SUMMARYReproductive biology knowledge of species is essential for in situ conservation, propagation and breeding. Consequently, we describe the first detailed phenology of Nothofagus alpina, mentioned until now only in general terms, and provide the morphological description of the male and female flowers. The study was conducted in 2011 in the Clonal Seed Orchard "Huillilemu" located in the locality of San José Mariquina, Region Los Ríos, Chile, at 23 meters of altitude. Observations were made in six clones of the orchard. The phenology model proposed presents ten states from the leaf bud in latency to the initial fruit. The first phenophase characterizes the leaf bud state in rest, while the second shows the morphological changes associated with getting out of dormancy. Five states described below develop male flowers and female inflorescences that appear later. The fall of male flowers mark the transition status into the female flower. The last two phenological stages describe the initial infructescence and development of the dome, respectively. These changes took place over a period of 58 days in individuals with early budding and in 40 days in late budding individuals with accumulated heat requirement of 560-623 GDD. The start date break was related to the altitudinal origin of trees, suggesting a genetic control of this feature. The overall overlap rate of flowering was calculated by 0.22, indicating low reproductive synchrony among analyzed individuals.Key words: characterization morphology, reproduction, dichogamy, overlapping index, flowering. RESUMENEl conocimiento de la biología reproductiva de las especies, es fundamental para la conservación in situ, reproducción y mejoramiento genético. Por ello, se describe por primera vez la fenología detallada de Nothofagus alpina, hasta ahora mencionada sólo de forma general y se presenta la descripción morfológica de las flores masculinas y femeninas. El estudio se realizó durante el año 2011, en el huerto semillero clonal "Huillilemu", ubicado en la comuna San José de la Mariquina, región de Los Ríos, Chile, a una altitud de 23 m snm. Las observaciones se realizaron en seis clones del huerto. El modelo fenológico propuesto, presenta 10 estados, desde la yema en latencia hasta el fruto inicial. El primer estado caracteriza a la yema en reposo, mientras que el segundo, ilustra los cambios morfológicos asociados a la salida del reposo. Cinco estados describen a continuación, el desarrollo de las flores masculinas y de las inflorescencias femeninas, que aparecen después de estas. La caída de las flores masculinas, marca en la flor femenina la transición al estado de fruto. Los dos últimos estados fenológicos, describen la infrutescencia inicial y desarrollo de la cúpula, respectivamente. Estos cambios se desarrollaron durante un lapso de 58 días, en los individuos con brotación temprana y en 40 días en los individuos con brotación tardía, con requerimiento de calor acumulado de 560 a 623 grados días de desarrollo. La fecha de inicio de la brotación, está relacion...

Long-term pollen storage is important so that producers and breeders may be able to utilise stored pollen to cross individuals that bloom at different times or in different places. The objective of the current study was to determine the moisture content and optimal temperature for the storage of Nothofagus alpina (P. et E.) Oerst pollen. Pollen samples used for this study were obtained from established trees at the 'Huillilemu' clonal seed orchard, located in the commune of San José de la Mariquina, in southwestern Chile. Pollen extraction was conducted by placing it in a climatic chamber (25°C and 50% relative humidity) for 24 hours. The obtained pollen was subjected to various drying times and stored at three temperatures: 4°C; -13°C; and -80°C. Longevity of pollen stored at 4°C and -13°Cwas short, although it was possible to increase longevity by storing it at -80°C.

SUMMARYUnderstanding the biology of pollen is necessary to optimize the reproduction of plants and the successful implementation of conservation and breeding programs. In order to have information on the morphological characteristics and pollen viability in Nothofagus nervosa, male flowers of trees established in a clonal seed orchard were collected. Morphological characterization of pollen was carried out from observations under light microscope and scanning electron microscope. Motic Images Plus 2.0 software was used for measurements. Viability was assessed by in vitro germination, germination in vivo and staining with p-phenylenediamine. The pollen size of these species ranges from medium to large and the shape is from sub-oblate to oblate spheroid. Initial viability by in vitro germination ranged from 17.1 to 84.9 %; with the staining method, it ranged from 70.7 to 99.3 %; using in vivo germination it was calculated between 77.4 and 97.1 %. There was a strong correlation between in vitro germination and in vivo germination, but there was no correlation between viability calculated by the staining method and that calculated by germination in vivo; whereby the in vitro method is recommended to assess both viability and longevity of pollen of this species. According to assessments by in vitro germination, pollen lost its viability within a period of eight days stored at room temperature.Key words: viability, morphology, longevity, germination, staining. RESUMENEntender la biología del polen es necesario para mejorar la reproducción de las especies vegetales y para la implementación exitosa de programas de mejoramiento genético y conservación. Con el fin de contar con información sobre las características morfológicas y de viabilidad del polen en Nothofagus nervosa, se colectó flores masculinas de árboles establecidos en un huerto semillero de clones. La caracterización morfológica del polen se realizó a partir de observaciones en microscopio de luz y electrónico de barrido. Para las mediciones se utilizó el software Motic Images Plus 2,0. El polen de esta especie es de tamaño mediano a grande, de forma sub-oblada a oblada esferoidal. La viabilidad se evaluó mediante germinación in vitro, germinación in vivo y tinción con p-fenilendiamina. Su viabilidad inicial, mediante la germinación in vitro varió desde 17,1 a 84,9 %; con el método de tinción osciló de 70,7 a 99,3 % y mediante germinación in vivo se calculó entre 77,4 y 97,1 %. Hubo una fuerte correlación entre la germinación in vitro y la germinación in vivo, más no hubo correlación entre la viabilidad calculada por el método de tinción y la calculada mediante germinación in vivo, por lo cual se recomienda el método in vitro para realizar evaluaciones tanto de viabilidad como de longevidad del polen de esta especie. Según evaluaciones mediante germinación in vitro, el polen perdió su viabilidad dentro de un lapso de ocho días, almacenado bajo condiciones ambientales (± 20 °C y ± 40 % HR).

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Background: Understanding pathways of genetic information transfer from one generation to another is particularly important when open-pollinated seeds are collected for reforestation or for enrichment of native forests. Nothofagus alpina (Poepp. & Endl.) Oerst. is native to temperate forests of southern Chile and Argentina. However, lack of knowledge of its reproductive biology has hampered its management in forest plantations and conservation of natural populations. Therefore, a clonal seed orchard of N. alpina in southern Chile was used to analyse the mating system and to estimate the number of pollen donors in this species. Methods: The mating system was investigated using manual pollination tests on six clones, open-pollinated seeds were collected from four other clones to study gene flow and paternity analysis was done on progenies using microsatellite genetic markers. Pollen donors were inferred for 194 offspring of four adult trees using five microsatellite loci and an exclusion process. Results: This species is exogamous and largely self-incompatible. The effective number of pollen donors found was 5, which was less than the 19 expected theoretically. Results indicated that pollen flow in the seed orchard is deficient. This could be a result of limitations in the wind-pollination system, coupled with phenological dissynchrony among clones present. Conclusions: N. alpina is an exogamous, highly self-incompatible species. It does not reproduce through agamospermy, since there was no seed formation in the absence of pollen. Only a few trees contributed to the paternity of the progeny evaluated. This finding indicates that there exists a limited gene flow within the orchard due to reproductive isolation among genotypes.