Microaspiration of Solanum tuberosum root cells at early stages of infection by Globodera pallida

Process of microaspiration and RNA isolation from Globodera pallida infected Solanum tuberosum root cells. a Schematic diagram of the workflow of nematode inoculation, microaspiration of infected cells and RNA isolation. b S. tuberosum on a glass plate (7.5 cm L × 5.0 cm W) ready for microaspiration, the left arrow shows the glass plate lined with agarose; right arrow shows the needle and the root; the inset is of a close view of the root perforated by the capillary needle. c PKH26 stained G. pallida located under a fluorescent microscope with capillary needle inside the root cell; the bright field illumination was turned on to a low setting to allow the capillary needle to be seen; the inset shows the image of G. pallida through a rhodamine filter without bright field illumination; the left arrow indicates the head region of the nematode; right arrow shows the root perforated by the capillary needle. d The head region of G. pallida (lower arrow) inside the root cell with the aspirator needle (upper arrow). The needle contains extraction buffer to prevent RNA degradation. Bar = 20 μ