864
Over the past decades the study of several parameters of the chlorophyll fluorescence emission has
become a rapid, sensitive and non-destructive laboratory method to study numerous aspects of the
photosynthetic function [4,5]. As the possibility of airborne detection of laser-induced fluorescence from trees,
bushes, and grasses has been demonstrated [ 6 ], the application of one of these fluorescence techniques in remote
sensing is under discussion [7]. In addition to chlorophyll fluorescence, recent developments take into account
the additional emission bands at 440 nm and 520 nm, which appear upon excitation with UV lasers [ 8 ]. Within
the frame of the EUREKA project LASFLEUR (EU 380), several institutes and companies are investigating the
use of different fluorescence parameters for the remote detection of vegetation stress.
However the intensity of the fluorescence signal depends on several factors including ambient light,
chlorophyll content, fluorescence re-absorption and fluorescence quantum yield. The later parameter is essential
in order to define plant status, as it can be related to photochemistry and carbon assimilation [9]. This paper
describes a new method to determine the chlorophyll fluorescence quantum yield, based on the measurement of
the lifetime. In addition recent progress in our knowledge of blue fluorescence are presented.
3. CHLOROPHYLL FLUORESCENCE LIFETIME IN-VIVO
The absorption of a light by a chlorophyll molecule leads to an excited state. The excess of energy of this state
can be dissipated by a number of competing reactions. One of these ways is the re-emission of a fluorescence
photon. The fluorescence lifetime is the mean time of duration of the excited state. It depends of all the
competing processes including radiationless transitions and has only a statistical meaning (see Fig. 1).
Laser
pulse
CHL*
Mean lifetime X =
I
t. F(t) dt
I
F(t) dt
FLUORESCENCE
FIG. 1 . Definition of the mean lifetime
1600
ÌL
;
—
—
-
Control !
.
t :
«
*. 1
•. Tv*
Q« qu
enching
V
1
• •
•
• •
!
—
—
0.4 0.6 0.8
Relative fluorescence yield
FIG. 2. Relationship between mean fluorescence lifetime of chl_a and fluorescence yield during Qe quenching
in an attached leaf of barley. The fluorescence yield is changed by varying the pre-illumination (After Y. Goulas
I20JI
It is well documented from a large number of reports that chlorophyll fluorescence in vivo is a heterogeneous
emission [10-17], containing at least 3 lifetime components, the origin of which is still a matter of discussion.
However it has also been shown in previous works [12, 18, 20] that the mean chlorophyll fluorescence lifetime