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.

Total Soluble Sugar Quantification from Ethanolic Plant Extracts v1

2021 ◽  
Author(s):  
Lynn Doran ◽  
Amanda P. De Souza
Keyword(s):  
Plant Extracts ◽  
Plant Tissue ◽  
Soluble Sugar ◽  
Soluble Sugars ◽  
Total Soluble Sugars ◽  
Total Soluble Sugar ◽  
Tissue Extracts

Quantification of total soluble sugars (as glucose) in plant tissue extracts via the sulfuric phenol method adapted for 96 well plates.

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.

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).

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.

Estimation of nitric nitrogen and total nitrogen in plant tissue extracts

1923 ◽  
Vol 13 (1) ◽  
pp. 63-68 ◽  
Author(s):  
Patrick Hugh Gallagher
Keyword(s):  
Sulphuric Acid ◽  
Total Nitrogen ◽  
Plant Extracts ◽  
Plant Tissue ◽  
Nitrate Nitrogen ◽  
Reducing Agents ◽  
Amino Groups ◽  
Tissue Extracts ◽  
Colloidal Matter ◽  
Nitric Nitrogen

Several methods for estimating nitrates by reduction to ammonia were examined. The error involved in the use of acid reducing agents in presence of amino groups was emphasised.A method based on the use of Devarda's alloy and magnesia is recommended.In the estimation of nitrates in plant extracts, etc., by this method an equal volume of rectified spirit is first added to the solution to precipitate colloidal matter. The precipitate is filtered off and an aliquot part of the filtrate containing the equivalent in nitrate nitrogen of about 0.1 gm. of KNO3 is distilled with steam for 45 minutes with 1 gm. of Devarda's alloy and ½ gm. of freshly ignited magnesia. A blank determination is essential, in which the same volume of filtrate is distilled for the same length of time with ½ gm. of magnesia.In Kjeldahl estimations in presence of nitrates, where the separate estimation of nitrates is not required, 2 c.c. of 25 per cent. NaOH may be substituted for the magnesia. Relatively this is very much less soda than Devarda recommends. The mixture in this case may be distilled directly for half-an-hour. The nitrates are thus reduced and the ammonia produced is collected before beginning the digestion with sulphuric acid.This work was carried out under the supervision of Mr F. W. Foreman to whom the author's best thanks are due for much valuable criticism and advice.

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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