Laser Induced Fluorescence Transient (LIFT):
remote measurement of light use efficiency
in ecosystems
Roland Pieruschka, Uwe Rascher, Denis Klimov, Zbigniew S. Kolber, Joseph A. Berry
February, 08. 2007
Objectives
• Inaccessible outer canopies - need for remote
measurement of light use efficiency
• Evaluation of the LIFT approach by PAM
fluorometry and gas exchange
• Measurement of cold/light stress of outer canopies
in the field
• Implementation of the data into canopy models
to reduce their uncertainty
LIFT – Fluorometer• remote measurement: from a distance up to 50 m • excitation signal: 665 nm laser diode
• fluorescence emission at 690 nm: collected
by Cassegrian telescope and detected
by avelange photodiode
Fast Repetition Rate Fluormetry
ETRCSA RRt
Q
][
AQ
AAPSIIPSIIA
QQnIQ
t
][])[(**][
Cn
Qwith
PSII
A ][
(Kolber et al. 1998; Kolber et al. 2005. Ananyev et al. 2005)
AQPSII
C
pC
CIC
t
1
1**
CFFmFtf *)'()( time [ms]
0.0 0.5 1.0 1.5 2.0 2.5
fluor
esce
nce
yiel
d [a
.u.]
0
1
2
3
4
5
What are we measuring?single vs. multiple turnover (Govindjee, 1995)
FRR: sequence of single turnover fleshlets:
• corresponds to O-J phase
(photochemical phase)
• kinetics of QA- accumulation/re-oxidation
with minimal effect on PQ pool
PAM: multiple photochemical turnover:
• corresponds to O-J-I-P phase (photochemical and thermal phase)
• well defined final state with fully reduced PQ pool
Laboratory experiments: bench top FRR vs. PAM
Combined measurements of:
• gas exchange (LI-6400)
• chlorophyll fluorescence
using PAM and a bench top FRR
• measurements under non-
photorespiratory conditions
PAR [µmol photons m-2 s-1]
0 200 400 600 800 1000 1200 1400
Fs
& F
m' (
no
rmal
ize
d)
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
Fm'Fs
PAR [µmol photons m-2 s-1]
0 200 400 600 800 1000 1200 1400
Fs
& F
m' (
norm
aliz
ed)
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
Fm'Fs
Laboratory experiments: bench top FRR vs. PAM
FRR
PAM
PAR [µmol photons m-2 s-1]
0 200 400 600 800 1000 1200
Fv
Fm
-1'
0.0
0.2
0.4
0.6
0.8
PAMFRR
Laboratory experiments: bench top FRR vs. PAM
PAR [µmol photons m-2 s-1]
0 200 400 600 800 1000 1200
ET
R [µ
mol
m-2
s-1
]
0
20
40
60
80
100
APAM
PAR [µmol photons m-2 s-1]
0 200 400 600 800 1000 1200
ET
R [µ
mol
m-2
s-1
]
0
20
40
60
80
100
AFRR
Electron Transport Ratesgas exchange vs. fluorescence
PAM FRR
5.0*'*/ PARaFmFvETR (Genty et al. 1989)
ETR (A) [µmol m-2 s-1]
0 20 40 60 80 100 120 140 160 180
ET
R (
FR
R)
[µm
ol m
-2 s
-1]
0
20
40
60
80
100
120
140
Electron Transport Ratesfrom FRR fluorescence and gas exchange
ETR (A) [µmol m-2 s-1]
0 20 40 60 80 100 120 140 160 180
ET
R (
FR
R)
[µm
ol m
-2 s
-1]
0
20
40
60
80
100
120
140
250 µmol CO2 mol-1
400 µmol CO2 mol-1
PSII quantum efficiency vs. quantum yieldof CO2 assimilation
Fv Fm'-10.1 0.2 0.3 0.4 0.5 0.6
C
O2
0.02
0.03
0.04
0.05
0.06
250 µmol CO2 mol;
y=0.08*x+0.02; r ²=0.94400 µmol CO2 mol;y=0.09*x+0.01; r ²=0.93
Estimate of underestimationof ETR by FRR
Fv Fm'-10.1 0.2 0.3 0.4 0.5 0.6
C
O2 )
*
0.05
0.10
0.15
0.20
0.25 250 µmol CO2 mol:y=0.29*x+0.07; r ²=0.94400 µmol CO2 mol:y=0.37*x+0.03; r ²=0.95
4*(A+Rd) /PAR vs. Fv/Fm
ETR(A) [µmmol m-2 s-1]
0 20 40 60 80 100 120 140 160 180E
TR
(FR
R) C
[µ
mm
ol m
-2 s
-1]
0
20
40
60
80
100
120
140
160
180
250 µmol CO2 mol:
y=0.77*x-2.37; r ²=0.98400 µmol CO2 mol:
y=0.77*x-3.47; r ²=0.99
Corrected ETR
Summary I
• The LIFT/FRR is working with low excitation intensities
resulting in lower maximum fluorescence then
the PAM approach in particular under high light intensities
• However, the LIFT approach provides an advantage of remote
measurement of Fv Fm-1’ as an index of stress correlated
to ETR within undisturbed microenvironment of leaves
Field experiments
Impact of cold stress on photosynthesis- Capsicum annuum- Ficus spec.- Lycopersicon esculentum- Persea americana- grass community dominated by Lolium spec.
Predawn measurements on Lycopersicon esculentum
time
11/6
/200
6
11/1
3/20
06
11/2
0/20
06
11/2
7/20
06
12/4
/200
6
12/1
1/20
06
Fv
Fm
-1
0.0
0.2
0.4
0.6
0.8
1.0
FRR (Plant 1)FRR (Plant 2)PAM (Plant 1)PAM (Plant 2)
TM
IN [°
C]
0
5
10
15
PA
RM
AX
[µm
ol photons m-2 s
-1]
0
200
400
600
800
1000
1200
Diurnal course: Lycopersicon esculentum
time
11/2
6/ 1
2:00
AM
11/2
7/ 1
2:00
AM
11/2
8/ 1
2:00
AM
11/2
9/ 1
2:00
AM
11/3
0/ 1
2:00
AM
12/1
/ 12:
00 A
M
12/2
/ 12:
00 A
M
Fv
Fm
`-1
0.0
0.2
0.4
0.6
T [
°C]
0
5
10
15
20
25 PA
R [µ
mo
l ph
oto
ns m
-2 s-1]
200
400
600
800
1000
1200
T
PAR
1
2
43
Predawn measurements on Persea americana
time
11/6
/200
6
11/1
3/20
06
11/2
0/20
06
11/2
7/20
06
12/4
/200
6
12/1
1/20
06
Fv
Fm
-1
0.0
0.2
0.4
0.6
0.8
TM
IN [
°C]
0
2
4
6
8
10
12
14
16
18
PA
RM
AX
[µm
ol p
ho
ton
s m-2 s
-1]
0
200
400
600
800
1000
1200
time
11/2
7/ 1
2:00
AM
11/2
8/ 1
2:00
AM
11/2
9/ 1
2:00
AM
11/3
0/ 1
2:00
AM
12/1
/ 12:
00 A
M
12/2
/ 12:
00 A
M
12/3
/ 12:
00 A
M
Fv
Fm
`-1
0.0
0.2
0.4
0.6
2D Graph 2
Fv
Fm
`-1
0.0
0.2
0.4
0.6
T [°
C]
0
5
10
15
20
25
PA
R [µ
mol
pho
tons
m-2
s-1
]
200
400
600
800
1000
1200
T
PAR
target 1
target 3
Diurnal cours: Persea americana
Steady state fluorescence
TM
IN [
°C]
0
2
4
6
8
10
12
14
16
18
PA
RM
AX
[µm
ol ph
oto
ns m
-2 s-1]
0
200
400
600
800
1000
1200
Fs
(me
an
12
-13
:00
)
0
2
4
6
8
10
12 Capsicum annuum
time
11/1
5/-4
607
11/1
9/-4
607
11/2
3/-4
607
11/2
7/-4
607
12/1
/-460
7
12/5
/-460
7
12/9
/-460
7
Fs
(me
an
12
-13
:00
)
0
2
4
6
8
10
12 Lycopersicon esculentum
time
11/2
6/ 1
2:00
AM
11/2
7/ 1
2:00
AM
11/2
8/ 1
2:00
AM
11/2
9/ 1
2:00
AM
11/3
0/ 1
2:00
AM
12/1
/ 12:
00 A
M
12/2
/ 12:
00 A
M
Fs
0
2
4
6
8
10
Fs/
PA
R
0.00
0.02
0.04
0.06
Capsicum annuum
Lycopersicon esculentum
T [°
C]
0
5
10
15
20
25
PA
R[µ
mol photons m
-2 s-1]200
400
600
800
1000
1200TPAR
Fs
0
2
4
6
8
10
Fs/
PA
R
0.00
0.02
0.04
FsFs/PAR
Summary II
• The LIFT system can sensitively detect stress
correlated to ETR
• In order to quantify photosynthetic CO2 uptake
the LIFT system has to be extended by measurements of:
- light
- stomatal conductance
- temperature
Acknowledgement
Marie Curie Outgoing International Fellowships(Nr: 041060 – LIFT)
Joe Berry
Zbigniew Kolber
Uwe Rascher
Denis Klimov
Larry Giles
Bob Haxo