C-MORE 2: Observing life, measuring the phytoplankton Oscar Schofield (RU COOL)

Picture by Chris Gotschalk

Phytoplankton growth and nutrient assimilation is tied to ambient light levels.

Enough energy to excite (vibrate a molecule)

Enough energy move electrons

Enough energy make something new (rearrange a molecule)

alar visible irradiance

Photosynthesis = PAR * aph *

Quantum efficiency of ..

Absorption

Spectrally averaged absorption

700nm
Irradiance 0.00 20.00 0.00 0.20 0.40 0.60 0.80 1.00

a(l ) * Eo(l )dl

a ph = 400nm
700nm

Fluorescence or charge separation oxygen carbon fixation

Eo(l )dl
400nm

Dep t h (m )

40.00 60.00 80.00 ~ 1% 100.00

Energy that is into the cell Varies cell pigmentation, light history, and size
dE d ( ) = K d ( ) E d ( ) dz

growth

rgy into the ocean ies with depth according to the IOPs s with radiative transfer eqns describe the AOPS

Efficiency of converting energy int End product (electrons, oxygen, carbo Varies with end product and physiol

Low

High

Every day, the ocean changes colour or rather, it passes though a variety of hues between the morning, noon and night of a single day. The subtle shapes of clouds, the glittering light of the sun, and the shifts in atmospheric pressure tint the sea with deep tones, cheerful tomes, plaintive tones that would cause any painter to pause in wonder. from The Samurai by Shusaku Endo (1980)

Eyeball Optics
The Secchi Disk: First systematic usage reported in 1866, but observed and remarked upon much earlier. Early experiments carried out by Commander Cialdi, head of the Papal Navy, and Professor Secchi onboard the SS LImmacolata Concezione (Cialdi, 1866). Used operationally for establishing aids to navigation over shallow water.
Thanks to Marlon Lewis

Measuring the light into the system

From John T. O. Kirks billabongs

Reflectance( ) = G* bb( )/{bb( ) +a( )}

What are some of the classical DIMS?

Local weather

= P/(Qpar(0+)<Chl>)
Morel and Platt show * variability Of 50% around a value of at specific chl values

seasonal

Claustre et al.

Diatoms 0.114 + 0.051 Cryptophytes 0.053 + 0.011

Penetration of light is determined by the material in the water which is determined by the overall inherent optical properties (IOPs)

Absorption (a) color

Photos by S. Etheridge

Scattering (b) clarity a + b = c c = attenuation

From Collin Roesler

WetLabs

detector

detector

Absorption (aAttenuation (b) )

Scattering Case I waters

cp(660) bp(660)

c p (660) = A Chl B

Positively correlated Loisel and Morel 1998 Nonlinear, B 0.7 High

Particle backscattering
West Florida Shelf

Karenia brevis bloom

Cannizzaro et al. 2002

POC Scattering (Case I waters)

Subtropical Pacific, North Atlantic

c p (660) = A POC B
B1 Contrast with non-linear dependence on Chl POC-Chl variations are important

Loisel and Morel 1998

POC-Scattering
Bottom layer cp(660)
New England Continental Shelf

Surface layer

Gardner et al. 2001

Under specific conditions, the cp POC relationship varies

From beautiful work of Morel, Bricaud, and Kirk

E.m. radiation propagating as plane waves; g geometric cross section (its shadow ) EFFICIENCY FACTORS Energy absorbed within Energy scattered out by.. Dividedby Energy impinging on g

Qa and Qb , respectively

Durand et al. 2002

Positively correlated Chl: 0.02 25 mg m-3

(eutrophic, mesotrophic, and oligotrophic waters) Non-linear dependence
Thanks to Heidi Sosik Bricaud et al. 1995

Chl-specific phytoplankton absorption Second-order

variability in aph ( ) a ( ) =
* ph

a ph ( ) Chl

Negatively correlated

a * ( ) = A( ) Chl B ( ) ph
A( ) and B( ) statistically determined

This reflects effects of changing growth conditions and community structure with trophic status Note: unexplained variability

High light

Low light

Photosynthetic pigments

Low light

Photo-protective pigments

High light

chlorophyte alga Haematococcus pluvialis

mol photons (m-2 s-1 )

2000

0 2 6 8

2000 1500 1000

1 .2 0 .9 0 .6

1.2 0.9 0.6

surface 1m 2m 5m 13m

4 1500
1000

1500 0 1 2

D e p t ( ) h m

0 500 2 2 0 2 0 4 0 6 0 8 1 0 1 2 1 4 2 2 2 2 2

0 .3

C
0 4 0 0

0.3 0
4 5 0

Calendar D a y J u l n Day a i
0 0
D D D p ptp (t ( )( ) e e e h t mmm hh

1 . 2 0 . 9 0 . 6 0 . 3

0
0 5 0 1 00 1 5 0 0 0 0

4 4 0 050 0 60 5 60 0 70 5 0 0 0 0 400 55
D D Dp e t pm () () ) e e p t ( tm m h h h

Wavelength (nm)
450
5 0 0

Depth (m)

4 4
0 4 8

Depth (m)

2 2
6 2 6

2 4

0 2 4 8

500

5 5 0

550

6 0 0

600

65

6 5 0

0 6

1 0 10 1 2

8 6

8 4

2 6

1 8 0
2 0 2 2 0 4 2 0 6 2 0 8 2 1 02 1 2 2 1 4
2 0 2 2 0 4 2 0 6

6 1 0

1 1 22

1 0 2 1

8 1 0

1 2 2 1

2 0 22 0 42 0 62 0 82 1 02 1 22 1 4
2 0 2 0 2 22 0 4 42 0 6 62 0 8 82 1 0 02 1 2 22 1 4 4 2 0 2 0 2 0 2 1 2 1 2 1

2 2 2 2 0 2 0 42 0 u2 l008 82 1a0 y0 112 22 114 4 2 62 a J u l in n a y 1 2 D J a i D Calendar Day J u l n D a y a i

J u lln n a ya y u ii D D a J a

J u l n D a y a i
-1

Calendar Day

Oliver et al. 2004

6 3 1 2 61 79 0 0 3 mol photons m-2 s-1

6 3 31 2 6 7 9 0 0 1

0 . 0 . 6 . 1 . 7 0 0 1 0 3 0 4

0 . 0 0 . 6 0 . 1 0 . 7 0 1 3 4

6 3 13 2 6 7 0 0 1 9

(m 0 . 0 a.ph 6 . )1 . 7 0 1 0 3 0 4

Vertical migration of Karenia brevis

Individual pigments can be measured on discrete samples biochemically

absorption coefficient (m2 mg-1 )

0.08 0.06 0.04 0.02 0.0

chl a chl b chl c PSC PPC

400

450

500

550

600

650

700

wavelength (nm)
From Bidigare

chl a
1.25

chl c chl b

phycobilins carotenoids

chl b chl a

Spectral Irradiance (

chl a-chl c-carotenoids 20 chl a-chl b-carotenoids chl a-phycobilins

15

Relative Absorption

1.0 0.75

10 0.50 0.25 0 400 450 500 550 600 650 700 5

W cm-2 nm-1 )

Wavelength (nm)

Spectral irradiance ( Wcm-2 s-1 )

10 1 1 0.1 0.01 0.001 0.0001 90 25

Sun stimulated fluorescence

0 20

B)

Depth (m)

Chl a Chl b PSC Chl c

40 60 80 100
Decreasing efficiency Increasing efficiency

A) Wavelength (nm)
500 600 700

0.00001 400

Relative pigment-specific spectrally weighted absorption

Heat

Absorbed photon

Charge stabilization & photosynthesis

Fluorescence

Chlorophyll a - Fate of photons absorbed by an isolated molecule

Diagram of energy states in chlorophyll and possible transitions

Light Absorption

Heating

Thermal Expansion Alexander Graham Bell developed spectrophone, essentially an ordinary spectroscope equipped with a hearing tube instead of an eyepiece listening to light induced changes in the thermal sound.

Pressure Wave

Photoacoustic signal

Strong light flash

Weak light flash

Quantum yield

max

Light

Fluorescence

Chlorophyll fluorescence

An

l ea id

ld or w

Stress (light, nutrients)

Chlorophyll concentration

0 5

monospecific G. breve community

0 5

Depth (m)

10 15 20

Depth (m)

10 15 20 Chl a

mixed chromophyte community

CDOM
8:00 12:00 18:00 22:00

8:00

12:00

18:00

22:00

Local DaylightTime

Local DaylightTime

Fluorescence: The Basics

Fm

F0 = a ph PAR k p + k f + kd
Fm Ft

kf

Fluorescence intensity

Fm = a ph PAR k f + kd Fv = a ph PAR k p (Q) + k f + kd kp Fm - F0 o = = f IIe Fm k p + k f + kd kf

kf

F0

Saturating flash

time

Integrated area is reflection of the absorption cross-section

Other useful indices FLUORESCENCE INDUCTION

Photons

Low light cells

High light cells

Fluorescence rise

Photo-acclimation

Time Flash is on RC2

Other useful indices FLOURESCENCE DECAY CONSTANTS

Fluorescence

Time D2 Qa Pheo RC

PQ Qb D1

Fluorescence decay constants

Light Flash Turned Off

Local time of day

Carbon Quantum Yield (mol C mol photons absorbed-1 )

0.1

10

Productivity (mg C mg Chl a-2 h-1 )

0.8
max

0.08

0.7

0.06

Fv/Fm

Pmax
Environmental stress

0.6

0.04

0.02

Ik
0.1

0.5

0 1 10 100

0.4 1000

Irradiance ( mol photons m-2 s-1 )

Is a cell a puddle or lake?

From Jassby and Platt 1976

Predicting oxygen evolution from fluorescence (model assuming FII = 0.5)

6

measured oxygen evolution

5 4 3 2 1 0 0 1 2 3 4

ne 1 li 1:

R2=0.92, P<<0.0001
5 6

fluorescence-based predictions of oxygen evolution

Photosynthesis a chain of cascading reactions: Each step sets the upper limit efficiency for each following step down the line mpsii (0.65) > m02 (on the order 0.125)> mco2 (on the order 0.07) For each use of energy go to one process, it is the expense of another reaction, this impacts the overall efficiency Nutrient source Ammonium Nitrate mco2 0.09 0.07

Simplest expression for photosynthesis is P = * aph * PAR

While chlorophyll specific absorption varies 3-4 fold quantum yields vary by an order of magnitude
Even in 1980s was treated as a constant

From Babin et al.

From Behrenfeld et al.

The conversion efficiency can varies between end products

Ratio of Oxygen to Carbon Quantum Yields
Light-limited photosynthesis
12 10 8 6 4 2 0

Light-saturated photosynthesis

0.5

1.5

2.5

3.5

4.5

Absorbed Quanta by phytoplankton

These instruments can be carried on remote platforms

Physiological response

Environmental Stress
PAR UVA + PAR UVB + UVA + PAR 0.6
Visible light downregulation

0.75

1600

800

EPS

1200 0.50 800 0.25

PAR ( mol m-2 s-1 )

PAR ( mol m-2 s-1 )

600 0.4

Fv/Fm

UVB damage

400

Fv/Fm

0.2 200
Ik>PAR Ik>PAR

400

0.00 5:00 9:00 13:00 17:00

0 21:00

0 6:00 10:00 14:00 18:00 Local Daylight Time

Local Daylight Time

-1 m (mol C mol photons absorbed ) ax

Rough seas
0.125 0.105 0.085 0.065 0.045
11 8 9
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 Arabian Sea (SW Monsoon) Arabian Sea (Inter Monsoon) Arabian Sea (NE Monsoon) Antarctic (Bransfield-Bellingshausen water) Antarctic (Bransfield-Weddell water) Antarctic (Ice-Edge water) Antarctic (Weddell-Scotia Confluence waters) Antarctic (Bellingshausen Cold water) Antarctic (Bellingshausen Warm water) Antarctic (Transitional Weddell Water) Antarctic (Palmer Station) Canary Islands (Fall) NE Atlantic Subtropical Gyre (Fall) NW Atlantic Subtropical Gyre (Fall) Gulf Stream (Fall) NW Atlantic Continental Shelf (Fall) Canary Islands (Spring) NE Atlantic Subtropical Gyre (Spring) NW Atlantic Subtropical Gyre (Spring) Gulf Stream (Spring) NW Atlantic Continental Shelf (Spring) Southern California Bight New Jersey Coastal Region

10

4 5 6

0.025
21

23 13 15 7 16 14 19 18 17

22

20

1 2

12

Oligotrohic seas
250 300 350

0.005

50

100

150

200

Ek(PAR) (mol photons m-2 s-1 )

High light

Low light

-carotene globules

thylakoids

NUTRIENT LIMITATIONS
Iron-Ex

V=Vmax*S/(Ks+S)
Austin Powers Fat Bastard System

Nutrient Uptake (V)

Vmax

Miss Manner System

Ks (usually at Vmax/2) Nutrient concentration (can be nitrogen, phosphorus)

Diatoms
High Vm High Ks

Coccolithophores
Low Vm Low Ks

High or fluctuating nutrients High mixing, upwelling Low average irradiance, light fluctuations High turbulence

Chronically oligotrophic Stratified conditions High average irradiance Low turbulence

V m ax 1 0 .8 y

12 10

light Ek

depth Chlorophyll