Abstract

Main conclusionThree types of nematode-feeding sites (NFSs) caused by M. graminicola on rice were suggested, and the NFS polarized expansion stops before the full NFS maturation that occurs at adult female stage.Root-knot nematodes, Meloidogyne spp., secrete effectors and recruit host genes to establish their feeding sites giant cells, ensuring their nutrient acquisition. There is still a limited understanding of the mechanism underlying giant cell development. Here, the three-dimensional structures of M. graminicola-caused nematode-feeding sites (NFSs) on rice as well as changes in morphological features and cytoplasm density of the giant cells (GCs) during nematode parasitism were reconstructed and characterized by confocal microscopy and the Fiji software. Characterization of morphological features showed that three types of M. graminicola-caused NFSs, type I–III, were detected during parasitism at the second juvenile (J2), the third juvenile (J3), the fourth juvenile (J4) and adult female stages. Type I is the majority at all stages and type II develops into type I at J3 stage marked by its longitudinal growth. Meanwhile, NFSs underwent polarized expansion, where the lateral and longitudinal expansion ceased at later parasitic J2 stage and the non-feeding J4 stage, respectively. The investigation of giant cell cytoplasm density indicates that it reaches a peak at the midpoint of early parasitic J2 and adult female stages. Our data suggest the formation of three types of NFSs caused by M. graminicola on rice and the NFS polarized expansion stopping before full NFS maturation, which provides unprecedented spatio-temporal characterization of development of giant cells caused by a root-knot nematode.

Highlights

  • Root-knot nematodes (RKNs), Meloidogyne spp. are soilborne pathogens of crops causing tremendous yield losses, and have been determined to be one of top ten plant parasitic nematodes based on their scientific and economic importance (Jones et al 2013)

  • Different from the nematode-feeding sites (NFSs) caused by cyst nematodes that are achieved by cell wall dissolution of neighboring cells (Sobczak and Golinowski 2009), giant cells (GCs) caused by Meloidogyne spp. originate from karyokinesis without cytokinesis evidenced by previous investigations of cell wall stubs, expression of marker genes involved in mitosis, assessment of DNA content in GCs as well as the observation of spindles formed by microtubules in GCs

  • Sedentary parasitic nematodes such as Meloidogyne spp. invade plant roots to induce NFSs by secreting a number of effectors through their stylets leading to the induction of GCs

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Summary

Introduction

Root-knot nematodes (RKNs), Meloidogyne spp. are soilborne pathogens of crops causing tremendous yield losses, and have been determined to be one of top ten plant parasitic nematodes based on their scientific and economic importance (Jones et al 2013). The pre-parasitic second-stage juveniles of M. incognita invade plant roots in the elongation zone by penetrating and destroying epidermal cells. They migrate intercellularly down to the meristem, turning round to enter into the vascular tissue. Nematodes induce root swellings, named galls, characterized by the induction of vascular parenchyma asymmetric cell division (de AlmeidaEngler et al 2015). These nematode-feeding sites (NFSs), composed of asymmetrically shaped giant cells (GCs) embedded in the gall, are likely induced by injection of a cocktail of secreted effectors (Wyss et al 1992; Berg et al 2008). Different from the NFSs caused by cyst nematodes that are achieved by cell wall dissolution of neighboring cells (Sobczak and Golinowski 2009), GCs caused by Meloidogyne spp. originate from karyokinesis without cytokinesis evidenced by previous investigations of cell wall stubs, expression of marker genes involved in mitosis, assessment of DNA content in GCs as well as the observation of spindles formed by microtubules in GCs (de Almeida-Engler et al 1999; Banora et al 2011; de AlmeidaEngler and Gheysen 2013; Escobar et al 2015)

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