Abstract
A simple method for the detection of DNA and RNA from small tissue samples (<-1 mg) is described. Stabs of fresh leaves from wheat, clover, tobacco, broad bean, grape, tomato, lettuce, or asparagus were taken using glass microcapillaries, combined with RNase inhibitor and subjected to RT-PCR either directly or after a DNase treatment. This method was used to successfully detect the presence of (1) virus in infected plants, (2) transgenes such asneomycin phosphotransferase II in transformed plants and (3) various plant genes including RubiscoL and 1,3-β-D-glucanase. With the described DNase treatment, the technique was shown to effectively detect RNA and, therefore, can be used to localize and study patterns of gene expression, virus distribution, and/or transgene expression. The benefits of the described tissue stab technique are that it is quick, it requires little manipulation, and is applicable to a variety of plants.
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Abbreviations
- AMV RT:
-
avian myeloblastosis virus reverse transcriptase
- npt II:
-
Neomycin phosphotrasferase II
- RT-PCR:
-
reverse transcription PCR
- RubiscoL:
-
large subunit of Rubisco
- WSMV:
-
wheat streak mosaic virus
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Myslik, J.T., Nassuth, A. Rapid detection of viruses, transgenes, and mRNAs in small plant leaf samples. Plant Mol Biol Rep 19, 329–340 (2001). https://doi.org/10.1007/BF02772831
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DOI: https://doi.org/10.1007/BF02772831