Spiroplasmas: serological grouping of strains associated with plants and insects

1979 ◽  
Vol 25 (8) ◽  
pp. 861-866 ◽  
Author(s):  
R. E. Davis ◽  
I.-M. Lee ◽  
L. K. Basciano
Keyword(s):  
Growth Inhibition ◽  
Liriodendron Tulipifera ◽  
Spiroplasma Citri ◽  
Cross Reactions ◽  
Corn Stunt ◽  
Corn Stunt Spiroplasma ◽  
Deformation Tests ◽  
Tulip Tree

Spiroplasma strains from plant and arthropod hosts, and from surfaces of flowers, were classified into three serological groups (designated I, II, and III) based on results from growth-inhibition tests. No significant cross reactions were observed among groups. The groupings were confirmed by ring-interface precipitin and microprecipitin tests, using membrane preparations as test antigens, and by organism-deformation tests. Serogroup I contained three subgroups: subgroup A (Spiroplasma citri strains Maroc R8A2 and C 189), subgroup B (strain AS 576 and closely related strains from honeybee or flowers), and subgroup C (corn stunt spiroplasma strains). Serogroup II contained strains 23-6 and 27-31 isolated from flowers of the tulip tree (Liriodendron tulipifera L.) growing in Maryland. Serogroup III contained strains SR 3 and SR 9 isolated from flowers of the tulip tree growing in Connecticut. The subgroups of serogroup I were based on organism deformation, microprecipitin, and ring-interface precipitin tests. The data are consistent with the hypothesis that the three serogroups represent no less than three distinct spiroplasma species.

An in vitro comparison of some biochemical and biological properties of California and Morocco isolates of Spiroplasma citri

1979 ◽  
Vol 25 (10) ◽  
pp. 1125-1132 ◽  
Author(s):  
E. C. K. Igwegbe ◽  
Clauzell Stevens ◽  
John J. Hollis Jr.
Keyword(s):  
Biological Properties ◽  
Tetrazolium Chloride ◽  
Potassium Tellurite ◽  
Spiroplasma Citri ◽  
Antimicrobial Substances ◽  
Solid Media ◽  
Corn Stunt ◽  
Optimum Ph ◽  
Corn Stunt Spiroplasma

California (CI) and Morocco (MI) isolates of Spiroplasma citri incubated at 4, 24, or 32 °C yielded typical mycoplasma colonies on agar at each passage. MI but not CI incubated at 37 °C yielded a few colonies at first passage only, but both isolates did not survive 42 °C for 3 days. In liquid and solid media, best growth of both isolates occurred between pH 7.0 and pH 8.5 and the optimum pH for growth in liquid medium was 7.5 (CI) and 8.5 (MI). The sensitivity of CI or MI to 21 antimicrobial substances was identical in most cases. Corn stunt spiroplasma (CSS) and CI had similar antimicrobial substance spectrum, although CSS was much more sensitive to any given substance than CI. CI fermented the following with acid production: cellobiose, dextrose, galactose, trehalose, fructose, mannitol. maltose, and sucrose. MI fermented all these in addition to arabinose, mannose. sorbitol, and raffinnose. Arginine but not urea was hydrolyzed by both isolates. CI or MI failed to reduce potassium tellurite, methylene blue, or tetrazolium chloride.

Presence of spiroplasma-inhibitory substances in plant tissue extracts

1980 ◽  
Vol 26 (7) ◽  
pp. 807-811 ◽  
Author(s):  
C. H. Liao ◽  
T. A. Chen
Keyword(s):  
Plant Extracts ◽  
Plant Tissue ◽  
Cytoplasmic Membrane ◽  
Cynodon Dactylon ◽  
Culture Media ◽  
Cell Replication ◽  
Spiroplasma Citri ◽  
Tissue Extracts ◽  
Corn Stunt ◽  
Corn Stunt Spiroplasma

Plant tissue extracts prepared from corn, periwinkle, celery, and lettuce contain substance(s) which are inhibitory for spiroplasma growth in culture media. The inhibition titer, measured as the highest dilution of plant extracts which block spiroplasma growth, is inversely proportional to the spiroplasma concentration used for testing. At a concentration of 104 cells/mL, the growth of four spiroplasma isolates investigated (corn stunt spiroplasma, Spiroplasma citri, honey bee spiroplasma, and Bermuda grass (Cynodon dactylon) spiroplasma) was suppressed by the addition of corn and periwinkle extracts up to dilutions of 1/160 and 1/1280, respectively. Spiroplasmas remained viable in extract-supplemented media for at least 5 days and continued to multiply shortly after the plant extract was removed. These results suggest that (1) the inhibitory action of plant extracts is spiroplasmastatic, and (2) inhibitory factors may temporarily bind to cytoplasmic membrane and subsequently suppress the process of cell replication. The anti-mycoplasma activity was greatly reduced when the extract was placed in boiling water for 10 min or at 37 °C for 5 days. No loss of activity was detected when the extract was kept at −20 or 4 °C for 5 days.

Electron Microscopy of A New Plant-Pathogenic Spiroplasma Isolated From Opuntia

1976 ◽  
Vol 34 ◽  
pp. 56-57 ◽  
Author(s):  
F. KONDO ◽  
A. H. MCINTOSH ◽  
S. B. PADHI ◽  
K. MARAMOROSCH
Keyword(s):  
Electron Microscopy ◽  
Thin Section ◽  
Liquid Media ◽  
Serological Tests ◽  
Spiroplasma Citri ◽  
Corn Stunt ◽  
Section Electron ◽  
Agar Media ◽  
Darkfield Microscopy ◽  
Corn Stunt Spiroplasma

Thin section electron micrographs of phloem from the ornamental cactus Opuntia tuna monstrosa revealed the presence of wall-less prokaryotic cells earlier described as mycoplasma-like bodies (1). Isolation and growth of the microorganism in liquid media yielded cultures of a spiroplasma, as ascertained by darkfield microscopy. Negatively stained spiroplasmas (Fig. 1) were reminiscent of Spiroplasma citri (2) and of corn stunt spiroplasma (3). The spirals observed in negatively stained preparations seemed attached to blebs. PAGE analysis showed distinct differences between the three species of microorganisms. Serological tests confirm ed that the spiroplasma isolated from the ornamental cactus differed from S. citri and from corn stunt spiroplasma. On agar media, “fried-egg” colonies were formed (Fig. 2).

Spiroplasma associated with flowers of the tulip tree (Liriodendron tulipifera L.)

1978 ◽  
Vol 24 (8) ◽  
pp. 954-959 ◽  
Author(s):  
Robert E. Davis
Keyword(s):  
Honey Bee ◽  
Solid Surface ◽  
Liriodendron Tulipifera ◽  
Glass Slide ◽  
Agar Culture ◽  
Solid Agar ◽  
Viscous Media ◽  
Freely Suspended ◽  
Deformation Tests ◽  
Tulip Tree

Spiroplasmas were isolated and cultivated from nonsurface-sterilized petals and bracts excised from flowers of Liriodendron tulipifera L. (tulip tree) in Anne Arundel, Charles, and Prince George's Counties in Maryland, and East Lyme County, Connecticut. All isolates grew at 30 and at 37 °C. Morphology of colonies on solid agar (1%) medium containing serum differed among isolates: some isolates formed highly diffuse, barely visible colonies; others formed distinct colonies with granular centers surrounded by minute surface or submerged 'satellite' colonies. Cellular morphology and motility of organisms in broth and in agar culture were typical of the spiroplasmas pathogenic in plants and insects. In viscous media containing methylcellulose, spiroplasmas freely suspended in the medium swam, and organisms in contact with glass slide or coverslip "crawled" across the solid surface. Growth inhibition, metabolic inhibition, and organism deformation tests failed to reveal a serologic relationship between spiroplasma strain 23-6 from tulip tree flowers and spiroplasma strain AS 576 from honey bee.

Serum-free media for cultivation of spiroplasmas

1989 ◽  
Vol 35 (12) ◽  
pp. 1092-1099 ◽  
Author(s):  
I.-M. Lee ◽  
R. E. Davis
Keyword(s):  
Good Growth ◽  
Supporting Capacity ◽  
Spiroplasma Citri ◽  
Serum Free ◽  
Spiroplasma Kunkelii ◽  
Corn Stunt ◽  
Agar Media ◽  
Corn Stunt Spiroplasma ◽  
Free Media ◽  
Mycoplasma Spp

Serum-free media that contained bovine serum albumin and lipids (fatty acids, cholesterol, phosphatidic acid, and phospholipids) to replace serum were formulated for the culture of some fastidious strains of corn stunt spiroplasma, Spiroplasma kunkelii. These media also supported good growth of other spiroplasmas, including epiphytic, entomopathogenic, and phytopathogenic strains. The growth rates attained in these serum-free media were comparable to those in media which contained serum. The serum-free broth and agar media were of major importance for culture of several fastidious strains of corn stunt spiroplasma because the serum-containing media supported little, if any, of their growth. Colonies of these fastidious strains were readily formed on serum-free agar media in aerobic, candle jar, and anaerobic environments, but were not readily formed on the serum-containing media. The serum-free media were less subject to deterioration during storage or incubation, and the growth-supporting capacity of serum-free media diminished more slowly than that of serum-containing media.Key words: Spiroplasma kunkelii, Spiroplasma citri, Mollicutes, Mycoplasma spp. culture, entomopathogens.

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