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Plant Cell Reports

, Volume 30, Issue 8, pp 1555–1569 | Cite as

Immunohistochemical analysis of cell wall hydroxyproline-rich glycoproteins in the roots of resistant and susceptible wax gourd cultivars in response to Fusarium oxysporum f. sp. Benincasae infection and fusaric acid treatment

  • Dasen Xie
  • Li Ma
  • Jozef Šamaj
  • Chunxiang XuEmail author
Original Paper

Abstract

Hydroxyproline-rich glycoproteins (HRGPs) play a defensive role in host–pathogen interactions. However, specific roles of individual HRGPs in plant defense against pathogen are poorly understood. Changes in extracellular distribution and abundance of individual cell wall HRGPs were investigated on root sections of two wax gourd (Benincasa hispida Cogn.) cultivars (Fusarium wilt resistant and susceptible, respectively), which were analyzed by immunolabelling with 20 monoclonal antibodies recognizing different epitopes of extensins and arabinogalactan proteins (AGPs) after being inoculated with Fusarium oxysporum f. sp. Benincasae or treated with fusaric acid (FA). These analyses revealed the following: (1) The levels of JIM11 and JIM20 interacting extensins were higher in the resistant cultivar. Either treatment caused a dramatic decrease in signal in both cultivars, but some new signal appeared in the rhizodermis. (2) The AGPs or rhamnogalacturonan containing CCRCM7-epitope were enhanced in the resistant cultivar, but not in the susceptible one by either treatment. (3) Either treatment caused a slight increase in the levels of the AGPs recognized by LM2 and JIM16, but there were no differences between two cultivars. (4) The MAC204 signal nearly disappeared after FA treatment, but this was not the case with pathogen attack. (5) The LM14 signal slightly decreased after both treatments in both cultivars, but a less decrease was observed with the resistant cultivar. These results indicate that the CCRCM7 epitope likely contributed to the resistance of wax gourd to this pathogen, and JIM11 and JIM20 interacting extensins as well as LM2, LM14, MAC204 and JIM16 interacting AGPs were involved in the host–pathogen interaction.

Keywords

HRGPs Monoclonal antibodies Immunolabelling Plant–pathogen interaction 

Abbreviations

AG

Arabinogalactan

AGP

Arabinogalactan protein

BSA

Bovine serum albumin

DAPG

2,4-Diacetylphloroglucinol

DAPI

4′-6-Diamidino-2-phenylindole

EGTA

Ethylene glycol-bis(2-aminoethylether)-N,N,N,N′-tetraacetic acid

FA

Fusaric acid

HRGP

Hydroxyproline-rich glycoprotein

PBS

Phosphate buffered saline

PIPES

Piperazine-N,N′-bis (2-ethanesulfonic acid)

ROS

Reactive oxygen species

Notes

Acknowledgments

This study was supported by the special fund for Agro-industry (nyhyzx07-007), Guangdong Science and Technology Project (2009B020201004), Guangzhou Science and Technology Project (GZCQC0802FG06001) and by grant no. ED0007/01/01 Centre of the Region Haná for Biotechnological and Agricultural Research. Development and distribution of CCRCM7 antibody, distribution of MAC and some JIM antibodies were supported in part by NFS grants DBI-0421683 and RCN-0090281.

Supplementary material

299_2011_1069_MOESM1_ESM.tif (938 kb)
Fig. 10 The negative controls where only the second antibody was used (results from second antibody F6258 were selected as representatives). a-b The control of the non-treated resistant cultivar ‘B184’ (a) and susceptible cultivar ‘B214’ (b); c-d ‘B184’ (c) and ‘B214’ (d) after inoculation with the pathogen (Fusarium oxysporum f. sp. Benincasae); e-f ‘B184’ (e) and ‘B214’ (f) treated with fusaric acid. Bars represent 100 μm (TIFF 937 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.Institute of Vegetable ResearchGuangdong Academy of Agricultural SciencesGuangzhouChina
  2. 2.College of HorticultureSouth China Agricultural UniversityGuangzhouChina
  3. 3.Centre of the Region Haná for Biotechnological and Agricultural ResearchFaculty of Science, Palacký UniversityOlomoucCzech Republic

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