DOI QR코드

DOI QR Code

Microwave Irradiation-assisted RNA Extraction from Woody Tissues for Plant Virus Detection

  • Duong, Thanh Van (Department of Biotechnology, Catholic University of Daegu) ;
  • Shin, Dong-Il (Department of Biotechnology, Catholic University of Daegu) ;
  • Park, Hee-Sung (Department of Biotechnology, Catholic University of Daegu)
  • Received : 2010.07.07
  • Accepted : 2010.08.03
  • Published : 2010.09.01

Abstract

Plant tissue homogenization using a mortar or mechanical equipment has been the preferred method for obtaining high yields of total RNA; this method, however, is both time-consuming and expensive. Additionally, homogenization may generate excessive endogenous RNases, polyphenolics, and other substances that reduce the quality and quantity of RNA. In this study, we describe the microwave irradiation-assisted RNA extraction (MIRE) technique which, without tissue disruption and homogenization, allows for the cost-effective and rapid generation of intact RNA from apple cane shavings and the reliable detection of apple virus by RT-PCR.

Keywords

References

  1. Besancon, B., Bencsik, A., Voutsinos, B., Belin, M. F. and Fevre-Montange, M. 1995. Rapid in situ hybridization using digoxigenin probe and microwave oven. Cell Mol. Biol. 41:975-977.
  2. Campbell, A. I. 1963. The effect of some latent virus infections on the growth and cropping of apples. J. Hort. Sci. 38:15-19.
  3. Ekuni, D., Firth, J. D. and Putnins, E. E. 2006. RNA integrity and in situ RT-PCR in dento-alveolar tissues after microwave accelerated demineralization. Arch. Oral Biol. 51:164-169. https://doi.org/10.1016/j.archoralbio.2005.06.010
  4. Hirata, H., Lu, X., Yamaji, Y., Kagiwada, S., Ugaki, M. and Namba, S. 2003. A single silent substitution in the genome of Apple stem grooving virus causes symptom attenuation. J. Gen. Virol. 84:2579-2583. https://doi.org/10.1099/vir.0.19179-0
  5. James, D. 1999. A simple and reliable protocol for the detection of apple stem grooving virus by RT-PCR and in a multiplex PCR assay. J. Virol. Methods. 83:1-9. https://doi.org/10.1016/S0166-0934(99)00078-6
  6. John, M. E. 1992. An efficient method for isolation of RNA and DNA from plants containing polyphenolics. Nucleic Acids Res. 20:2381. https://doi.org/10.1093/nar/20.9.2381
  7. Lazarowitz, S. G. and Beachy, R. N. 1999. Viral movement proteins as probes for intracellular and intercellular trafficking in plants. Plant Cell. 11:535-548. https://doi.org/10.1105/tpc.11.4.535
  8. McMahon, J. and McQuaid, S. 1996. The use of microwave irradiation as a pretreatment to in situ hybridization for the detection of measles virus and chicken anaemia virus in formalinfixed paraffin-embedded tissue. Histochem. J. 28:157-164. https://doi.org/10.1007/BF02331439
  9. Menzel, W., Jelkmann, W. and Maiss, E. 2002. Detection of four apple viruses by multiplex RT-PCR assays with coamplification of plant mRNA as internal control. J. Virol. Methods. 99:81-92. https://doi.org/10.1016/S0166-0934(01)00381-0
  10. Picard, C., Ponsonnet, C., Paget, E., Nesme, X. and Simonet, P. 1992. Detection and enumeration of bacteria in soil by direct DNA extraction and polymerase chain reaction. Appl. Environ. Microbiol. 58:2717-2722.
  11. Saini, H. S., Shepherd, M. and Henry, R. J. 1999. Microwave extraction of total genomic DNA from barley grains for use in PCR. J. Inst. Brew. 105: 185-190. https://doi.org/10.1002/j.2050-0416.1999.tb00018.x
  12. Shia, S. R., Cotea, R. J., Wub, L., Liua, C., Datara, R., Shia, Y., Liua, D., Lima, H. and Taylora, C. R. 2002. DNA extraction from archival formalin-fixed, paraffin-embedded tissue sections based on the antigen retrieval principle: heating under the influence of pH. J. Hist. Cyto. 50:1005-1011. https://doi.org/10.1177/002215540205000802
  13. Tabanifar, B., Salehi, R., Asgarani, R., Faghihi, M., Heidarpur, M. and Allame, T. S. 2008. An efficient method for DNA extraction from paraffin wax embedded tissues for PCR amplification of human and viral DNA. Iran. J. Pathol. 3:173-178.

Cited by

  1. Using microwaves to prepare gastropods for DNA barcoding vol.14, pp.4, 2014, https://doi.org/10.1111/1755-0998.12231