun sequences (MK521830, MK521833, MK521834, and MK521835). The lowest aa sequence identity of 98.45% was found in a 2018 Daejeon isolate (MK521836); the distance between Buyeo-gun and Daejeon is about 45 km. Phylogenetic analysis indicated that the currently reported CP sequences are most closely related to Korean sequences from Masan (HM130912), Goseong (JN680149), Busan (GQ141873), Boseong (GU325634), and the 2017 isolate TYLCV-N (MG787543) in the 'Japan' cluster of TYLCV isolates and distinct from the 'China' cluster isolates from Nonsan (GU325632), Jeonju (HM130913) and Jeju (GU325633, HM130914). Our survey data from 2017 and 2018 suggest that TYLCV has become established in Korea and may be spread by whitefly vectors from weed reservoirs within the farm environment.

Fusarium oxysporum Protects Douglas-fir (Pseudotsuga menziesii) Seedlings from Root Disease Caused by Fusarium commune

  • Dumroese, R. Kasten;Kim, Mee-Sook;James, Robert L.
    • The Plant Pathology Journal
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    • v.28 no.3
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    • pp.311-316
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    • 2012
  • Fusarium root disease can be a serious problem in forest and conservation nurseries in the western United States. Fusarium inoculum is commonly found in most container and bareroot nurseries on healthy and diseased seedlings, in nursery soils, and on conifer seeds. Fusarium spp. within the F. oxysporum species complex have been recognized as pathogens for more than a century, but attempts to distinguish virulence by correlating morphological characteristics with results of pathogenicity tests were unsuccessful. Recent molecular characterization and pathogenicity tests, however, revealed that selected isolates of F. oxysporum are benign on Douglas-fir (Pseudotsuga menziesii) seedlings. Other morphologically indistinguishable isolates, which can be virulent, were identified as F. commune, a recently described species. In a replicated greenhouse study, inoculating Douglas-fir seedlings with one isolate of F. oxysporum prevented expression of disease caused by a virulent isolate of F. commune. Moreover, seedling survival and growth was unaffected by the presence of the F. oxysporum isolate, and this isolate yielded better biological control than a commercial formulation of Bacillus subtilis. These results demonstrate that an isolate of nonpathogenic F. oxysporum can effectively reduce Fusarium root disease of Douglas-fir caused by F. commune under nursery settings, and this biological control approach has potential for further development.

Deep Sequencing Analysis of Apple Infecting Viruses in Korea

  • Cho, In-Sook;Igori, Davaajargal;Lim, Seungmo;Choi, Gug-Seoun;Hammond, John;Lim, Hyoun-Sub;Moon, Jae Sun
    • The Plant Pathology Journal
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    • v.32 no.5
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    • pp.441-451
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    • 2016
  • Deep sequencing has generated 52 contigs derived from five viruses; Apple chlorotic leaf spot virus (ACLSV), Apple stem grooving virus (ASGV), Apple stem pitting virus (ASPV), Apple green crinkle associated virus (AGCaV), and Apricot latent virus (ApLV) were identified from eight apple samples showing small leaves and/or growth retardation. Nucleotide (nt) sequence identity of the assembled contigs was from 68% to 99% compared to the reference sequences of the five respective viral genomes. Sequences of ASPV and ASGV were the most abundantly represented by the 52 contigs assembled. The presence of the five viruses in the samples was confirmed by RT-PCR using specific primers based on the sequences of each assembled contig. All five viruses were detected in three of the samples, whereas all samples had mixed infections with at least two viruses. The most frequently detected virus was ASPV, followed by ASGV, ApLV, ACLSV, and AGCaV which were withal found in mixed infections in the tested samples. AGCaV was identified in assembled contigs ID 1012480 and 93549, which showed 82% and 78% nt sequence identity with ORF1 of AGCaV isolate Aurora-1. ApLV was identified in three assembled contigs, ID 65587, 1802365, and 116777, which showed 77%, 78%, and 76% nt sequence identity respectively with ORF1 of ApLV isolate LA2. Deep sequencing assay was shown to be a valuable and powerful tool for detection and identification of known and unknown virome in infected apple trees, here identifying ApLV and AGCaV in commercial orchards in Korea for the first time.

Role of Intergenic and 3'-Proximal Noncoding Regions in Coat Protein Expression and Replication of Barley yellow dwarf virus PAV

  • Moon, Jae-Sun;Nancy K. McCoppin;Leslie L. Domier
    • The Plant Pathology Journal
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    • v.17 no.1
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    • pp.22-28
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    • 2001
  • Barley yellow dwarf virus PAV (BYDV-PAV) has a 5.7-kb positive-sense single-stranded RNA genome that contains six open reading frames (ORFs). BYDV-PAV produces three subgenomic RNAs (sgRNAs). The largest of which encodes the coat, 17-kDa, and readthrough proteins from two initiation codons. To investigate the role of intergenic and 3'-proximal noncoding regions (NCRs) in coat protein (CP) expression and BYDV-PAV replication, a full-length infectious cDNA of the RNA genome of an Illinois isolate of BYDV-PAV was constructed downstream of the Cauliflower mosaic virus-35S promoter. Linear DNA molecules of these cDNAs were infectious, expressed the 22-kDa CP, and produced both genomic RNA sgRNAs in ratios similar to those observed in protoplasts inoculated with viral RNA. The portion of 5'NCR of sgRNA1 between ORFs 2 and 3 was not required for, but enhanced translation of CP from ORF3. Mutants containing deletions in the NCR downstream of ORF5 failed to replicate in oat protoplasts. These results indicate that an intact 3$^1$NCR is required for BYDV-PAV replication.

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Isolation of porcine reproductive and respiratory syndrome virus(PRRSV) in Korea (돼지생식기 및 호흡기증후군(Porcine Reproductive and Respiratory Syndrome ; PRRSV) 바이러스의 국내분리주 작성에 관한 연구)

  • Kweon, Chang-hae;Kwon, Byung-joon;Lee, Han-jung;Cho, Jae-jin;Hwang, Eui-kyung;Shin, Jin-ho;Yoon, Yong-dhuk;Kang, Yung-bai;An, Soo-hwan;Kim, Yong-hee;Huh, Won;Jun, Moo-hyung;Wensvoort, G.
    • Korean Journal of Veterinary Research
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    • v.34 no.1
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    • pp.77-83
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    • 1994
  • Three viral strains, causing CPE in porcine alveolar macrophage cell, were isolated from aborted fetus, serum from young pig showing blue-ear sign and lung of suspected pig, respectively. The differential diagnostic results showed no characteristics of Aujeszky's disease virus(ADV), hog cholera virus (HCV), Japanese encephalitis virus(JEV), porcine parvovirus(PPV) and encephalomyocarditis virus (EMCV). However, positive reactions were demonstrated by IFA using monospecific porcine antibodies against Lelystad virus. When the paired sera of experimentally inoculated swine with one of isolate, KPRRSV-l were tested by IPMA, the result indicated that the isolate was related to United States isolate than European LV.

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Immunohistochemical detection of viral antigen and pathological lesion in piglets experimentally infected with encephalomyocarditis virus (뇌심근염 바이러스의 실험적 감염자돈에 대한 병리학적 소견과 바이러스 항원의 면역조직화학적 검출)

  • Cho, Sung-hwna;Joo, Han-soo;Kim, Hyun-soo
    • Korean Journal of Veterinary Research
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    • v.33 no.2
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    • pp.301-308
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    • 1993
  • Three or 7day old piglets were infected experimentally with different encephalomyocarditis virus isolates to detect the viral antigen by the immunoperoxidase technique and to observe strain difference in their pathogenecity in newborn pigs by comparing clinical signs and pathologic lesions. Clinical signs of the infected pigs were different depending on the virus strain, pig age and infection route. Encephalomyocarditis virus(EMCV) NVSL-PR isolate was more pathogenic than MN-25 and MN-30 isolate. Three day old piglets showed more severe illness than 7 day old piglets. Predominant clinical signs were sudden death without noticeable clinical signs and dyspnea manifested as heavy abdominal breathing. Contact-infection from infected piglets to controls was observed in the oro-nasally infected group but not the intramuscular group. Common necropsy findings of dead piglets in both age groups infected with MN-25 and NVSL-PR were accumulation of excessive fluid in the body cavities and mild to diffuse necrotic areas observed in the hearts and occasionally in the livers. Microscopically, myocarditis with inflammatory cell infiltration, necrosis of the myocardial muscle fibers and occasional mineralization were observed along with interstitial pneumonia and centrolobular necrosis in the liver. Using an immunoperoxidase technique, viral antigen was detected in myocardial muscle fibers of piglets infected with EMCV.

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Light and Electron Microscopy Studies Elucidating Mechanisms of Tomato Leaf Infection by Pseudocercospora fuligena

  • Zelalem Mersha;Girma Birru;Bernhard Hau
    • The Plant Pathology Journal
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    • v.39 no.2
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    • pp.181-190
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    • 2023
  • The fungal pathogen Pseudocercospora fuligena, known to affect tomatoes in the tropics and subtropics, has been reported from temperate climates including the United States and Turkey in recent years. In this study, an isolate from fresh tomatoes and the disease it causes were characterized and infection mechanisms investigated. Macroscopically, both sides of tomato leaves show indistinct effuse patches but prolific production of fuliginous lesions is conspicuous on the abaxial side first but also on the adaxial side later on as infection progressed. Microscopically, fascicles of conidiophores (11-128 ㎛ × 3.5-9 ㎛) arising from stromata and conidia with up to 12 septations were observed. Molecular characterization of the isolate revealed high homology (99.8%) to other P. fuligena isolated from tomatoes in Turkey. Out of the 10 media tested, P. fuligena grew significantly well and sporulated better on unsealed tomato oatmeal agar and carrot leaf decoction agar, both supplemented with CaCO3. Direct transfer of conidia from profusely sporulating lesions was the easiest and quickest method of isolation for in-vitro studies. Light and scanning electron microscopy on cleared and intact tomato leaves further confirmed stomatal penetration and egress as well as prevalence of primary and secondary infection hyphae. In situ, blocked stomatal aperture areas of 154, 401, and 2,043 ㎛2 were recorded at 7, 12, and 17 days after inoculation, respectively. With the recent expanded horizon of the pathosystem and its consequential impact, such studies will be useful for a proper diagnosis, identification and management of the disease on tomato worldwide.