Paleoclimatic and taxonomic significance of alkenone isomer

ratios from northern Alaskan freshwater lakes !

William M. Longo1*, Susanna Theroux2, Anne E. Giblin3, Yinsui Zheng1, James T. Dillon1, Yongsong Huang1*!
1Department of Earth, Environmental and Planetary Sciences, Brown University, Providence, RI 02912, USA

* William_longo@brown.edu,!
Yongsong_huang@brown.edu!

E1147

2The Ecosystems Center, Marine Biological Laboratory, Woods Hole, MA 02543, USA
3Department of Energy Joint Genome InsOtute, Walnut Creek, CA 94598, USA

Introduction!

Samples, Study Area and Methods!

Samples and study area

Alkenones are a class of unsaturated long-chain

ketones used to reconstruct sea surface
temperature and, more recently, continental
temperature, through the UK37 and UK37 indices.

Analytical Methods

Surface sediments were collected from a series of lakes

(n=35) in northern Alaska (MAAT = -8 to -12 C).

Alkenones were analyzed with an Agilent

6890N GC-FID system with VF-200ms
capillary column, for separation of isomers.10

SPM was collected from Toolik Lake for in situ temperature

Available genetic data indicate that freshwater

lakes host a distinct phylogenetic clade of
alkenone-producing haptophyte algae the
Group I or Greenland phylotype. Group I
haptophytes produce alkenones with a distinct
distribution, characterized by tri-unsaturated
alkenone isomers.

(UK37=0.021*T-0.68; r2=0.85;

calibration
RMSE=1.37 C)
and evaluation of the temperature dependence of triunsaturated isomers.
150 W

19

?
34

!
(
15

22
!!
21(
?(
32

!
(

18 17
Toolik Lake
(
23 !

69 N

C39:3aEt

(
09 !
(
08 !
!
(

68.57 N

150 W

C39:4Et

(
10 !

12

06

O
O

O
O
O
O

O
O
O
O

68.6 N

!
(

!
(

C38:3bMe
C38:2Me

11

03
(
? 04
!
(
!
(
? !
(
!
(
?
!
(!
!
(
?
!
(
!
26
(
!
(
05

C38:4Me
C38:3aMe

(
13 !

69 N

25

!
(
16 ?
28

68.6 N

Kilometers
0 25 50

152.5 W

02

C38:3bEt

C38:2Et

!
(

?
31

27 29
??
(?
19 !
33 24

Alaska

!
(

*
68.63 N

70 N

?
35

C38:3aEt

01

North Slope of Alaska

Salinity ranges for alkenone-producing haptophyte algae by species and

phylogeny. Shaded areas represent salinity ranges where haptophytes from
specific phylogenetic groups are commonly found in nature. Black bars
represent the salinity ranges in haptophyte culture conditions, or for
operational taxon units based on 18S rRNA sequences from environmental
samples. Filled black circles represent reported salinities of lakes in which
specific phylogenetic clades are the dominant alkenone producers.

Because species effects present challenges for

lacustrine alkenone temperature reconstruction,
we also developed isomer ratios as taxonomic
indicators for the Group I haptophytes.

?
30

70 N

Our objectives were to describe the abundance

and distribution of these novel isomers in northern
Alaskan lakes and evaluate isomer ratios as new
temperature proxies using suspended particulate
matter (SPM) from Toolik Lake.

!
(

68.63 N

C37:2Me

!
(

y.

71 N

Hw

Beaufort Sea

!
(
14

* C37:3bMe

149.5 W

71 N

149.67 W

lto

152.5 W

C37:3aMe

C38:4Et

Da

155 W

C37:4Me

!
(

07

Kilometers
0
1
2

149.67 W

68.57 N

149.5 W

Map of study region. Green filled circles represent freshwater lakes in which LCAs were
detected, blue filled circles represent saline lakes (n=1) in which LCAs were detected, and
circles with an x represent lakes in which LCAs were not detected.

C39:3bEt

O
O
O
O

C39:2Et

Chemical Structures of the 16 LCAs that commonly

occur in northern Alaskan Lakes. Double bond
positions are indicated above. ( refers to the number of
carbon atoms between the double bond and the
carbonyl). Tri-unsaturated isomers are indicated (*).

Results and Discussion!

Average FA

Abundance and distribution

0.3

0.3

(a)

4
3a
3b
2

0.2
0.1

(b)

4
3a
3b
2

0.2
0.1

C37Me

C38Et

C38Me

C39Et

C37Me

C38Et

C38Et

Relative Intensity

C37Me

C38Me

C39Et

C37Me

4
3a
3b

(c)
C38Me

3b
3a

C38Et

3a
4

3b 2

C38Me

3a 3b

3a

C39Et

4 3a 3b 2

(e)

3b

3a

3b 2

C39Et
4 3a 3b
2

4
3a
4

3a
3b

3a

(f)

3b
2

23

3a

(d)
2

3a
3b 2

4 3a 3b 2

24
25
Retention Time (min)

23

3b 2

24
25
Retention Time (min)

We devised Ratios of Isomeric Ketones

(RIK indices) to examine the significance
of the tri-unsaturated alkenone isomers.

Average fractional
abundances of
alkenones in (a)
Toolik Lake SPM
(n=52) and (b)
surface sediments
in which all 16 LCAs
were detected
(n=9). Partial
chromatograms of
(c) 12.3 oC and (d)
3.1 oC samples of
Toolik Lake SPM.
Partial
chromatograms of
(e) Lake E5 and (f )
Lake IKP-003 (the
only saline lake in
the transect).

RIK !"
RIK !"!
RIK !"!
RIK !"!

Alkenones were detected in 22 of 34 freshwater lake

Alkenone distributions were consistent in freshwater lakes.

(%C37:4 = 25.3 3.3; C37/C38 = 0.88 0.16; Me/Et =1.37 0.28).

C!":!"
=!
!
C!":!" + C!":!"

RIK38E and in situ temperature

highlights its potential utility as a
temperature proxy.

RIK37 showed a only a modest

correlation with little overall trend or

variability in the index.

The methyl isomer ratios were both

negatively correlated with

temperature, whereas the ethyl
isomer ratios were positively and
more strongly correlated with
temperature, a pattern that potentially
reflects features of biosynthesis.

Ratio of Isomeric Ketones

The strong correlation between

RIK37

(a)

C!":!" Et
=!
!
C!":!! Et! + ! C!":!" Et

C37:4

C37:4

C37:3b

Ternary diagrams illustrate key taxonomic differences in C37 distributions. The three components represent

end members for Group III marine distributions (C37:3a+C37:2), Group II distributions (C37:4), and Group I
distributions (C37:3b). Northern Alaskan freshwater lakes plot with the Group I cluster. Alkenone distributions
in sediments from several locations follow predicted patterns based on Salinity and phylotype.

r2 = 0.20

We propose to use the RIK37 index as a taxonomic indicator for Group I and, more importantly, as an

indicator of the multi-species effect. All known Group I samples, fall within an RIK37 range of 0.50 to 0.60.
All Group II and III samples have RIK37 values of 1.0. We hypothesize that samples containing mixed
phylotypes will exhibit RIK37 values between 0.60 and 1.0.

r2 = 0.76

RIK39E

r2 = 0.49

r2 = 0.57
o

Observed Temperature ( C)
RIK indices from Toolik Lake SPM plotted vs. observed lake water temperature.
All RIK indices are shown with a common y-range for comparison. All correlations are
significant (p<0.01; Pearsons correlation coecient and Spearmans Rank).

Conclusions!
Group I haptophytes are the primary alkenone-producers in
freshwater lakes and they produce alkenones with a consistent
distribution, characterized by tri-unsaturated isomers.

Alkenones

commonly occur in northern Alaskan freshwater lakes

and they feature typical Group I alkenone distributions.

Newly

defined Ratios of Isomeric Ketones (RIK) indices are

correlated with temperature. RIK38E demonstrated the strongest
correlation, showing potential as a new temperature proxy.

The

C37:3b

Ternary diagram showing relative proportions of C37 LCAs in various samples. C37 distributions separate by phylogenetic
groupings (a) and by lake salinity (b). NAK, northern Alaskan Lakes; CAN, interior Canada Lakes; CHN, Chinese Lakes.

1!

(b)

C!":!" Me
=!
!
C!":!" Me! + ! C!":!" Me

RIK38E

RIK38M

C37:3a + C37:2

C!":!" Et
=!
!
C!":!" Et! + ! C!":!" Et

Ratios of isomeric alkenones as temperature indicators

correlated with in situ temperature
providing evidence that temperature
affects alkenone double bond
position.

C37:3a + C37:2

RIK indices are a metric of the abundance

of isomers relative to each other. RIK
values of 1.0 indicate the presence of only
the 7,14,21 alkenone, whereas a value of
0.0 indicates the presence of only the
isomeric 14,21,28 alkenone.

sediments. Concentrations were as high as 357 g gTOC-1.

All RIK indices were significantly

Ratios of isomeric alkenones as chemotaxonomic indicators

Defining indices

C37:3b alkenone is a chemotaxonomic indicator for Group I

haptophytes. RIK37 can be used in sediment samples to test for
species effects on alkenone temperature estimates.

Samples

BrayaS sediment
Toolik Lake SPM
Northern Alaskan freshwater lakes
Northern Alaskan saline lakes
Lenore Lake, Canada
Fishing Lake, Canada
Shannon Lake, Canada
Humboldt Lake, Canada
Canadian saline lakes
Chinese saline Lakes
Lake George sediment
Group II cultures
Group III cultures
Combined ocean sediment sample

1
52
22
1
1
1
1
1
7
11
1
4
2
1

Reported
Salinity
0.9
Fresh
Fresh
3.5
3
2.2
2.4
1.5
5.6 - 52.7
0.7 - 146.3
9.7
-

RIK37 (2SD)
0.57
0.53 (0.012)
0.54 (0.048)
1
0.56
0.61
0.69
0.78
1
1
1
1
1
1

RIK37 inferred
phylotype
Group I (known)
Group I (known)
Group I
Group II
Group I
Group I + Group II
Group I + Group II
Group I + Group II
Group II
Group II
Group II (known)
Group II (known)
-

Reference
Longo et al., 2013; DAndrea et al., 2011
This Study
This Study
This Study
Toney et al., 2011
Toney et al., 2011
Toney et al., 2011
Toney et al., 2011
Toney et al., 2011
Zhao et al., 2014
Longo et al., 2013; Toney et al., 2010
This study; Longo et al., 2013; Theroux et al., 2013
This study; Longo et al, 2013
Longo et al., 2013

Acknowledgements and References!

This work would not have been possible without the support and limnological monitoring of the Arctic Long Term Ecological
Research (Arctic LTER) program. We thank J. Stuckey and the Toolik Field Station GIS office for providing morphometric data on
the Arctic LTER lakes. We Thank B. Gaglioti and the Circum-Arctic Lakes Observation Network (CALON) for providing sediments
and environmental data and F. Hu for providing surface sediment from Perch Lake. We also thank J. Toney for previous work on
the interior Canada Lake sediments and for providing samples for new analyses. A. Carter, D. White and W. Daniels provided field
assistance and J. Crowther and R. Tarozo provided laboratory assistance. Funding for this research was provided by grants NSFPLR-1503846 and NSF-DEB-1026843.
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