, Volume 254, Issue 6, pp 2107–2115 | Cite as

Spatiotemporal deep imaging of syncytium induced by the soybean cyst nematode Heterodera glycines

  • Mina OhtsuEmail author
  • Yoshikatsu Sato
  • Daisuke Kurihara
  • Takuya Suzaki
  • Masayoshi Kawaguchi
  • Daisuke Maruyama
  • Tetsuya Higashiyama
Original Article


Parasite infections cause dramatic anatomical and ultrastructural changes in host plants. Cyst nematodes are parasites that invade host roots and induce a specific feeding structure called a syncytium. A syncytium is a large multinucleate cell formed by cell wall dissolution-mediated cell fusion. The soybean cyst nematode (SCN), Heterodera glycines, is a major soybean pathogen. To investigate SCN infection and the syncytium structure, we established an in planta deep imaging system using a clearing solution ClearSee and two-photon excitation microscopy (2PEM). Using this system, we found that several cells were incorporated into the syncytium; the nuclei increased in size and the cell wall openings began to be visible at 2 days after inoculation (DAI). Moreover, at 14 DAI, in the syncytium developed in the cortex, there were thickened concave cell wall pillars that resembled “Parthenon pillars.” In contrast, there were many thick board-like cell walls and rarely Parthenon pillars in the syncytium developed in the stele. We revealed that the syncytia were classified into two types based on the pattern of the cell wall structures, which appeared to be determined by the position of the syncytium inside roots. Our results provide new insights into the developmental process of syncytium induced by cyst nematode and a better understanding of the three-dimensional structure of the syncytium in host roots.


Plant-parasitic nematode Heterodera Syncytium development Two-photon excitation microscopy (2PEM) Deep imaging 



We thank Dr. S. Aiba (National Agriculture and Food Research Organization) and Prof. Dr. K. Kawakita (Nagoya University) for kindly providing cyst of H. glycines and Prof. Dr. M.G.K. Jones (Murdoch University) for kindly providing methods of plant-parasitic nematodes. This work was supported by a grant from the Integrative Graduate Education and Research Program in Green Natural Sciences of Nagoya University, the Japan Society for the Promotion of Science (JSPS) Fellowship (15J04623 for M. O.), the Japan Advanced Plant Science Research Network, the JSPS Grant-in-Aid for challenging Exploratory Research (15K14542 for Y. S., 15K14541 for D. M.), the Japan Science and Technology Agency (ERATO project to T. H.), Toyoaki Scholarship Foundation (for D. M.), the JSPS Grant-in-Aid for Young Scientists (A) (16H06173 for D. M.), and the JSPS KAKENHI (16H06465 for T. H.).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

709_2017_1105_MOESM1_ESM.pdf (3 mb)
ESM 1 (PDF 2.98 MB)
709_2017_1105_MOESM2_ESM.avi (3.6 mb)
Movie S1 Z-stack of optical sections of syncytial nuclei in 14 DAI syncytium (Fig. 1a). (AVI 3648 kb).
709_2017_1105_MOESM3_ESM.avi (6.6 mb)
Movie S2 Z-stack of optical sections of cell walls in 14 DAI syncytium induced in the cortex (Fig. 2b). (AVI 6714 kb)
709_2017_1105_MOESM4_ESM.avi (137.3 mb)
Movie S3 Three-dimensional (3D) projections of serial optical sections of cell wall in 14 DAI syncytium induced in the cortex (Fig. 2d). (AVI 140629 kb) (1.9 mb)
Movie S4 X-Y, X-Z and Y-Z projections of serial optical sections of cell wall in 14 DAI syncytium induced in the cortex (Fig. 2c). (MOV 1922 kb). (20.2 mb)
Movie S5 Z-stack of optical sections of cell walls in 14 DAI syncytium induced in the stele at 14 DAI. (MOV 20660 kb). (1.7 mb)
Movie S6 X-Y, X-Z and Y-Z projections of serial optical sections of cell wall in 14 DAI syncytium induced in the stele at 14 DAI. (Fig. S4a). (MOV 1718 kb)


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Copyright information

© Springer-Verlag Wien 2017

Authors and Affiliations

  1. 1.Graduate School of ScienceNagoya UniversityNagoyaJapan
  2. 2.Institute of Transformative Bio-Molecules (ITbM)Nagoya UniversityNagoyaJapan
  3. 3.JST ERATO Higashiyama Live-Holonics ProjectNagoya UniversityNagoyaJapan
  4. 4.Graduate School of Life and Environmental SciencesUniversity of TsukubaTsukubaJapan
  5. 5.Division of Symbiotic SystemsNational Institute for Basic Biology (NIBB)OkazakiJapan
  6. 6.Kihara Institute for Biological ResearchYokohama City UniversityYokohamaJapan

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