scholarly journals Natural Infection of Alstroemeria brasiliensis with Lily Mottle Virus

Plant Disease ◽  
2000 ◽  
Vol 84 (1) ◽  
pp. 103-103 ◽  
Author(s):  
I. Bouwen ◽  
R. A. A. van der Vlugt
Keyword(s):  
Polymerase Chain Reaction Amplification ◽  
Natural Infection ◽  
Tobacco Rattle Virus ◽  
Chenopodium Quinoa ◽  
Flower Color ◽  
Systemic Vein ◽  
Leaf Chlorosis ◽  
Local Lesions ◽  
Funded Project ◽  
Reaction Amplification

During a survey for a European Union-funded project on viruses of Alstroemeria, two A. brasiliensis plants were found expressing virus-like symptoms, including leaf chlorosis with deep-green oval spots and flower color breaking. In enzyme-linked immunosorbent assays (ELISA), no positive reaction was obtained with antisera to Alstroemeria mosaic, Alstroemeria carla, Cucumber mosaic, Freesia mosaic, or Tobacco rattle virus or potyvirus-specific monoclonal antibodies (Agdia, Elkhart, IN). ELISA reactions were positive with antisera to Lily mottle (LMoV) and Rembrandt tulip breaking viruses (1). In electron microscopy preparations of A. brasiliensis, potyvirus-like particles were observed. Using sap-inoculation, the virus was transferred to a range of host species. Chenopodium quinoa, Nicotiana occidentalis accession 37B, and N. occidentalis subsp. obliqua (P1) expressed local lesions; N. clevelandii expressed local and systemic mottle; and N. benthamiana expressed local lesions, systemic vein yellowing, and leaf crinkling. Isolated total RNA from infected N. benthamiana was used for initial cDNA synthesis and polymerase chain reaction amplification with a potyvirus-specific primer set (2). The amplicon (≈670 bp) was cloned and sequenced. The sequence showed 92% homology with the corresponding region of LMoV RNA (GenBank accession no. S44147). The results confirm the infection of A. brasiliensis with LMoV. This is the first report of natural infection of Alstroemeria by LMoV. References: (1) E. L. Dekker et al. J. Gen. Virol. 74:881, 1993. (2) R. A. A. van der Vlugt et al. Phytopathology 89:148, 1999.

scholarly journals Natural Infection of Alstroemeria caryophyllea with Ornithogalum mosaic virus

Plant Disease ◽  
2000 ◽  
Vol 84 (2) ◽  
pp. 202-202 ◽  
Author(s):  
I. Bouwen ◽  
R. A. A. van der Vlugt
Keyword(s):  
Electron Microscopy ◽  
Mosaic Virus ◽  
Natural Infection ◽  
Tobacco Rattle Virus ◽  
Flower Color ◽  
Chenopodium Amaranticolor ◽  
Funded Project ◽  
Necrotic Spots ◽  
Polymerase Chain ◽  
Dark Green

During a survey for a European Union-funded project on the viruses of Alstroemeria, an A. caryophyllea plant was found expressing virus-like symptoms, including dark green vein banding, necrotic spots, and flower color breaking. In enzyme-linked immunosorbent assays (ELISA), no positive reaction was obtained with antisera to Alstroemeria mosaic, Alstroemeria carla, Cucumber mosaic, Freesia mosaic, or Tobacco rattle virus. A positive ELISA reaction was obtained with potyvirus-specific monoclonal antibodies (Agdia, Elkhart, IN) and antiserum to Ornithogalum mosaic virus (OrMV) (1). In electron microscopy leaf dip preparations of A. caryophyllea, potyvirus-like particles were observed. Using sapinoculation, the virus was transferred to Chenopodium amaranticolor and C. quinoa, resulting in local lesions 6 days postinoculation. The presence of OrMV in both Chenopodium spp. was confirmed by electron microscopy and ELISA with antiserum to OrMV. Sequence alignment of DNA fragments (740 bp) obtained in immunocapture-reverse transcription-polymerase chain reaction on RNA isolated from the suspect virus, using a potyvirus-specific primer set (2), showed 91% homology with the corresponding region of OrMV RNA (GenBank accession no. D00615). The results confirm the infection of A. caryophyllea by OrMV. This is the first report of natural infection of Alstroemeria by OrMV. References: (1) J. T. Burger and M. B. von Wechmar. Phytopathology 79:385, 1989. (2) R. A. A. van der Vlugt et al. Phytopathology 89:148, 1999.

scholarly journals Lutzomyia longipalpis naturally infected by Leishmania (L.) chagasi in Várzea Grande, Mato Grosso State, Brazil, an area of intense transmission of visceral leishmaniasis

2010 ◽  
Vol 26 (12) ◽  
pp. 2414-2419 ◽  
Author(s):  
Nanci A. Missawa ◽  
Érika Monteiro Michalsky ◽  
Consuelo Latorre Fortes-Dias ◽  
Edelberto Santos Dias
Keyword(s):  
Visceral Leishmaniasis ◽  
Polymerase Chain Reaction Amplification ◽  
Natural Infection ◽  
Rflp Analysis ◽  
Lutzomyia Longipalpis ◽  
Sand Flies ◽  
Mato Grosso ◽  
Phlebotomine Sand Flies ◽  
Leishmania Spp ◽  
Reaction Amplification

The American visceral leishmaniasis (AVL) is caused by parasites belonging to the genus Leishmania (Trypanosomatidae) and is transmitted to humans through the bite of certain species of infected phlebotomine sand flies. In this study, we investigated the natural infection ratio of Lutzomyia longipalpis, the main vector species of AVL in Brazil, in Várzea Grande, Mato Grosso State. Between July 2004 and June 2006, phlebotomine sand flies were captured in peridomestic areas using CDC light-traps. Four hundred and twenty (420) specimens of Lu. longipalpis were captured. 42 pools, containing 10 specimens of Lu. longipalpis each, were used for genomic DNA extraction and PCR (polymerase chain reaction) amplification. Leishmania spp. DNA was detected in three out of the 42 pools tested, resulting in a minimal infection ratio of 0.71%. Restriction fragment length polymorphism (RFLP) analysis indicated that Leishmania (L.) chagasi was the infective agent in the positive pools.

scholarly journals First Report of Tobacco rattle virus in Spinach in California

Plant Disease ◽  
2010 ◽  
Vol 94 (1) ◽  
pp. 125-125 ◽  
Author(s):  
S. T. Koike ◽  
T. Tian ◽  
H.-Y. Liu
Keyword(s):  
Sugar Beet ◽  
Spinacia Oleracea ◽  
Sandwich Elisa ◽  
Tobacco Rattle Virus ◽  
Chenopodium Quinoa ◽  
Mechanical Transmission ◽  
Rt Pcr ◽  
First Report ◽  
Local Lesions ◽  
Rigid Rod

In 2009 in coastal California (Santa Barbara County), commercially grown spinach (Spinacia oleracea) in two nearby fields exhibited symptoms of a previously unrecognized virus-like disease. Symptoms consisted of general chlorosis and bright yellow blotches and spots. Necrotic spots were also associated with the disease. In affected fields, disease occurred in limited, irregularly shaped patches that ranged from one to several meters in diameter. Symptomatic plants were unmarketable and these small patches of spinach were not harvested. With a transmission electron microscope, rigid, rod-shaped particles with a clear central canal were observed from plant sap of the symptomatic spinach. Analysis by a double-antibody sandwich-ELISA assay (Agdia Inc., Elkhart, IN) for Tobacco rattle virus (TRV) showed that the symptomatic plants were positive. Symptomatic spinach from the field was used for mechanical transmission to Chenopodium quinoa, C. murale, C. capitatum, spinach, and sugar beet (Beta vulgaris). All inoculated plants showed chlorotic local lesions and sugar beet showed chlorotic local lesions with rings. To further confirm the presence of TRV, reverse transcription (RT)-PCR was conducted. Total RNA was extracted from the mechanically inoculated symptomatic spinach plants using an RNeasy Plant Kit (Qiagen Inc., Valencia, CA) and used as a template in RT-PCR with forward (5′-TACATCACATCTGCCTGC-3′) and reverse (5′-CTTCATTCACACAACCCTTG-3′) primers specific to the movement protein gene from the spinach isolate of TRV (GenBank Accession No. AJ007294). Amplicons of the expected size (approximately 562 bp) were obtained. The RT-PCR products were sequenced (GenBank Accession No. GU002156) and compared with TRV sequences in GenBank to confirm the identity of the products. Sequences obtained had 96% nucleotide identity and 97% amino acid identity with TRV sequences available under the GenBank Accession Nos. FJ357571 and AJ007294. On the basis of the data from electron microscopy and serological and molecular analyses, the virus was identified as TRV. Soil samples collected from one of the fields were assayed for nematodes; however, Paratrichodorus or Trichodorus species were not recovered. To our knowledge, this is the first report of TRV in spinach in California. TRV has also been reported in spinach in England (1) and Germany (2). References: (1) A. Kurppa et al. Ann. Appl. Biol. 98:243, 1981. (2) K. Schmidt and R. Koenig. Arch. Virol. 144:503, 1999.

scholarly journals First Report of Artichoke yellow ringspot virus in Globe Artichoke in Turkey

Plant Disease ◽  
2013 ◽  
Vol 97 (10) ◽  
pp. 1388-1388 ◽  
Author(s):  
I. C. Paylan ◽  
M. Ergun ◽  
S. Erkan
Keyword(s):  
Natural Infection ◽  
Chenopodium Quinoa ◽  
French Bean ◽  
Ringspot Virus ◽  
Growth And Yield ◽  
Globe Artichoke ◽  
Rt Pcr ◽  
First Report ◽  
Plant Samples ◽  
Local Lesions

Turkey is one of the main globe artichoke (Cynara cardunculus L. subsp. scolymus (L.) Hayek) producers in the world. Cultivation of this crop is done mainly in the Aegean and Eastern Marmara regions with asexually propagated cultivars such as Bayrampasa and Sakiz. More than half of total globe artichoke production in Turkey is obtained from the provinces of Izmir, Aydin, and Mugla in the Aegean region. Surveys in 2011 and 2012 were carried out to look for the presence of Artichoke yellow ringspot virus (AYRSV), Tobacco mosaic virus (TMV), and Tomato spotted wilt virus (TSWV) in the globe artichoke production areas in these three provinces. Double antibody sandwich (DAS)-ELISA and reverse transcriptase (RT)-PCR assays conducted for TMV and TSWV showed that the samples were not infected with these two viruses. Due to the lack of commercial ELISA kits against AYRSV, RT-PCR and biological indexing were used for its identification. Leaf tissues from 35 symptomatic and 25 symptomless plants were sampled and analyzed by RT-PCR using as template total RNAs extracted by a silica gel method (1). RT-PCR was conducted as previously reported (1). A PCR product of the expected size (about 530 bp) was obtained from five plant samples that were collected from Izmir province and had symptoms of bright yellow spots and line patterns on the leaves. The incidence of diseased plants in the fields ranged from 1 to 5%. In previously conducted studies, these symptoms were defined as typical symptoms of AYRSV on artichokes (2,3,4). One of the PCR products was cloned and sequenced. BLASTn analysis of the obtained sequence (GenBank Accession No. KC622054) showed 92% nucleotide identity with the partial RNA1 sequence of an AYRSV isolate from Allium cepa (AM087671.2). Furthermore, selected test plants were mechanically inoculated with sap from plant samples that were positive in RT-PCR. Chlorotic local lesions and systemic mottling symptoms were observed on Chenopodium quinoa; chlorotic lesions, mosaic, and deformation on Cucumis sativus; and systemic mosaic, reddish necrotic local lesions, and malformation on Phaseolus vulgaris (French bean). Results of the biological tests were confirmed by RT-PCR. AYRSV has a wide host range including artichoke and six other cultivated plant species and can be easily transmitted by seed, plant sap, and vegetative propagation (3). To our knowledge, this is the first report of natural infection of globe artichoke by AYRSV in Turkey. AYRSV infections can have a detrimental effect on the growth and yield of artichoke plantings. This assay will be useful for further epidemiological studies. References: (1) X. Foissac et al. Acta Hortic. 550:37, 2001. (2) D. Galliitelli et al. Adv. Virus Res. 84:289, 2012. (3) P. E. Kyriakopoulou et al. Ann. Inst. Phytopathol. Benaki 14:139, 1985. (4) V. I. Maliogka et al. Phytopathology 96:622, 2006.

Polymerase chain reaction amplification and typing of rotavirus in environmental samples

1995 ◽  
Vol 31 (5-6) ◽  
pp. 371-374 ◽  
Author(s):  
R. Gajardo ◽  
R. M. Pintó ◽  
A. Bosch
Keyword(s):  
Polymerase Chain Reaction ◽  
Environmental Samples ◽  
Polymerase Chain Reaction Amplification ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Group A ◽  
Reaction Amplification ◽  
Stool Specimens ◽  
Polymerase Chain

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.

Detection of enteroviruses in marine waters by direct RT-PCR and cell culture

1995 ◽  
Vol 31 (5-6) ◽  
pp. 323-328 ◽  
Author(s):  
K. A. Reynolds ◽  
C. P. Gerba ◽  
I. L. Pepper
Keyword(s):  
Cell Culture ◽  
Polymerase Chain Reaction Amplification ◽  
Cost Effective ◽  
The United States ◽  
Water Runoff ◽  
Rt Pcr ◽  
Marine Waters ◽  
Storm Water Runoff ◽  
Routine Monitoring ◽  
Reaction Amplification

Sewage outfalls and storm water runoff introduces pathogenic human enteric viruses into marine coastal waters, which may pose a potential public health risk. Although members of the enterovirus group have been suggested as possible indicators of sewage pollution in marine waters, the lack of rapid, sensitive and cost effective methods have prevented routine monitoring in the United States. This study compared traditional cell culture and direct RT-PCR (reverse transcriptase-polymerase chain reaction) amplification for detection of an enterovirus. Poliovirus could be recovered from 100 L of artificial seawater with an average efficiency of 77%, using adsorption and elution from electronegative filters. Viruses were eluted from the filters with 1.5% beef extract for viruses (BEV) adjusted to pH 9.5 and reconcentrated by organic flocculation to a volume of 30 mL. Substances which interfered with detection by RT-PCR were removed by treatment of the concentrates with sephadex and chelex resins. Direct RT-PCR could detect 2.5 and 0.025 PFU (plaque forming units) for single (25 cycles) and double PCR (2 × 25 cycles) in 10 μL of pure culture poliovirus samples, respectively. These methods are currently being applied to assess the occurrence of enteroviruses at marine bathing beaches influenced by sewage discharges.

scholarly journals Molecular Characterization of hobo-Mediated Inversions in Drosophila melanogaster

Genetics ◽  
1996 ◽  
Vol 144 (2) ◽  
pp. 647-656
Author(s):  
William B Eggleston ◽  
Nac R Rim ◽  
Johng K Lim
Keyword(s):  
X Chromosome ◽  
Polymerase Chain Reaction Amplification ◽  
Polytene Chromosomes ◽  
Chromosomal Inversions ◽  
Hobo Element ◽  
Reaction Amplification ◽  
Notch Locus ◽  
Polymerase Chain

Abstract The structure of chromosomal inversions mediated by hobo transposable elements in the Uc-1 X chromosome was investigated using cytogenetic and molecular methods. Uc-1 contains a phenotypically silent hobo element inserted in an intron of the Notch locus. Cytological screening identified six independent Notch mutations resulting from chromosomal inversions with one breakpoint at cytological position 3C7, the location of Notch. In situ hybridization to salivary gland polytene chromosomes determined that both ends of each inversion contained hobo and Notch sequences. Southern blot analyses showed that both breakpoints in each inversion had hobo-Notch junction fragments indistinguishable in structure from those present in the Uc-1 X chromosome prior to the rearrangements. Polymerase chain reaction amplification of the 12 hobo-Notch junction fragments in the six inversions, followed by DNA sequence analysis, determined that each was identical to one of the two hobo-Notch junctions present in Uc-1. These results are consistent with a model in which hobo-mediated inversions result from homologous pairing and recombination between a pair of hobo elements in reverse orientation.

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