scholarly journals Characterization of a Severe Strain of Cucumber Mosaic Cucumovirus Seedborne in Cowpea

Plant Disease ◽  
1998 ◽  
Vol 82 (4) ◽  
pp. 419-422 ◽  
Author(s):  
A. G. Gillaspie ◽  
M. R. Hajimorad ◽  
S. A. Ghabrial

A new seedborne strain of cucumber mosaic cucumovirus (CMV) that induces severe symptoms on many cowpea genotypes was detected in Georgia in 1994. This strain, designated CMV-Csb, is asymptomatic on tobacco, but it produces more severe cowpea stunt symptoms when present in combination with blackeye cowpea mosaic potyvirus than do the more prevalent CMV isolates. The new strain is seedborne in cowpea (1.5 to 37%), has no associated satellite RNA, and is classified as a member of subgroup I of CMV strains based on nucleic acid hybridization assays.

1991 ◽  
Vol 37 (9) ◽  
pp. 1506-1512 ◽  
Author(s):  
E F Templeton ◽  
H E Wong ◽  
R A Evangelista ◽  
T Granger ◽  
A Pollak

Abstract A new nonisotopic detection method based on time-resolved fluorescence for nucleic acid hybridization assays with alkaline phosphatase labels has been developed: enzyme-amplified lanthanide luminescence (EALL). EALL combines the amplification of an enzyme label with the sensitivity and background elimination of time-resolved fluorescence detection of lanthanide ion luminescence. The detection system for alkaline phosphatase makes use of a phosphorylated salicylic acid derivative that, upon dephosphorylation, gives a product capable of forming a luminescent terbium chelate. We demonstrate DNA hybridization assays by using two substrates, one for membrane and one for solution-based formats. Using the substrate that produces a more adhesive product allows performance of dot-blot and Southern blot assays on nylon membranes; results can be recorded with a time-resolved photographic camera system, or with an ultraviolet transilluminator-based system. Less than 4 pg of target sequence can be detected in a dot-blot assay after incubation with substrate for 2-4 h. DNA microwell-plate hybridization assays with the more soluble substrate/product pair can be quantified with time-resolved fluorescence plate readers, giving a similar detection sensitivity. EALL is thus a practical time-resolved fluorescence-based alternative to other detection systems for DNA hybridization assays.


1993 ◽  
Vol 56 (10) ◽  
pp. 834-840 ◽  
Author(s):  
JENNIFER L. JOHNSON ◽  
CHARLES P. LATTUADA

A comparison was made of various biochemical and nucleic acid hybridization assays used for the confirmation of Listeria monocytogenes. Identification and confirmation tests included traditional carbohydrate utilization tests, several types of hemolysis assays, an immunocompromised mouse-pathogenicity test, the API® LISTERIA identification system, the Biolog GP MicroPlate™, a prototype Biolog Listeria MicroPlate™, and two nucleic acid hybridization assays (a slot-blot assay and the commercially available AccuProbe™ L. monocytogenes culture confirmation test). A panel of 44 Listeria isolates, including representatives from all six existing Listeria species, was tested. A considerable number of the L. monocytogenes isolates tested gave ambiguous reactions on the CAMP-hemolysis assay. In many instances, the Biolog GP MicroPlate™ was unable to differentiate L. monocytogenes from other Listeria species. Reactions on the prototype Listeria MicroPlate™ mirrored those of traditional biochemical tests, but additional analyses were necessary to differentiate between L. monocytogenes and Listeria innocua. The API® LISTERIA identification system could readily differentiate between the latter two Listeria species without the need for additional tests. Atypical L. monocytogenes isolates (nonhemolytic and nonpathogenic or hemolytic and nonpathogenic) were correctly identified by the API® LISTERIA system. A perfect correlation was observed between results from the slot-blot format hybridization assay and the AccuProbe™ test. The probes used in the two assays hybridized with all L. monocytogenes isolates, regardless of hemolytic reaction or mouse pathogenicity. The commercially available assays examined in this study were all easy to use and capable of providing results within 24 hours.


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