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Related Experiment Videos

Detection of double-stranded RNA by serologically specific electron microscopy.

K S Derrick, R C French, C A Clark

    Journal of Virological Methods
    |December 1, 1984
    PubMed
    Summary

    A new electron microscopy staining method simplifies the detection of double-stranded RNA (dsRNA) in plant virus infections. This technique allows for routine dsRNA assays in crude extracts, improving diagnostic accuracy for viral diseases.

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    Plant disease·2019

    Area of Science:

    • Plant Pathology
    • Molecular Virology
    • Microscopy Techniques

    Background:

    • Detecting viral double-stranded RNA (dsRNA) is crucial for diagnosing plant virus infections.
    • Traditional electron microscopy methods for dsRNA detection can be variable and require heavy metal shadowing.

    Purpose of the Study:

    • To develop and describe a novel staining procedure for visualizing dsRNA in infected plant tissues.
    • To establish a reliable and routine method for dsRNA detection using electron microscopy without heavy metal shadowing.

    Main Methods:

    • A staining method based on in situ formation of uranyl phosphate was employed.
    • Serologically specific electron microscopy was used to examine dsRNA in virus-infected tissue extracts.
    • The procedure was applied to sorghum infected with sugarcane mosaic virus and cowpea infected with cowpea mosaic virus.

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    Main Results:

    • The uranyl phosphate staining method allowed consistent examination of dsRNA by electron microscopy without shadowing.
    • Routine dsRNA assays in crude extracts were achieved with reduced variability compared to shadowing procedures.
    • Accumulation of dsRNA was observed in older leaves of sorghum infected with sugarcane mosaic virus.
    • In contrast, dsRNA was not detected in older cowpea leaves infected with cowpea mosaic virus but was present in inoculated and young systemically infected leaves.

    Conclusions:

    • The described staining method offers a robust and simplified approach for dsRNA detection in plant virology.
    • This technique facilitates routine diagnosis of viral infections by enabling consistent dsRNA visualization.
    • Differential dsRNA accumulation patterns were observed between the two virus-host systems studied.