Koninklijk Nederlands Instituut voor ZeeonderzoekRoyal Netherlands Institute for Sea Research

1NIOZ is an institute of the Netherlands Organisation for Scientific Research (NWO)

Automated FRRF measurements provide an alternative means to obtain seasonal and

annual primary production estimatesJacco Kromkamp, Greg Silsbe, Jethro Waanders &

Jan Peene

2

Partners in PROTOOLAcknowledgements: EU-FP7-Env program for financing the project

2

Dave SuggettEvelyn Lawrenz

Denise Smythe-WrightDiane Purcell/Adrian

Rüdiger RöttgersRodney ForsterEliza Capuzzo Rüdiger Heuermann

Karin Munderloh

Stefan SimisPasi Ylostalo Martin Trtilek

Michal Sicner

Jacco/Greg/JethroJan Peene

Ondrej PrasilOndrej Komarek

Why PROTOOL (PROductivity TOOLs)

Understanding aquatic ecosystems is not possible without knowledge of primary production

Biomass (chla) is no good measure of primary production (due to high –but varying - turnover rates)

Carrying capacity higher trophic levels depend on primary production, not on chla

Currently no simple PP method, hampering development of long term time series of primary production, certainly by water management agencies

Active fluorescence techniques (PAM-FRRF) are optical techniques, so “easy” to automate

FRRF-based approach can standardize measurements3

Chl can be measured from space, but turning this into primary production is risky (lack of PI-data, uncertainty ~ 100%)

4

0 150 300 450 g C/m2/yr

Composite of annual NPP

http://marine.rugers.edu/opp/Production

Solution: use ships of opportunity and equip it with automated sensors to measure GPP etc?

5 1 year of shipping routes

FRRF basics

Fv/Fm = indicator of physiological condition of the algae Relative (!) ETR = PAR x ΔF/Fm’

6

2000

3000

4000

5000

6000

7000

8000

9000

0 50 100 150 200

Fluo

resc

ence

time (µsec)

dark HL

m

mmv F

FFFF 0/

'

''/

mPSIIm F

FFFF m

Max PSII efficiency

Effective PSII efficiencyFo (proxy for [chla]

Fm

Fm’ΔF FV

Rise in F to Fm gives σPSII

FRRF-basics: the functional and optical cross sections

7

effective = functional PSII cross section:

PSIIPSIPSIIaa

na

kkkk

a

PSIIph

PSIIPSIIPSII

hfp

pPSIIPSII

**

*

aσPSII In dark:

aPSII

P680

QA QB

2H2O O2

4e-

P680 PQPQH2 cytb6f

4e- PC

P700

Fd

O2

NO3-

NADP CO2-fixation

ETR ≠ C-fixation

8

Linear ETR via PSI to NADPH Alternative electron sinks :

Mehler reaction (water-water cycle) NO3 reduction PSI cyclic transport PSII cycle PTOX activity

8

Investigate Φe,C for different water bodies to

develop stochastic prediction model

Requirements for PROTOOL:1. convert ETR into C-fixation2. integrate ETR of whole water column over time

9

• Can be measured with FRRF• R-module to measure [chl], kd (zP),E• Unknowns at start project:

• Φe,C (mol C/mol electrons) =0.25 mol C/e-

• nPSII: can now be measured with new Oxborough sigma-algorithm = 0.002 PSII/chla

]Φ[][ PSII0

e

Zp sunset

sunrisePSIIPSII nEchlaPP

ETR

Algorithms used

10

𝑃 𝐵=𝐸× ∆𝐹𝐹𝑚′ ×𝑛𝑃𝑆𝐼𝐼×𝜎 𝑃𝑆𝐼𝐼×Φ𝑒 ,𝐶

[𝑛𝑃𝑆𝐼𝐼 ]=𝐾𝑅

𝐸𝐿𝐸𝐷×

𝐹 𝑜

𝜎 𝑃𝑆𝐼𝐼

𝐸𝑇𝑅𝑉=𝐹𝑚×𝐹 𝑜

𝐹𝑚−𝐹 𝑜× ∆𝐹𝐹𝑚 ′

×𝐾 𝑅

𝐸𝐿𝐸𝐷×𝐸

❑

K&F algorithm

NEW:Sigma algorithm

Absorption algorithm(volumetric)

2 new algorithms allow, after proper calibration of nPSII (O2-flash yields) measurement of absolute ETR

Poster Oxborough et al: recent advancements in the methods used to analyse Fast Repetition rate Fluorometry (FRRf) data….

11

Poster Silsbe et al: Highly resolved measures of

photosynthetic electron transport in European

coastal waters

Sampling stations monitoring program

12

Oosterschelde = Eastern Scheldt

Westerschelde = Western Scheldt

MesotrophicMarine < 2 PSU

Secchi:3-5m

EutrophicSecchi: 2-0.2mTrue estuary:

0-30 PSU

Fv/Fm as stress indicator

Low Fv/Fm nutrient limitation (red arrows) and in winter 13

Fv/Fm Eastern Scheldt mouth to east

distance from mouth (storm surge barrier, km)5 10 15 20 25

Dat

e

1/1/2005

7/1/2005

1/1/2006

7/1/2006

1/1/2007

7/1/2007

1/1/2008

7/1/2008

1/1/2009

7/1/2009

1/1/2010

7/1/20100.1 0.2 0.3 0.4 0.5 0.6 0.7

Eastern Scheldt, station OS1, new algorithm no calibration for Φe,C (0.25)

Some problems with winter values (underestimations), but in general good agreement

14

OS2

1/1/2005 1/1/2006 1/1/2007 1/1/2008 1/1/2009 1/1/2010 1/1/2011

PP

(mgC

m-2

d-1)

1

10

100

1000 14CK&FAbsSigma

All data Eastern Scheldt, Φe,C = 0.25

15

Eastern Scheldt, all data

GPP (mg C m-2 day-1) obtained from 14C-fixation data

0.1 1 10 100 1000

GP

P (m

g C

m-2

day

-1) f

rom

FR

RF

dat

a

0.1

1

10

100

1000

K&FAbsSigmaK&F fitAbs fitSigma fit

For the Western Scheldt, Φe,C=0.25

16

WS all

GPP-14C (mgC m-2d-1)

0.1 1 10 100 1000

GP

P-F

RR

F (m

gC m

-2d-1

)

0.1

1

10

100

1000

K&FAbsSigma

“calibration” of electron requirement for C-fixation based on comparison of daily water column primary production: Eastern Scheldt

Seasonality (?) in quantum requirement (QR=1/Φe,C)

17

Next step: use annual GPP for QR and average QR over all years. Station specific: example OS2

QR varies between years for some stations K&F algorithms produced too low QR (<4)

18

Estimates of annual primary productionEastern Scheldt

Annual GPP >90% accurate for new Oxborough algorithms!! “old” K&F algorithm less reliable 19

Quantum requirements Western Scheldt K&F algorithm

no clear seasonality, but sigma and absorption algorithms show seasonality

Lowest QR April-Sept

Minimal QR<4

20

Western Scheldt: annual GPP as % 14C-GPP

Most estimates 75-125% of measured GPP Cycle in QR? Something odd with station WS4 21

conclusions Automated application of FRRF and spectral reflectance

makes automated primary production measurements possible FRRF measurements accurately predict seasonal dynamics in

GPP Quantum requirements (QR) for C-fixation seem rather

constant (5-7 in main growth season), but higher in winter (related to low Fv/Fm?).

QR are similar for each station, but year to year variation does exist. Reason??? (2006 was odd year in all measurements, also in Westerschelde)

More need to be done to understand variability in QRs Using autonomous FRRF measurements on SOOPs can

significantly improve global GPP estimates Miniaturize for use on gliders

22

Thank you for your attention

suggested reading:

23

Chla is not a good predictor for primary production

Westerschelde estuary: high SPM, eutrophic Oosterschelde estuary: low SPM, mesotrophic

24

Westerschelde

year1991 2001 2006 2007 2008 2009

P:B

-rat

io

0

20

40

60

80

100

120

140 WS1WS6WS14

Oosterschelde

year1991 1996 2000 2006 2007 2008 2009

P:B

-rat

io0

20

40

60

80

100

120

140 OS1OS2OS8

25

Map of chl-a derived through continuous reflectance measurements. The height of the green line is

proportional to the chl-a concentration

Project Spectral Reflectance Measurements

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High spatial resolution (~100 m) characterization of the optical properties and its driving constituents in European Coastal waters.

The fluorometer

Different flow through systems: direct connection to water inflow Via storage tank (for dark acclimation or fixed sample)

27

Automated ETR from flow-through

28

Baltic Sea

North Atlantic Cruise (England – Iceland)

Acknowledgments:

EU-FP7 program for financing the project Jan Peene for assistance with the 14C measurements Partners in PROTOOL

29

Dave SuggettEvelyn Lawrenz

Denise Smythe-Wright

Rüdiger RöttgersRodney ForsterEliza Capuzzo Rüdiger Heuermann

Karin Munderloh

Ondrej PrasilOndrej Komarek

Stefan SimisPasi Ylostalo Martin Trtilek

Michal Sicner

Jethro Waanders

PSICAM (point source integrated cavity absorption meter)

30Data Rüdiger Röttgers, HZG

Example: using a priori assumptions (4 electrons/C and nPSII=0.002 units/mg chla

FRRF accurately captures seasonal dynamics and C-fixation estimates are close to measured one, even using a priori assumptions 31

OS2

1/1/

2005

1/1/

2006

1/1/

2007

1/1/

2008

1/1/

2009

1/1/

2010

daily

GP

P (m

g C

m-2

d-1

)

1

10

100

1000 OS5

1/3/

2005

1/6/

2005

1/9/

2005

1/12

/200

5

1/3/

2006

1/6/

2006

1/9/

2006

1/12

/200

6

FRRF14C

OS9

1/4/

2007

1/7/

2007

1/10

/200

7

1/1/

2008

1/4/

2008

1/7/

2008

1

10

100

1000

FRRF14C

Central north arm North Sea

Western Scheldt, Φe,C=0.25

Good performance in both marine and freshwater areas32

WS14

1/1/

2006

7/1/

2006

1/1/

2007

7/1/

2007

1/1/

2008

7/1/

2008

1/1/

2009

7/1/

2009

1/1/

2010

7/1/

2010

GP

P (m

gC m

-2d-1

)

0.1

1

10

100

1000

14CK&FAbsSigma

WS1

1/1/

2006

7/1/

2006

1/1/

2007

7/1/

2007

1/1/

2008

7/1/

2008

1/1/

2009

7/1/

2009

1/1/

2010

7/1/

2010

GP

P (m

gC m

-2d-1

)

0.1

1

10

100

1000

14CK&FAbsSigma

33

Estuaries worldwide

• Linear relationship between PP and

biomass macrobenthos

Herman et al. 1999; Kemp et al. 2005