Robles in Lagunas de Epulauquen, Argentina: previous and recent evidence of their distinctive character
© Azpilicueta et al.; licensee Springer. 2014
Received: 2 June 2014
Accepted: 22 October 2014
Published: 2 December 2014
The northernmost ‘robles’ of Argentina, located in Lagunas de Epulauquen (northwest Patagonia), are considered a population of Nothofagus obliqua, even though they differ in some respects from individuals of other populations of the species. In order to analyse a possible genetic basis of their distinctive character, we revised previous information and added new evidence based on biochemical (isozyme) and molecular (chloroplast DNA and nuclear microsatellite) markers, as well as quantitative trait variation (seed traits, leaf morphology, plant architecture and field performance). The comparison of Lagunas de Epulauquen specimens with specimens from other areas in Argentina demonstrated their genetic distinction. Glacial history, in conjunction with hybridisation processes, provides support for the main hypothesis to explain these results. Future research lines are proposed, aimed at identifying the taxonomic status of the Lagunas de Epulauquen population. The conservation value of these forests is highlighted.
Los robles más septentrionales de la Argentina, ubicados en Lagunas de Epulauquen (noroeste de Patagonia) son considerados una población de la especie Nothofagus obliqua, pero presentan características diferenciales respecto al resto de las poblaciones argentinas de esa especie. El presente trabajo reúne información generada previamente y otra original que demuestra el carácter genéticamente distintivo de esa población. Se presentan los resultados logrados a través de análisis genéticos con marcadores bioquímicos (isoenzimas) y moleculares (de ADN de cloroplasto y marcadores de microsatélites nucleares), con caracteres cuantitativos seminales, de morfología foliar, de arquitectura de las plantas y de aptitud a campo. La historia glacial de la región, en conjunción con procesos de hibridación pasados, se postula como la causa de esta diferenciación. Se propone la realización de nuevas líneas de investigación que permitan confirmar o re-definir el estatus taxonómico de los árboles de esta población. Adicionalmente se presenta una breve consideración sobre el valor de conservación de los bosques en Lagunas de Epulauquen, en relación a los resultados aquí presentados.
Nothofagus obliqua characteristics
Nothofagus obliqua (Mirb.) Oerst. (Nothofagaceae, Hill and Jordan ), known as ‘roble’, ‘pellín’, ‘roble pellín’ or ‘hualle’, is a tree species endemic to South American temperate forests. It belongs to the Lophozonia sub-genus [recently upgraded to genus in the classification by Heenan and Smissen ()] along with other six species (Hill and Read ; Martin and Dowd ; Heenan and Smissen ), three of which are also South American: Nothofagus alpina (= N. nervosa) (Poepp. & Endl.) Oerst., Nothofagus glauca (Phil.) Krasser (Veblen et al. ) and Nothofagus macrocarpa (A. DC.) F. M. Vázquez and R. Rodr. (Vázquez and Rodríguez ).
Natural hybridisation is common among species of the Nothofagus genus. N. obliqua hybridises with N. alpina, especially in sympatric forests (Donoso et al. ; Gallo et al. ; Marchelli and Gallo ; Gallo ). In addition, N. × leonii Espinosa (Donoso and Landrum ; Grant and Clement ) is recognised as the hybrid between N. obliqua and N. glauca. Finally, hybridisation between N. obliqua and N. macrocarpa has been suggested (Vázquez and Rodríguez ).
Robles from Lagunas de Epulauquen
Forests in Lagunas de Epulauquen, growing at 36° 49′ S, 71° 04′ W and 1,500 masl, in an area of 1,501 ha (Sabatier et al. ), are considered the northernmost distribution of N. obliqua in Argentina. The nearest co-specific Argentinean forests are located at a distance of 220 km, at Moquehue lake (Sabatier et al. ), and are separated by mountain chains, which contribute to the geographical isolation of Lagunas de Epulauquen.
The main goal of this study is to review previous results and present new evidence - based on biochemical, molecular, morphological and architectural data - in order to postulate the hypothesis that the origin of the Lagunas de Epulauquen roble population is different to that of other Argentinean populations of N. obliqua.
Previous evidence supporting the distinctiveness of robles from Lagunas de Epulauquen
In Chile, N. obliqua forests grow approximately 50 km west of the Lagunas de Epulauquen population, with high mountains in between. Based on pollen fossil analysis, Markgraf () postulated the occurrence of a post-glacial introgression within the Lagunas de Epulauquen region, coming from the west. Therefore, a historical connection between the two sides of the Andes Mountains at this latitude can be inferred.
Because of its maternal inheritance in the majority of angiosperm species (Harris and Ingram ), chloroplast DNA allows direct estimation of seed-mediated dispersal. Furthermore, the low mutation rate of the chloroplast (Wolfe et al. ), which determines its highly conservative nature, helps in the inference of glacial refugia and post-glacial migration routes of glaciation-affected species. Azpilicueta et al. () identified two different cpDNA haplotypes in the Lagunas de Epulauquen population. One of these haplotypes was found in two Chilean populations from similar latitudes (Altos de Vilches at 35° 34′ S and Embalse Bulilleo at 36° 22′ S), supporting the connection postulated by Markgraf (). The other cpDNA haplotype found at Lagunas de Epulauquen was fixed in all the Argentinean populations of N. obliqua growing north of Lanin volcano (39° 30′ S) (Azpilicueta et al. ).
The occurrence of natural hybridisation between N. obliqua and other congener species (see N. obliqua, previous section) suggested a possible hybrid origin for Lagunas de Epulauquen individuals, and their identification as possible N. × leonii hybrids was even proposed (Gallo et al. ). The proximity of potentially hybridising Nothofagus forests in the west (Chile) reinforced this hypothesis. N. obliqua, together with N. glauca (Donoso and Landrum ; San Martín and Donoso ; Le-Quesne and Sandoval ), N. alpina and N. macrocarpa (Vázquez and Rodríguez ), grow in Chile in Embalse Bulilleo, less than 60 km from Lagunas de Epulauquen. Accordingly, species-specific isozyme markers revealed a very high proportion of N. alpina alleles in the robles of Lagunas de Epulauquen. Adh locus (alcohol dehydrogenase, E.C.220.127.116.11) showed the highest frequency for Adh-2 allele - up to then considered as species-specific for N. alpina (Gallo et al. ; Marchelli and Gallo ) - suggesting the hybrid origin of trees in this population (Azpilicueta and Gallo ). This allele had a frequency of 25%, in contrast with frequencies of <14% in sympatric populations where hybrids are more likely to occur (Azpilicueta and Gallo ). This result was unexpected given the absence of N. alpina within the area. In addition, Lagunas de Epulauquen showed the highest number of private microsatellite alleles of all Argentinean populations (five; Azpilicueta et al. ), two of which were shared with N. alpina. In conclusion, both isozyme and microsatellite markers supported the possible ancestral hybrid origin of Lagunas de Epulauquen individuals with N. alpina. The other Nothofagus species growing in the Lagunas de Epulauquen population - Nothofagus antarctica (G. Forst.) Oerst. and Nothofagus pumilio (Poepp. & Endl.) Krasser (Di Martino et al. ) - belong to the sub-genus Nothofagus, and no reports of hybridisation with N. obliqua exist.
New evidence supporting the distinctiveness of robles from Lagunas de Epulauquen
Biochemical markers: isozymes
Geographical location and allele frequency at Adh locus
Lagunas de Epulauquen
Geographical and altitudinal characteristics of the origins of roble populations evaluated
Lagunas de Epulauquen
Quillén Fondo de Lago
Quillén Corral de Bueyes
Quillén Casa Guardaparque
Trunk growth and branching were compared for a total of 244 8-year-old roble individuals from Lagunas de Epulauquen and a southern Argentinean population (Quila Quina 40° 10′ S, 71° 26′ W), grown from germination in a common garden (Table 2). For Lagunas de Epulauquen, seven open-pollination families were differentiated (L1 to L7), consisting of 21 to 25 individuals per family. For Quila Quina, two families (QQ1 and QQ2) and two groups of families (QQ3 and QQ4) were differentiated, which had 13 to 43 individuals per family or group of families. For each individual, the length (with a measuring tape), basal diameter (with digital callipers) and number of green leaves of the last but one annual shoot of the trunk (i.e. the stem portion extended in the year preceding the year of measuring) were registered. For each node of each annual shoot, the phyllotactic pattern and the presence/absence of axillary branch were recorded. In Nothofagus spp., phyllotaxis may adopt one of two conditions: alternate distichous (leaves arranged in two lines along the stem) or alternate tristichous (leaves arranged along three lines). Regarding branching, we recorded the following: presence/absence of an immediate branch (i.e. a branch developed as the annual shoot extended) or a delayed branch (i.e. a branch developed 1 year after the extension of the bearing annual shoot). Analysis of variance for unbalanced designs was applied to all annual shoots so as to compare origins (Lagunas de Epulauquen and Quila Quina, fixed factor) and families (random factor, nested within each origin). Comparisons between origins regarding phyllotaxis and branching were performed graphically. For each stem node, numbered from the stem's proximal end, we computed the proportions of nodes with distichous and tristichous phyllotaxis and the proportions of nodes with immediate and delayed branches.
Morphological attributes of annual shoots of 7-year-old roble plants
Basal diameter (mm)
Lagunas de Epulauquen
Total length (cm)
Lagunas de Epulauquen
Number of leaves
Lagunas de Epulauquen
Lagunas de Epulauquen
Mean internode length (cm)
Lagunas de Epulauquen
Survival and height growth in field trials
During 2004, a provenance field trial - including the Lagunas de Epulauquen population - was installed at Lácar lake, Argentina (40° 07′ S to 71° 28′ W), beneath a mixed forest of N. obliqua, N. alpina and Nothofagus dombeyi (Mirb.) Oerst. The site is characterised by volcanic soils and a mean precipitation of 2,200 mm/year. The essayed populations and their geographical locations are shown in Table 2. We used a randomised complete block design (RCBD) with a plot size of nine individuals. Survival and height were measured at the first and fourth years from trial installation. Both height measures were used to calculate growth rate relative to the measurement at the first year and later transformed with the logarithm 10 for variance analysis; for multiple mean comparisons, we used a Tukey test.
Lagunas de Epulauquen had the lowest total height (at both the first and fourth years) and presented significant differences to the other analysed populations, except for Pulmarí (PUL) and Quillén Fondo de Lago (QF), which correspond to the species' northern distribution areas. On the other hand, Lagunas de Epulauquen exhibited the highest relative growth rate, significantly different from all the other populations except Chumpiru (CH) and Quillén Corral de Bueyes (QCO).
The evidence for the genetically distinctive character of robles in the Lagunas de Epulauquen population compared to other Argentinean N. obliqua forests is supported in the present revision by both previous and new information. Lagunas de Epulauquen individuals show differences at biochemical, molecular, morphological and architectural levels. Molecular (Azpilicueta et al. ) and palynological (Markgraf ) data suggested that the Lagunas de Epulauquen population could have originated as an introgression from the western side of the Andean Cordillera during post-glacial re-colonisation. The presence in Lagunas de Epulauquen of a chloroplast DNA haplotype private to western populations reinforces this hypothesis. In agreement with this, Markgraf et al. () dated the forests of this region as 5,000 years BP, while an older origin was postulated for western refugia (Heusser ; Heusser et al. ; Villagrán ; Villagrán et al. ). The co-existence of N. glauca, N. macrocarpa, N. alpina and N. obliqua to the west suggests the possibility of a hybrid origin for robles from Lagunas de Epulauquen. Therefore, post-glacial history together with ancient hybridisation processes could stand as the main reasons for the present distinctiveness of the roble population in Lagunas de Epulauquen.
The geographical isolation of the Lagunas de Epulauquen population predicts genetic drift and inbreeding processes. However, a high level of genetic variation was detected with all genetic markers (isozymes: Azpilicueta and Gallo ; chloroplast DNA: Azpilicueta et al. ; microsatellites: Azpilicueta et al. ). It is possible that past hybridisation increased genetic diversity by providing new genetic variants. Moreover, long-distance pollen gene flow can rapidly restore genetic diversity (Hampe et al. ). Although, at least for N. alpina, most pollen disperses at short distances, a high potential for long-distance dispersal was reported (Marchelli et al. ).
The high frequency of the allele Adh-2 found in coastal and northern Chilean N. obliqua populations where N. macrocarpa grows casts doubt on the origin of this allele. Neither N. alpina nor N. glauca grow within this northern area; therefore, the Adh-2 allelic variant could come from N. macrocarpa. The phylogenetic affinity between N. macrocarpa and N. alpina can explain the occurrence of common alleles in these two species. Vázquez and Rodríguez () reported a closer morphological affinity between N. macrocarpa and N. alpina than between these two species and N. obliqua. Apparently, N. macrocarpa has been wrongly associated with N. obliqua, since both morphological and ecological (altitudinal niche) data support a closer relationship between N. macrocarpa and N. alpina (Vázquez and Rodríguez ). However, an alternative interpretation must be considered. Among the drawbacks of isozymes is the inability to detect variants in the DNA sequence that produces isozymes with no changes in electromorph mobility. Thus, the Adh-2 allelic variant observed in gels of N. obliqua, N. macrocarpa and N. alpina, although identical, could be the expression of different DNA sequences. The sequencing of the gene coding for the Adh enzyme should help in the precise identification of Adh-2 allele and its species origin.
The large seed size observed in Lagunas de Epulauquen could be a consequence of genecological variation, as found for N. obliqua in Chile (Donoso ). The largest seed size of the northernmost N. obliqua populations was explained as an adaptation to xeric environments within a wide latitudinal species cline. However, since N. macrocarpa was only later recognised as a separate species, it is possible that the sample in Donoso's study consisted of seeds from the two species (or included hybrids with N. macrocarpa). The results of our study could then be explained by clinal variation together with ancient hybridisation with N. macrocarpa.
The differences regarding leaf characters (leaf area and specific leaf area) between robles from Lagunas de Epulauquen and N. obliqua plants from other Argentinean populations reinforced the distinctiveness of the former population. Previous non-systematic field and nursery observations have now been confirmed based on these results.
Clear differences in length, number of leaves, mean internode length and the branching pattern of annual shoots were found between robles from Lagunas de Epulauquen and those from Quila Quina. With regard to these traits, Quila Quina plants resembled those from other areas within the distribution of N. obliqua in Argentina (e.g. Puntieri et al. ). In annual shoots of N. alpina, as in those of roble plants from Lagunas de Epulauquen, the frequency of development of immediate branches is relatively low and internodes are long compared to those of annual shoots in similar architectural positions on N. obliqua plants (Barthélémy et al. ). Recent observations confirmed that the architectural traits that distinguished nursery-grown Lagunas de Epulauquen plants from Quila Quina ones also occur in field conditions. Architectural studies incorporating the closely related Nothofagus entities considered in the present study (N. macrocarpa, N. glauca and N. × leonii), little known in this regard, may provide a clearer picture of the evolution of morpho-architectural traits in plants.
Height growth analysis showed that Lagunas de Epulauquen individuals exhibited low stem height - at both the first and fourth years - but, notwithstanding, the highest height growth rate compared to N. obliqua plants from other origins. All results coming from quantitative trait analyses may be associated with environmental conditions. In this regard, it must be highlighted that the roble population at Lagunas de Epulauquen is located at a relatively high altitudinal range compared to other roble populations from Argentina: from 1,500 to 1,700 masl, as opposed to the 600 to 1,200 masl range for other roble populations in Argentina. This environmental feature, together with the current genetic isolation of this population, leads us to put forward a hypothesis involving adaptation processes. New quantitative traits, especially phenological ones, should be considered in order to explore this alternative.
The glacial history of Patagonia - with valley-type glaciers at northern latitudes leaving ice-free areas and re-colonised southern regions - probably promoted the high genetically divergent evolution of N. obliqua forests as proposed by Donoso et al. (). The profusion of races, varieties, ecotypes and sub-species found nowadays in N. obliqua forests supports this idea. The results of the present study also provide evidence supporting the idea that the isolated roble population at Lagunas de Epulauquen shared (unlike other roble populations in Argentina) a common glacial history with Nothofagus populations from low-ice-impact areas in Chile, which could have resulted in a distinctive evolutionary process.
Future analyses of vegetative and reproductive characters in N. alpina, N. glauca and N. macrocarpa together with other N. obliqua populations could probably help in determining the taxonomic identity of the Nothofagus trees in Lagunas de Epulauquen. Architectural variation during early ontogenetic stages as well as phenological traits should be included in these studies. Sequencing of ITS nuclear ribosomal gene (internal transcript unit) and comparison with the results obtained by Manos () and Acosta and Premoli () should provide additional and valuable information for taxonomic identification through phylogenetic reconstruction.
The conservation value of Lagunas de Epulauquen
The Lagunas de Epulauquen area is of remarkable conservation value due to the diversity of its flora, which includes species from xeric environments (e.g. Mulinum spinosum), species typical of Valdivian forests (e.g. Lomatia ferruginea) and endemic species (e.g. Puya alpestris) (Alfonso and Prinna ; Di Martino et al. ). Most of the Lagunas de Epulauquen ecosystem - with a surface area of 7,450 ha - has been included within a provincial protected area called Reserva Lagunas de Epulauquen (Neuquén province), created in 1973. However, the limits of this reserve do not follow ecological criteria and, as a consequence, some N. obliqua forests lie outside the protected area, on private land. The limits of the protected area should be re-considered and sustainable management policies should be applied so as to promote forest regeneration. Conservation of this population's genetic pool could be the key to securing its adaptative capacity and, consequently, its persistence, especially considering the ongoing global climate change.
Thanks are due to Sebastián Di Martino for allowing the collection of samples at Lagunas de Epulauquen and to Soledad Ghirardi, Abel Martínez, Mario Huentú, Liliana Lozano and Sebastián Zuki for their valuable field and nursery work. This work was financially supported by projects PIP 112 200801 01026 (CONICET, Argentina), PNFOR044321 (INTA, Argentina), PNFOR044001 (INTA, Argentina) and Subprograma Nothofagus (PROMEF - Ministerio Agricultura Ganadería y Pesca de Argentina).
- Acosta MC, Premoli AC: Evidence of chloroplast capture in South American Nothofagus (subgenus Nothofagus , Nothofagaceae). Mol Phylogenet Evol 2010, 54: 235–242. 10.1016/j.ympev.2009.08.008View ArticlePubMedGoogle Scholar
- Alfonso G, Prinna A: Catálogo florístico de la Reserva lagunas de Epu Lauquen, Departamento Minas, Provincia de Neuquén, Argentina. Ernstia 2009, 19: 109–136.Google Scholar
- Allnut TR, Newton AC, Premoli A, Lara A: Genetic variation in the threatened South American conifer Pilgerodendron uviferum (Cupressaceae), detected using RAPD markers. Biol Conserv 2003, 114: 245–253. 10.1016/S0006-3207(03)00044-2View ArticleGoogle Scholar
- Araya L, Oyarzún MV (2000) Descripción de los bosques de N. alpina y N. obliqua en Chile. In: Ipinza Carmona R, Gutiérrez Caro B, Emhart Schmidt V (eds) Domesticación y mejora genética de raulí y roble. UACH/INFOR, ValdiviaGoogle Scholar
- Azpilicueta MM, Gallo LA: Shaping forces modelling genetic variation patterns in the naturally fragmented forests of a South American beech. Biochem Sys Ecol 2009, 37: 290–297. 10.1016/j.bse.2009.05.004View ArticleGoogle Scholar
- Azpilicueta MM, Marchelli P, Gallo LA: The effects of Quaternary glaciations in Patagonia as evidenced by chloroplast DNA phylogeography of Southern beech Nothofagus obliqua . Tree Genet Genomes 2009, 5: 561–571. 10.1007/s11295-009-0209-xView ArticleGoogle Scholar
- Azpilicueta MM, Gallo LA, van Zonneveld M, Thomas E, Moreno C, Marchelli P: Management of Nothofagus genetic resources: definition of genetic zones based on a combination of nuclear and chloroplast marker data. For Ecol Manag 2013, 302: 414–424. 10.1016/j.foreco.2013.03.037View ArticleGoogle Scholar
- Barthélémy D, Puntieri JG, Brion C, Raffaele E, Marino J, Martinez P: Morfología de las unidades estructurales y modo de desarrollo básico de especies Patagónicas de Nothofagus (Fagaceae). Bol Soc Argent Bot 1999, 34: 29–38.Google Scholar
- Bekessy SA, Allnut TR, Premoli AC, Lara A, Ennos RA, Burgman MA, Cortes M, Newton AC: Genetic variation in the vulnerable and endemic monkey puzzle tree, detected using RAPDs. Heredity 2002, 88: 243–249. 10.1038/sj.hdy.6800033View ArticlePubMedGoogle Scholar
- Di Martino S, Maletti EJ, Mazieres AV: Plan General de Manejo del Área Natural Protegida Epu Lauquen. Dirección General de Áreas Naturales Protegidas, Ministerio de Producción y Turismo, Provincia de Neuquén, Argentina; 2005.Google Scholar
- Donoso C: Genecological differentiation in Nothofagus obliqua (Mirb.) Oerst. in Chile. For Ecol Manag 1979, 2: 53–66. 10.1016/0378-1127(79)90036-7View ArticleGoogle Scholar
- Donoso C, Landrum LR: Nothofagus leonii Espinosa, a natural hybrid between Nothofagus obliqua (Mirb.) Oerst. and Nothofagus glauca (Phil.) Krasser. N Z J Bot 1979, 17: 353–360. 10.1080/0028825X.1979.10426908View ArticleGoogle Scholar
- Donoso C, Morales J, Romero M: Hibridación natural entre roble ( Nothofagus obliqua (Mirb.) Oerst.) y raulí ( N. alpina (Poepp. et Endl.) Oerst.) en bosques del sur de Chile. Revista Chilena de Historia Natural 1990, 63: 49–60.Google Scholar
- Donoso C, Gallo LA, Donoso P, Azpilicueta MM (2004) Variación en Nothofagus obliqua (Mirb.) Oerst. (Roble, Coyán, Hualle o Pellín). In: Donoso C, Premoli A, Gallo L, Ipinza R (eds) Variación intraespecífica en las especies arbóreas de los bosques templados de Chile y Argentina. Editorial Universitaria, Santiago de ChileGoogle Scholar
- Gallo LA (2004) Modelo conceptual sobre hibridación natural interespecífica entre Nothofagus nervosa y N. obliqua. In: Donoso C, Premoli A, Gallo L, Ipinza R (eds) Variación intraespecífica en las especies arbóreas de los bosques templados de Chile y Argentina. Editorial Universitaria, Santiago de ChileGoogle Scholar
- Gallo LA, Marchelli P, Breitembücher A: Morphological and allozymic evidence of natural hybridization between two Southern beeches ( Nothofagus spp.) and its relation to heterozygosity and height growth. For Genet 1997, 4: 15–23.Google Scholar
- Gallo L, Marchelli P, Crego P, Oudkerk L, Breitembücher A, Peñalba M, Chauchard L, Maresca L: Distribución y variación genética en características seminales y adaptativas de poblaciones y progenies de Raulí en Argentina. In Domesticación y mejora genética de raulí y roble. Edited by: Ipinza Carmona R, Gutiérrez Caro B, Emhart Schmidt V. UACH/INFOR, Valdivia; 2000.Google Scholar
- Grant ML, Clement EJ: Clarification of the name Nothofagus alpina and a new epithet for a Nothofagus hybrid. Bot J Linn Soc 2004, 146: 447–451. 10.1111/j.1095-8339.2004.00346.xView ArticleGoogle Scholar
- Hampe A, Pemonge M-H, Petit RJ: Efficient mitigation of founder effects during the establishment of a leading-edge oak population. Proc Biol Sci B 2013, 280: 1070. 10.1098/rspb.2013.1070View ArticleGoogle Scholar
- Harris SA, Ingram R: Chloroplast DNA and biosystematics: the effects of intra-specific diversity and plastid transmission. Taxon 1991, 14: 393–412. 10.2307/1223218View ArticleGoogle Scholar
- Heenan PB, Smissen RD: Revised circumscription of Nothofagus and recognition of the segregate genera Fuscospora , Lophozonia , and Trisyngyne (Nothofagaceae). Phytotaxa 2013, 146: 1–31. 10.11646/phytotaxa.146.1.1View ArticleGoogle Scholar
- Heusser CJ: Quaternary pollen record from Laguna de Tagua Tagua, Chile. Science 1983, 219: 1429–1432. 10.1126/science.219.4591.1429View ArticlePubMedGoogle Scholar
- Heusser CJ, Heusser LE, Lowell TV: Paleoecology of the southern Chilean lake district-Isla de Chiloé during middle-late Llanquihue glaciation and deglaciation. Geogr Ann 1999, 81A: 231–284. 10.1111/j.0435-3676.1999.00058.xView ArticleGoogle Scholar
- Hill RS, Jordan GJ: The evolutionary history of Nothofagus (Nothofagaceae). Aust Syst Bot 1993, 6: 111–126. 10.1071/SB9930111View ArticleGoogle Scholar
- Hill RS, Read J: A revised infrageneric classification of Nothofagus (Fagaceae). Bot J Linn Soc 1991, 105: 37–72. 10.1111/j.1095-8339.1991.tb00199.xView ArticleGoogle Scholar
- Le-Quesne C, Sandoval L: Extensión del límite sur para Nothofagus glauca (Phil.) Krasser. Gayana Bot 2001, 58: 139–142. 10.4067/S0717-66432001000200005View ArticleGoogle Scholar
- Manos PS: Systematics of Nothofagus (Nothofagaceae) based on rDNA spacer sequences (ITS): taxonomic congruence with morphology and plastid sequences. Am J Bot 1997, 84: 1137–1155. 10.2307/2446156View ArticlePubMedGoogle Scholar
- Marchelli P, Gallo LA: Genetic analysis of isozyme variants in open pollinated families of southern beech Nothofagus nervosa (Phil.) Dim. et Mil. Silvae Genetica 2000, 49: 90–98.Google Scholar
- Marchelli P, Gallo LA, Scholz F, Ziegenhagen B: Chloroplast DNA markers reveal a geographical divide across Argentinean southern beech Nothofagus nervosa (Phil.) Dim. et Mil. distribution area. Theor Appl Genet 1998, 97: 642–646. 10.1007/s001220050940View ArticleGoogle Scholar
- Marchelli P, Baier C, Mengel C, Ziegenhagen B, Gallo LA: Biogeographic history of the threatened species Araucaria araucana (Molina) K. Koch and implications for conservation: a case study with organelle DNA markers. Conserv Genet 2010, 11: 951–963. 10.1007/s10592-009-9938-5View ArticleGoogle Scholar
- Marchelli P, Smouse P, Gallo LA: Short-distance pollen dispersal for an outcrossed, wind-pollinated southern beech ( Nothofagus nervosa (Phil.) Dim. et Mil.). Tree Genet Genomes 2012, 8: 1123–1134. 10.1007/s11295-012-0500-0View ArticleGoogle Scholar
- Markgraf V: Paleoenvironmental changes at the northern limit of the subantarctic Nothofagus forests, lat. 37°S, Argentina. Quat Res 1987, 28: 119–129. 10.1016/0033-5894(87)90037-8View ArticleGoogle Scholar
- Markgraf V, MacGlone M, Hope G: Neogene paleoenvironmental and paleoclimatic change in southern temperate ecosystems - a southern perspective. Trends Ecol Evolut 1995, 10: 143–147. 10.1016/S0169-5347(00)89023-0View ArticleGoogle Scholar
- Markgraf V, Whitlock C, Anderson RS, Garcia A: Late Quaternary vegetation and fire history in the northernmost forest region: Mallín Vaca Lauquen, Neuquén province, Argentina. J Quat Sci 2009, 24: 248–258. 10.1002/jqs.1233View ArticleGoogle Scholar
- Martin PG, Dowd JM: Using sequences of rbcL to study phylogeny and biogeography of Nothofagus species. Aust Syst Bot 1993, 6: 441–447. 10.1071/SB9930441View ArticleGoogle Scholar
- Pastorino MJ, Gallo LA: Quaternary evolutionary history of Austrocedrus chilensis , a cypress native to the Andean-Patagonian forest. J Biogeogr 2002, 29: 1167–1178. 10.1046/j.1365-2699.2002.00731.xView ArticleGoogle Scholar
- Poole AL: Studies of New Zealand Nothofagus species. 2 Nut and cupule development. T Roy Soc NZ 1950, 78: 502–508.Google Scholar
- Premoli AC, Kitzberger T, Veblen TT: Isozyme variation and recent biogeographical history of the long-lived conifer Fitzroya cupressoides . J Biogeogr 2000, 27: 251–260. 10.1046/j.1365-2699.2000.00402.xView ArticleGoogle Scholar
- Puntieri JG, Grosfeld J, Stecconi M, Brion C, Azpilicueta MM, Gallo LA: Desarrollo temprano del roble ( Nothofagus obliqua ): un análisis arquitectural de procedencias de Argentina. Bosque 2006, 27: 44–51.Google Scholar
- Rabassa J, Clapperton CM: Quaternary glaciations in the southern Andes. Quat Sci Rev 1990, 9: 153–174. 10.1016/0277-3791(90)90016-4View ArticleGoogle Scholar
- Ravenna P: Nothofagus macrocarpa y Nothofagus rutila (Fagaceae), dos especies diferentes. Onira 2002, 7: 57–60.Google Scholar
- Sabatier Y, Azpilicueta MM, Marchelli P, González-Peñalba M, Lozano L, García L, Martinez A, Gallo LA, Umaña F, Bran D, Pastorino MJ: Distribución natural de Nothofagus alpina y Nothofagus obliqua (Nothofagaceae) en Argentina, dos especies de primera importancia forestal de los bosques templados norpatagónicos. Bol Soc Argent Bot 2011, 46: 131–138.Google Scholar
- San Martín J, Donoso C: Estructura florística e impacto antrópico del bosque maulino de Chile. In Ecología de los bosques nativos de Chile. Edited by: Armesto JJ, Villagrán C, Arroyo MK. Editorial Universitaria, Santiago de Chile; 1995.Google Scholar
- Vázquez FM, Rodríguez RA: A new subspecies and two new combinations of Nothofagus Blume (Nothofagaceae) from Chile. Bot J Linn Soc 1999, 129: 75–83.Google Scholar
- Veblen TT, Donoso C, Kitzberger T, Rebertus AJ (1996) Ecology of southern Chilean and Argentinean Nothofagus forests. In: Veblen TT, Hill RS, Read J (eds) The ecology of biogeography of Nothofagus forests. Yale University Press, New Haven and LondonGoogle Scholar
- Veit H, Garleff K: Evolución del paisaje cuaternario y los suelos en Chile central-sur. In Ecología de los bosques nativos de Chile. Edited by: Armesto JJ, Villagrán C, Kalin Arroyo M. Editorial Universitaria, Santiago de Chile; 1995.Google Scholar
- Villagrán C: Historia de los bosques templados del sur de Chile durante el Tardiglacial y Postglacial. Rev Chil Hist Nat 1991, 64: 447–460.Google Scholar
- Villagrán C, Moreno P, Villa R: Antecedentes palinológicos acerca de la historia cuaternaria de los bosques chilenos. In Ecología de los bosques nativos de Chile. Edited by: Armesto JJ, Villagrán C, Kalin Arroyo M. Editorial Universitaria, Santiago de Chile; 1995.Google Scholar
- Wolfe KH, Li W-H, Sharp PM: Rates of nucleotide substitution vary greatly among plant mitochondrial, chloroplast and nuclear DNAs. Proc Natl Acad Sci U S A 1987, 84: 9054–9058. 10.1073/pnas.84.24.9054PubMed CentralView ArticlePubMedGoogle Scholar
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