Imaging Fluorescently Tagged Phytophthora Effector Proteins Inside Infected Plant Tissue

Author(s):  
Petra C. Boevink ◽  
Paul R. J. Birch ◽  
Stephen C. Whisson
Author(s):  
Russell L. Steere ◽  
Eric F. Erbe

It has been assumed by many involved in freeze-etch or freeze-fracture studies that it would be useless to etch specimens which were cryoprotected by more than 15% glycerol. We presumed that the amount of cryoprotective material exposed at the surface would serve as a contaminating layer and prevent the visualization of fine details. Recent unexpected freeze-etch results indicated that it would be useful to compare complementary replicas in which one-half of the frozen-fractured specimen would be shadowed and replicated immediately after fracturing whereas the complement would be etched at -98°C for 1 to 10 minutes before being shadowed and replicated.Standard complementary replica holders (Steere, 1973) with hinges removed were used for this study. Specimens consisting of unfixed virus-infected plant tissue infiltrated with 0.05 M phosphate buffer or distilled water were used without cryoprotectant. Some were permitted to settle through gradients to the desired concentrations of different cryoprotectants.


2021 ◽  
Author(s):  
Cameron Stephens ◽  
Travis W Gannon ◽  
Marc Cubeta ◽  
Tim L. Sit ◽  
Jim Kerns

Take-all root rot is a disease of ultradwarf bermudagrass putting greens caused by Gaeumannomyces graminis (Gg), Gaeumannomyces sp. (Gx), Gaeumannomyces graminicola (Ggram), Candidacolonium cynodontis (Cc), and Magnaporthiopsis cynodontis (Mc). Many etiological and epidemiological components of this disease remain unknown. Improving pathogen identification and our understanding of the aggressiveness of these pathogens along with growth at different temperatures will advance our knowledge of disease development to optimize management strategies. Take-all root rot pathogens were isolated from symptomatic bermudagrass root and stolon pieces from 16 different golf courses. Isolates of Gg, Gx, Ggram, Cc, and Mc were used to inoculate ‘Champion’ bermudagrass in an in planta aggressiveness assay. Each pathogen was also evaluated at 10, 15, 20, 25, 30, and 35C to determine growth temperature optima. Infected plant tissue was used to develop a real-time PCR high resolution melt assay for pathogen detection. This assay was able to differentiate each pathogen directly from infected plant tissue using a single primer pair. In general, Ggram, Gg, and Gx were the most aggressive while Cc and Mc exhibited moderate aggressiveness. Pathogens were more aggressive when incubated at 30C compared to 20C. While they grew optimally between 24.4 and 27.8C, pathogens exhibited limited growth at 35C and no growth at 10C. These data provide important information on this disease and its causal agents that may improve take-all root rot management.


1982 ◽  
Vol 28 (6) ◽  
pp. 600-604 ◽  
Author(s):  
Mary L. Smidt ◽  
Anne K. Vidaver

The production and activity of syringacin W-1, a particulate bacteriocin made by Pseudomonas syringae PsW-1, was studied in plant tissue. The bacteriocin is rod shaped, approximately 20 nm wide and 75 nm long, and composed of an outer sheath and inner core. Both the producing strain, PsW-1, and a sensitive strain, 16, grew within red kidney bean stems. Strains PsW-1 and 16, or mutants derived from them, were injected into bean stems singly or in mixtures. All singly inoculated strains grew well. However, when the bacteriocin-producing strain was co-inoculated with the sensitive strain, the latter grew poorly, if at all. This was not due to competition for available nutrients, since the sensitive strain grew as well in the presence of a bacteriocin-nonproducing mutant as it did alone. Also, a bacteriocin-resistant mutant grew as well in the presence of the producing strain as it did alone. Bacteriocin activity and particles were recovered from infected plant tissue.


1981 ◽  
Vol 11 (3) ◽  
pp. 321-338
Author(s):  
A. Poison ◽  
K. J. van der Merwe
Keyword(s):  

2013 ◽  
Vol 104 (2) ◽  
pp. 117-136 ◽  
Author(s):  
A. Sharma ◽  
A.N. Khan ◽  
S. Subrahmanyam ◽  
A. Raman ◽  
G.S. Taylor ◽  
...  

AbstractMany hemipteroids are major pests and vectors of microbial pathogens, infecting crops. Saliva of the hemipteroids is critical in enabling them to be voracious feeders on plants, including the economically important ones. A plethora of hemipteroid salivary enzymes is known to inflict stress in plants, either by degrading the plant tissue or by affecting their normal metabolism. Hemipteroids utilize one of the following three strategies of feeding behaviour: salivary sheath feeding, osmotic-pump feeding and cell-rupture feeding. The last strategy also includes several different tactics such as lacerate-and-flush, lacerate-and-sip and macerate-and-flush. Understanding hemipteroid feeding mechanisms is critical, since feeding behaviour directs salivary composition. Saliva of the Heteroptera that are specialized as fruit and seed feeders, includes cell-degrading enzymes, auchenorrhynchan salivary composition also predominantly consists of cell-degrading enzymes such as amylase and protease, whereas that of the Sternorhyncha includes a variety of allelochemical-detoxifying enzymes. Little is known about the salivary composition of the Thysanoptera. Cell-degrading proteins such as amylase, pectinase, cellulase and pectinesterase enable stylet entry into the plant tissue. In contrast, enzymes such as glutathione peroxidase, laccase and trehalase detoxify plant chemicals, enabling the circumvention of plant-defence mechanisms. Salivary enzymes such as M1-zinc metalloprotease and CLIP-domain serine protease as inAcyrthosiphon pisum(Aphididae), and non-enzymatic proteins such as apolipophorin, ficolin-3-like protein and ‘lava-lamp’ protein as inDiuraphis noxia(Aphididae) have the capacity to alter host-plant-defence mechanisms. A majority of the hemipteroids feed on phloem, hence Ca++-binding proteins such as C002 protein, calreticulin-like isoform 1 and calmodulin (critical for preventing sieve-plate occlusion) are increasingly being recognized in hemipteroid–plant interactions. Determination of a staggering variety of proteins shows the complexity of hemipteroid saliva: effector proteins localized in hemipteran saliva suggest a similarity to the physiology of pathogen–plant interactions.


2001 ◽  
Vol 58 (6) ◽  
pp. 247-258 ◽  
Author(s):  
Y.F. Zhao ◽  
W.T. Jones ◽  
P. Sutherland ◽  
D.A. Palmer ◽  
R.E. Mitchell ◽  
...  
Keyword(s):  

2010 ◽  
Vol 76 (21) ◽  
pp. 7061-7067 ◽  
Author(s):  
Jacob M. Scherf ◽  
Annett Milling ◽  
Caitilyn Allen

ABSTRACT Most Ralstonia solanacearum strains are tropical plant pathogens, but race 3, biovar 2 (R3bv2), strains can cause bacterial wilt in temperate zones or tropical highlands where other strains cannot. R3bv2 is a quarantine pathogen in North America and Europe because of its potential to damage the potato industry in cooler climates. However, R3bv2 will not become established if it cannot survive temperate winters. Previous experiments showed that in water at 4°C, R3bv2 does not survive as long as native U.S. strains, but R3bv2 remains viable longer than U.S. strains in potato tubers at 4°C. To further investigate the effects of temperature on this high-concern pathogen, we assessed the ability of R3bv2 and a native U.S. strain to survive typical temperate winter temperature cycles of 2 days at 5°C followed by 2 days at −10°C. We measured pathogen survival in infected tomato and geranium plants, in infected potato tubers, and in sterile water. The population sizes of both strains declined rapidly under these conditions in all three plant hosts and in sterile water, and no culturable R. solanacearum cells were detected after five to seven temperature cycles in plant tissue. The fluctuations played a critical role in loss of bacterial viability, since at a constant temperature of −20°C, both strains could survive in infected geranium tissue for at least 6 months. These results suggest that even when sheltered in infected plant tissue, R3bv2 is unlikely to survive the temperature fluctuations typical of a northern temperate winter.


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