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Photosynthetica

, Volume 46, Issue 3, pp 441–451 | Cite as

Conventional and combinatorial chlorophyll fluorescence imaging of tobamovirus-infected plants

  • M. Pineda
  • J. Soukupová
  • K. Matouš
  • L. Nedbal
  • M. BarónEmail author
Original Papers

Abstract

We compared by chlorophyll (Chl) fluorescence imaging the effects of two strains of the same virus (Italian and Spanish strains of the Pepper mild mottle virus — PMMoV-I and-S, respectively) in the host plant Nicotiana benthamiana. The infection was visualized either using conventional Chl fluorescence parameters or by an advanced statistical approach, yielding a combinatorial set of images that enhances the contrast between control and PMMoV-infected plants in the early infection steps. Among the conventional Chl fluorescence parameters, the non-photochemical quenching parameter NPQ was found to be an effective PMMoV infection reporter in asymptomatic leaves of N. benthamiana, detecting an intermediate infection phase. The combinatorial imaging revealed the infection earlier than any of the standard Chl fluorescence parameters, detecting the PMMoV-S infection as soon as 4 d post-inoculation (dpi), and PMMoV-I infection at 6 dpi; the delay correlates with the lower virulence of the last viral strain.

Additional key words

biotic stress Nicotiana benthamiana non-photochemical quenching Pepper mild mottle virus 

Abbreviations

AS

asymptomatic

CCD

charge-coupled device

Chl

chlorophyll

Chl-F

chlorophyll fluorescence

Chl-FI

chlorophyll fluorescence imaging

dpi

days post-inoculation

F0

minimum chlorophyll fluorescence in the dark adapted state

FM

maximum chlorophyll fluorescence in the dark adapted state

FM

maximum chlorophyll fluorescence in the light adapted state

FM’’

maximum chlorophyll fluorescence after switching off the “actinic light” (protocol 2)

FP

maximum chlorophyll fluorescence reached at the induction curve without saturating flashes

Ft

chlorophyll fluorescence value prior to the saturating pulse in the light-adapted state

FV

variable fluorescence

HI

high irradiance

LDA

Linear Discriminant Analysis

LED

light-emitting diode

LI

low irradiance

PAR

photosynthetically active radiation

NPQ

non-photochemical quenching

NPQt

non-photochemical quenching at referred kinetics times, t

PN

net photosynthetic rate

PMMoV-I and-S

Italian and Spanish strains of the Pepper mild mottle virus

PS

photosystem

R

image resulting from the combinatorial imaging

Re

recovered

S

symptomatic

SFFS

Sequential Forward Floating Search

ΦPS2

photosystem 2 quantum yield

ΦPS2-t

photosystem 2 quantum yield at referred kinetics times, t [s]

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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • M. Pineda
    • 1
  • J. Soukupová
    • 2
    • 3
  • K. Matouš
    • 2
    • 3
  • L. Nedbal
    • 2
    • 3
  • M. Barón
    • 1
    Email author
  1. 1.Department of Biochemistry, Molecular and Cell Biology of PlantsEstación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas (CSIC)GranadaSpain
  2. 2.Institute of Systems Biology and EcologyAcademy of Sciences of the Czech RepublicNové HradyCzech Republic
  3. 3.Institute of Physical BiologyUniversity of South BohemiaNové HradyCzech Republic

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