Revised version to Wetlands
Genetic diversity and functional traits of Pterocarpus officinalis Jacq.
associated with symbiotic microbial communities in Caribbean swamp
forests in relation to insular distribution, salinity and flooding
Amadou M. Bâ 1* and Elsie Rivera-Ocasio 2
1
Laboratoire
de
biologie
et
physiologie
végétales,
LSTM-
UMR113/IRD/CIRAD/UM2/INRA/SupAgro, UFR Sciences exactes et naturelles, Université
Antilles-Guyane, Campus Fouillole, BP. 592, 97159, Pointe-à-Pitre, Guadeloupe (Lesser
Antilles), France, 2Department of Biology, University of Puerto Rico-Bayamon, Parque
Industrial Minillas Carr 174, Bayamón PR, 00959-1911
*Corresponding author: E-mail: amadou.ba@univ-ag.fr
Electronic Supplementary materials
Table 1. Checklist of vertebrates and macroinvertebrates species frequently found in
Pterocarpus officinalis forests in Puerto Rico. Mammal, reptile, amphibian, fish, and
macroinvertebrate data from Puerto Rico taken from Ríos-López (2007) and Neftalí RíosLópez (personal observation); density data in individuals/ha between February 2001 and
March 2002. Bird data from Puerto Rico taken from Acevedo and Aide (2008); density data
expressed as the range in annual mean number of individuals per 20-m fixed-radius point
count between August 2002 and July 2004. NA = not assessed; NCN = no common name.
Taxonomic Group
Family
Common name
Scientific name
Density
Muridae
Black Rat
Rattus rattus
NA
Ardeidae
Green Heron
Butorides virescens
0.42–0.57
Falconidae
American Kestrel
Falco sparverius
0.01–0.01
Columbidae
Zenaida Dove
Zenaida aurita
0.26–0.37
Columbidae
Common Ground-Dove
Columbina passerine
0.00–0.02
Cuculidae
Mangrove Cuckoo
Coccyzus minor
0.21–0.69
Cuculidae
Smooth-billed Ani
Crotophaga ani
0.02–0.07
Picidae
Puerto Rican Woodpecker
Melanerpes portoricensis
0.20–0.49
Tyrannidae
Gray Kingbird
Tyrannus dominicensis
0.36–0.17
Tyrannidae
Loggerhead Kingbird
Tyrannus caudifasciatus
0.00–0.02
Tyrannidae
Puerto Rican Flycatcher
Myiarchus antillarum
0.05–0.20
Vireonidae
Black-whiskered Vireo
Vireo altiloquus
0.00–0.23
Turdidae
Red-legged Thrush
Turdus plumbeus
0.05–0.14
Mimidae
Pearly-eyed Thrasher
Margarops fuscatus
0.01–0.02
Parulidae
Northern Waterthrush
Seiurus noveboracensis
0.79–0.82
Parulidae
Ovenbird
Seiurus aurocapilla
0.04–0.13
Parulidae
Black-and-white warbler
Mniotilta varia
0.06–0.12
Parulidae
Prarie Warbler
Dendroica discolor
0.02–0.13
Parulidae
Common Yellowthroat
Geothlypis trichas
0.03–0.06
Parulidae
American Redstart
Setophaga ruticilla
0.05–0.05
Parulidae
Prothonotary Warbler
Protonotaria citrea
0.00–0.04
Parulidae
Northern Parula
Parula Americana
0.00–0.01
Parulidae
Yellow Warbler
Dendroica petechial
0.00–0.01
Coerebidae
Bananaquit
Coereba flaveola
3.24–4.97
Mammals (non-volant)
Birds
Thraupidae
Puerto Rican Spindalis
Spindalis portoricensis
0.01–0.03
Emberizidae
Black-faced Grassquit
Tiaris bicolor
0.00–0.02
Icteridae
Greater Antillean Grackle
Quiscalus niger
0.11–0.62
Icteridae
Shiny Cowbird
Molothrus bonariensis
0.01–0.07
Estrildidae
Oranged-cheeked Waxbill
Estrilda melpoda
0.00–0.01
Dactyloidae
Puerto Rican Crested
Anole
Anolis cristatellus
1097.2
Dactyloidae
Puerto Rican Saddle Anole
Anolis stratulus
197.2
Dactyloidae
Puerto Rican Grass Anole
Anolis pulchellus
30.6
Iguanidae
Green Iguana
Iguana iguana
NA
Emydidae
Antillean Slider
Trachemys stejnegeri
2.8
Eleutherodactylidae
Common Coqui
Eleutherodactylus coqui
Bufonidae
Marine Toad
Rhinella marina
Leptodactylidae
White-lipped Frog
Leptodactylus albilabris
Hylidae
Red-snouted Treefrog
Scinax ruber
5.6
Ranidae
Pig Frog
Lithobates grylio
38.9
Ranidae
Bull Frog
Lithobates catesbianus
NA
Eleotridae
Fat Sleeper
Dormitator maculatus
NA
Poecilidae
Platyfish
Xiphophorus sp.
NA
Poecilidae
Guppy
Poecilia vivipara
NA
Eleotridae
Bignouth Sleeper
Gobiomorus dormitator
NA
Eleotridae
Atlantic Guavina
Guavina guavina
NA
Cichlidae
African Cichlids
Tilapia sp.
NA
Gobiidae
Sirajo Goby
Sicydium plumieri
NA
Locariidae
Spotted Pleco
Hypostomus plecostomus
NA
Gecarcinidae
Blue Land Crab
Cardisoma guanhumi
NA
Palaemonidae
NCN
Macrobrachiumcrenulatum
NA
Atyidae
Yellow-nose Shrimp
Xiphocaris elongate
NA
Ampullariidae
Applesnail
Pomacea sp.
NA
Ampullariidae
NCN
Marisa cornuarietis
NA
Planorbidae
Bloodfluke Planorb
Biomphalaria glabrata
NA
Scolopendridae
Centipede
Scolopendra alternans
NA
Termitidae
Termite
Nasutitermes costalis
NA
Reptiles
Amphibians
1613.9
2.8
127.8
Fishes
Macro invertebrates–
aquatic
Macroinvertebrates–
terrestrial
Figure 2. Unrooted neighbor-joining tree of (A) nuclear and (B) chloroplast microsatellites of
different Pterocarpus officinalis populations drawn with Phylips 6.1 (Felsenstein 1993).
Bootstrap values are presented at the base of branches (Muller et al. 2009).
A
B
Figure 3. Non-metric multidimensional scaling ordination plots based on AFLP data for the
three spatial scales considered; (a) watershed (Luquillo, Puerto Rico), (b) island (Puerto Rico)
and (c) regional scales (Caribbean). Each point represents an individual of Pterocarpus
officinalis. All of the NMDS ordinations plots were based on a Sorensen (Bray-Curtis)
distance measure. Significance of groups representing individuals with similar within
variation in ordination space are represented graphically by ellipses (Rivera-Ocasio et al.
2006).
A) Watershed
B) Island
C) Region
Figure 4. BioNJ phylogenetic tree based on 16S-23S rRNA intergenic sequences of 17
Bradyrhizobium spp. strains (in bold) isolated from Pterocarpus officinalis (of the 66 strains,
17 strains were represented; Underlined strains with similar sequences were not represented in
tree; PR, Puerto Rico; G, Guadeloupe; M, Martinique, FG, French Guiana). Only bootstrap
probability values higher than 80% (100 replications) are given at the branching points. Gaps
were not considered. Scale indicated 2% sequence divergence. Bradyrhizobium denitrificans
was chosen as the outgroup (Le Roux et al. 2014).
0.02
B. liaoningense genosp. III LMG 18230T (AJ279301)
B. betae LMG 21987T (AJ631967)
B. canariense BTA-1T (AY386708)
B. japonicum genosp. I LMG 6138T (AJ279264)
B. huanghuaihaiense CCBAU 23303T (HQ428043)
B. arachidis CCBAU 051107T (HM107198)
B. iriomotense LMG 24129T (AB300993)
B. japonicum genosp. IV LMG 8321 (AJ279317)
Bradyrhizobium sp. CIRADAc12 (DQ311111)
B. yuanmingense CCBAU 10071T (AY386734)
B. daqigense CCBAU15774T (HQ231312)
Bradyrhizobium genosp. V LMG 11955 (AJ534590)
96 PR 16_3
G 1844
G 1852
Gbpom 6
G 1866
88
M 1b1
M 1a3
G 1861
Gbp 9_2
FG 7_1
86
FG 9_1
FG 15_2
99
93
FG 7_10_10
82
FG 5_1
100 FG 7_10_13
Bradyrhizobium sp. Tv2a-2 (AY187818)
89
FG 11_10_1
87
PR 1_3
Bradyrhizobium sp. STM3567 (FJ002413)
B. jicamae PAC68T (AY628094)
100
B. lablabi CCBAU 23086T (GU433583)
B. pachyrhizi PAC48T (AY628092)
89
80
Bradyrhizobium genosp. VI LMG8293 (AJ279311)
Bradyrhizobium genosp. XI LMG11951 (AJ534594)
98
Bradyrhizobium genosp. X LMG10705 (AJ534592)
94
B. elkanii genosp. II LMG6134T (AJ279308)
Bradyrhizobium genosp. VII LMG10671 (AJ279272)
95
Bradyrhizobium genosp. IX LMG10677 (AJ534597)
B. denitrificans LMG8443T (AF338176)
97
85
Table 2. Effects of cross inoculation of three Pterocarpus officinalis provenances with three
bradyrhizobial strains1 on plant dry weight, shoot height, nodule number and leaf N content
after 4 months of plant growth (Le Roux et al. 2014).
Factor tested 2
Treatment 3
P. officinalis
Marie-Galante
provenance
(MG)
French Guiana,
Plant dry
Shoot
Nodule
Leaf N
weight
height
number
content
(g.plant-1)
(cm)
(/plant)
(g.kg-1)
1.85 a
34.8 b
30.9 a
13.4 b
1.36 b
40.0 a
20.8 b
21.5 a
1.29 c
38.6 a
20.9 b
13.0 b
Crique A. Jacques
(FG)
Guadeloupe, Le
Moule (Glmo)
P<0.001
P<0.001
FG 5_1
1.58 a
37.9 a
25.0 a
15.5 b
Glmo 3_2
1.49 b
38.6 a
26.6 a
15.1 b
MG 17_1
1.42 c
37.0 a
21.1 a
17.3 a
P<0.001
NS
NS
MG x FG 5_1
1.81
34.8
14.8
MG x Glmo 3_2
1.87
36.2
35.0
9.0
MG x MG 17_1
1.86
33.5
43.0
22.0
FG x FG 5_1
1.48
40.4
40.3
26.7
FG x Glmo 3_2
1.40
40.0
11.2
18.7
FG x MG 17_1
1.19
39.5
10.8
19.2
Glmo x FG 5_1
1.46
38.5
19.8
10.5
Glmo x Glmo 3_2
1.19
39.6
33.5
17.7
Glmo x MG 17_1
1.21
37.9
9.5
10.8
P<0.001
NS
P<0.001
P<0.001
Significance level
Bradyrhizobium strain
Significance level
Interaction
P<0.05
P<0.001
P<0.05
9.3
provenance x strain
Significance level
1
Origins of Bradyrhizobium strains: FG 5_1 (French Guiana), MG 17_1 (Marie-Galante, Guadeloupe), Glmo
3_2 (Le Moule, Guadeloupe)
2
The effects of Bradyrhizobium strain and P. officinalis provenance factors were tested through two-way
analyses of variance (6 replicates were used in the 9 strain x provenance combinations tested). For each
parameter, the means per Bradyrhizobium strain or per P. officinalis provenance followed by different letters are
significantly different according to the Newman & Keuls multiple range test at P=0.05.
3
In bold characters are indicated the homologous Bradyrhizobium strain x P. officinalis provenance associations.
When homologous associations gave higher values than heterologous ones for each parameter analyzed, these
values were indicated in bold characters.
Table 3. Comparison of forest structure from P. officinalis swamps from Puerto Rico (PR)
and Guadeloupe (G).
Locality
Sabana Seca (PR)
Golconde Swamp (G)
Stem density
(stem ha-1)
1,843
Pterocarpus
Stem density %
97
Basal area
(m2 ha-1)
54.1
Canopy
height (m)
10.3
2,567
71
50.14
17
Table 4. Pterocarpus officinalis stand dynamics at different salinity levels at Sabana Seca
locality in Puerto Rico.
Variable
Litterfall a
Measurement
Units
Mg ha-1 yr-1
Low salinity
(10 ‰)
4.8
High salinity
(15 ‰)
1.8
Flower productiona
g m2yr-1
3.2
0.3
Fruit productiona
g m2yr-1
13.3
0
% Increment basal areab
m2 ha-1
37
8
Adult density b
Individuals/m2
0.16
0.14
Adult mortalityb
% (Dead/total)
4
12
Juvenile densityb
Individuals/m2
0.19
0.02
Juvenile mortalityb
% (Dead/total)
27
48
New individuals/m2
2
0.3
Juvenile recruitmentb
a
Eusse and Aide 1999, bRivera-Ocasio et al. 2007
Table 5. Soil salinity, mycorrhizal soil infectivity (Most Probable Number and confident
limits) and mycorrhizal colonization of Pterocarpus officinalis at the end of the wet and dry
seasons (Saint-Etienne et al. 2006).
Plots
Salinity (‰)
(0-20 cm)
Water depth
(cm)
MPN*
(100 g)
Confident
limits
(p<5%)
Root**
colonization
(%)
Dry season
1
26
4
75 a
35-160
14.5 a
2
20
3
150 a
70-320
29.4 b
3
15,5
21
560 b
262-1196
47.5 c
4
10
-12
2400 bc
1123-5126
66.0 d
5
3
-3
3402 c
1592-7266
76.9 e
6
2
-7
3800 c
1779-8166
82.8 e
Wet season
1
22
16
20 a
9-42
22.1 a
2
15
15
31 a
15-67
40.3 b
3
13.5
23
90 bc
42-191
48.8 b
4
8
0
1157 d
542-2472
65.1 c
5
3
9
688 d
322-1469
68.9 c
6
5
5
301 cd
141-644
75.1 c
* Values followed by the same letters are not significantly different (Confident limits, p<5%)
** Values followed by the same letters are not significantly different (Newman-Keuls test,
p<5%)
Figure 7. Mean root nodule a) diameter (mm) and b) mass (g) in relation to depth from the
surface. c) Nodule mass comparison between montane and coastal individuals of P. officinalis
(Pérez and Heartsill 2008).
Table 6. Correlation coefficients (r) between mycorrhizal soil infectivity (Most Probable
Number values), soil salinity (S‰) and mycorrhizal colonization of roots (% AM
colonization) (Saint-Etienne et al. 2006).
Dry season
S
AM (%)
MPN
-0.96*
0.95*
S
-0.99*
*Significant (p<0.05).
MPN
MPN
Wet season
S
AM (%)
-0.86*
0.75*
-0.97*