Unravelling the evolutionary history and promoting conservation genetics of Anadenanthera colubrina var. cebil (Leguminosae), a paradigmatic species in Seasonally Dry Tropical Forests

Name: Unravelling the evolutionary history and promoting conservation genetics of Anadenanthera colubrina var. cebil (Leguminosae), a paradigmatic species in Seasonally Dry Tropical Forests
Keywords: plastid genome

Datos de investigación

Unravelling the evolutionary history and promoting conservation genetics of Anadenanthera colubrina var. cebil (Leguminosae), a paradigmatic species in Seasonally Dry Tropical Forests

Autores: Zerda Moreira, Andrea; García, María Victoria Icon

; Barrandeguy, Maria Eugenia Icon

Publicador: Consejo Nacional de Investigaciones Científicas y Técnicas

Fecha de depósito: 14/06/2024

Fecha de creación: 01/2024-06/2024

Clasificación temática:

Genética y Herencia

Resumen

Seasonally Dry Tropical Forests (SDTFs) exhibit a patchy distribution throughout the Neotropics and are considered one of the most threatened tropical forests in the world. Nonetheless, their conservation has often been neglected in comparison to efforts directed towards other biomes. The distribution pattern of Anadenanthera colubrina (Vell.) Brenan var. cebil (Griseb) Altschul allows an adequate mapping of this biome. The main objectives of this work are to study the historical processes that determined the current distribution of A. colubrina var. cebil under the hypothesis that the demographic history of the remaining fragments of SDTFs in Argentina leads back to a single event of ancestral divergence, and identify Argentinean localities with priority for conservation. Plastid genetic diversity of A. colubrina var. cebil from northern Argentina is characterised using three plastid microsatellite loci. From this, genetic characterization, population genetics, and Bayesian inference were carried out. The results from approximate Bayesian computations show that the Misiones and Piedmont Subandean nuclei diverged from an ancestral population in the early Neogene while the Formosa population derived from the Misiones nucleus in the late Neogene. Two out of five locations from the Misiones nucleus and three of nine locations from the Subandean Piedmont nucleus are proposed as priority conservation areas.

Métodos

Sample collection Anadenanthera colubrina belongs to order Fabales, family Leguminosae, subfamily Caesalpinioideae (ex Mimosoideae, LPWG 2017). Two varieties were described: A. colubrina var. colubrina and A. colubrina var. cebil (Altschul 1964) while Martinez et al. (2013) confirmed the presence of both varieties in Argentina and described the morphological traits for distinguishing them. Anadenanthera colubrina var. cebil is locally known as ‘curupay’ or ‘cebil colorado’ in Argentina (Cialdella 2000). This species is fast-growing and trees can reach 35 m in height. Its bark is greyish, with longitudinal striations and conical protuberances or ‘mamelons’. The crown has a feathery appearance due to its fine foliage (López Villalba et al. 2002) with bipinnate, pale green compound leaves, up to 25 cm long with 8–20 pairs of pine cones with nectaries between each pair and numerous fine leaflets. The hermaphrodite flowers are yellow, grouped in axillary raceme, and the fruit is a flat, reddish-brown legume up to 25 cm long and 3 cm wide with a wavy edge and compressed oblong seeds (Alstchul 1964, López Villalba et al. 2002, Tortorelli 2009). The seeds have been used by numerous native populations since ancient times for their hallucinogenic power (Arenas 1992). It has hard, resistant, and highly valuable reddish-brown wood, with rural and naval uses, in buildings, posts, good quality beams, door and window frames, floors, firewood, and charcoal, among others. The bark has a high concentration of tannins that are used in leather factories (Tortorelli 2009). Recently, Feres et al. (2021) reported a mixed mating system and low effective pollination neighbourhood sizes for A. colubrina var. cebil. The analysed localities from SDTFs nuclei are characterised by woody flora dominated by deciduous species belonging to the families Leguminosae, Euphorbiaceae, Anacardiaceae, and Bignoniaceae (Pennington et al. 2000). A total of 119 adult individuals of A. colubrina var. cebil from 14 localities within the Subandean Piedmont and Misiones nuclei, and one isolated population from Formosa, were analysed (Supporting Information, Table S1 and Fig. 1B). The distances between sampled localities range from 18 to 1300 km. The analysed individuals were identified as A. colubrina var. cebil according to the morphological description given by Martinez et al(2013). Young leaves of adult trees were collected and preserved in silica gel in labelled sealed bags until processing in the laboratory. Every collected individual was georeferenced through the global positioning system. Five dried leaves per individual are kept in the herbarium of the Laboratory of Landscape and Population Genetics (Facultad de Ciencias Exactas, Químicas y Naturales, Universidad Nacional de Misiones, Argentina). Also, one individual from Tucumán (MCNS11943), one individual from Formosa (MCNS11945), and two individuals from Jujuy (MCNS11941 and MCNS11942) are stored in the herbarium of Universidad de Salta (Argentina). DNA extraction, amplification, and fragment analysis Total DNA was extracted using the ROSE extraction protocol (Steiner et al. 1995) modified by García et al. (2007). Agarose gels at 1% were used to check the DNA concentration and quality using different concentrations of λ DNA as reference. Three plastid microsatellite (cpSSR) loci (Ccmp4, Ccmp5, and Ccmp7) developed by Weising and Gardner (1999) were analysed. Each forward primer was modified by addition of the sequence of the universal primer M13 (tgtaaaacgacggccagt) to its 5ʹ end for the subsequent genotyping of the individuals using the method proposed by Schuelke (2000) and modified by Goncalves et al. (2013). Polymerase chain reactions (PCRs) were performed in a 14 μL final volume with 0.5 ng of DNA;1 X of KCl buffer; 2.5 mM Cl2Mg (Thermo Scientific—Fermentas); 0.2 mM of each dNTP (Thermo Scientific—Fermentas); 0.75 U of Taq polymerase (Thermo Scientific—Fermentas); 0.33 μM of Reverse primer; 0.20 μM of M13-Forward primer and 0.33 μM of primer M13-FAM (Ccmp4, Ccmp5) or primer M13-HEX (Ccmp7). The amplification reactions were carried out in a BIOMETRA thermocycler following a Touchdown type protocol where the annealing temperature was −1°C in each cycle. Touchdown was implemented with the following conditions: 95°C for 4 min, six cycles of 45 s at 95°C, 30 s annealing with corresponding temperature regimes (Ccmp4: 55–50°C, Ccmp5: 60–55°C, and Ccmp7: 56–49° C), and 45 s at 72°C followed by 23 similar cycles using each fixed temperature (Ccmp4: 50°C, Ccmp5: 55°C, and Ccmp7: 49°C) for annealing. Eight cycles of 45 s at 95°C, 30 s at 53°C, and 45 s at 72°C were performed to include fluorescently labelled universal primer (M13 with FAM or HEX), and a final elongation at 72°C for 20 min. Amplifications were checked by electrophoresis in 1.5% agarose gels with a 1× TBE buffer stained with 1% Gel Red using a DNA ladder of 100 bp as a molecular size marker (Productos Biológicos). Fragment sizes were determined by the Applied Biosystems 3730/3730xl DNA analyser (Macrogen) through the use of size standards GeneScan 400 HD ROX and GeneScan 500 LIZ (Applied Biosystems). Identification of fragment sizes was performed using Peak Scanner v.1.0 (Applied Biosystems).

Palabras clave: plastid genome, microsatellite markers, curupay, genetic diversity conservation

Alcance geográfico

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Identificador del recurso

URI: http://hdl.handle.net/11336/238123

Colecciones

Datos de Investigación(IBS)
Datos de Investigación de INSTITUTO DE BIOLOGIA SUBTROPICAL

Datos de Investigación(INBIONATEC)
Datos de Investigación de INSTITUTO DE BIONANOTECNOLOGIA DEL NOA

Citación

Zerda Moreira, Andrea; García, María Victoria; Barrandeguy, Maria Eugenia; (2024): Unravelling the evolutionary history and promoting conservation genetics of Anadenanthera colubrina var. cebil (Leguminosae), a paradigmatic species in Seasonally Dry Tropical Forests. Consejo Nacional de Investigaciones Científicas y Técnicas. (dataset). http://hdl.handle.net/11336/238123

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