Journal of Plant Research

, Volume 126, Issue 6, pp 811–821 | Cite as

Chitosan and a fungal elicitor inhibit tracheary element differentiation and promote accumulation of stress lignin-like substance in Zinnia elegans xylogenic culture

  • Chisato Takeuchi
  • Kouji Nagatani
  • Yasushi Sato
Regular Paper


We investigated the effect of elicitors on xylem differentiation and lignification using a Zinnia elegans xylogenic culture system. Water-soluble chitosan and a fungal elicitor derived from Botrytis cinerea were used as elicitors. Elicitor addition at the start of culturing inhibited tracheary element (TE) differentiation in a concentration-dependent manner, and 30 μg mL−1 of chitosan or 16.7 μg mL−1 of the fungal elicitor strikingly inhibited TE differentiation and lignification. Addition of chitosan (at 50 μg mL−1) or the fungal elicitor (at 16.7 μg mL−1) during the culturing period also inhibited TE differentiation without inhibiting cell division, except for immature TEs undergoing secondary wall thickening. Elicitor addition after immature TE appearance also caused the accumulation of an extracellular lignin-like substance. It appears that elicitor addition at the start of culturing inhibits the process by which dedifferentiated cells differentiate into xylem cell precursors. Elicitor addition during culturing also appears to inhibit the transition from xylem cell precursors to immature TEs, and induces xylem cell precursors or xylem parenchyma cells to produce an extracellular stress lignin-like substance.


Botrytis cinerea Chitosan Fungal elicitor Stress lignin Tracheary element differentiation Zinnia elegans 



Microbe-associated molecular pattern


Tracheary element



This work was supported in part by project grants to Y. S from Ehime University, Japan (Rudimentary Research Support) and from the Faculty of Science, Ehime University (Research Support).

Supplementary material

10265_2013_568_MOESM1_ESM.tif (4.2 mb)
Fig. S1 The method developed for measuring lignified areas. A 120-h culture with fungal elicitor addition at 72 h was subjected to the analysis with Image J. a Original RGB image. b G image of split RGB image. c The image with subtracted background. d The image with threshold adjusted to 0-223, TE and the extracellular lignin-like substance separated by a white line, and non-lignified particles removed. e Analysis of particle area. f Total of the area of lignified TEs or extracellular lignin-like substances (TIFF 4318 kb)
10265_2013_568_MOESM2_ESM.doc (28 kb)
Fig. S2 Macro program of Image J (version 1.45s) used for measuring lignified area.//, comment line; *, these parameters were adjusted for this research (DOC 28 kb)


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Copyright information

© The Botanical Society of Japan and Springer Japan 2013

Authors and Affiliations

  • Chisato Takeuchi
    • 1
  • Kouji Nagatani
    • 1
  • Yasushi Sato
    • 2
  1. 1.Department of Biology, Faculty of ScienceEhime UniversityMatsuyamaJapan
  2. 2.Biology and Environmental Science, Graduate School of ScienceEhime UniversityMatsuyamaJapan

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