Comparison of extracellular polysaccharide of Xanthomonas campestris from culture and from infected cabbage leaves

1970 ◽  
Vol 48 (3) ◽  
pp. 645-651 ◽  
Author(s):  
John C. Sutton ◽  
Paul H. Williams

The extracellular polysaccharide fraction of Xanthomonas campestris was precipitated with hexadecyltrimethylammonium bromide from aqueous extracts of cabbage leaves infected with the black rot pathogen, and further purified by repeated precipitation with ethanol. The polysaccharide fraction was shown to be similar to extracellular polysaccharide fraction purified from culture fitlrates of X. campestris. Sugars present in samples of acid-hydrolyzed polysaccharide were identified by gas–liquid chromatography. Chromatograms showed that polysaccharide purified from infected cabbage leaves contained glucose, mannose, and glucuronolactone in ratios similar to those of the polysaccharide from culture filtrates. Polysaccharide purified from noninfected cabbage leaves contained only glucose and galactose. The polysaccharide from infected cabbage leaves was serologically related to polysaccharide from culture filtrates. In immunodiffusion tests both the polysaccharide fractions from culture filtrates and that from infected leaves reacted to give two precipitin bands with an antiserum to the polysaccharide from culture filtrates. No precipitin bands formed with polysaccharide from noninfected leaves. Antibodies against the culture polysaccharide were completely removed from the antiserum by the polysaccharides isolated from infected cabbage leaves. A weakly virulent isolate of X. campestris did not produce extracellular polysaccharide in culture filtrates.

1970 ◽  
Vol 48 (2) ◽  
pp. 391-401 ◽  
Author(s):  
John C. Sutton ◽  
Paul H. Williams

Black rot lesion development in cabbage leaves was correlated with accumulation of plugging material in the xylem vessels as Xanthomonas campestris multiplied and spread. Plug formation preceded a partial disorganization of the xylem, and spillage of bacteria from the vessels. The movement of eosin and 32PO43− was blocked within the vessels, and plugs caused water shortage in affected cells. Vein blackening, the first symptom of the disease, occurred after plugs had formed, and was associated with an accumulation of melanins among collapsing cells of the vascular bundle. The population of viable bacteria within the veins declined rapidly as the veins blackened.Advance of the bacteria in the xylem vessels accounted for the relationship between venation and lesion spread. A weakly virulent isolate spread extensively in the xylem vessels if inoculated with a virulent isolate, but not when inoculated alone. Acid mucopolysaccharide plugging material was not formed in vessels infected with the weakly virulent isolate.Water stress in cells of interveinal tissues of inoculated leaves resulted in chlorosis, loss in turgor, and loss of electrolytes. If net water loss from infected leaves was prevented by high atmospheric relative humidity, affected cells remained green and turgid and maintained their electrolyte balance.


1982 ◽  
Vol 65 (6) ◽  
pp. 1388-1394 ◽  
Author(s):  
Roger C Snyder ◽  
William C Brumley ◽  
Charles V Breder ◽  
Thomas Fazio

Abstract The confirmation of 2,4- and 2,6-toluenediamine (TDA) in aqueous extracts from boil-in-bags and retortable pouches is described. The extracts were initially analyzed by a high performance liquid chromatographic procedure and any apparent 2,4- and/or 2,6-TDA were quantitated. The liquid chromatographic effluent corresponding to any apparent 2,4- or 2,6-TDA was collected. TDA was then partitioned into ethyl acetate and reacted with trifluoroacetic anhydride (TFAA). The TDA-TFAA derivative formed was confirmed by gas-liquid chromatography (GLC) using a 1.2 m × 0.32 cm nickel column packed with 6% OV-17 on Superpak-20M. Results obtained from analyzing extracts of several retortable pouches and boil-in-bags showed levels of TDA migration ranging from <0.1 to 2.2 ppb (μg/L). Additional confirmation of the TDA-TFAA derivative from retortable pouches by multiple ion detection GC/mass spectrometry is also described.


1984 ◽  
Vol 30 (9) ◽  
pp. 1157-1162 ◽  
Author(s):  
P. Rupérez ◽  
B. Gomez-Miranda ◽  
J. A. Leal

Penicillium allahabadense grown statically in liquid medium formed an extracellular polysaccharide which was collected by precipitation with ethanol (fraction A). The mycelium mat retained a considerable amount of polysaccharide extractable with water, which did not precipitate with ethanol but precipitated with FeCl3 (fraction B). Both fractions were formed mainly by glucose and malonic acid. Their optical rotation values and infrared spectra indicated β configuration. From fraction A was obtained the polysaccharide in the acid form (fraction C), whose equivalent weight is 425, and the demalonylated polysaccharide (fraction D). The main linkage type among glucose residues in the demalonylated polysaccharide, determined after Smith degradation, was 1 → 6. The main component obtained by methylation analysis was 2,3,4-tri-O-methyl-glucitol.


2001 ◽  
Vol 14 (6) ◽  
pp. 768-774 ◽  
Author(s):  
Adrian A. Vojnov ◽  
Holly Slater ◽  
Michael J. Daniels ◽  
J. Maxwell Dow

The gum gene cluster of Xanthomonas campestris pv. campestris comprises 12 genes whose products are involved in the biosynthesis of the extracellular polysaccharide xanthan. These genes are expressed primarily as an operon from a promoter upstream of the first gene, gumB. Although the regulation of xanthan synthesis in vitro has been well studied, nothing is known of its regulation in planta. A reporter plasmid was constructed in which the promoter region of the gum operon was fused to gusA. In liquid cultures, the expression of the gumgusA reporter was correlated closely with the production of xanthan, although a low basal level of β-glucuronidase activity was seen in the absence of added carbon sources when xanthan production was very low. The expression of the gumgusA fusion also was subject to positive regulation by rpfF, which is responsible for the synthesis of the diffusible signal factor (DSF). The expression of the gumgusA fusion in bacteria recovered from inoculated turnip leaves was maximal at the later phases of growth and was subject to regulation by rpfF. These results provide indirect support for the operation of the DSF regulatory system in bacteria in planta.


Microbiology ◽  
2007 ◽  
Vol 153 (12) ◽  
pp. 4284-4294 ◽  
Author(s):  
Guang-Tao Lu ◽  
Zheng-Jiu Yang ◽  
Fang-Yin Peng ◽  
Yi-Ning Tan ◽  
Yong-Qin Tang ◽  
...  

2005 ◽  
Vol 95 (12) ◽  
pp. 1381-1390 ◽  
Author(s):  
Mahfuzur Rahman ◽  
Zamir K. Punja

The fungus Cylindrocarpon destructans (Zins) Scholten is the cause of root rot (disappearing root rot) in many ginseng production areas in Canada. A total of 80 isolates of C. destructans were recovered from diseased roots in a survey of ginseng gardens in British Columbia from 2002-2004. Among these isolates, 49% were classified as highly virulent (causing lesions on unwounded mature roots) and 51% were weakly virulent (causing lesions only on previously wounded roots). Pectinase and polyphenoloxidase enzymes were produced in vitro by C. destructans isolates when they were grown on pectin and phenol as a substrate, respectively. However, highly virulent isolates produced significantly (P < 0.001) higher enzyme levels compared with weakly virulent isolates. Histopathological studies of ginseng roots inoculated with a highly virulent isolate revealed direct hyphal penetration through the epidermis, followed by intracellular hyphal growth in the cortex. Subsequent cell disintegration and accumulation of phenolic compounds was observed. Radial growth of highly and weakly virulent isolates on potato dextrose agar was highest at 18 and 21°C, respectively and there was no growth at 35°C. Mycelial mass production was significantly (P ≤ 0.01) lower at pH 7.0 compared with pH 5.0. To study the effects of pH (5.0 and 7.0) and wounding on disease development, ginseng roots were grown hydroponically in Hoagland's solution. Lesions were significantly larger (P < 0.001) at pH 5.0 compared with pH 7.0 and wounding enhanced disease by a highly virulent isolate at both pHs. In artificially infested soil, 2-year-old ginseng roots were most susceptible to Cylindrocarpon root rot among all root ages tested (1 to 4 years) when evaluated using a combined scale of disease incidence and severity. Root rot severity was significantly (P < 0.002) enhanced by increasing the inoculum density from 3.45 × 102 CFU/g of soil to 1.86 × 103 CFU/g of soil. Disease severity was higher at 20°C compared with 15 and 25°C and at -0.02 MPa soil moisture compared with -0.005 and -0.001 MPa. A significant interaction between soil moisture and temperature was observed for root rot severity.


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