PHOTOBIOLOGY 1-PS and Fluorescence
Transcript of PHOTOBIOLOGY 1-PS and Fluorescence
PHOTOBIOLOGY PHOTOBIOLOGY
1Nadine Schubert
Instituto de Ciencias del Mar y Limnología de la UNAM 1Unidad de Sistemas Arrecifales, Puerto Morelos, México
WHAT DOES PHOTOBIOLOGY MEAN?
Photosynthesis Photomorphogenesis Cirvadian Rhythm Ultraviolet Radiation
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PHOTOBIOLOGY PHOTOBIOLOGY
Part 1: Photosynthesis and FluorescencePart 1: Photosynthesis and Fluorescence
Part 2: Photoacclimation/Part 2: Photoacclimation/‐‐adaptationadaptation
Part 3: PhotoprotectionPart 3: Photoprotection
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pp
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Part 1:Part 1:h h d lh h d lPhotosynthesis and FluorescencePhotosynthesis and Fluorescence
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PHOTOSYNTHESIS
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LIGHT ABSORPTIONLIGHT ABSORPTION
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THE PHOTOSYNTHETIC APPARATUSTHE PHOTOSYNTHETIC APPARATUS
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THE PHOTOSYNTHETIC APPARATUSTHE PHOTOSYNTHETIC APPARATUS
PSIILHCII Cyt bf PSI LHCIATPase
y
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LIGHT ABSORPTION
The absorbed light energy is
Antenna pigmentsAntenna pigments
The absorbed light energy is
funneled by excitation transfer
into the RC’s, where energy , gy
conversion by charge
separation takes place.
9Photochemistry
PS II
Photochemistry
PS II
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LIGHT ABSORPTIONLIGHT ABSORPTION
photonexcited state
p
molecule b b g
ener
gy
absorbs photon
crea
sing
10ground state
In
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EXCITATION ENERGY TRANSFER
Excitation transfer Electron transfer
Light Acceptore-
Reaction C tCenter
11AntennaDonor
e-
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ELECTRON TRANSFER
ADP + Pi ATP
2H+ Fd
NADP + H+ NADPH
PSIILHCII Cyt bf PSI LHCIPQH2 ATPase
yPQ
122H2O O2+ 4H+
2H+
PC H+
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LIGHT ABSORPTION AND ENERGY TRANSFER
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PHOTOSYNTHESIS AND FLUORESCENCE
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PHOTOSYNTHESIS AND FLUORESCENCE
photonexcited state
pexcited state
molecule b b
Photochemistry
absorbs photon Fluorescence
15ground state
ground state
Heat
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PHOTOSYNTHESIS AND FLUORESCENCE
Antenna pigments
Heat Fluorescence
PS II
16Photochemistry
PS II
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PHOTOSYNTHESIS AND FLUORESCENCE
Antenna pigments
Non-light -tress conditions
Heat Fluorescence
PS II
17Photochemistry
PS II
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PHOTOSYNTHESIS AND FLUORESCENCE
Photochemistry = 1Fluorescence = 0
Photochemistry = 0Fluorescence = 1Fluorescence 0 Fluorescence 1
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Whitmarsh & Govindjee (2002)
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CHLOROPHYLL FLUORESCENCE MEASUREMENT
PS = 0PS 0NPQ = 0
Fv/Fm = (Fm‐Fo)/Fm
Fm = maximum fluorescence (RC’s closed)Fo = minimum fluorescence (RC’s open)
PS = 1NPQ = 0
(higher plants – 0.85, macroalgae usually lower)
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Fv/Fm – MAXIMUM QUANTUM YIELD
Quantum yield: Probability that the energy of a photon absorbed will be used for photosynthesis (i.e. enters in the
e‐ ‐ transport chain)
⇓⇓Indicator of photosynthetic efficiency
M i i ld
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Maximum quantum yield: requires complete relaxation of the competing mechanisms with the photochemical energy conversion
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Fv/Fm – Diurnal and spatial variationChondrus crispus
Dep
th (m
)
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Macrocystis pyrifera
Colombo-Pallotta (2007)21
Hanelt et al. (1992)
Fv/Fm – Comparison of stress responses between species
Littoral Sublittoral Sublittoral
Littoral SublittoralSublittoral
22van de Poll et al. (2001)
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CHLOROPHYLL FLUORESCENCE MEASUREMENT
Fv/Fm ∆F/Fm’PS 0Fv/Fm ∆F/FmPS = 01 > NPQ > 0
1 > PS > 01 > NPQ > 0
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∆F/Fm’ – EFFECTIVE QUANTUM YIELD
Used to describe the variation in the photochemical ffi i f PSII d ill i t d ditiefficiency of PSII under illuminated conditions.
Measurement of this parameter at certain irradianceMeasurement of this parameter at certain irradiance value. ⇓⇓
Indicator of the ability of an organism to move electrons beyond PSII (ETR)
24∆F/Fm’ = (Fm’‐F)/Fm’
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ELECTRON‐TRANSPORT RATE (ETR)– CURVES
ETR = Irradiance ⋅ ∆F/Fm’ ⋅ 0,5 ⋅ Absorptance (Genty et al. 1989)
∆F/Fm’ = effective quantum yield (under light)
0,5 = Assumption that 50% of these quanta are absorbed by PSII
Absorptance = fraction of incident light that is absorbed by the photosynthetic tissue. Not the p g y p y
same as absorbance (quantifies how much of the incident light is absorbed by an object).
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ELECTRON‐TRANSPORT RATE (ETR)– CURVES
ETR = Irradiance ⋅ ∆F/Fm’ ⋅ 0,5 ⋅ Absorptance
Relative ETR = Irradiance ⋅ ∆F/Fm’ ⋅ 0,5 (Ralph et al. 2002)
ETR h b ti h t i ti h b t i li ti-ETR: when absorption characteristics change between species, acclimations, seasons…
- rel. ETR: use only when it is sure that there are no differences in the absorption characteristics
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ETR– CURVES AS AN ANALOGUE TO P‐E‐ CURVES
Macrocystis pyrifera
27Colombo-Pallotta et al. (2006)
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CHLOROPHYLL FLUORESCENCE
EXTENSIVELY USE DUE TO:
NON DESTRUCTIVE• NON‐DESTRUCTIVE• NON‐INVASIVE
• RAPID• RAPID• SENSITIVE
• IN REAL‐TIMEIN REAL‐TIME
Since 1995 the number of articles published applyinghl h ll fl th l i f th
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chlorophyll fluorescence on the analysis of thephotosynthetic performance in macroalgae and seagrasseshas increased more than five times.
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FLUOROMETERS
The Chl fluorometer should be capable of measuring the fluorescence yield ina non-intrusive way:
very low measuring light (i.e. exciting light) intensity for assessment of thefluorescence yield of a dark-adapted sample
the detection system has to be very selective to distinguish betweenfluorescence excited by the measuring light and the much stronger signalscaused by ambient and actinic light (full sun light saturating light pulses forcaused by ambient and actinic light (full sun light, saturating light pulses forassessment of maximum fluorescence)
fast time response to resolve the rapid changes in fluorescence yield upon
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fast time response to resolve the rapid changes in fluorescence yield upondark-light and light-dark transitions
PAM fluorometers: Pulse-Amplitude-Modulated fluorometers 29p
Pulse‐Amplitude‐Modulated Fluorometers
Distinguish between fluorescence and ambient light
→ Allows measurement of fluorescence in the presence of actinic light (light absorbed by the photosynthetic apparatus to drive g ( g y p y pp
photosynthesis)
How? – Measuring light is modulated and the fluorescenceHow? Measuring light is modulated and the fluorescence amplifier is highly selective for the modulated signal (yield of
chlorophyll fluorescence)
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- pulse-modulated measuring light can be generated either by a light-emitting diode (LED; most PAM fluorometers) or a flash
discharge lamp (i e XE-PAM) 30discharge lamp (i.e. XE PAM)
Pulse‐Amplitude‐Modulated Fluorometers
DUAL-PAMIMAGE-PAM
MINI-PAM
31DIVING PAM XE-PAM 31DIVING-PAM XE-PAM