Multiplication of Cupressus guadalupensis Using the RITA® Temporary Immersion System.

The Guadalupe cypress (Cupressus guadalupensis S. Watson) is an endangered species included in the list of the NOM-059-SEMARNAT-2010. The presence of wild goats in the habitat has been the greatest threat to the propagation and survival of this species. Therefore, there is a need to generate propagation protocols that facilitate the regeneration of the species. Plant tissue culture offers various possibilities that can facilitate the regeneration of species under some risk. Temporary immersion systems have proven to be an option with various advantages in plant tissue culture, such as increasing the number of seedlings generated and reducing production times, compared to semisolid media. The objective of this chapter is to describe a protocol to propagate Guadalupe cypress tissues in a RITA® temporary immersion system.

Cupressus macrocarpa leaves for biomonitoring the environmental impact of an industrial complex: The case of Puchuncaví-Ventanas in Chile.

The Punchuncaví Valley is one of the most polluted areas in central Chile affected by anthropogenic emissions from the Ventanas Industrial Complex (IC) where the most important industry is the copper smelter and refinery. In this context, this research aims were to assess the usefulness of the Cupressus macrocarpa as a biomonitor. The leaf samples were taken from five selected sites, located between 0.8 and 15 km away from the source. A total of 34 elements were analyzed in leaf samples by ICP-MS and examined by enrichment factor (EF), hierarchical cluster analysis (HCA), and principal component analysis (PCA). Leaf concentration of As, Ca, Cd, Cu, Dy, Er, Gd, K, Li, Mg, Mn, Mo, Na, Nd, P, Pb, Pr, S, Sb, Sr, Ti, Yb and Zn showed statistically significant differences between sampling sites (p-value < 0.05). A clear trend to increase the concentration of Cu, Sb, S, As, Cd and Pb with the proximity to the IC. Besides, high values of Cu (93.4-369 mg kg-1) and As (7.6-12.7 mg kg-1) were observed near to industrial area exceed the phytotoxic levels reported in plants with EF > 3000% for Cu and >1300% for As. The application of PCA and HCA identified 6 factors related to the industrial complex, traffic and geogenic sources, providing the greatest variance the component related to industrial activity mainly with copper smelter and refinery. According to the results, the C. macrocarpa leaves are a good biomonitor to evaluate the high pollution load for anthropogenic elements in industrial areas.

Spontaneous abortion in cattle after consumption of Hesperocyparis (Cupressus) macrocarpa (Hartw.) Bartel and Cupressus arizonica (Greene) needles in Uruguay.

An outbreak of cattle abortion associated with the consumption of Hesperocyparis (Cupressus) macrocarpa and Cupressus arizonica needles has been reported in Uruguay. Twenty-five of 125 pregnant heifers in late-term pregnancy aborted after being introduced into a paddock containing numerous H. macrocarpa, C. arizonica and Casuarina cunninghamiana trees. There were no significant macroscopic or histological findings in the two fetuses analyzed. Abortifacient labdane acids were detected in the needles of H. macrocarpa (1.68%, D.W.) and C. arizonica (0.36%, D.W.) but none were found in C. cunninghamiana.

Differential metabolism of diastereoisomeric diterpenes by Preussia minima, found as endophytic fungus in Cupressus lusitanica.

The plant diastereoisomeric diterpenes ent-pimara-8(14)-15-dien-19-oic acid, obtained from Viguiera arenaria, and isopimara-8(14)-15-dien-18-oic acid, isolated from Cupressus lusitanica, were distinctly functionalized by the enzymes produced in whole cell cultures of the fungus Preussia minima, isolated from surface sterilized stems of C. lusitanica. The ent-pimaradienoic acid was transformed into the known 7ß-hydroxy-ent-pimara-8(14)-15-dien-19-oic acid, and into the novel diterpenes 7-oxo-8 ß-hydroxy-ent-pimara-8(14)-15-dien-19-oic and 7-oxo-9ß-hydroxy-ent-pimara-8(14)-15-dien-19-oic acids. Isopimara-8(14)-15-dien-18-oic acid was converted into novel diterpenes 11α-hydroxyisopimara-8(14)-15-dien-18-oic acid, 7ß,11α-dihydroxyisopimara-8(14)-15-dien-18-oic acid, and 1ß,11α-dihydroxyisopimara-8(14)-15-dien-18-oic acid, along with the known 7ß-hydroxyisopimara-8(14)-15-dien-18-oic acid. All compounds were isolated and fully characterized by 1D and 2D NMR, especially 13C NMR. The diterpene bioproduct 7-oxo-9ß-hydroxy-ent-pimara-8(14)-15-dien-19-oic acid is an isomer of sphaeropsidin C, a phytotoxin that affects cypress trees produced by Shaeropsis sapinea, one of the main phytopathogen of Cupressus. The differential metabolism of the diterpene isomers used as substrates for biotransformation was interpreted with the help of computational molecular docking calculations, considering as target enzymes those of cytochrome P450 group.

Effects of Post-Fire Plant Cover in the Performance of Two Cordilleran Cypress (Austrocedrus chilensis) Seedling Stocktypes Planted in Burned Forests of Northeastern Patagonia, Argentina.

Cordilleran cypress (Austrocedrus chilensis [D.Don] Pic. Serm. et Bizarri) forests occupy 140,000 ha along a sharp environmental gradient of central Andean-Patagonia in Argentina. Every summer, about 3200 ha of these forests are affected by wildfires, taking thereafter long time to recover. To accelerate forest recovery, we determined in xeric and mesic cypress stands burned 5 and 2 year before whether survival and growth of two planted cypress seedling stocktypes are affected by plant cover and contrasting precipitation conditions. Two experiments were conducted on each site, involving 100 replicates of two seedling stocktypes, having each significantly different morphological attributes. The experiments comprised a dry and humid growing season on each site. Both stocktypes performed similarly within stands, but differently between stands. In the xeric stand, plant cover had neutral effects on seedling survival, favored seedling height growth in the dry season, and was negative on collar diameter and stem growth. In the mesic site, high plant cover favored survival and height growth, but was inconsequential for collar diameter and stem growth. In this short-term post-fire period, and independent of precipitation received during both seasons (dry or humid), plant cover appears as playing a facilitative role, having neutral or even positive effects on survival and growth of planted seedlings. During the early post-fire successional stages, and besides seedling stocktype, there was a synergistic balance between light and soil moisture that seems to benefit planted seedling performance in burned cypress forests, and especially in mesic sites.

Improving survival and growth of planted Austrocedrus chilensis seedlings in disturbed patagonian forests of Argentina by managing understory vegetation.

This study was aimed at determining, under field conditions, early interactions between planted cypress seedlings and their associated shrubs in a mesic area of Andean Patagonia and, in a nursery, the effects of increasing light availability on cypress performance when soil water was not a limiting factor. The field experiment was performed in a former cypress-coihue mixed forest (42°02'S, 71°33'W), which was replaced in the 1970s by a plantation of radiata pine. In 2005, 800 cypress seedlings were planted under maqui shrubs in a clear-cut area of the pine stand. In 2007, two treatments were set: no-competition treatment ([NCT] i.e., the surrounding aboveground biomass was removed) and competition treatment ([CT] i.e., without disturbance). The nursery experiment (42°55'S, 71°21'W) consisted of two groups: "shade" (grown under shade cloth) and "sun" (grown at full sun) cypress seedlings. After one growing season, seedling survival and stem growth (in height and diameter) were determined at both sites. Furthermore, the growth rate of leaves, stems, and roots was determined in the nursery. In the field experiment, height growth and survival in NCT were significantly greater than in CT, and a competition process occurred between cypress and surrounding shrubs. In the nursery, sun plants grew more in diameter and increased root weight more than shade plants. Results also showed that in mesic areas of Patagonia, decreasing competition and increasing light levels produced stouter seedlings better adapted to support harsh environmental conditions. Therefore, the removal of protecting shrubs could be a good management practice to improve seedling establishment.

Morphometric Variation on the Cypress Aphid Cinara cupressi (Buckton) (Hemiptera: Aphididae) Associated to Urban Trees.

Cinara cupressi (Buckton) is an important aphid pest of the Cupressaceae family, originally reported in Chile in 2003. Since then, it has spread over 4,000 km, contributing to conservation issues, mostly associated with native and urban trees of the Cupressaceae. In the present work, the morphometric variation of C. cupressi was examined to determine if the species present in Chile corresponds to a specific entity, and to identify variations among specimens from different localities in the study area. Colonies were collected from urban trees from northern, central, and southern Chile. Morphometric data for 14 characters in aphids from 63 localities in all the distribution ranges were measured and analyzed by multivariate analysis. Results showed that the species present in Chile corresponds to C. cupressi like a single specific entity, showing no morphological variation across the regions sampled. Our data will be discussed within the context of correct taxonomic identification for the implementation of effective biological control strategies.

Neozygites osornensis sp. nov., a fungal species causing mortality to the cypress aphid Cinara cupressi in Chile.

An entomophthoralean fungus causing epizootics in populations of the cypress aphid, Cinara cupressi Buckton, in Chile is described as a new species, Neozygites osornensis Montalva et Barta. The aphid pathogen is described based on morphological characters. An exhaustive description, illustrations and a comparison with closely related species are provided. The fungus differs from similar Neozygites species by smaller hyphal bodies, nuclei, primary conidia, capilliconidia and capilliphores and by noticeably different shape of capilliconidia. A key to aphid-pathogenic species of Neozygites is also included.

Fomitiporia cupressicola sp. nov., a parasite on Cupressus arizonica, and additional unnamed clades in the southern USA and northern Mexico, determined by multilocus phylogenetic analyses.

Fomitiporia cupressicola sp. nov., found in living Cupressus arizonica, is described on the basis of several collections originating from a high altitude forest in the northern Sierra Madre Occidental, Mexico. The species forms a monophyletic clade, basal to a larger lineage comprising species originating mainly from temperate to Mediterranean areas of the northern hemisphere. The phylogenetic approach in Fomitiporia also revealed multiple unnamed clades within the F. robusta complex in the southern USA and northern Mexico, representing potential species. The status of the F. robusta complex in North America is discussed briefly.

Batch and dynamic biosorption of basic dyes from binary solutions by alkaline-treated cypress cone chips.

A simple alkaline pre-treatment of Cupressus sempervirens cone chips was performed to improve their biosorption capacity towards methylene blue and rhodamine B from aqueous solutions, in batch and continuous modes. Biosorption kinetics were determined from single and binary dyes solutions, and properly described by the pseudo-second-order rate model. Experimental single-dye equilibrium isotherms fitted the Langmuir-Freundlich model, with maximum biosorption capacities of 0.68mmol/g for methylene blue and 0.50mmol/g for rhodamine B. Single-dye dynamic biosorption showed that breakthrough time for methylene blue biosorption was almost four times longer than for rhodamine B and that the alkaline modification of the chips greatly improved the biosorption performance. Competitive dynamic biosorption demonstrated the preference of the modified cone chips for biosorbing methylene blue, confirmed by the exit concentration overshoots obtained in the breakthrough curves of rhodamine B.

Evaluación de la virulencia de hongos entomopatógenos del pulgón del ciprés, de dos regiones ecológicas de Chile. II. / Evaluation of the virulence of entomopathogenic fungi of cypress aphid, from two ecological regions of Chile. II

El pulgón del ciprés Cinara cupressi, se considera uno de los insectos invasores más dañinos del mundo, ha provocado una gran mortalidad en especies Cupresáceas nativas y exóticas en varios países. En Chile, fue detectado el año 2003 y para el año 2008 esta plaga ya se encontraba distribuida en todo el país, afectando especies cupresáceas forestales exóticas como Cupressus macrocarpa y especies nativas, tales como, Austrocedrus chilensis (Ciprés de la Cordillera) y Fitzroya cupressoides (Alerce), que están categorizadas en el estado de conservación vulnerable y en peligro respectivamente. El área forestal de Chile ha potenciado y privilegiado el uso de controladores biológicos como parte del manejo integrado de plagas. Es por esto que se ha utilizado el parasitoide Pauesia juniperorum. Sin embargo, se han detectado bajos niveles de parasitismo. Se propone el uso de hongos entomopatógenos como alternativa y complemento al control biológico. Se prospectaron hongos entomopatógenos en la Región ecológica mediterránea per-húmeda y en la Región oceánica con influencia mediterránea. Como resultado se han identificado seis géneros de hongos descritos como patógenos de insectos. Sin embargo, bajo condiciones de laboratorio sólo cepas del género Verticillium fueron virulentas, causando sobre el 80 por ciento de mortalidad acumulada a los 7 días. No hubo diferencias significativas entre las cepas de Verticillium y un producto químico (pirimicarb) aplicado como control, aunque el hongo fue más lento. Las cepas más virulentas fueron identificadas como Verticillium lecanii, Ve 1 y Ve 2, con un TL50 de: 3.2 y 3.1 días y un DL50 : 1.24 y 1.37 conidias mL-1 respectivamente, sugiriendo el uso de estos hongos para controlar C. cupressi en Chile.

The cypress aphid, Cinara cupressi, is considered one of the most important invasive species causing high mortality in exotic and native species of Cupressaceae in several countries in the world. In Chile it was detected in 2003 and in 2008 was distributed throughout the country affecting the exotic forest species Cupressus macrocarpa and the native forest species Austrocedrus chilensis (Ciprés de la Cordillera) and Fitzroya cupressoides (Alerce), both classified as vulnerable and endangered species respectively. Efforts to their management have focused on biological control by using the parasitoid Pauesia juniperorum but until now it has not reached satisfactory control. We propose using entomopathogenic fungi, as alternative and complementary biocontrol. Entomopathogenic fungi were prospected in the ecological region Mediterranean per-humid and in the ecological region Oceanic with mediterranean influence in both colonies of C. cupressi. There were identified six genera of fungi described as insect pathogens. However, in laboratory assays only Verticillium strains were virulent, causing about 80 percent cumulative mortality at seven days. There were not significant differences among strains of Verticillium and chemical (pirimicarb) applied as control, although fungi were slower. The most virulent strains were two Verticillium lecanii, Ve 1 and Ve 2, with an LT50 of: 3.2 and 3.1 days and LD50 : 1.24 and 1.37 conidia mL-1 respectively, suggesting the use of these fungi to control C. cupressi in Chile.

Linkage mapping of the Mediterranean cypress, Cupressus sempervirens, based on molecular and morphological markers.

Gene mapping for a Cupressus species is presented for the first time. Two linkage maps for the Mediterranean cypress (Cupressus sempervirens) varieties, C. sempervirens var. horizontalis and C. sempervirens var. pyramidalis, were constructed following the pseudo-testcross mapping strategy and employing RAPD, SCAR and morphological markers. A total of 427 loci (425 RAPDs, two SCARs) representing parents and F(1) progeny were screened for polymorphism with 32 random decamer and two SCAR primers. A morphological marker defined as "crown form" was also included. Of 274 polymorphic loci, the 188 that presented Mendelian inheritance formed the mapping dataset. Of these loci, 30% were mapped into seven linkage groups for the horizontalis (maternal) and four linkage groups for the pyramidalis (paternal) map. The putative "crown form" locus was included in a linkage group of both maps. The horizontalis and the pyramidalis maps covered 160.1 and 144.5 cM, respectively, while genome length was estimated to be 1696 cM for the former variety and 1373 cM for the latter. The four RAPD markers most tightly linked to crown form were cloned and converted to SCARs. Each of the cloned RAPD markers yielded two to three different sequences behaving as co-migrating fragments. Two SCAR markers, SC-D05(432) and SC-D09(667), produced amplified bands of the expected sizes and maintained linkage with the appropriate phenotype, but to a lesser extent compared to their original RAPD counterparts. These linkage maps represent a first step towards the localization of QTLs and genes controlling crown form and other polygenic traits in cypress.

Plastid DNA diversity is higher in the island endemic Guadalupe cypress than in the continental Tecate cypress.

BACKGROUND: Callitropsis guadalupensis (Guadalupe cypress) is endemic to Guadalupe Island, Mexico, where it is the dominant species of the only forest. The species has suffered declining numbers following the introduction of goats to the island over 150 years ago. Callitropsis guadalupensis is closely related to Callitropsis forbesii (Tecate cypress), distributed in small isolated populations in mainland Baja California and southern California. The objective of the present study was to compare the genetic diversity of the island endemic to the continental species. METHODOLOGY/PRINCIPAL FINDINGS: We measured genetic diversity in Callitropsis guadalupensis (n =54) from Guadalupe Island and in Callitropsis forbesii (n = 100) from five populations in mainland Baja California. The plastid DNA trnS-trnG spacer and the trnL-trnF region were chosen for characterization. Thirty-four haplotypes were observed, of which six were shared between both species. One of these haplotypes was also shared with three other species, Callitropsis lusitanica, Callitropsis montana, and Callitropsis stephensonii. Haplotype diversity (h) and nucleotide diversity (π) were significantly higher for Callitropsis guadalupensis (h = 0.698, π = 0.00071) than for Callitropsis forbesii (h = 0.337, π = 0.00024). CONCLUSIONS/SIGNIFICANCE: Callitropsis guadalupensis shows no evidence of a founder effect or of a genetic bottleneck, and can be added to a growing list of insular species with higher genetic diversity than their mainland relatives.

Prospección de hongos entomopatógenos del pulgón del ciprés en dos regiones ecológicas de Chile: I / Prospection of entomopathogenic fungi of cypress aphid in two ecological regions of Chile: I

El pulgón del ciprés (Cinara cupressi), es una plaga exótica distribuida en todo el territorio continental chileno que ha provocado daños importantes tanto enespecies forestales introducidas como nativas. Se han estudiado algunos aspectos de la biología de estepulgón, mediante control biológico y productos químicos, los cuales no presentan actualmente resultadossatisfactorios. Durante el tiempo que esta plaga se ha establecido en Chile, no se ha realizado un estudio sistemático de hongos entomopatógenos asociados aeste pulgón, aunque se cuenta con antecedentes de un control promisorio en pulgones con estos agentes enotros países. Con el objetivo inicial de aislar en una primera fase del presente estudio hongos patogénicospotenciales para el control biológico de C. cupressi, iniciamos su búsqueda en dos regiones ecológicas del sur de Chile (Región ecológica Mediterránea Per-Húmeda y Región ecológica Oceánica con influencia Mediterránea), seleccionándose en cada una de ellas 6sitios de muestreo donde en cada uno se recolectaron 10 ramas con colonias de pulgones que fueron mantenidas en una cámara bioclimática (20 +/- 2 °C, 16:8 h) por 7 días, para estimular el desarrollo fúngico. Se aislaron integrantes de lo géneros Verticillium, Paecilomyces(ambos con mayor frecuencia de presencia), Fusarium y un entomophthoral en estudio.

The cypress aphid (Cinara cupressi) is an exotic plague distributed throughout the Chilean continental territory which has caused significant damage both inintroduced forest species as well as in native ones. Some characteristics as to the biology of this aphid have beenstudied by means of biological control and chemicals yet they have not revealed any satisfactory results up tonow. During the time of occurrence of this plague in Chile systematic study on entomopathogenic fungi associated to this aphid has failed to be accomplishedalthough information about a promissory control in aphids with these agents has been reported in other countries. In order to carry out an isolation of potential pathogenic fungi for the biological control of C.cupressiin the first phase of this present research, we began to look for them in two ecological regions from southernChile (Ecological Mediterranean Per-humid Region and Ecological Oceanic Region having Mediterraneaninfluence). Six sampling sites were selected in each of them while 10 branches infected with aphid colonies were kept in a bioclimatic chamber (20+2ºC, 16:8 h) for 7 days to stimulate fungal growth. Fungi of generaVerticillium, Paecilomyces (both showing the highest occurrence frequency), Fusarium and an entomophtoral under study were isolated.

Effectiveness of Cupressus sempervirens cones as biosorbent for the removal of basic dyes from aqueous solutions in batch and dynamic modes.

The feasibility of using cypress cone chips from Cupressus sempervirens as a low-cost biosorbent for the removal of two representative basic dyes, methylene blue (MB) and rhodamine B (RhB), from aqueous solutions was investigated in batch and continuous modes. Dyes biosorption was strongly dependent on the solution's pH. Sorption kinetics was determined and properly described by the pseudo-second-order rate model. Experimental equilibrium isotherms fitted the Langmuir model, showing maximum biosorption capacities of 0.62 mmol/g for MB and 0.24 mmol/g for RhB. Competitive experiments from a binary solution of the dyes demonstrated the preference of the cone chips for biosorbing MB. Very low desorption efficiencies were obtained for both dyes. Dynamic experiments showed that the breakthrough time was three times higher for MB biosorption than for RhB for the same conditions. Breakthrough curves were properly represented by a mathematical model.

[Prevalence of hypersensitivity to Cupressus pollen in allergic patients in Monterey, Mexico]. / Prevalencia de hipersensibilidad al polen de Cupressus en pacientes alérgicos en Monterrey, México.

BACKGROUND: Cupressus pollen is one of the main causes of allergic respiratory disease in North America. In a previous study of pollen concentration in Monterrey, Cupressus occupied the 3rd place in frequency. OBJECTIVE: To know the sensitization to Cupressus in patients treated at our center. METHODS: We conducted an observational, cross-sectional and prospective study, with a questionnaire to assess potential exposure and allergic symptoms to pollen, skin tests with allergen extract of Cupressus 1:20 dilution, and skin tests with others 36 aeroallergens in our hospital. RESULTS: We did Cupressus skin test in 256 patients (136 women), 130 younger than 18 years old and 126 adults. The skin test was positive in 39 patients (15.2%). Of the 36 aeroallergens tested, Cupressus occupied the 7th place in frequency. Of the 130 under 18 years, only 10 (7.7%) had a positive skin test. Among the other 129 adults, 29 (22.5%) had a positive skin test (p = 0.001). Patients with a positive skin test Cupressus had at least one positive skin test to other aeroallergen in 97% of cases (p = 0.001). CONCLUSIONS: Cupressus sensitization in our study group was high (15.2%). The routine use of allergen extract of Cupressus should be considered in patients with respiratory allergy. However, it is necessary to determine precisely the prevalence of sensitization and its possible clinical implications.

High genetic variation in marginal fragmented populations at extreme climatic conditions of the Patagonian Cypress Austrocedrus chilensis.

Knowledge about current patterns of genetic structure of populations together with the evolutionary history of a species helps to understand and predict the adaptation of populations to future climate change. We assayed variation at nuclear microsatellite markers among peripheral vs. continuous populations of the temperate South American species Austrocedrus chilensis, to investigate the role of historical vs. demographical forces in shaping population genetic structure. This species occurs in continuous populations in the west and central distribution range, but becomes highly fragmented at the eastern limit, which comprised ice-free areas during Quaternary glaciations and has extreme climatic conditions at present times. Bayesian analysis methods identified two contrasting patterns of genetic structure; (I) populations from humid, mesic and peri-glacial regions formed a single deme with relatively low genetic differentiation and high admixture levels whereas (II) a highly heterogeneous genetic structure with low level of admixture was found in the steppe, towards the east and northeast limit of the distribution range. In the steppe, population fragmentation, restricted gene flow and isolation-by-distance were also inferred. In addition, several small steppe populations showed high genetic diversity and divergent gene pools, suggesting that they constitute ancient refuges from pre-Holocene glaciations with just a subgroup of them contributing significantly to post-glacial spread. These results are discussed in relation to patterns of genetic variation found for other temperate species and the contribution of the particular southern Andes topography and climate to post-glacial spread.

Airborne pollen of Carya, Celtis, Cupressus, Fraxinus and Pinus in the metropolitan area of Monterrey Nuevo Leon, Mexico.

The concentration of pollen grains in the atmosphere over the metropolitan area of Monterrey, Nuevo Leon, Mexico, was analyzed throughout a year from March 2003-February 2004, focused on the genus Carya, Celtis, Cupressus, Fraxinus and Pinus owing to their interest as etiological pollinosis agents in diverse regions of the world. A 7-day Hirst type volumetric spore and pollen trap was located on a building roof of the city at 15 m from ground level for continuous sampling. The total quantity of pollen recorded for the study period was 21,083 grains/m(3), corresponding to 49.75 % of the taxa of interest. February and March were the months with higher pollen amounts in the air with 7,525 and 2,781 grains/m(3), respectively, and amounted to 49 % of total year through pollen. Fraxinus was the genus which contributed to the largest amount of pollen with 28 % of total grains (5,935 grains/m(3)) followed by Cupressus with 13 % (2,742 grains/ m(3)). Celtis, Pinus and Carya contributed with 5.3 % , 2.7 % , and 0.6 % of total pollen, respectively. These results indicate that Fraxinus and Cupressus are present in the area in sufficient quantity to indicate likely involvement in the origin of allergic disorders in the human population.

Mating system in a low-density natural population of the dioecious wind-pollinated Patagonian cypress.

The mating system of a species is an essential factor that determines the genotypic frequencies of its populations in each generation. Thus, knowledge in this regard is important in the application of practical measures in conservation and forest tree breeding. Austrocedrus chilensis (D.DON) PIC. SER. ET BIZZARRI: is a member of the Cupressaceae native to the Andean-Patagonian Forest of Argentina and Chile with regional economic importance. It is a dioecious tree with pollen and seeds distributed by wind. Isozyme markers were utilized to study the mating system of this species through the survey of individual effective pollen clouds in a natural low-density population. If random mating occurs, effective pollen clouds that pollinate single seed trees are expected to be genetically homogeneous. The classical HardyWeinberg test of panmixia was also performed. Although it failed to reject random mating, the analysis of the pollen clouds showed that this mating pattern does not occur. Evidence of locally distributed heterogeneous pollen clouds is presented. The two methods are critically compared.