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T5. Sources and methodology

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Authors 7

Heloisa Dantas Brum1*, Alexandre F. Souza2

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1 - Programa de Pós-Graduação em Ecologia, CB, Universidade Federal do Rio Grande do 9

Norte, Campus Universitário, Lagoa Nova, Natal 59072-970, RN, Brazil. E-mail: 10

[email protected] 11

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2 - Departamento de Ecologia, CB, Universidade Federal do Rio Grande do Norte, Campus 13

Universitário, Lagoa Nova, Natal 59072-970, RN, Brazil. E-mail: 14 [email protected] 15 16 *Corresponding author 17

Programa de Pós-Graduação em Ecologia, CB, Universidade Federal do Rio Grande do 18

Norte, Campus Universitário, Lagoa Nova, Natal 59072-970, RN, Brazil. 19

Telephone number: +55 (84) 99915-4336 20

[email protected] 21

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Abstract

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Euterpe precatoria (açaí) is the most abundant plant species in the Amazon basin and one 23

of the main non-timber forest products of the continent. A thorough understanding of

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the ecology of this species is needed to support sustainable management initiatives.

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Resource availability, disturbance regime, and human management are some of the main

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factors influencing population structure. Here we described life stages of E. precatoria,

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evaluated their allometric relationships, and assessed the effects of habitat type

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(floodplain and unflooded upland) and proximity of human settlements on population

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size distribution in Central Amazon near the Purus River. The height:diameter relationship

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increased from Seedlings to Juvenile 2 but decreased from Juvenile 2 to Reproductive 2,

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indicating changing height investment for any given diameter along these life stages.

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There was a marked habitat dependency in both the density and population size

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distribution, with populations in upland forests dominated by juveniles while populations

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in the floodplains were dominated by reproductive palms. Nearness of human

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settlements was not related to population structure parameters. The patterns we

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uncovered have implications for our interpretation of widespread Amazon forest species

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that inhabit habitats with contrasting disturbance regimes and resource levels like

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flooded várzea and upland terra firme.

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Key-words: Várzea, Terra firme, Population structure, Forest Management, Skewness, 41 Allometry. 42 43 44 45

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Introduction

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Euterpe precatoria Mart. is an iconic palm tree popularly known as açaí or assaí in South

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America. It is the most abundant species in the Amazon basin, being one of the few 48

hyperdominant species in four of the broad five forest types in the region (ter Steege et 49

al. 2013). It is also one of the main non-timber forest products of the continent (Stoian 50

2004). The species is traditionally used for the extraction of fruits, heart of palm, and 51

timber by indigenous peoples (Anderson 1977; Albert and Tourneau 2007). It produces 52

large quantities of dark single-seeded fruits that, as in other Euterpe species, is dispersed 53

by many birds and mammals (Leite et al. 2012). The fruit is commonly collected and a 54

regional market developed around its use to produce a creamy beverage known as ‘açaí 55

wine’ that is very nutritious and bears superior antioxidant and anti-inflammatory 56

properties (Kang et al. 2012) that have been shown to bring health benefits like memory 57

protection when used as a dietary supplementation (Carey et al. 2017). Euterpe 58

precatoria fruit consumption may be a sustainable alternative to this species’ use in

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countries like Bolivia, where the traditional consumption of palm hearth implies that 60

palms are killed for resource extraction (Rocha 2004; Velarde and Moraes 2008). Most 61

recently prices for E. precatoria products have increased dramatically due to the global 62

commodization of palm hearts and raw, processed, or lyophilized fruit pulp as nutritional 63

supplement that are increasingly consumed in Brazil as well as exported to countries 64

outside South America, mainly Europe, Canada and USA (Bussmann and Zambrana 2012). 65

Market sales for açaí berry (E. oleracea and E. precatoria combined) amount USD 126.3 66

millon/yr in Brazil alone (Martinot et al. 2017). With increasing trade volumes, E. 67

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precatoria and many other exploited native species cannot meet demand in a sustainable

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manner (Mostacedo and Fredericksen 1999; Stoian 2004; Vallejo et al. 2016). A thorough 69

understanding of the biology and ecology of this species is thus needed, in order to 70

support sustainable management initiatives (Velarde and Moraes 2008). 71

One of the most easily measurable population parameter in the field is population 72

structure, which integrates in itself a wealth of demographic information (Avalos et al. 73

2013; Peltzer et al. 2014). Yet, it may be a puzzling tool for the assessment of natural 74

populations, because of two reasons. The first is that the interpretation of population 75

structure may be easier if biologically relevant classes are clearly identified, e.g. through 76

the identification of life stages instead of arbitrary size classes (Gatsuk et al. 1980; Souza 77

et al. 2000, 2003; Caswell 2001). The second is that the patterns of relative abundances 78

of juveniles and adults they portray are no reliable proxy of future population growth and, 79

therefore, of population persistence in any given habitat or set of environmental or 80

human-imposed conditions (Johnson et al. 1994; Condit et al. 1998; Souza 2007; Virillo 81

et al. 2011; Bin et al. 2012). Attempts to infer population persistence from static 82

population structure and the relative abundance of juveniles relative to adults are, 83

therefore, flawed. In fact, population size structures result from the realized growth, 84

mortality, and fecundity rates across different size classes (Caswell 2001; Wright et al. 85

2003). A more fruitful use of population structure is the comparison of population size 86

distributions across habitats with contrasting disturbance histories or between co- 87

occurring species in the same habitat, which allows the detection of functional groups of 88

species with similar life histories (Swaine et al. 1990; Poorter et al. 1996; Wright et al. 89

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2003; Souza 2007; Souza et al. 2008, 2010). Following the fast-growing resource acquiring 90

vs slow growth stress tolerant life history gradient repeatedly found worldwide (Grime 91

and Pierce 2012; Reich 2014; Díaz et al. 2016), left-skewed size distributions dominated 92

by large individuals characterize chronic recruitment failure of gap-dependent species 93

with large fecundities, high seedling mortality, and high sapling growth, while right- 94

skewed size distributions dominated by small individuals characterize shade-tolerant 95

species with the opposite traits (Lorimer and Krug 1983; Swaine et al. 1990; Poorter et al. 96

1996; Wright et al. 2003; Souza 2007; Souza et al. 2008; Vlam et al. 2014). 97

Among the factors that influence population structure are the effects of resource 98

availability, the disturbance regime, and human management (Mostacedo and 99

Fredericksen 1999; Souza and Martins 2004; Souza 2007; Avalos et al. 2013; Peltzer et al. 100

2014). Habitat differentiation between várzea floodplains and upland terra firme are 101

among the main drivers of both light resource and disturbance levels in the Amazon 102

basin. The Amazon basin harbors a great variety of floodplains that cover nearly 500,000 103

Km2 (Junk et al. 2011) that includes a great variety of habitats such as seasonally

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inundated forests and swamps, with plant communities adapted to differing flooding 105

regimes (Junk et al. 2012). These hydro-edaphic conditions exert strong control on tree 106

species distributions, connecting floodplain floras even when rivers drain distinct climatic 107

regions (Wittmann and Junk 2003). Environmental variation linked to flooding regime 108

affects demographic parameters of neighboring populations, and may be detectable 109

even over short geographic distances (Otárola and Avalos 2014). The disturbance caused 110

by seasonal flooding involves oxygen deprivation, sedimentation, and mechanic damage 111

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that frequently kills younger plants (Parolin 2008). It has triggered the evolution of 112

metabolic pathways promoting either endurance or escape strategies (Parolin 2008; 113

Voesenek and Bailey-serres 2015). Contrary to flooded forests, which present lower and 114

more irregular canopies allowing more light penetration (Souza and Martins 2005; 115

Sawada et al. 2015)⁠, in uplands the main limitation to plant growth and establishment is 116

the deep shade cast by tall canopies, which severely reduce understory light levels 117

(Svenning 2001, 2002; Myster 2016). 118

The shady understory environments have induced the evolution of ecological strategies 119

that fall along the low growth/size shade stress tolerant – fast growth/height light 120

acquisition trade-off (Grime 1977; Forgiarini et al. 2013; Reich 2014). In palms, the lack 121

of secondary meristems precludes exploitation of light through lateral growth, restricting 122

possible plant responses to shading to increases in allometric height: diameter 123

increments (Avalos and Otárola 2010). The height growth of Euterpe precatoria is 124

supported by stilt roots whose size scales with plant height rather than with topographic 125

variation (Avalos and Otárola 2010), but the effects of habitat variation between upland 126

and floodplains on palm shape and allometry are poorly known. Even less studied are the 127

effects of human management on E. precatoriapopulation ecology. These effects area 128

potentially sizable because the species is considered domesticated or semi-domesticated 129

by indigenous people since pre-Columbian times (Clement et al. 2015; Levis et al. 2017)⁠. 130

The intensive management of preferred species by indigenous and current locals has 131

taken place mainly along the floodplain margins of large rivers (McMichael et al. 2012), 132

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and is thus expected to influence the abundance of the species across different habitat 133

types (Ticktin et al. 2012; Baldauf and Maës dos Santos 2013). 134

Here we describe the macromorphological life stages of E. precatoria, evaluate their 135

allometric relationships, and assess the effects of habitat type (floodplain and unflooded 136

upland) and proximity of human settlements on population size distribution in Central 137

Amazon. Specifically, we tested the following hypotheses: 1) Due to the higher and more 138

closed canopies in upland than in floodplain forests (Myster 2016), established palms 139

should present taller and slenderer trunks in upland forests. Therefore, there should be 140

a significant interaction between height vs. diameter allometric relationship and habitat; 141

2) Because E. precatoria forms seedling banks in upland shaded understory (Peña-Claros 142

and Zuidema 2000; Rocha 2004; Isaza et al. 2017), we infer that the species present a 143

moderate degree of shade tolerance (Condit et al. 1998; Mostacedo and Fredericksen 144

1999) that allows it to wait for canopy opening increases (Avalos et al. 2013; Otárola and 145

Avalos 2014). We thus expect right-skewed size distributions in upland habitats where 146

adult recruitment and productivity are light-limited. Considering that in the floodplains 147

flooding disturbs seedling recruitment by killing seedlings (Parolin 2008), we also expect 148

reduced size distribution skewness or even symmetrical size distributions. This would 149

reflect the prevalence of larger plants in brighter and more productive floodplains, which 150

would be more favorable to adult growth and survivorship once the juvenile recruitment 151

bottleneck is surpassed (Avalos et al. 2013; Otárola and Avalos 2014). 3) Given the 152

rationale above, we expect to find greater seedling and juvenile densities in upland but 153

greater adult densities in floodplains (Rocha 2004; Velarde and Moraes 2008; Otárola and 154

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Avalos 2014); 4) Due to negative effects of fruit harvesting on population recruitment 155

(Mostacedo and Fredericksen 1999), we expect that proximity to human settlements 156

depress the number of juveniles. Hence, we expect to find a positive relationship 157

between the distance from the nearest human settlement and the skewness of 158

population size distribution (where more positive skewness reflects increased juvenile 159

abundance). The outcomes of these tests can help stakeholders and local users to deal 160

with management decisions. 161

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Methods

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Study area and species

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Euterpe precatoria is a single-stemmed palm (but see Avalos and Schneider 2011)that

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may reach up to 20 m height occurring from Central America to Bolivia (Stoian 2004). 166

Individuals are obligate outcrossing monoecious and present a long flowering time that 167

provide easily accessed resources to thousands of insect flower visitors (Kuchmeister et 168

al. 1997). It’s large seeds (ca. 11 cm diameter Aguiar and Mendonça 2003) germinate 169

immediately after harvesting (Costa et al. 2018). The ability of E. precatoria to germinate 170

in darkness and its preference for relatively mild germination temperatures (20ºC) may 171

favor its establishment in seasonally flooded habitats, while its tolerance to moderate 172

desiccation (Costa et al. 2018) allows germination in upland forests. As show for other 173

Euterpe species, germination is likely facilitated by frugivorous dispersal (Leite et al. 174

2012). Seedlings tolerate shade and do not respond promptly to increased irradiance 175

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(Coelho et al. 2015). The species attains higher densities, sizes, and fruit productivity in 176

floodplains than in unflooded uplands, where it shows etiolation signs (Velarde and 177

Moraes 2008). 178

The study was carried out in the Piagaçu-Purus Sustainable Development Reserve (SDR- 179

PP), Amazonas State, located in the Purus-Madeira and Purus-Juruá interfluve (Figure 1). 180

The reserve covers 834,245 ha and allows human habitation and the sustainable use and 181

commercialization of natural resources. It harbors 65 riverine communities and more 182

than 5000 people, who live mainly from family agriculture, fishing, hunting, and the 183

extraction of non-timber forest products like açaí. The SDR-PP is located in the 184

municipalities of Coari and Codajás, which together account for 96% of the 50,000 tons 185

of açaí produced by the Amazonas state annually (IBGE 2018). Açaí is collected in the 186

study region non-destructively by climbing. Although the Purus river presents up to 10 m 187

annual level variation (ANA 2018), the várzea we studied is located at a high várzea area, 188

with seasonal flooding of up to 3 m height and during less than 50 days.year-1 (Wittmann

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et al. 2010). 190

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Figure 1. Lower Purus river area in Central Amazon, showing the location of riverine 193

communities and the 10 blocks where individuals of E. precatoria (açaí) were marked and 194 measured. 195 196 Data collection 197

Sampling was directed at sites selected after a quick participatory mapping with local 198

residents, who indicated areas used for açaí fruit harvesting with differing intensities. 199

Data were collected in 200 20 x 20m permanent plots (total 8 ha) distributed in 10 blocks, 200

five in the floodplain and five in the upland (Figure 1). In each block, plots were arranged 201

in four 100-m long transects, each one subdivided into five plots, and stratified into two 202

topographic positions. In each block, two transects were located in low-lying terrain, thus 203

subjected to more frequent and long-lasting flooding, and two were located in elevated 204

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terrain, thus subjected to less frequent and short-lasting flooding (approximately with 205

one month of difference). Because the number of newly germinated seedlings was very 206

large in the floodplain plots, we subsampled this life stage only through four 1 x 1 m 207

subplots located at the corners of each 20 x 20m plot. From February to March 2016, all 208

individuals of E. precatoria were tagged and measured for diameter at soil level, total 209

height, and number of leaves in all plots. Individuals in early development that lacked the 210

aerial stem had diameter and height measured at the base, which correspond to the 211

group of leaf sheaths. For seedlings, tags were tied to a wooden (on uplands) or metal (in 212

the floodplains) stick, in order to avoid damaging the plants. For plants with stem base 213

enlargement or with aerial roots, diameter measurement was made above these 214

structures. Presence and height of reproductive structures were also measured. We used 215

the distance to the nearest human settlement as a proxy to the effects of human activities 216

(Duvall 2007; Vandam et al. 2013; Sumarga 2017). Distance to the nearest human 217

settlement was measured as sum of linear distances actually traveled by land and water 218

by the inhabitants of each settlement to reach the location of each transect using a 219

Garmin GPSmap 62s and the QGIS software (QGIS Development Team 2009). 220

Statistical analysis

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Euterpe precatoria life stages were established based on macromorphological traits

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(Gatsuk et al. 1980). Further subdivisions within life stages were established based on 223

breakpoints in the relationship between stem diameter and height. This was assessed 224

through a loess regression between diameter and height after iterative adjustment of 225

alpha (α) and lambda (λ) parameters (Jacoby 2000). All statistical analyses were 226

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performed in the open software R (R Core Team 2017). We performed an analysis of 227

covariance to assess the effect of life stages on the allometric relationship between 228

diameter and height. We used the natural logarithm of height to quantify palm size, and 229

the coefficient of skewness (g1, Zar 1996) to summarize the skewness of the height 230

distributions and, thus, population structure (Wright et al. 2003) at the transect (i.e., 231

blocks of 5 20 x 20 m plots) scale (n = 40). When g1 > 0, it indicated right-skewed height 232

distributions with few tall and many short palms, g1 = 0 indicated symmetrical height 233

distributions, and g1 < 0 indicated left-skewed height distributions with many tall and few 234

short palms (Zar 1996). Seedlings were excluded from population structure analyses 235

because their short-lived duration, mainly in the floodplains, could distort the results 236

(personal observation). 237

We used Generalized Linear Models (GLM) to evaluate habitat (floodplain x upland) and 238

human (distance to the nearest human settlement) on the skewness of population height 239

distribution. Following Zuur et al. (2009) and Plant (2012), different models were fitted in 240

order to decide which model structure best described the error structure in the data. In 241

order to decide whether a random term was necessary, we used the second-order Akaike 242

Information Criterion (AICc, used for small sample sizes) and ANOVA to compare models. 243

Models whose ΔAIC < 2 were regarded as equally plausible model (Burnham and 244

Anderson 2002). We fitted a GLM without a random term (i.e., containing fixed terms 245

only), a Generalized Linear Mixed Model (GLMM) using blocks as a random intercept 246

term, and a GLMM using both blocks and habitat types as random terms (Zuur et al. 247

2009). All models were fit by maximizing the restricted log-likelihood using the functions 248

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gls or lme of the nlme package (Pinheiro et al. 2017). The random effect in the GLMM 249

accounted for the possible lack of independence of the skewness values estimated for 250

the blocks. The 20 x20 m plot-level effects of habitat type and distance to human 251

settlement on palm density were assessed through a GLMM using Poisson error and 252

blocks and transects as nested random terms (Zuur et al. 2009). 253

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Results

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We marked and mapped 3613 individuals of açaí in different life stages in both the 256

floodplains and upland habitats. Observation of external macromorphological structures 257

allowed the distinction of two pre-reproductive stages: seedlings and juveniles. Seedlings 258

were defined as individuals without the presence of an aerial stem. Seedling leaves 259

present fan-shaped leaves (Figure 2). Juveniles were all individuals with an aerial stem 260

and diameter at soil level below the minimum diameter found for reproductive palms, 261

which was 10.0 cm (Figure 2). Juveniles presented a marked discontinuity in their height 262

vs. diameter relationship at ca. 1.5 m height (Figure 3A) and were thus subdivided into 263

Juvenile 1 (height < 1.5 m) and Juvenile 2 (1.5 ≤ height < 10 m). The diameter x height 264

relationship was nonlinear (loess regression: α = 0.35, λ = 2, Figure 3) with two marked 265

slope changes. The first change was located at the transition between Juveniles 1 and 2, 266

and the second among reproductive palms at ca. 23.0 cm diameter, a threshold from 267

which higher diameter values were not accompanied by higher height values (Figure 3). 268

Reproductive palms were thus divided into Reproductive 1 (height ≥10 m and diameter 269

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< 23 cm) and Reproductive 2 (height ≥ 10 m and diameter ≥ 23 cm). The dependency of 270

the relationship between diameter and height on life stages was further confirmed 271

through an ANCOVA (R2 = 0.99; F = 2.64× 104; df = 3598; P = 2.2 × 10-16) (Figure 3B). On 272

the one hand, the declivity of the linear fit increased from Seedlings to Juvenile 1, and 273

from these to Juvenile 2, indicating increasing heights for any given diameter in these life 274

stages. On the other hand, the same declivity decreased from Juvenile 2 to Reproductive 275

1, and from these to Reproductive 2, indicating decreasing height investment for any 276

given diameter in these life stages. A second ANCOVA tested for the effects of habitat 277

type (upland x flooded forest) on the height:diameter relationship, this time excluded 278

both seedlings and Reproductive 2 palms because these two life stages did not occur in 279

upland forests. Habitat type, however, did not alter the height:diameter relationship 280

significantly either as an isolated factor (F = 2.66, df = 1, P = 0.103) or in interaction with 281

life stages (F = 1.479, df = 1, P = 0.2241). 282

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Figure 2. Life stages of Euterpe precatoria (Arecaceae). A. Fruit bunch; B. Detail of fruits; 285

C. Detail of the seed with initial development of seedling; D. Seedling; E. Juvenile 1; F. 286

Reproductive. See text to description of stages characteristics. Drawing by Eliziane Garcia. 287

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Figure 3. A) Overall relationship between height and diameter of 3613 açaí palms 290

(Euterpe precatoria) measured in upland and floodplain forests, Central Amazon. The 291

A B

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fitted line is a Loess regression. B) Height × diameter relationship on a log scale with fitted 292

linear model adjusted by life stage through ANCOVA. 293

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Seedlings were by far the denser life stage, with average 470.75 ± 486.21 plants ha-1 in

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the unflooded areas and 4025.0 ± 5086.31 plants ha-1 in the floodplains. Seedlings apart,

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overall palm density was greater in unflooded forests (66.88 ± 129.38 plants ha-1) than in

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flooded forests (31.13 ± 79.65 plants ha-1). Palm density changed significantly with

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habitat as well as with life stage. The best GLMM model included the interaction between 299

habitat and life stage, as well as blocks and transects as random terms (Table 1). This 300

indicates that population density varied between transects within blocks as well as 301

between blocks in a random yet sizable way. The habitat vs.× life stage interaction term 302

indicates that the density of different stages varied between flooded and unflooded 303

forests in different ways. Unflooded forests showed to be dominated by juveniles, with 304

Juvenile 1 and Juvenile 2 palms having the highest densities. On the contrary, flooded 305

forests were dominated by Reproductive 1, which was the densest life stage (Fig. 4).

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