Application of Elisa to Spiroplasma Detection and Classification

Saillard, Colette; Bové, J. M.

doi:10.1016/b978-0-12-583801-6.50077-9

Cited by 19 publications

(9 citation statements)

References 6 publications

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“…BSR medium (B medium supplementcd with sorbitol and reduced) was derived from B medium of Ern0 and Stipkovits (14) by adding 70g of sorbitol per liter, decreasing the concentration of horse serum from 20 to lo%, and omitting fresh yeast extract and DNA. The strains were identified as S. citn' strains by an enzyme-linked immunosorbent assay in which a polyclonal antibody directed against S. citri R8A2T was used (40).…”

Section: Methodsmentioning

confidence: 99%

Chromosomal Heterogeneity among Various Strains of Spiroplasma citri

Laigret

Carle

et al. 1995

International Journal of Systematic Bacteriology

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The genomes of various Spiroplasma citri strains were digested with several restriction enzymes, and the fragments were analyzed by pulsed-field gel electrophoresis. Polymorphism of the restriction patterns was observed. The genome sizes of the strains obtained when we added the restriction fragment sizes ranged from 1,650 to 1,910 kbp. Physical and genetic maps of 12 strains were constructed by using several DNA probes as genetic markers. The relative positions of mapped loci were conserved in most of the strains; the main differences were differences in the order and number of restriction sites and differences in the sizes of certain fragments. The distribution of viral sequences, which occur at multiple sites in the S. c h i genome and are homologous to the sequences of S. citri virus SpVl strains RSA2B and S102, was also studied.Since the first culture of Spiroplasma citri (38), the helical mollicute that causes stubborn disease of citrus, several hundred isolates of this agent have been obtained. These organisms have been isolated from infected citrus trees and herbaceous hosts and from insect vectors of the spiroplasmas, such as the leafhopper Circulifer haematoceps (15). The major characteristics of S. citri include helical morphology, motility, small genome size, and low genome G + C content (26 mol%).Phylogenetic studies have shown that spiroplasmas, like all other organisms in the class Mollicutes, arose from the phylum of gram-positive bacteria by successive elimination of genomic DNA (46,53,54).Physical and genomic maps of several mollicutes, including S. citri R8A2' (T = type strain), have been published previously (2, 12, 21, 22, 29, 31, 43, 47-50, 55, 56). Polymorphism has been observed among strains of the same species with several organisms, including Mycoplasma mycoides, Mycoplasma hominis, Mycoplasma hyopneumoniae, and Ureaplasma urealyticum (16,22,31,36), suggesting that these organisms belong to a group of prokaryotes that exhibits a high evolution rate, as suggested previously by 16s rRNA sequence data (54).Several previous studies revealed that polymorphism could be found in several S. citri strains isolated from various hosts in different geographical areas; these studies were studies in which the researchers used serology, protein electrophoretic profiles, restriction fragment length polymorphism, and spiralin gene nucleotide sequences (4,5,25). A striking feature of the S. citri genome is that it contains multiple copies of sequences homologous to the spiroplasma virus SpVl sequence (34,35,56) and that the number and distribution of these sequences are variable from one strain to another (34).In order to investigate further the chromosomal heterogeneity of various S. citri strains, we analyzed the genomic DNAs of these organisms by performing restriction enzyme digestion followed by pulsed-field gel electrophoresis (PFGE). Physical and genomic maps of 12 strains were constructed, and the relative positions of restriction sites and genetic markers, including virus-like sequences, were co...

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Section: Methodsmentioning

confidence: 99%

Chromosomal Heterogeneity among Various Strains of Spiroplasma citri

Laigret

Carle

et al. 1995

International Journal of Systematic Bacteriology

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“…Techniques for deformation tests, metabolism inhibition tests, and ELISAs have already been published (48,49,67,69). Most metabolism inhibition test antigens were prepared from cultures grown for 24 h in SP-4 broth; most test procedures were performed in this medium.…”

Section: Methodsmentioning

confidence: 99%

“…The ELISA, performed on crude plant extracts by techniques described earlier (48,49), was used for the detection of strain P40T in plants infected by grafting with shoots of naturally infected periwinkles.…”

Section: Hyman Et Al (28)mentioning

confidence: 99%

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Spiroplasma phoeniceum sp. nov., a New Plant-Pathogenic Species from Syria

Saillard

Vignault

Bové

et al. 1987

International Journal of Systematic Bacteriology

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Sixteen spiroplasma isolates, recovered over a 2-year period from symptomatic periwinkle plants (Catharanthus roseus) collected in eight different locations in Syria, were compared with other established Spiroplasma species or serogroups. Serological analysis of selected representatives of the new isolates revealed sharing of some antigenic components with several spiroplasmas currently classified within subgroups of group I of the genus. Strain P40T was selected as the type strain and examined, meeting the criteria proposed by the International Committee on Systematic Bacteriology Subcommittee on the Taxonomy of Mollicufes. The organism was shown to belong to the class Mollicufes by its morphology, ultrastructure of its limiting membrane, colony characteristics, and filtration patterns. The helicity and motility of the cells indicated its placement within the family Spiroplasmafaceae. Although some serological cross-reactions could be observed with representatives of group I subgroups, strain P40T could be readily distinguished from other plant or insect pathogenic spiroplasmas in subgroup 1-1 (Spiroplasma cif7-2'9, subgroup 1-2 ( S . melliferum), or subgroup 1-3 (S. kunkeliq and from spiroplasmas assigned to subgroups 1-4 through 1-7 and groups I1 through XI. Cholesterol was required for growth. Glucose was fermented, and arginine was hydrolyzed. The base composition (guanine plus cytosine) of the deoxyribonucleic acid of strain P40T was found to be 26 mol%. Deoxyribonucleic acid-deoxyribonucleic acid hybridization comparisons between strain P40T and other subgroup I representatives revealed approximately 60% relatedness to S. cifri and S . kunkelii and 50% relatedness to S. mellverum. Experimental transmission of two of the new isolates (P40T and P354) occurred through inoculation of spiroplasma broth cultures into leafhoppers (Macrosfeles fascifrons), multiplication of the organism in the insects, and subsequent transmission of the organism by insect feeding on aster or periwinkle plants. The organism was also successfully recovered from broth cultures of symptomatic tissues of experimentally infected periwinkle plants, thus fulfilling Koch's postulates. We propose that such strains be named Spiroplasma phoeniceum. Strain P40T has been deposited 'in the American Type Culture Collection (= ATCC 43115T) Although many Spiroplasma species have been isolated during the last 10 years from insects and plants, only twoSpiroplasma citri (47), the etiological agent of citrus stubborn disease (38), and S. kunkelii (60), the spiroplasma that causes corn stunt disease (11, 68) -ar e plant pathogenic. Citrus stubborn disease is widespread in the Mediterranean region as a result of the natural dissemination of S. citri by leafhoppers (2, 24). In surveys designed to demonstrate natural transmission of this disease in Syria, 2-month-old, healthy periwinkles (Catharanthus roseus) were exposed during summer months in the vicinity of orchards with high rates of stubborn disease infection. In October, many periwinkles showed sym...

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“…Between 1 and 20 individuals of each species were triaturated in O,2 ml of grinding buffer and filled up to 0,6 ml. The ELISA was conducted by using polyclonal antibodies against SPA strain "Israel" as described elsewhere (SAILLARD and BOVE 1983;.…”

Section: Introductionmentioning

confidence: 99%

Detection of insect vectors of the citrus stubborn disease pathogen, Spiroplasma citri Saglio et al., in the citrus growing area of South Turkey¹

Kersting¹,

ŜEngonca²

1992

J Applied Entomology

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Citrus stubborn disease (CSD), caused by Spiroplasma cztn Saglio et al., is one of the most important vector transmittable citrus diseases in the East Mediterranean Area and the Near East.In the present study 39 leafhopper species were determined, and 31 analyzed for S. cztri by ELISA. In 19 leafhopper species the CSD pathogen was detected. The infection rate of the leafhoppers strongly oscillate within one year, but only slight differences were obscrved between the years. The ELISA results were confirmed by culture. Three new leafhopper hosts of S. citri, namely Balclutha hebe (Kirkaldy), Cicadulina bipunctella (Matsamura) and Orosius orientalis (Matsamura) were found during this study. Using transmission experiments with field collected leafhoppers it was shown that only Circulifer opacipennis (Lethierry) is able to transmit S. cztri to Curharanthus ro~eus L. However, two different origins of C. opacipennis occured in the Cukurova region but only one of them transmit the citrus stubborn disease pathogen.

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Application of Elisa to Spiroplasma Detection and Classification

Cited by 19 publications

References 6 publications

Chromosomal Heterogeneity among Various Strains of Spiroplasma citri

Chromosomal Heterogeneity among Various Strains of Spiroplasma citri

Spiroplasma phoeniceum sp. nov., a New Plant-Pathogenic Species from Syria

Detection of insect vectors of the citrus stubborn disease pathogen, Spiroplasma citri Saglio et al., in the citrus growing area of South Turkey¹

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Application of Elisa to Spiroplasma Detection and Classification

Cited by 19 publications

References 6 publications

Chromosomal Heterogeneity among Various Strains of Spiroplasma citri

Chromosomal Heterogeneity among Various Strains of Spiroplasma citri

Spiroplasma phoeniceum sp. nov., a New Plant-Pathogenic Species from Syria

Detection of insect vectors of the citrus stubborn disease pathogen, Spiroplasma citri Saglio et al., in the citrus growing area of South Turkey1

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Detection of insect vectors of the citrus stubborn disease pathogen, Spiroplasma citri Saglio et al., in the citrus growing area of South Turkey¹