Amadou M. Bâ 1* and Elsie Rivera-Ocasio 2

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

1Laboratoire 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:

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 Ríos-López (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 / Country
Mammals (non-volant)
Muridae / Black Rat / Rattus rattus / NA / Puerto Rico
Birds
Ardeidae / Green Heron / Butorides virescens / 0.42–0.57 / Puerto Rico
Falconidae / American Kestrel / Falco sparverius / 0.01–0.01 / Puerto Rico
Columbidae / Zenaida Dove / Zenaida aurita / 0.26–0.37 / Puerto Rico
Columbidae / Common Ground-Dove / Columbina passerine / 0.00–0.02 / Puerto Rico
Cuculidae / Mangrove Cuckoo / Coccyzus minor / 0.21–0.69 / Puerto Rico
Cuculidae / Smooth-billed Ani / Crotophaga ani / 0.02–0.07 / Puerto Rico
Picidae / Puerto Rican Woodpecker / Melanerpes portoricensis / 0.20–0.49 / Puerto Rico
Tyrannidae / Gray Kingbird / Tyrannus dominicensis / 0.36–0.17 / Puerto Rico
Tyrannidae / Loggerhead Kingbird / Tyrannus caudifasciatus / 0.00–0.02 / Puerto Rico
Tyrannidae / Puerto Rican Flycatcher / Myiarchus antillarum / 0.05–0.20 / Puerto Rico
Vireonidae / Black-whiskered Vireo / Vireo altiloquus / 0.00–0.23 / Puerto Rico
Turdidae / Red-legged Thrush / Turdus plumbeus / 0.05–0.14 / Puerto Rico
Mimidae / Pearly-eyed Thrasher / Margarops fuscatus / 0.01–0.02 / Puerto Rico
Parulidae / Northern Waterthrush / Seiurus noveboracensis / 0.79–0.82 / Puerto Rico
Parulidae / Ovenbird / Seiurus aurocapilla / 0.04–0.13 / Puerto Rico
Parulidae / Black-and-white warbler / Mniotilta varia / 0.06–0.12 / Puerto Rico
Parulidae / Prarie Warbler / Dendroica discolor / 0.02–0.13 / Puerto Rico
Parulidae / Common Yellowthroat / Geothlypis trichas / 0.03–0.06 / Puerto Rico
Parulidae / American Redstart / Setophaga ruticilla / 0.05–0.05 / Puerto Rico
Parulidae / Prothonotary Warbler / Protonotaria citrea / 0.00–0.04 / Puerto Rico
Parulidae / Northern Parula / Parula Americana / 0.00–0.01 / Puerto Rico
Parulidae / Yellow Warbler / Dendroica petechial / 0.00–0.01 / Puerto Rico
Coerebidae / Bananaquit / Coereba flaveola / 3.24–4.97 / Puerto Rico
Thraupidae / Puerto Rican Spindalis / Spindalis portoricensis / 0.01–0.03 / Puerto Rico
Emberizidae / Black-faced Grassquit / Tiaris bicolor / 0.00–0.02 / Puerto Rico
Icteridae / Greater Antillean Grackle / Quiscalus niger / 0.11–0.62 / Puerto Rico
Icteridae / Shiny Cowbird / Molothrus bonariensis / 0.01–0.07 / Puerto Rico
Estrildidae / Oranged-cheeked Waxbill / Estrilda melpoda / 0.00–0.01 / Puerto Rico
Reptiles
Dactyloidae / Puerto Rican Crested Anole / Anolis cristatellus / 1097.2 / Puerto Rico
Dactyloidae / Puerto Rican Saddle Anole / Anolis stratulus / 197.2 / Puerto Rico
Dactyloidae / Puerto Rican Grass Anole / Anolis pulchellus / 30.6 / Puerto Rico
Iguanidae / Green Iguana / Iguana iguana / NA / Puerto Rico
Emydidae / Antillean Slider / Trachemys stejnegeri / 2.8 / Puerto Rico
Amphibians
Eleutherodactylidae / Common Coqui / Eleutherodactylus coqui / 1613.9 / Puerto Rico
Bufonidae / Marine Toad / Rhinella marina / 2.8 / Puerto Rico
Leptodactylidae / White-lipped Frog / Leptodactylus albilabris / 127.8 / Puerto Rico
Hylidae / Red-snouted Treefrog / Scinax ruber / 5.6 / Puerto Rico
Ranidae / Pig Frog / Lithobates grylio / 38.9 / Puerto Rico
Ranidae / Bull Frog / Lithobates catesbianus / NA / Puerto Rico
Fishes
Eleotridae / Fat Sleeper / Dormitator maculatus / NA / Puerto Rico
Poecilidae / Platyfish / Xiphophorus sp. / NA / Puerto Rico
Poecilidae / Guppy / Poecilia vivipara / NA / Puerto Rico
Eleotridae / Bignouth Sleeper / Gobiomorus dormitator / NA / Puerto Rico
Eleotridae / Atlantic Guavina / Guavina guavina / NA / Puerto Rico
Cichlidae / African Cichlids / Tilapia sp. / NA / Puerto Rico
Gobiidae / Sirajo Goby / Sicydium plumieri / NA / Puerto Rico
Locariidae / Spotted Pleco / Hypostomus plecostomus / NA / Puerto Rico
Macro invertebrates–aquatic
Gecarcinidae / Blue Land Crab / Cardisoma guanhumi / NA / Puerto Rico
Palaemonidae / NCN / Macrobrachiumcrenulatum / NA / Puerto Rico
Atyidae / Yellow-nose Shrimp / Xiphocaris elongate / NA / Puerto Rico
Ampullariidae / Applesnail / Pomacea sp. / NA / Puerto Rico
Ampullariidae / NCN / Marisa cornuarietis / NA / Puerto Rico
Planorbidae / Bloodfluke Planorb / Biomphalaria glabrata / NA / Puerto Rico
Macroinvertebrates–terrestrial
Scolopendridae / Centipede / Scolopendra alternans / NA / Puerto Rico
Termitidae / Termite / Nasutitermes costalis / NA / Puerto Rico

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).

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).

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).

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 / Plant dry weight (g.plant-1) / Shoot height
(cm) / Nodule
number (/plant) / Leaf N
content (g.kg-1)
P. officinalis provenance / Marie-Galante (MG) / 1.85 a / 34.8 b / 30.9 a / 13.4 b
French Guiana, Crique A. Jacques (FG) / 1.36 b / 40.0 a / 20.8 b / 21.5 a
Guadeloupe, Le Moule (Glmo) / 1.29 c / 38.6 a / 20.9 b / 13.0 b
Significance level / P<0.001 / P<0.001 / P<0.05 / P<0.001
Bradyrhizobium strain / 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
Significance level / P<0.001 / NS / NS / P<0.05
Interaction provenance x strain / MG x FG 5_1 / 1.81 / 34.8 / 14.8 / 9.3
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
Significance level / P<0.001 / NS / P<0.001 / P<0.001

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 / Stem density
(stem ha-1) / Pterocarpus
Stem density % / Basal area
(m2 ha-1) / Canopy height (m)
Sabana Seca (PR) / 1,843 / 97 / 54.1 / 10.3
Golconde Swamp (G) / 2,567 / 71 / 50.14 / 17

Table 4. Pterocarpus officinalis stand dynamics at different salinity levels at Sabana Seca locality in Puerto Rico.

Variable / Measurement
Units / Low salinity
(10 ‰) / High salinity
(15 ‰)
Litterfall a / Mg ha-1 yr-1 / 4.8 / 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
Juvenile recruitmentb / New individuals/m2 / 2 / 0.3

aEusse 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 / Wet season
MPN / S / AM (%) / MPN / S / AM (%)
MPN / -0.96* / 0.95* / -0.86* / 0.75*
S / -0.99* / -0.97*

*Significant (p<0.05).