Impaginato 283 Adv. Hort. Sci., 2019 33(2): 283-294 DOI: 10.13128/ahs-24300 First insight into Araucaria araucana (Molina) K. Koch under its southern- most European growing condition: a proposed descriptor list for morphological characterization M. Antonetti, S. Nin, G. Burchi CREA, Centro di Ricerca Orticoltura e Florovivaismo,Via dei Fiori, 8, 51017 Pescia (PT), Italy. Key words: germplasm, monkey puzzle, phenology, phenotyping, climate, Pistoia’s nurseries. Abstract: Araucaria araucana is a south American endemic Conifer of conserva- tion concern. After its introduction to Europe, this species has been often plant- ed for ornamental purposes in parks and gardens, where its unusual appear- ance was admired. After the mid-1970s this tree received increasing attention from the nursery growers of Pistoia (Tuscany) and became of considerable eco- nomic importance. However, being one of the iconic threatened trees listed in CITES Appendix I, the international trade of these species is rigorously regulat- ed. This study was aimed at developing a first morphological descriptor list for further phenotyping of in loco produced plant material. A first step of the research focused on the description and a better understanding of A. araucana phenological phases inferred from Mediterranean climate conditions. The sec- ond phase regarded the analysis of observed or measured morphological char- acteristics of the tree, branches, scales, inflorescences, fruits and seeds observed on a subset of 4 selected putative populations over a 3-year period of vegetative growth. The results allowed to select 39 most discriminant descrip- tors, which are presented together with their range of variability and classes. The achieved descriptor list represents a suitable tool for the selection of geno- types and for the breeding of A. araucana. 1. Introduction Araucaria araucana (Molina) K. Koch, commonly known as ‘monkey puzzle tree’ or ‘pehuèn’, is an endangered conifer species native to south- central Chile and south-western Argentina, where it has a relatively limit- ed distribution, split between the main area spanning both sides of the Andes and two other disjunct small subpopulations in the Coastal Cordillera of Chile (Donoso, 1993, 2006; Donoso et al., 2008; Drake et al., 2009). The present distribution is a remnant of a more extensive former distribution, which has been severely diminished by logging, human-set (*) Corresponding author: maurizio.antonetti@crea.gov.it Citation: ANTONETTI M., NIN S., BURCHI G., 2019 - First insight into Araucaria araucana (Molina) K. Koch under its southernmost European growing condi- tion: a proposed descriptor list for morphological characterization. - Adv. Hort. Sci., 33(2): 283-294 Copyright: © 2019 Antonetti M., Nin S., Burchi G. This is an open access, peer reviewed article published by Firenze University Press (http://www.fupress.net/index.php/ahs/) and distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper and its Supporting Information files. Competing Interests: The authors declare no competing interests. Received for publication 10 December 2018 Accepted for publication 11 March 2019 AHS Advances in Horticultural Science http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ Adv. Hort. Sci., 2019 33(2): 283-294 284 fires and land clearance since European colonization in the mid-19th century (Veblen, 1982; Burns, 1993; Rechene, 2000). In particular, the intense human seed collecting and animal grazing have led to a lack of natural reproduction by seed, and when any regeneration occurs it is principally asexual with tree sprouting from roots (Schilling and Donoso, 1976; Gallo et al., 2004). Since 1976, this species has been protected in Chile under the status of a Chilean National Monument and, since 1997, it is also pro- tected internationally under the Convention on International Trade in Endangered Species of Wild Fauna and Flores (CITES) (Farjon and Page, 1999; Herrmann, 2006). A r a u c a r i a a r a u c a n a w a s f i r s t i n t r o d u c e d i n England by the Scottish naturalist Archibald Menzies in 1795. Its unusually straight cylindrical bole and whorled branches as well as its 10-15 cm thick tor- toise-shell-like bark made it internationally popular as an ornamental plant. The following decades saw a rapid spread of this impressively large and long-lived conifer throughout all the European continent. It was introduced in Italy from Paris in 1822 and the first A. araucana tree was planted in the garden of the Marquis Pucci’s favourite property in Florence, Tuscany. This Italian region represents the southern- most limit of its distribution area. Thanks to its per- fect adaptability to the Tuscan soil and climate condi- tions, this majestic slow-growing tree was included in the important ornamental horticulture district of Pistoia (NW from Florence) starting from the II post- world war period. Thereafter, there was a notable and rapid increase of A. araucaria commercial propa- gation in the Pistoia district in line with the steadily- growing demand. This tree became of considerable economic importance and the period between the 1970s till the early 2003 saw the maximum expansion of commercialization rate and tree planting in private properties and public gardens. Due to its constantly declining distribution, together with its slow growth and its limited dispersal ability, in 2003, according to the Regulation (CE) n. 1497, the listing of this species was transferred from Appendix II to Appendix I of CITES (http://www.cites.org/eng/app/appendices. html; valid from October 4th, 2017), which strictly regulates the trade in its timber and seeds, and listed in the 2008 IUCN Red List of Threatened Species (http://www.iucnredlist.org; March 2017) as an endangered species currently on risk of extinction. As a consequence, plant nurseries had to adopt a mandatory stock register of alive and dead specimen, where both entries and exits (including origin, quan- tity, causes of death, etc.) had to be specified. A pro- gressive and ongoing reduction of monkey puzzle tree propagation in the Pistoia district has resulted from increased regulation, complex management, high risk of penalties, increased costs for staff train- ing, and risks from plant diseases associated with cli- mate change. Ultimately, the number of nurseries holding and propagating Araucaria plants conspicu- ously declined during the last fifteen years. The identification of a new Tuscan variety could offer the opportunity to disengage from the proce- dural constraints imposed by the CITES Convention. Hence, the production of a descriptor list for the characterization of Tuscan selected germplasm is a necessary first step towards the definition of genetic diversity based on morphological variation and for varietal identification in Araucaria. The descriptor list might represent the first attempt at achieving an uni- fied documentation system thereby enabling through a standardized format an easier exchange of informa- tion between researchers and collection curators. Although there is a high demand for new descriptor lists to be developed for many forest conifer species, up to our knowledge there is only an UPOV descrip- t o r l i s t a v a i l a b l e f o r P i c e a a b i e s L . ( https:// www.upov.int/test_guidelines/en/fulltext_tgdocs.jsp?q= Picea; copyright © 2011, UPOV). Despite the conservation interest in this species, little is known of its phenotypic and genetic variation. The genetic diversity of monkey puzzle between Andean and coastal Chilean populations has been investigated in previous studies by Delmastro and Donoso (1980) and Rafii and Dodd (1998). More recently, advanced biotechnologies, such as RAPDs, Isozymes, microsatellite and RFLP analysis, were used to characterize genetic heterogeneity within and among some South American populations (Bekessy et al., 2002; Ruiz et al., 2007; Marchelli et al., 2010; Martín et al., 2012). However, no reports were found in the world literature on morphological traits. T h i s stu d y was p art o f th e C ARAV IV p ro j ect ‘Characterization of Araucaria araucana germplasm selected by the nursery industries of the Pistoia’s dis- trict for commercial development’, supported by the Ministero delle Politiche Agricole Alimentari e Forestali (MiPAAF-OIGA), aimed at contributing to a better understanding of Araucaria growing and to enhance the commercial exploitation of local genetic resources. This paper provides a brief account of Araucaria araucana phenological phases under Italian climatic conditions and is focused on the development of a first descriptor lists in order to Antonetti et al. - Auracaria araucana. First insight 285 characterize in loco produced plant material and make information available to other growers in a sys- tematic and unambiguous form. 2. Materials and Methods Plant material The plant material used in this study was the A. araucana germplasm available in the Pistoia’ nursery district, covering an area of approx. 965 sq km, rang- ing from 50 m to 550 m above sea level, located in northern Tuscany. Phenology Throughout three-year growing cycles (2015- 2017), the phenological phases (onset of flowering, full bloom, fertilization, fruit ripening and seed pro- duction) of A. araucana trees belonging to 8 putative populations were observed every two weeks from March to June (during the flower maturation), and monthly in the rest of the year. Taking into account the peculiar structure of the reproductive buds of this species, we decided to consider as onset of flow- ering the inflorescence appearance and as full bloom the inflorescence maturity, i.e. the pollination phase. Four out of the selected populations belong to Pistoia’s hinterland in a plain area (43°53’ N; 10°55’ E; 60 m a.s.l.), while the remaining four are located in high hills (44°0’ N; 10°52’ E; 550 m a.s.l.). The number of plants for each population ranged from 10 to more than 200, varying in age, gender and sexual maturity. Morphological descriptor list Twenty/twenty-five-year-old specimens belonging to a subset of 4 putative populations were randomly defined in order to perform a morphological descrip- tion. The considered populations were derived from seeds of different origin (unknown, Dutch fair, Spanish fair, local selected progeny) and grown in three private nurseries, located very close to each other in a plain area under the same organic regime. Local selected progeny refers to seeds collected from a couple of old trees located in Villa Lodolo (S. Marcello Pistoiese, 44°03’ N; 10°47’ E; 623 m a.s.l.). These trees were introduced from Argentina in 1920 and represent the main genetic source of A. arau- cana local germplasm. Climatic data, i.e. atmospheric pressure (AP), medium (AVG T), maximum (MAX T) and minimum (MIN T) air temperatures, relative air humidity (RH), wind run (WR), global horizontal irra- diation (GHI), rainfall (RAIN), evaporation (EV), for the same area were collected monthly from January 2015 to December 2017 (Table 1). Minimum, maximum, average, standard deviation and coefficient of variability (%) values of each morpho- metric character were calculated for the whole set of plants by using one-way analysis of variance (ANOVA). This in turn led to the definition of classes for all the measured traits by subdividing the total range into intervals 2 times the standard deviation, i.e. whether above or below the average value of each parameter as described in Bassi (2003). Statistical analysis was per- formed using SPSS 20 software (Chicago, IL, USA). Table 1 - Seasonal averages (2015-2017) of main meteorological data collected in Pistoia’s nursery areaa AP= atmospheric pressure; AVG T= medium air temperature; MAX T= maximum air temperature; MIN T= minimum air temperature; RH= relative air humidity; WR= wind run; GHI= global horizontal irradiation; RAIN= rainfall; EV= evaporation. a Processed from Ce.Spe.Vi database, Pistoia (http://www.cespevi.it/meteo.htm; Paolo Marzialetti © 1996/2018 Ce.Spe.Vi. - Pistoia). Year Season AP (mbar) AVG T (°C) MAX T (°C) MIN T (°C) RH (%) WR (km) TSI (kWh/m²) RAIN (mm) EV (mm) 2015 Winter 1009.33 8.33 14.73 3 68.33 86.7 1.53 2.7 1.6 Spring 1011 18.7 26.67 11.03 62.33 73.37 3.93 1.73 4.9 Summer 1009 24.77 33.13 16.87 57 69.6 4.17 1.23 6.5 Fall 1016.33 11.5 18.07 6.97 78.33 33.73 1.17 3.67 1.07 2016 Winter 1007.33 9.1 14.9 4.47 74 77.6 1.37 5.9 1.4 Spring 1007 16.83 25.2 9.83 65 77.9 3.7 2 5 Summer 1010.67 24.2 33.03 16.37 57.67 66.9 4.2 1.57 6.5 Fall 1014.67 10.43 18.3 5.1 78 24 1.3 3.57 1.03 2017 Winter 1012.33 8.47 15.77 2.43 67.33 66.4 1.67 3.53 1.77 Spring 1009.67 19.5 27.7 11.53 60 76.03 4.07 1.4 6.3 Summer 1009.33 23.67 32.47 15.43 57 65.6 4.03 1.7 6.43 Fall 1011.33 9.93 18 4.3 76 28.3 1.27 5.13 1.1 Adv. Hort. Sci., 2019 33(2): 283-294 286 3. Results Phenology The results of our observations throughout the growing cycles of male and female A. araucana trees in the Pistoia district are shown in figure 1. The main collected phenological data (flowering onset, flower maturation, fertilization, fruit ripening and seed pro- duction) were compared with those found in previ- ous published surveys on native Andean populations (Table 2). Morphological descriptor list Starting from field observations over a three-year growing cycle, a total number of 39 descriptors were developed for further germplasm phenotyping, divid- ed into 6 sections: 1) tree, 2) branches, 3) leaves, 4) male inflorescences, 5) female strobiles, 6) seeds and p r o d u c t i v i t y . T h e s e d e s c r i p t o r s a p p l y t o twenty/twenty-five-year-old trees (i.e. approx. the age of first fruit bearing) grown in nurseries as orna- mental plants. The descriptor list has been enriched by images and drawings for a better understanding Fig. 1 - Araucaria araucana male (♂) and female (♀) phenological stages under Tuscan climate conditions. Table 2 - Differences between the maturation stages of female and male inflorescences and seed production in Araucaria araucana populations in their origin area and in Italy I = first year; II second year; III = third year. a Tortorelli, 1956; Montaldo, 1974; Donoso and Cabello, 1978; Rodriguez et al., 1983; Marticorena, 1995. Phenological phases Chile/Argentinea Italy ♀ ♂ ♀ ♂ Flowering onset Nov. I Aug. I / Sep. I Mar. I Jul. /Aug. I Maturation Dec. I Dec. I May I May II Fertilization Jan. II - Apr. II - Seed production Dec. II / Feb. III - Sep. / Oct. II - Antonetti et al. - Auracaria araucana. First insight 287 of the less familiar and discernible traits. Descriptor list EXAMPLES a 288 Adv. Hort. Sci., 2019 33(2): 283-294 4. Discussion and Conclusions Very few studies have examined botanical aspects in A. araucaria following individual plants over their lifetimes, and all are rather dated and referred to plants grown under South America climatic condi- tions (Montaldo, 1974; Donoso and Cabello, 1978; Hoffman, 1982; Rodríguez et al., 1983). No data con- cerning the phenology and growing of this species under both European and Italian environmental con- ditions have been reported elsewhere, except for a very old contribution to the understanding of cytol- ogy and sexual reproduction in A. araucana plants grown in Northern France (Favre-Duchartre, 1960). Søndergaard (2003) reported about new introduc- tions of monkey puzzle to Scandinavia and the West coast of Norway, while Kubus et al. (2014) evaluated hardiness of A. araucana trees grown in open ground in Poland. However only data on annual shoot growth, tree height and degree of frost damages were given. On the other hand, phenological obser- vations are some of the most sensitive data in identi- fying how plant species respond to regional climate conditions and to climatic changes. L i k e a l l G y m n o s p e r m , m o n k e y p u z z l e h a s extremely simple flowers without any ornamental value. The male and female reproductive structures are carried by ‘cones’ and are as a rule separated, in a Araucaria draws have been taken and modified from the web- site http://www.eryprihananto.com (© Ery Prihananto, Indonesia). b Mean of 25 seeds. Antonetti et al. - Auracaria araucana. First insight 289 fact monkey puzzle is usually dioecious (Martinez, 1957; Bekessey et al., 2002). Nevertheless, although being basically female, 4 out of 15 flowering trees were found to produce both male and female cones during recording in southwestern of Norway; more- over, on completely isolated male and female trees, a couple of cones of the other sex were observed in some years in the northernmost area (Søndergaard, 2003). The Author suggested a possible correlation between stress (by isolation and climatically exposed situations) and monoecious behavior in A. araucana. In our census, only one monoecious tree was found among all considered specimen (approx. 700 plants) of the Pistoia province. The ratio of females to males in the examined populations grown in plain area was 1:4 (on a total of approx. 25% differentiated trees) being biased towards the male sex, while a rather balanced sex ratio was observed in older (approx. forty-five-year-old) totally differentiated trees grown in high hills (Fig. 2). This finding is almost in accor- dance with sex occurrence reported by Søndergaard (1975), who determined half female and half male trees in a population with 76% flowering trees in the West coast of Norway. No other comparison with lit- erature was possible for sex ratio, which has never been reported in elsewhere published data. Since the relationship between tree growth and climate appeared to be sex-dependant, in that male trees were more sensitive to land precipitation and female Fig. 3 - Araucaria araucana mature catkins during pollination. Fig. 2 - Totally differentiated Araucaria araucana female (A) and male (B) trees grown in Pistoia’s high hills, showing stro- biles and catkins formed in consecutive years. trees appeared more sensitive to air surface temper- ature during the prior period of growth (Hadad and Roig Juñent, 2016), it is probably realistic to assume that gender imbalance in favor of male or female at the beginning of sexual differentiation might vary according to different climatic environments, such as those found in Norway and Italy. On the contrary, day length did not seem to have a strong influence on the growth and development of the monkey puz- zle (Søndergaard, 2003); as a matter of fact, day l e n g t h d e p e n d a n t t r e e s w o u l d n e v e r s u r v i v e Scandinavian latitudes, since growth would begin too early and cease too late causing extensive frost dam- age and eventually killing the plant. Under the Tuscan environmental conditions, male catkins start out erect in July-August, stop growing during the winter season and then become elongated in shape, pendant and reddish-brown at maturity in May of the following year. Formed by many small scale-shaped leaflets, called microsporophyllus, they gave rise to a large quantity of pollen. In barely adult trees male caktins are found mostly in groups of 1-3 cones (Fig. 3), while older adult trees have groups of 2 up to 7-8 cones. First differences in the apex of potentially vegeta- tive or female flower buds become visible at the end of March (Fig. 1D). In flower buds the apex develops into a round dome of 3-4 cm, with more elongated and less tight scales provided with long yellow- orange appendixes. Female inflorescences, having f e r t i l e s c a l e s w h i c h c o n t a i n t h e o v u l e s , c a l l e d macrosporophylls, are distinguishable in April (Fig. 1E), grouped in light green strobiles at the extremity of the new sprouts. Greatest frequency of full bloom Adv. Hort. Sci., 2019 33(2): 283-294 290 and pollination was observed in May (Fig. 1C and 1F). Some differences were found in flower appearance among growing areas. Generally, flower buds devel- oped two up to three weeks later when trees were grown at higher altitudes and experienced rigid win- ters. After fruit set, that occurs in April of year II (Fig. 1G), the sessile and generally solitary and immature female strobiles are erect, globular, with a symmetri- cal shape, and green colored. They take usually about 4-5 months to develop into ripe globular dark brown mature cones (Fig. 1I) and remain closed until the complete maturation of the seeds. Scales usually fall off at maturity in August-September of the same year, although an ongoing trend towards the post- ponement of fruit drop towards November-late December was observed as a consequence of the gradual rise in mean temperatures (Fig. 4). As already assessed in previous studies (Søndergaard, 2003; Sanguinetti, 2014), cones occurrence was found to vary in relation with sun exposure; more in details, cones were most abundant in the South part of the crown. Almost all trees showed a marked alternate bearing; especially in males, yearly fluctuation in cone number seemed too large to be imputable only to climatic variations among years, particularly evi- dent for rainfall (Fig. 5). Alternate bearing was found to be much more pronounced in barely sexually dif- ferentiated trees compared to plants older than fifty years, suggesting that in young plants alternate bear- ing represents a strategic mechanism to save nutri- ent reserves for significant vegetative growth. Approx. 200-250 seeds, reddish to brown and oblong to obconical in shape, were released from each strobile; these range of seeds is fully included in t h o s e f o u n d o u t f ro m t h e l i t era t u re ( 150 - 3 0 0 ) (Montaldo, 1974; Donoso and Cabello, 1978; Salazar et al., 2000). Negative effects of heavy rainfall on pol- lination and seed production have been reported (Sanguinetti et al., 2002). Fructification began when plants were twenty- twenty-five-year-old, partially in agreement with reproductive organs appearance reported by Salazar et al. (2000) in Chilean A. araucana plants, while trees in Neuquén-Argentina have been reported to become sexually mature after thirty years of age, once the trunk has reached a diameter larger than 20 cm (Muñoz Ibáñez, 1984). Conversely, according to Søndergaard (2003), flowering was not initiated before the trees were forty/fifty-year-old in northern Europe and this discrepancy might be related to the altitude and latitude difference. The ontogenetic stages presented here (Fig. 6) were in accordance with the A. araucaria biological Fig. 4 - Minimum, maximum and average temperature values a n d t r e n d s i n P i s t o i a ( y e a r s 1 9 5 1 - 2 0 1 7 ) (http://www.cespevi.it/meteo.htm; Paolo Marzialetti © 1996/2018). Fig. 5 - Annual rainfall values and trends in Pistoia (years 1951- 2 0 1 7 ) ( h t t p : / / w w w . c e s p e v i . i t / m e t e o . h t m ; P a o l o Marzialetti © 1996/2018) Fig. 6 - Schematic representation of the ontogenetic cycle of Araucaria araucana. Solid lines represent diploid phases (female and male gametogenesis) and embryogenesis after fertilization, while dashed lines represent haploid phases (ovules and pollen). (From Favre-Duchartre, 1960, modified). Antonetti et al. - Auracaria araucana. First insight 291 cycle highlighted by Favre-Duchartre (1960) in north- ern France; moreover, differences in springtime tem- peratures between France and Norway corresponded fairly well to the differences in time (about one month) for release of pollen and seedfall at the two sites, as reported by Søndergaard (2003). On the con- trary, as shown in Table 2, A. araucana phenology was found to differ strongly from that observed in its n a t u r a l d i s t r i b u t i o n i n t h e A n d e s M o u n t a i n s . Obviously, differences between months were princi- pally due to the reversal of the hemispheres and con- sequentially of the seasons: when it is spring in Europe, it is autumn in the Andes and vice versa. However, changes seemed probably to be associated also to other factors, such as altitude, climate changes, growing conditions, etc. The effect of cli- mate on the phenology, and in particular on the tim- ing of reproduction, is well known for plants and extensively documented (Beebee, 1995; Stenseth and Mysterud, 2002). These studies have demon- strated that onset of reproduction in spring may have advanced by a week or two due to recent changes in climate over much of Europe, but to a much lesser extend in South America (Walther et al., 2002; Stenseth et al., 2002). Contrary to expectations, it was noticed that in males the appearance of the catkins takes place approximately in the same months (August-September in Chile and July-August in Italy), which, however, correspond to the end of summer in Italy and the end of winter in Chile. Moreover, in Chile the male catkins reach full maturi- ty within 3-4 months, whereas in our environmental conditions full bloom occurred within 9-10 months. This could be explained by the fact that in Italy the f u r t h e r d e v e l o p m e n t o f m a l e i n f l o r e s c e n c e i s stopped during the winter, while in Chile, where the season turns towards spring, caktins keep growing without interruption. On the other hand, maturation seems to be related to the photoperiod coinciding in both hemispheres with long days in late spring (beginning of December in Chile and end of May in Italy). Similar considerations can be drawn for females as well. The time between female flower appearance (March in Italy and November in Chile) and matura- tion (end of May in Italy and beginning of December in Chile, as for males) is about 3 months in our coun- try and 1 month in Chile. In both cases, the entry of the pollen tube into the ovary takes place within a month from the pollination. On the other hand, in our experimental conditions the true fertilization has been reported to occur after approx. 11 months (April of year II) from pollination (Favre-Duchartre, 1960), whereas timing of fecundation has never been mentioned in the literature under Andean environ- ments. There are contradictory records about the time of seed maturation in Chile, as it is unclear if fruits release the seeds after 11-13 months (Donoso and Cabello, 1978) or 16-18 months (Tortorelli, 1956; Montaldo, 1974) from pollination. Data herein obtained showed that gravity seed dispersal, which u s u a l l y t a k e s p l a c e a t t h e e n d o f t h e S u m m e r (approx. 12-13 months after pollination), moved towards Autumn (14-15 months after pollination) in 2016, and then towards early Winter (16-18 months after pollination) in 2017. Descriptor lists include the basic description of the traits, and the different classes of their expression (characterization) or how to measure the range of their variation (evaluation). Most of the descriptors for characterization and evaluation are species-spe- cific, but should be preferentially evaluated under homogeneous growing conditions in order to obtain comparable data and to avoid potential environmen- tal influence on the phenotype. In our study, in order to find the most appropriate descriptors able for dis- tinguishing effectively between individual pheno- types without loss of discriminating power, repeated observations were made on all plant material avail- able in the Pistoia’s nursery district. It immediately became clear that there were no evident differences among the expression of growing characters of young potted seedlings (Fig. 7), despite the very high number of individuals within the various collections, whereas a high degree of between- and within-popu- lation variability was noted among older trees (Fig. 8). A first analysis of the available specimen clearly evidenced that plant habitus as well as various other vegetative traits consistently varied with plant age, Fig. 7 - Araucaria araucana seedlings in Pistoia’s nurseries (A: two-year-old seedlings; B: six-year-old seedlings). Adv. Hort. Sci., 2019 33(2): 283-294 292 being this species a long-lived and massive tree up to 50 m tall and 2 m in diameter and attaining maxi- mum ages of at least 1300 years (Montaldo, 1974). Moreover, different climatic patterns and growing conditions, such as plant density and weed control, might have influenced plant growth and habitus as well. The variability observed here is consistent with the outbreeding dioecious reproductive habit of this species and suggests that these populations should continue to be viable and able to respond to moder- ate levels of environmental change (Hadad and Roig Juñent, 2016). However, recent phytopathological analysis in Tuscany have revealed an increase in mor- tality, especially of female individuals, without a spe- cific pathogen responsible (Rizzo, 2017, personal communication). The most widely accepted theory is that weaker plants, due to climate-related stress, are more vulnerable to damage caused by aspecific pathogens. well as plants grown in nearby locations. With respect to the 39 descriptors detailed in the descriptor list, some very peculiar characters, i.e. insertion angle of the scales on the trunk (N. 8), uni- formity of the apparent diameter on the primary and secondary branch (N. 19 and 20), catkin bending (N. 26), were individuated beside some of the most com- mon traits, such as shape and density of canopy (N. 3 and 4), maximum length and width of the leaves (N. 22 and 23). Undoubtedly, one of the tree’s most dis- tinguishing feature is its scales, which are stiff, dark green and glossy with a spiny tip and completely cover each branch, closely overlapping each other. But we found surprisingly interesting the size togeth- er with the insertion angle of the scales on primary and secondary branches, resulting in evident dissimi- lar apparent diameters of branches. On the contrary, female inflorescence didn’t show any distinctive fea- ture among populations, therefore it was not taken into account as a descriptor. In fact, while the true flower represents the main distinctive character in Magnoliophyta, the Pinophyta flowers, that are extremely simple, are not particularly relevant for the description of the species. In our study, morphological traits of A. araucana were observed, measured and documented for the first time under Tuscan growing condition. In particu- lar, the research was developed using trees from putative populations growing in the Pistoia’s nursery district. The comparison of the observed phenotypic characteristics showed a wide range of variability among and within the considered populations. The resulting data allowed to classify accessions, and to build a catalogue of specific descriptors with embed- ded biological information that is an essential step towards germplasm phenotyping (in particular new variety description), management or for direct use in agriculture. Limitations linked to the potential envi- ronmental influence on the phenotype are presented as well. Apart from this preliminary study, nothing is known of the patterns of morphological variation within this species. The development of this descrip- tor list will assist in the systematic and objective recording and exchange of information, which in turn will increase utilization of genetic resources along with a better screening and use of A. araucana biodi- versity for breeding programs. In order to analyze relationships between individuals or groups of speci- mens within locally grown populations, the morpho- metric characterization of A. araucana trees is in progress based on the defined descriptor list and suit- able statistical multivariate approaches. Genetic Fig. 8 - Habitus variability in Araucaria araucana trees grown in Pistoia’s hinterland in a plain area. A correct germplasm characterization should con- sider adult plants, being necessary the presence of flowers and fruits. However, as the aim of the pre- sent study is the phenotyping of young cultivated plants for sale, only trees sufficiently young for trade but at the beginning of sexual differentiation were suitable for characterization. This is why only twen- ty/twenty-five-year-old specimen belonging to a sub- set of 4 putative populations grown under similar environmental and agronomic conditions were con- sidered. Plants over 25 year-old were discarded as Antonetti et al. - Auracaria araucana. First insight 293 analysis of the same populations are being performed and will be processed in order to validate the discrim- inating efficiency of the presented descriptor list. Acknowledgements Authors acknowledge to the Vivai Bartolini, Azienda Macchia Tommaso and the kind collabora- tion of Andrea Tozzi (Villa Lodolo) for providing plant material and hystorical information. 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