Free and integrated plasmid DNA in spiroplasmas

Nur, Israel; Glaser, Gad; Razin, Shmuel

doi:10.1007/bf01568370

Cited by 19 publications

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“…DNA hybridization studies have identified certain S. citri plasmid sequences that are integrated into the chromosomal DNA of S. citri strains and other spiroplasma species unrelated to S. citri (3,25,26,29,30 Extrachromosomal DNA has also been identified in and cloned from the maize bushy stunt MLO (7), which causes stunting, foliar distortion, and chlorosis in maize (28). The maize bushy stunt MLO and AY-MLO do not appear to be related on the basis of plant symptoms, host range, and insect vector relationships.…”

Section: Resultsmentioning

confidence: 99%

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Identification and characterization of plasmids from the western aster yellows mycoplasmalike organism

Kuske

Kirkpatrick

1990

J Bacteriol

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Supercoiled double-stranded DNA molecules (plasmids) were isolated from plants infected with three laboratory strains of western aster yeiows mycoplasma-like organism (AY-MLO) by using cesium chlorideethidium bromide density gradients. Southern blot analysis, using plasmids from the severe strain of AY-MLO (SAY-MLO) Plant pathogenic mycoplasma-like organisms (MLOs) cause disease in a large number of higher plants (5,23). MLOs inhabit the phloem sieve elements of plants and are transmitted between plants by phloem-feeding insects (37). MLOs multiply in both plant and insect hosts (27). MLOs resemble culturable members of the genus Mycoplasma (class Mollicutes) in morphology and antibiotic sensitivity (22), but because they have not yet been successfully cultured in vitro they have remained largely unclassified. Very little is known about their genetic composition or the mechanisms by which they cause disease.The presence of extrachromosomal DNA in the Mollicutes is well established (10,21,34). Plasmid DNA from Spiroplasma (2,3,24,25,29,[33][34][35] and Mycoplasma (4, 40) species has been described. In addition, double-stranded (ds) and single-stranded (ss) DNA associated with spiroplasma, mycoplasma, and acholeplasma viruses have been described (8,21,34). The mollicute plasmids characterized to date are cryptic, and transfer of plasmid DNA between Mollicutes has not yet been demonstrated (34).Plasmid-borne genes are important in determining pathogenicity and virulence in several plant-pathogenic bacteria (6, 31). The possibility that plant-pathogenic MLOs possess plasmid DNA with the potential to affect MLO pathogenicity prompted the present investigation. We describe here the isolation and characterization of plasmids present in laboratory and field isolates of the western aster yellows MLO (AY-MLO). Isolation of DNA from plants and leafhoppers. Symptomatic shoot apices, leaf petioles, and midribs were cut into small pieces and ground in cold mortars in PS buffer (7) without fructose. Ground tissues were filtered through Miracloth (Behring Diagnostics), and the filtrate was centrifuged at 17,500 x g for 30 min at 4°C. The pellet was suspended in Tris-EDTA buffer (15 mM Tris, 3 mM EDTA, pH 8) containing 1% sodium dodecyl sulfate (SDS), incubated on ice for 10 min, and centrifuged at 9,260 x g for 5 min at 4°C. The clarified supernatant was extracted with an equal volume of neutralized phenol followed by chloroform-isoamyl alcohol (24:1, vol/vol), and the nucleic acids were ethanol precipitated (20). Chilled, live, or frozen leafhoppers were chopped in PS buffer and macerated in a tissue homogenizer. The extract was centrifuged, and DNA was isolated as described above. MATERIALS AND METHODSDNA isolation from spiroplasmas. Spiroplasma citri (Maroc isolate) and Spiroplasma kunkelii (California isolate 245) were cultured in a modified MlA medium (17). Cells were harvested by centrifugation, washed in cold phosphatebuffered saline, and centrifuged again. The pellet was sus-1628 on May 11, 2018 by guest

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Section: Resultsmentioning

confidence: 99%

“…Plasmid DNA from Spiroplasma (2,3,24,25,29,[33][34][35] and Mycoplasma (4, 40) species has been described. In addition, double-stranded (ds) and single-stranded (ss) DNA associated with spiroplasma, mycoplasma, and acholeplasma viruses have been described (8,21,34).…”

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confidence: 99%

Identification and characterization of plasmids from the western aster yellows mycoplasmalike organism

Kuske

Kirkpatrick

1990

J Bacteriol

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“…Unlike the other members of the Mycoplasmatales, spiroplasmas have a genome size of 1 x 10^ d. In addition, spiroplasmas have an abundance of plasmlds and episomes, unlike the other molllcutes (Nur et al, 1986;Ranhand et al, 1987;Razin, 1985). Like Mycoplasma spp.…”

Section: The Genus Hvcoolasmamentioning

confidence: 99%

Physiology of the wall-less Anaeroplasmataceae (Class Mollicutes) and related walled bacteria

Petzel

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Petzel, James P., "Physiology of the wall-less Anaeroplasmataceae (Class Mollicutes) and related walled bacteria " (1989). Retrospective Theses and Dissertations. 9077. https://lib.dr.iastate.edu/rtd/9077 INFORMATION TO USERSThe most advanced technology has been used to photo graph and reproduce this manuscript from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer.The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction.In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion.Oversize materials (e.g., maps, drawings, charts) are re produced by sectioning the original, beginning at the upper left-hand corner and continuing from left to right in equal sections with small overlaps. Each original is also photographed in one exposure and is included in reduced form at the back of the book. These are also available as one exposure on a standard 35mm slide or as a IT x 23" black and white photographic print for an additional charge.

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Spiroplasma

Williamson¹,

Gasparich²,

Regassa³

et al. 2015

Bergey's Manual of Systematics of Archaea and Bacteria

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Spi.ro.plas'ma. Gr. n. speira (L. transliteration spira ) a coil, spiral; Gr. neut. n. plasma something formed or molded, a form; N.L. neut. n. Spiroplasma spiral form. Tenericutes / Mollicutes / Entomoplasmatales / Spiroplasmataceae / Spiroplasma Cells are pleomorphic, varying in size and shape from helical and branched nonhelical filaments to spherical or ovoid. The helical forms, usually 100–200 nm in diameter and 3–5 µm in length, generally occur during the exponential phase of growth and in some species persist during stationary phase. The cells of some species are short (1–2 µm). In certain cases, helical cells may be very tightly coiled, or the coils may show continuous variation in amplitude. Spherical cells ~300 nm in diameter and nonhelical filaments are frequently seen in the stationary phase, where they may not be viable, and in all growth phases in suboptimal growth media, where they may or may not be viable. In some species during certain phases, spherical forms may be the replicating form. Helical filaments are motile, with flexional and twitching movements, and often show an apparent rotatory motility. Fibrils are associated with the membrane, but flagellae, periplasmic fibrils, or other organelles of locomotion are absent . Fimbriae and pili observed on the cell surface of insect‐ and plant‐pathogenic spiroplasmas are believed to be involved in host‐cell attachment and conjugation (Ammar et al., 2004; Özbek et al., 2003), but not in locomotion. Cells divide by binary fission, with doubling times of 0.7–37 h. Facultatively anaerobic. The temperature growth range varies among species, from 5 to 41°C. Colonies on solid media are frequently diffuse , with irregular shapes and borders, a condition that reflects the motility of the cells during active growth (Figure 111). Colony type is strongly dependent on the agar concentration. Colony sizes vary from 0.1 to 4.0 mm in diameter. Colonies formed by nonmotile variants or mutants, or by cultures growing on inadequate media are typically umbonate with diameters of 200 µm or less. Some species, such as Spiroplasma platyhelix , have barely visible helicity along most of their length and display little rotatory or flexing motility. Colonies of motile, fast‐growing spiroplasmas are diffuse, often with satellite colonies developing from foci adjacent to the initial site of colony development. Light turbidity may be produced in liquid cultures. Chemo‐organotrophic. Acid is produced from glucose. Hydrolysis of arginine is variable. Urea, arbutin, and esculin are not hydrolyzed. Sterol requirements are variable. An optimum osmolality, usually in the range of 300–800 mOsm, has been demonstrated for some spiroplasmas. Media containing mycoplasma broth base, serum, and other supplements are required for primary growth, but after adaptation, growth often occurs in less complex media. Defined or semi‐defined media are available for some species. Resistant to 10,000 U/ml penicillin. Insensitive to rifampicin, sensitive to erythromycin and tetracycline. Isolated from the surfaces of flowers and other plant parts, from the guts and hemolymph of various insects and crustaceans, and from tick triturates. Also isolated from vascular plant fluids ( phloem sap ) and insects that feed on the fluids . Specific host associations are common. The type species, Spiroplasma citri , is pathogenic for citrus (e.g., orange and grapefruit), producing “stubborn” disease. Experimental or natural infections also occur in horseradish, periwinkle, radish, broad bean, carrot, and other plant species. Spiroplasma kunkelii is a maize pathogen. Some species are pathogenic for insects. Certain species are pathogenic, under experimental conditions, for a variety of suckling rodents (rats, mice, hamsters and rabbits) and/or chicken embryos. Genome sizes vary from 780 to 2220 kbp (PFGE). DNA G + C content ( mol %): 24–31 ( T m , Bd). Type species : Spiroplasma citri Saglio, L'Hospital, Laflèche, Dupont, Bové, Tully and Freundt 1973, 202 AL .

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Free and integrated plasmid DNA in spiroplasmas

Cited by 19 publications

References 19 publications

Identification and characterization of plasmids from the western aster yellows mycoplasmalike organism

Identification and characterization of plasmids from the western aster yellows mycoplasmalike organism

Physiology of the wall-less Anaeroplasmataceae (Class Mollicutes) and related walled bacteria

Spiroplasma

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