Reveal diversity and temporal variability of marine

planktonic ciliates using metabarcoding

Wei-Ting Chen1,2*, Fabienne Rigaut-Jalabert3, Nathalie Simon3, Daniel Vaulot3, Kuo-Ping Chiang2, Fabrice Not3and Charles Bachy3
1
College of Ocean Science and Resource, National Taiwan Ocean University, Taiwan
2 Institute of Marine Environment and Ecology, National Taiwan Ocean University, Taiwan
3 UPMC Université Paris 06, CNRS, UMR 7144 Station Biologique de Roscoff, Roscoff, France

Introduction ENGLAND

Three orders of ciliates (Strombiddiida, Chreotrichida and Tintinnida)are major components of SOMLIT-Astan
marine microzooplankton and they function as important trophic links in microbial food webs. The
abundance of planktonic ciliates can be up to 106 cells．L-1 in the upper water layers (Chiang et al.
FRANCE
2003). In a previous large diversity study (Tara Ocean voyage, Gimmler et al. 2016), they represented a
total of 48% in OTUs and 35% in reads of the phylum Ciliophora.
We monitored the seasonal dynamics of planktonic ciliates community using metabarcoding in a
Figure 1. Location of sampling
temperate coastal site (SOMLIT-Astan, Lat 48 ° 46'18 '' N, Long 3 ° 58'6 '' W, Figure 1)in the western site, SOMLIT-Astan.
English Channel every two weeks over three years(2009-2011).

Results – Seasonal variations

Results – Overall community composition
• Obtained 4.6millions reads, 21251 OTUs in total • The seasonal changes of chlorophyll a was not
• Reads and OTUs of planktonic ciliates community are 19943 synchronous with ciliates communities except in
and 646. September 2010 and July 2011.
• These 2 peaks of chlorophyll a correspond to a low
shannon index caused by the relative strong
abundances of reads assigned to Tintinnopsis major
and Stombidiida (Figure 4).

3 3

Figure 2. Pie charts showing the contribution of all eukaryotes at (a) division-level taxa and planktonic ciliates at

Shaonnon Index
Chl a (µgL −1)

Parameter
2 2

(b) phylum-level taxa and (c) order-level taxa at SOMLIT-Astan time-series site. Affiliation was performed
Chl a
Shaonnon Index

according to the PR2 database (Guillou et al. 2013).

1 1

Result – Temporal pattern of planktonic ciliate 2009

AbunOTU of ciliates in Astan
2010
Date
2011 2012

communities
200 data11$cid
16119
18339

• The NMDS plot showed cyclic pattern of planktonic ciliate

19515
40367
reads

4043
43747
47854

diversity (more explicit in winter). The communities tend to be 100

67286
74635
81531

more similar between years when sampled during the same

season(Figure 3). Jan−09 Apr−09 Jul−09 Oct−09 Jan−10 Apr−10 Jul−10 Oct−10 Jan−11 Apr−11 Jul−11 Oct−11
Astan_n_2009_2011_Ciliate2size_NMDs_hellinger_68_apr3 date

May−09 Jul−11 Figure 4. Seasonal variations of (a) the Shannon index (b) abundant OTUs of ciliates
Month
Apr−10 communities at the SOMLIT-Astan time-series site. The green arrow indicates the peaks of
Mar−10
chlorophyll a and planktonic ciliates abundant OTUs.
May−10 Jun−10
0.4 Month

Apr−09 a Jan
Apr−10 Apr−11
May−09 a Feb
Apr−09 Jul−09 a Mar
Apr−11 Jun−09
Mar−10 Mar−09 a Apr

Conclusion & Future work

Jun−10 a May
Mar−11 Jun−09 a Jun
Feb−09 a Jul
Feb−09
NMDS2

Jul−09 a Aug
Feb−10
Mar−09 Feb−10 Mar−11 Aug−11
May−11 a Sep
Sep−09
0.0 a Oct
Oct−10
Feb−11 Jul−10 a Nov
Aug−11
Nov−11 Jul−10 a Dec
Oct−10
Jan−11 Sep−11 Sep−09
Aug−09 Jul−11
Oct−11
Jan−11 Oct−11 Jul−09 factor(Year)
Dec−11

This study showed seasonal variability in planktonic ciliate

Dec−11 2009
Feb−11 Nov−10 Jun−11
Aug−10 2010
Dec−09 Oct−09 Oct−09 Sep−10
Jan−09 Oct−10 2011

Aug−09

−0.4 Jan−10
Dec−10Dec−09 Nov−11

Sep−10
Aug−10
Year communities and high repeatability between years (more
Nov−09

obvious in winter). The next goal would be to combine with

Jan−09
Jan−10

Stress = 0.202
Nov−09

−0.5 0.0
NMDS1
0.5
another dataset to compare the temporal and spatial effects
on coastal planktonic ciliates communities.
Figure 3. Non-metric multidimensional scaling(NMDS) plot based on Bray–Curtis dissimilarities of
planktonic ciliates community at SOMLIT-Astan time-series site.

References & Acknowledgements

Chiang, K. P., C. Y. Lin, C. H. Lee, F. K. Shiah, and J. Chang. 2003. The coupling of oligotrich ciliate populations and hydrography in the East China Sea: spatial and temporal variations. Deep Sea Research Part II:
Topical Studies in Oceanography 50:1279-1293.
Gimmler, A., R. Korn, C. de Vargas, S. Audic, and T. Stoeck. 2016. The Tara Oceans voyage reveals global diversity and distribution patterns of marine planktonic ciliates. Scientific Reports 6:33555.
Guillou, L., D. Bachar, S. Audic, D. Bass, C. Berney, L. Bittner, C. Boutte, G. Burgaud, C. de Vargas, J. Decelle, J. del Campo, J. R. Dolan, M. Dunthorn, B. Edvardsen, M. Holzmann, W. H. C. F. Kooistra, E. Lara, N.
Le Bescot, R. Logares, F. Mahé, R. Massana, M. Montresor, R. Morard, F. Not, J. Pawlowski, I. Probert, A.-L. Sauvadet, R. Siano, T. Stoeck, D. Vaulot, P. Zimmermann, and R. Christen. 2012. The Protist Ribosomal
Reference database (PR2): a catalog of unicellular eukaryote Small Sub-Unit rRNA sequences with curated taxonomy. Nucleic Acids Research.
SOMLIT-Astan time-series sampling is part of the Roscoff coastal observatory and is supported by CNRS. The authors thank the captains and crew of the R.V. Neomysis (Gilles Maron, François Le Ven, Noël Guidal)
and the Service Mer et Observation.