Molecular characterization of Spiroplasma citri BR3 lines that differ in transmissibility by the leafhopper Circulifer tenellus

1996 ◽  
Vol 42 (2) ◽  
pp. 124-131 ◽  
Author(s):  
Jacqueline Fletcher ◽  
Mary E. Shaw ◽  
Ginger R. Baker ◽  
Karla J. Dugan ◽  
Fengchun Ye ◽  
...  
Keyword(s):  
Extrachromosomal Dna ◽  
Protein Patterns ◽  
Spiroplasma Citri ◽  
Dual Mobility ◽  
Beet Leafhopper ◽  
Circulifer Tenellus ◽  
Unique Restriction ◽  
Total Dna ◽  
Dna Profiles

Four lines of Spiroplasma citri strain BR3, derived from different maintenance conditions over several years and differing in their ability to be transmitted by the beet leafhopper Circulifer tenellus were characterized. The lines included BR3-T (transmissible), maintained in turnip by leafhopper transmission; BR3-G (now nontransmissible), maintained in plants by periodic graft transmission; BR3-P (now rarely transmissible), subcultured in artificial medium over 130 times; and BR3-M (transmissible), subcultured 43 times. Although all four lines had similar overall protein profiles, the two transmissible lines each contained two proteins missing in the non- or rarely transmitted lines. In addition, one protein was unique to BR3-M and another was unique to BR3-P. Spiralin, a major S. citri membrane protein, had dual mobility in line BR3-G only. Patterns of extrachromosomal DNA and restricted total DNA also were similar, although differences occurred among the four lines. The genome of line BR3-G was larger than those of the other lines and unique restriction bands occurred in this line. Protein and DNA profiles of six to eight individual clones of each line also were compared. Protein patterns within each clone were indistinguishable except for a difference in the migration rate of spiralin in clones of BR3-G. Restricted total DNA showed differential patterns among clones of each line, possibly reflecting differences in extrachromosomal DNA. Molecular differences among the spiroplasma lines may reflect the selection pressures of the different environments in which they were maintained and suggest genes and proteins that may be involved in the biological phenotypes of these lines.Key words: spiroplasma, Mollicutes, insect transmission.

scholarly journals Characterization of Beet curly top virus Strains Circulating in Beet Leafhoppers (Hemiptera: Cicadellidae) in Northeastern Oregon

Plant Disease ◽  
2016 ◽  
Vol 100 (8) ◽  
pp. 1586-1590 ◽  
Author(s):  
Silvia I. Rondon ◽  
Mary Sue Roster ◽  
Launa L. Hamlin ◽  
Kelsie J. Green ◽  
Alexander V. Karasev ◽  
...  
Keyword(s):  
Genomic Sequence ◽  
Beet Leafhopper ◽  
Beet Curly Top Virus ◽  
Circulifer Tenellus ◽  
Virus Incidence ◽  
Important Pest ◽  
Important Field ◽  
Virus Genomes ◽  
Virus Strains

The beet leafhopper, Circulifer tenellus, is an agriculturally important pest, particularly in the western United States. This insect transmits the Beet curly top virus (BCTV) to multiple crops, including bean, tomato, and pepper. In this study, we investigated the incidence of BCTV in individual leafhoppers collected at several sites in northeastern Oregon during the growing season in 2007, 2008, and 2009. Of the 800 insects tested, 151 (18.9%) were found positive for the virus. Percentage of virus incidence varied from 0% at one location in 2009 to a high of 55.6% for a location sampled in 2008. The complete virus genomes from one virus-positive insect collected in each of the 3 years were determined. BLAST analysis of the BCTV whole-genome sequences from 2007, 2008, and 2009 insects showed 98, 94, and 96% identities with the BCTV-Worland sequence (AY134867), respectively. The BCTV_2008 sequence showed the greatest identity (96%) with another BCTV genomic sequence (JN817383), and was found to be a recombinant between the BCTV-Worland type, representing the majority of the genome (approximately 2.2 kb), and the BCTV-CFH type that provided an approximately 0.8-kb fragment spanning replication-related genes C1 and C2. This area of the BCTV genome, between the C1 and C2 genes, was previously found to carry symptom determinants of the virus, and the data may suggest more severe effects of BCTV during the 2008 season. Results indicate that BCTV is common and widespread in C. tenellus in eastern Oregon and that there is substantial genetic diversity among the virus strains present in this important field and vegetable crop-growing region.

scholarly journals Characterization of clinically significant isolates ofStaphylococcus epidermidisfrom patients with cerebrospinal fluid shunt infections

1991 ◽  
Vol 106 (3) ◽  
pp. 467-475 ◽  
Author(s):  
J. Etienne ◽  
B. Charpin ◽  
J. Grando ◽  
Y. Brun ◽  
M. Bes ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Restriction Analysis ◽  
Cerebrospinal Fluid Shunt ◽  
Extrachromosomal Dna ◽  
Dna Restriction ◽  
Total Dna ◽  
Resistant Strains ◽  
Clinically Significant ◽  
Shunt Infections

SUMMARYBiotyping, slime production, antibiograms, extrachromosomal DNA banding and total DNA restriction analysis were used to characterizeStaphylococcus epidermidisstrains causing cerebrospinal fluid shunt infections in 11 patients. Infections considered to be community acquired and those acquired in the first 2 weeks of hospital admission were due to oxacillin-susceptible isolates. Multiply resistant strains were isolated from patients who were in hospital for more than 1 month before tube implantation. Slime was detected in staphylococci for 54% of cases, but its expression varied. Strains from different patients could be differentiated from one another by the extrachromosomal DNA bandings and total DNA restriction patterns, but isolates from the same patient were usually similar. During the period of external drainage, epidemiological markers were useful in differentiating persistence of infection from contamination or re-infection by a new strain.

scholarly journals Detection of Pathogens Associated with Psyllids and Leafhoppers in Capsicum annuum L. in the Mexican States of Durango, Zacatecas, and Michoacán

Plant Disease ◽  
2018 ◽  
Vol 102 (1) ◽  
pp. 146-153 ◽  
Author(s):  
K. D. Swisher ◽  
J. E. Munyaneza ◽  
R. Velásquez-Valle ◽  
J. Mena-Covarrubias
Keyword(s):  
Capsicum Annuum ◽  
Mixed Infections ◽  
Bactericera Cockerelli ◽  
Viral Pathogens ◽  
Spiroplasma Citri ◽  
Beet Leafhopper ◽  
Circulifer Tenellus ◽  
Capsicum Annuum L ◽  
Candidatus Liberibacter ◽  
Pepper Plants

In fall 2014, 5 to 75% percent of chili and bell pepper (Capsicum annuum L.) in commercial fields located in the Mexican states of Durango, Zacatecas, and Michoacán had symptoms of deformed, small, mosaic, curled, and chlorotic leaves; shortened internodes; plant dwarfing; or phyllody and rosetting leaf tips. At the same time, leafhoppers and psyllids were observed in the fields, and more than 50 beet leafhoppers (Circulifer tenellus) and nearly 300 potato psyllids (Bactericera cockerelli) were collected from the pepper plants and adjacent weeds. Based on the insect pressure and observed symptoms, nearly 400 pepper samples were collected across this region of Mexico and tested for the presence of leafhopper- and psyllid-associated pathogens. In all, 76% of the pepper samples were found to be infected with ‘Candidatus Liberibacter solanacearum’, beet leafhopper-transmitted virescence agent (BLTVA) phytoplasma, a strain of a curtovirus, or a combination of any two or three of these pathogens. Additionally, 77% of the collected leafhoppers and 40% of the psyllids were infected with one or more of these pathogens, in addition to Spiroplasma citri. Specifically, the leafhoppers were infected with BLTVA phytoplasma, S. citri, or a strain of curtovirus. Of particular interest, potato psyllids were not only infected with ‘Ca. L. solanacearum’ but also with phytoplasmas that belong to the groups 16SrVI subgroup A and 16SrI subgroup A. The presence of mixed infections in pepper plants and the insect vectors highlights the need for growers to effectively control both leafhoppers and potato psyllids from solanaceous crops in this region of Mexico in order to prevent the spread of these bacterial and viral pathogens.

Spiroplasma citri. [Descriptions of Fungi and Bacteria].

1991 ◽  
Author(s):  
J. F. Bradbury
Keyword(s):  
Geographical Distribution ◽  
Prunus Avium ◽  
Poncirus Trifoliata ◽  
Spiroplasma Citri ◽  
Beet Leafhopper ◽  
Circulifer Tenellus ◽  
Rough Lemon ◽  
Sweet Lime ◽  
Natural Hosts ◽  
Sisymbrium Irio

Abstract A description is provided for Spiroplasma citri. Information is included on the disease caused by the organism, its transmission, geographical distribution, and hosts. HOSTS: There are many natural hosts in the Rutaceae, including Citrus aurantifolia (sweet lime), C. aurantium (sour orange), C. clementina (clementine), C. jambhiri (rough lemon), C. limon (lemon), C. madurensis (syn. C. mitts, calamondin), C. maxima (syn. C. grandis, pummelo), C. paradisi (grapefruit), C. paradisi × C. reticulata (tangelo), C. sinensis (sweet orange), C. sinensis × Poncirus trifoliata (citrange), C. unshiu (satsuma) and Fortunella spp. (kumquats). Non-rutaceous natural hosts include Armoracia rusticana, Barbarea vulgaris, Brassica geniculata, B. kaber, B. nigra, B. oleracea var. botrytis, B. oleracea var. capitata, B. oleracea var. gemmifera, B. rapa, B. tournefortii, Capsella bursa-pastoris, Catharanthus roseus, Digitalis purpurea, Plantago ovata, Prunus avium, P. persica, Pyrus communis, Raphanus sativus, Sedum praealtum, Sisymbrium irio, S. orientale, Tagetes erecta, Viola cornuta, Zinnia elegans (63, 3239; 62, 2777; 59, 3154). A wide variety of plants has been infected artificially by transmission from inoculated leafhoppers. Symptoms are milder under cool conditions. Under warm conditions only Citrus spp. and hybrids survive infection more than a few months (59, 3154). DISEASE: Citrus stubborn or 'little leaf'. Affected trees are slightly to severely stunted and give low yields. Leaves are shorter, broader, upturned at the edges and may be mottled or chlorotic. Internodes of twigs are shorter and multiple axillary buds are produced. Fruits may be small, lopsided or acorn-shaped, caused by formation of very thin smooth rind at the blossom end. In horseradish, infection causes brittle root disease, in which leaves are stunted, chlorotic or necrotic and roots show a darkened ring of phloem (61, 2596). GEOGRAPHICAL DISTRIBUTION: Present in many citrus growing areas. Reports include: Algeria, Egypt, Libya, Madagascar, Morocco, Tunisia, Iran, Iraq, Israel, Lebanon, Pakistan, Saudi Arabia, Syria, Turkey, Corsica, Cyprus, Greece, Sardinia, Sicily, Mexico, United States (AR, CA), Argentina (Tucuman), Brazil (Sao Paulo), Peru, Surinam (IMl Distribution Map 375, ed. 2, 1970; 49, 1876b; 50, 1228a; 51, 1253b, 3247; 56, 5056; 63, 1797; 68, 367). TRANSMISSION: The insect vectors are leafhoppers and the following have been shown to transmit the disease: Neoaliturus tenellus[Circulifer tenellus] (sugar beet leafhopper), N. haematoceps (N. opacipennis), Scaphytopius nitridus and S. delongi. The disease is also spread in budwood, although not very consistently (68, 367).

scholarly journals Characterization of Xanthomonas axonopodis pv. phaseoli isolates

2008 ◽  
Vol 34 (3) ◽  
pp. 228-231 ◽  
Author(s):  
Willian Mário de Carvalho Nunes ◽  
Maria Júlia Corazza ◽  
Silvana Aparecida Crestes Dias de Souza ◽  
Siu Mui Tsai ◽  
Eiko Eurya Kuramae
Keyword(s):  
Xanthomonas Campestris ◽  
Random Amplified Polymorphic Dna ◽  
Hypersensitive Reaction ◽  
Chain Reaction ◽  
Xanthomonas Axonopodis ◽  
Paulo State ◽  
Total Dna ◽  
Polymerase Chain ◽  
Bacterial Plant Pathogen

A simple, quick and easy protocol was standardized for extraction of total DNA of the bacteria Xanthomonas axonopodis pv. phaseoli. The DNA obtained by this method had high quality and the quantity was enough for the Random Amplified Polymorphic DNA (RAPD) reactions with random primers, and Polymerase Chain Reaction (PCR) with primers of the hypersensitivity and pathogenicity gene (hrp). The DNA obtained was free of contamination by proteins or carbohydrates. The ratio 260nm/380nm of the DNA extracted ranged from 1.7 to 1.8. The hrp gene cluster is required by bacterial plant pathogen to produce symptoms on susceptible hosts and hypersensitive reaction on resistant hosts. This gene has been found in different bacteria as well as in Xanthomonas campestris pv. vesicatoria (9). The primers RST21 and RST22 (9) were used to amplify the hrp gene of nine different isolates of Xanthomonas axonopodis pv. phaseoli from Botucatu, São Paulo State, Brazil, and one isolate, "Davis". PCR amplified products were obtained in all isolates pathogenic to beans.

scholarly journals Absence of rDNA amplification in the uninucleolate oocyte of the cockroach Blattella germanica (Oorthoptera: Blattidae).

1976 ◽  
Vol 71 (1) ◽  
pp. 49-58 ◽  
Author(s):  
M D Cave
Keyword(s):  
Ribosomal Rna ◽  
General Pattern ◽  
Blattella Germanica ◽  
Nucleolar Organizer ◽  
Extrachromosomal Dna ◽  
Total Dna ◽  
Cockroach Blattella Germanica ◽  
Rdna Amplification ◽  
Large Nucleolus

Amplification of the genes coding for ribosomal RNA oocurs in the oocytes of a wide variety of organisms. In oocytes of various species of crickets (Orthoptera: Gryllidae) the amplified DNA is contained in a large extrachromosomal DNA body. Multiple nucleoli form about the periphery of the DNA body during the diplotene stage of meiosis I. In contrast to the general pattern of orthopteran oocytes, oocytes of the cockroach Blattella germanica demonstrate a single large nucleolus instead of many nucleoli. In order to determine whether the genes coding for rRNA are amplified in the oocytes of B. germanica, the relative amount of rDNA in oocytes was compared with the rDNA content of spermatocytes and somatic cells. An extrachromosomal DNA body similar to that present in crickets is not present in B. germanica. A satellite DNA band which contains nucleotide sequences complementary to rRNA accounts for approximately 3-5% of the total DNA in somatic and in male and female gametogenic tissues. Female cells contain approximately twice as much rDNA as do male cells. An XX-XO sex-determining mechanism is operative in B. germanica. In situ hybridization with rRNA indicates that the nucleolar organizer is located on one end of the X chromosome and that oocytes do not contain more than twice the amount of rDNA found in spermato cytes. The data indicate that rDNA is not amplified in the uninucleolate oocyte of B germanica.

Characterization of alpha-adrenoceptor gene expression in arterial and venous smooth muscle

1993 ◽  
Vol 265 (5) ◽  
pp. H1501-H1509 ◽  
Author(s):  
P. Ping ◽  
J. E. Faber
Keyword(s):  
Smooth Muscle ◽  
Messenger Rna ◽  
Vena Cava ◽  
Pcr Analysis ◽  
Relative Distribution ◽  
Alpha Adrenoceptor ◽  
Aortic Media ◽  
Unique Restriction ◽  
Polymerase Chain

Six genes coding for three unique alpha 1- (1A, 1B, 1C) and three unique alpha 2- (2A, 2B, 2C) adrenergic receptor (AR) subtypes have been cloned. Ligand binding and contractile studies have demonstrated that both alpha 1- and alpha 2-ARs can exist on vascular smooth muscle (VSM) cells, although less is known about the relative distribution and specific subtypes in different vascular segments. In the present study polymerase chain reaction (PCR) analysis was used to characterize the species of alpha-AR messenger RNA (mRNA) present in freshly isolated rat thoracic aortic media and vena cava and in cultured VSM cells (passage 2) derived from both sources. To prevent possible contamination of VSM mRNA, aortic media was separated from adventitia, and vessels were denuded of endothelial cells. Oligonucleotide primers specific for each of the six adrenergic genes were synthesized and used to probe for the presence of alpha-AR mRNA species after reverse transcription of total cellular RNA to cDNA. PCR-amplified AR transcripts were distinguished by the size of amplified DNA fragments and unique restriction endonuclease cleavage. Expression of alpha 1C- or alpha 2C-mRNA was not detected in vascular tissues or cultured VSM cells, although the alpha 2C-primers detected the expected alpha 2C expression in cerebral cortex. Only alpha 1A-mRNA was detected in aortic adventitia. VSM from aorta expressed alpha 1A-, alpha 1B-, and alpha 2A-mRNA, and this pattern was preserved in cultured aortic VSM. Vena cava also expressed both alpha 1A and alpha 1B; however only alpha 2B-mRNA was detected.(ABSTRACT TRUNCATED AT 250 WORDS)

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