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Laser photoacoustic detection allows in planta detection of nitric oxide in tobacco following challenge with avirulent and virulent Pseudomonas syringae Pathovars.

Authors
  • Mur, Luis A J
  • Santosa, I Edi
  • Laarhoven, Lucas J J
  • Holton, Nicholas J
  • Harren, Frans J M
  • Smith, Aileen R
Type
Published Article
Journal
PLANT PHYSIOLOGY
Publisher
American Society of Plant Biologists
Publication Date
Jul 01, 2005
Volume
138
Issue
3
Pages
1247–1258
Identifiers
PMID: 16009999
Source
Medline
License
Unknown

Abstract

We demonstrate the use of laser photoacoustic detection (LPAD) as a highly sensitive method to detect in planta nitric oxide ((*)NO) production from tobacco (Nicotiana tabacum). LPAD calibration against (*)NO gas demonstrated a linear relationship over 2 orders of magnitude with a detection threshold of <20 pmol h(-1) (1 part per billion volume [ppbv]). The specificity of the photoacoustic signal for (*)NO when adding gas or the (*)NO donor, sodium nitroprusside, on injection into plant leaves, was demonstrated by its abolition with O(3) ((*)NO + O(3) --> NO(2) + O(2)). The utility of the LPAD method was shown by examination of a nonhost hypersensitive response and a disease induced by Pseudomonas syringae (P. s.) pv phaseolicola and P. s. pv tabaci in tobacco. (*)NO was detected within 40 min of challenge with P. s. pv phaseolicola, some 5 h before the initiation of visible tissue collapse. The wildfire tobacco pathogen P. s. pv tabaci initiated (*)NO generation at 2 h postinfection. The use of (*)NO donors, the scavenger CPTIO ([4-carboxyphenyl]-4,5-dihydro-4,4,5,5-tetramethyl-3-oxide), and the mammalian nitric oxide synthase inhibitor l-NMMA (N(G)-monomethyl-l-arginine) indicated that (*)NO influenced the kinetics of cell death and resistance to both avirulent and virulent bacteria in tobacco. These observations suggest that (*)NO is integral to the elicitation of cell death associated with these two bacterial pathogens in tobacco.

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