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Nondestructive Imaging of Plant-Parasitic Nematode Development and Host Response to Nematode Pathogenesis

May 2014 , Volume 104 , Number  5
Pages  497 - 506

Phuong T. Y. Dinh, Michael Knoblauch, and Axel A. Elling

First and third authors: Department of Plant Pathology, and second author: School of Biological Sciences, Washington State University, Pullman 99164.

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Accepted for publication 21 November 2013.

The secluded lifestyle of endoparasitic plant nematodes hampers progress toward a comprehensive understanding of plant–nematode interactions. A novel technique that enables nondestructive, long-term observations of a wide range of live nematodes in planta is presented here. As proof of principle, Pratylenchus penetrans, Heterodera schachtii, and Meloidogyne chitwoodi were labeled fluorescently with PKH26 and used to infect Arabidopsis thaliana grown in microscopy rhizosphere chambers. Nematode behavior, development, and morphology were observed for the full duration of each parasite's life cycle by confocal microscopy for up to 27 days after inoculation. PKH26 accumulated in intestinal lipid droplets and had no negative effect on nematode infectivity. This technique enabled visualization of Meloidogyne gall formation, nematode oogenesis, and nematode morphological features, such as the metacorpus, vulva, spicules, and cuticle. Additionally, microscopy rhizosphere chambers were used to characterize plant organelle dynamics during M. chitwoodi infection. Peroxisome abundance strongly increased in early giant cells but showed a marked decrease at later stages of feeding site development, which suggests a modulation of plant peroxisomes by root-knot nematodes during the infection process. Taken together, this technique facilitates studies aimed at deciphering plant–nematode interactions at the cellular and subcellular level and enables unprecedented insights into nematode behavior in planta.

© 2014 The American Phytopathological Society