Bacterial Dynamics Within the Mucus, Tissue and Skeleton Ofthe Coral Porites Lutea During

Bacterial dynamics within the mucus, tissue and skeleton ofthe coral Porites lutea during different seasons

Jie Li1, Qi Chen1, Li-Juan Long1, Jun-De Dong1,Jian Yang1, Si Zhang1

1CAS Key Laboratory of Tropical Marine Bio-resources and Ecology, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, Guangdong, P. R. China

Corresponding author: Si Zhang

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Running title: Seasonal dynamics of coral-associated bacteria

Table S1 The abundant (>1%) bacteria genera distributed in each coral sample.

Class / Genus / Feb-M / Feb-S / May-M / May-T / May-S / Aug-M / Aug-T / Aug-S / Nov-M / Nov-T / Nov-S
Acidobacteria / Gp9 / 1.17 / 0.66 / 0.00 / 0.07 / 0.47 / 0.11 / 0.02 / 0.00 / 0.17 / 0.04 / 0.01
Actinobacteria / Mycobacterium / 0.05 / 3.80 / 0.00 / 0.00 / 0.13 / 0.02 / 0.01 / 0.01 / 0.02 / 0.09 / 0.08
Sphingobacteria / Sphingobacterium / 0.28 / 0.03 / 0.00 / 0.28 / 0.01 / 0.05 / 0.33 / 0.02 / 4.03 / 2.74 / 0.10
Flavobacteria / Chryseobacterium / 0.06 / 0.04 / 0.00 / 0.21 / 0.07 / 0.46 / 1.02 / 0.00 / 2.32 / 3.56 / 0.19
Marixanthomonas / 0.03 / 0.00 / 0.01 / 1.19 / 0.00 / 0.00 / 0.00 / 0.00 / 0.03 / 0.01 / 0.00
Tenacibaculum / 0.02 / 0.00 / 0.01 / 4.69 / 0.01 / 0.01 / 0.00 / 0.00 / 0.02 / 0.04 / 0.00
Chlorobia / Prosthecochloris / 0.49 / 0.30 / 0.19 / 3.95 / 25.60 / 0.76 / 11.02 / 0.28 / 4.57 / 12.69 / 8.13
Alphaproteobacteria / Brevundimonas / 1.29 / 0.14 / 0.00 / 0.17 / 0.25 / 1.06 / 0.44 / 0.94 / 1.95 / 1.47 / 3.72
Rhizobium / 0.03 / 0.01 / 0.00 / 0.60 / 0.13 / 0.35 / 0.32 / 1.52 / 1.04 / 1.75 / 7.97
Paracoccus / 0.11 / 0.09 / 0.00 / 0.14 / 0.19 / 0.07 / 0.08 / 0.50 / 0.17 / 0.33 / 1.63
Ruegeria / 0.19 / 0.39 / 0.04 / 1.34 / 0.84 / 0.02 / 0.00 / 0.18 / 0.03 / 0.01 / 0.52
Silicibacter / 1.86 / 0.62 / 0.05 / 5.65 / 0.50 / 0.34 / 0.05 / 0.01 / 0.37 / 0.28 / 0.07
Pelagibius / 1.02 / 0.20 / 0.00 / 0.01 / 0.13 / 0.02 / 0.03 / 0.02 / 0.32 / 0.11 / 0.05
Erythrobacter / 0.27 / 0.03 / 0.02 / 1.75 / 0.01 / 0.04 / 0.11 / 0.00 / 0.21 / 0.04 / 0.03
Novosphingobium / 0.02 / 0.01 / 0.00 / 0.01 / 0.07 / 0.06 / 0.02 / 0.01 / 1.97 / 0.35 / 0.05
Sphingobium / 0.63 / 0.06 / 0.01 / 1.31 / 0.10 / 8.46 / 1.36 / 6.60 / 6.82 / 7.37 / 25.03
Sphingomonas / 0.02 / 0.09 / 0.00 / 0.11 / 0.08 / 0.46 / 0.48 / 0.66 / 0.35 / 0.98 / 2.36
Betaproteobacteria / Aquabacterium / 0.19 / 0.01 / 0.00 / 0.07 / 0.01 / 0.01 / 0.06 / 0.00 / 2.33 / 0.31 / 0.02
Comamonas / 0.57 / 0.33 / 0.00 / 0.09 / 0.13 / 2.92 / 0.19 / 0.02 / 3.99 / 1.98 / 0.17
Delftia / 2.54 / 0.43 / 0.00 / 0.70 / 1.32 / 0.08 / 0.08 / 0.00 / 0.40 / 0.29 / 0.27
Massilia / 0.05 / 0.10 / 0.03 / 4.71 / 0.21 / 0.24 / 15.49 / 0.02 / 5.11 / 23.61 / 0.46
Methylophilus / 0.52 / 3.24 / 0.00 / 0.48 / 1.53 / 0.18 / 0.16 / 0.01 / 0.10 / 0.46 / 0.16
Dechloromonas / 0.02 / 0.00 / 0.00 / 0.10 / 0.00 / 0.01 / 0.03 / 0.01 / 3.53 / 0.34 / 0.01
Methyloversatilis / 0.25 / 0.77 / 0.00 / 0.21 / 0.40 / 0.07 / 0.05 / 0.01 / 1.16 / 0.17 / 0.05
Deltaproteobacteria / Desulfarculus / 0.87 / 1.51 / 0.00 / 0.03 / 0.21 / 0.04 / 0.01 / 0.00 / 0.10 / 0.20 / 0.06
Gammaproteobacteria / Listonella / 0.00 / 0.09 / 1.12 / 0.11 / 0.01 / 0.00 / 0.00 / 0.00 / 0.00 / 0.00 / 0.01
Photobacterium / 0.03 / 0.01 / 3.19 / 0.40 / 0.01 / 0.01 / 0.00 / 0.00 / 0.02 / 0.00 / 0.11
Vibrio / 0.05 / 4.74 / 78.18 / 7.38 / 0.54 / 0.15 / 0.00 / 0.77 / 0.02 / 0.05 / 1.94
Pseudoalteromonas / 0.22 / 2.12 / 0.16 / 4.51 / 0.29 / 0.63 / 0.06 / 2.24 / 0.08 / 0.12 / 1.18
Rheinheimera / 0.03 / 1.02 / 0.00 / 0.03 / 0.02 / 0.00 / 0.06 / 0.00 / 0.06 / 0.04 / 0.14
Acinetobacter / 0.22 / 0.37 / 0.04 / 7.99 / 0.51 / 5.63 / 1.76 / 0.26 / 2.51 / 2.84 / 2.17
Psychrobacter / 0.06 / 1.97 / 0.00 / 0.00 / 0.01 / 0.14 / 0.03 / 0.41 / 0.07 / 0.06 / 0.05
Pseudomonas / 0.44 / 6.73 / 0.00 / 1.28 / 1.63 / 0.63 / 56.19 / 21.52 / 1.60 / 0.79 / 1.08
Stenotrophomonas / 15.64 / 1.68 / 0.03 / 11.22 / 5.14 / 0.02 / 1.23 / 0.01 / 2.80 / 1.92 / 3.53
Mollicutes / Mycoplasma / 0.00 / 0.00 / 0.00 / 0.00 / 0.00 / 0.01 / 0.03 / 0.00 / 1.46 / 0.28 / 0.01
Cyanobacteria / GpIIa / 0.08 / 0.62 / 0.01 / 0.84 / 2.84 / 0.15 / 0.07 / 0.27 / 0.58 / 0.63 / 1.27
Bacilli / Bacillus / 0.19 / 6.33 / 0.03 / 0.68 / 3.95 / 1.17 / 0.16 / 3.78 / 2.15 / 2.84 / 0.64
Exiguobacterium / 0.16 / 0.05 / 0.00 / 0.16 / 0.05 / 53.39 / 0.26 / 0.00 / 0.00 / 0.15 / 0.05
Paenibacillus / 0.05 / 0.00 / 0.00 / 0.16 / 0.63 / 0.01 / 0.53 / 51.56 / 0.25 / 0.25 / 0.14
Planococcus / 0.02 / 0.00 / 0.00 / 0.31 / 0.01 / 9.50 / 0.02 / 0.25 / 0.00 / 0.00 / 0.09
Staphylococcus / 0.14 / 1.32 / 0.00 / 0.31 / 0.17 / 0.13 / 0.02 / 0.02 / 0.06 / 0.02 / 0.25

Table S2 The abundant (>1%) bacteria groups distributed in each seawater sample.

taxon / Feb-SW / May-SW / Aug-SW / Nov-SW
Polaribacter / 0.04 / 1.60 / 0.24 / 0.07
unclassified Flavobacteriaceae / 11.49 / 7.60 / 10.93 / 8.82
unclassified Flavobacteriales / 2.63 / 1.88 / 1.06 / 1.30
unclassified Bacteroidetes / 1.62 / 0.62 / 1.29 / 0.86
Oceanicola / 2.22 / 0.53 / 3.26 / 1.11
Ruegeria / 11.32 / 12.67 / 13.28 / 18.26
unclassified Rhodobacteraceae / 7.25 / 7.52 / 10.23 / 9.76
unclassified Alphaproteobacteria / 2.83 / 2.49 / 2.06 / 1.07
Limnobacter / 0.01 / 1.06 / 0.00 / 0.00
Alteromonas / 0.12 / 10.31 / 0.00 / 0.00
unclassified Gammaproteobacteria / 1.43 / 0.95 / 0.80 / 2.11
unclassified Proteobacteria / 23.32 / 10.60 / 4.97 / 12.62
GpIIa / 20.17 / 30.58 / 39.66 / 26.35

Table S3Summary ofthe redundancy analysis. Automatic forward selection was performed with Monta Carlo permutation tests.

Axes / 1 / 2 / 3 / 4
Eigenvalues / 0.208 / 0.147 / 0.108 / 0.088
Species-environment correlations / 0.998 / 0.894 / 0.927 / 0.935
Cumulative percentage variance
of species data / 20.8 / 35.5 / 46.3 / 55.1
of species-environment relation / 34.2 / 58.4 / 76.1 / 90.6
Sum of all canonical eigenvalues / 0.609

Table S4 Environmental parameters at the sampling location in different months

Sampling month / Temp
(o C) / Sal
(ppt) / DO
(mg L-1) / pH / Rainfall (mm) / UV
(MJ m-2)
Feb / 23.54±0.01 / 34.36±0.01 / 7.46±0.01 / 7.61±0.02 / 5.60 / 18.34
May / 27.35±0.01 / 33.97±0.01 / 6.62±0.01 / 8.51±0.01 / 104.20 / 29.88
Aug / 26.71±0.01 / 34.59±0.01 / 5.81±0.01 / 8.46±0.01 / 187.40 / 28.72
Nov / 26.89±0.01 / 33.66±0.01 / 6.13±0.02 / 8.18±0.01 / 39.00 / 21.29

Temperature (Temp), salinity (Sal), DO, pH, were measured with a YSI 6600V2

Data on ultraviolet radiation (UV) intensity and rainfall were collected from the Tropical Marine Biological Research Station at Sanya, Hainan.

Fig. S1 Rarefaction curves of bacterial communities from individual samples. OTU was determined by a cutoff value of 3% sequence similarity. M represents mucus; T represents tissue; S represents skeleton; SW represents seawater.

Fig. S2 This profile illustrates dominant bacteria classes (proportion >1%) and how bacterialcommunities changed in different months in different coral compartments. The y axis showsthe percentage of each group in each sample from different months. The x axis indicates the bacterial classes. Each vertical colour bar signifies the proportion (%) of a bacterial group in samples collected in different months including February, May, August and November.M represents mucus; T represents tissue; S represents skeleton; SW represents seawater.

Fig. S3 The population dynamics of Chlorobia, Alphaproteobateria,Betaproteobateria,GammaproteobacteriaandBacilli in different months in coral mucus, tissues and skeleton. The y-axis is the population proportion of the bacterial group in the community, the x-axis is the sampling time.

Fig. S4 This profile illustrates dominant bacteria families of Alphaproteobacteria (proportion >1%) and how bacterialcommunities changed in different months in different coral compartments. The y axis showsthe percentage of each group in each sample from different months. The x axis indicates the bacterial families or unclassified groups. Each vertical colour bar signifies the proportion (%) of a bacterial group in samples collected in different months including February, May, August and November.M represents mucus; T represents tissue; S represents skeleton.

Fig. S5 This profile illustrates dominant bacteria families of Gammaproteobacteria (proportion >1%) and how bacterialcommunities changed in different months in different coral compartments. The y axis showsthe percentage of each group in each sample from different months. The x axis indicates the bacterial families or unclassified groups. Each vertical colour bar signifies the proportion (%) of a bacterial group in samples collected in different months including February, May, August and November.M represents mucus; T represents tissue; S represents skeleton.

Fig. S6The environmental parameters at the sampling site from January to Decemberin 2012. The parameters measured wereultraviolet radiation (UV) intensityand rainfall.