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Seed Germination Characteristics of Phillyrea Angus- Tifolia L

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Seed Germination Characteristics of Phillyrea Angus- Tifolia L Ann. For. Res. 58(1): _-_ 2015 ANNALS OF FOREST RESEARCH www.afrjournal.org Seed germination characteristics of Phillyrea angus- tifolia L. and P. latifolia L. (Oleaceae), two Mediterra- nean shrub species having lignified endocarp S. Mira, L. Veiga-Barbosa, F. Pérez-García Mira S., Veiga-Barbosa L., Pérez-García F., 2015. Seed germination characteristics of Phillyrea angustifolia L. and P. latifolia L. (Oleaceae), two Mediterranean shrub species having lignifi ed endocarp. Ann. For. Res. 58(1): _-_ 2015. Abstract. The aim of this study was to determine the germination charac- teristics of Phillyrea angustifolia L. and P. latifolia L. seeds in order to develop an optimized propagation protocol for Phillyrea species. Seeds of P. angustifolia and P. latifolia were collected from wild plants growing in Cáceres province (CW Spain) and Andalucía (S Spain), respectively. Per- centage of water uptake for P. latifolia seeds was calculated. Seeds with and without endocarp were germinated at different constant and alternat- ing temperatures. Seeds without endocarp were soaked in distilled water or gibberellic acid, and then set to germinate. Seeds with endocarp of both species were stratified at 5 ºC for 30 or 90 days and then the endocarp was completely removed from the seeds before they were sowed. Chemi- cal scarification with sulfuric acid and mechanical scarification were tested on P. angustifolia seeds with endocarp. Phillyrea endocarp was permeable to water, since Phillyrea seeds with endocarp imbibed water, but water uptake was faster when the endocarp was removed. Moreover, the enco- darp could interfere mechanically in the emergence of the radicle, since seed germination of Phillyrea species was promoted by the complete re- moval of the lignified endocarp surrounding each seed. Optimal germina- tion temperature for both species was 15 ºC, and lower temperatures pro- duced secondary dormancy. Soaking in distilled water or gibberellic acid did not significantly enhance seed germination. Cold stratification and chemical scarification treatments were detrimental for seed germination. Keywords cold stratification, Phillyrea species, treatments before sowing, seed germination, seed scarification, lignified endocarp. Authors. Sara Mira ([email protected]), Félix Pérez-García - Department of Plant Biology, School of Agronomics Engineering, Technical University of Ma- drid, Madrid, Spain; Luciana Veiga-Barbosa - Department of General Biology, Federal University of Bahia, Salvador, Bahia, Brazil. Manuscript received September 29, 2014; revised January 26, 2015; accepted Ja- nuary 30, 2015; online fi rst February 05, 2015. 1 Ann. For. Res. 58(1): _-_ 2015 Research article Introduction bibition and thus seed germination (Baskin & Baskin 1998, Baskin et al. 2002). Due to the Seed germination is the most sensitive stage presence of a lignifi ed endocarp, it has been in the plant ability for spatial colonization suggested that Phillyrea species may presents (Baskin & Baskin 1998). In Mediterranean physical dormancy (García-Fayos et al. 2001, ecosystems, evergreens repeatedly face im- Takos & Efthimiou 2003), however, water ab- portant environmental constraints and cope sorption of the endocarp and dormancy break with extreme summer droughts, extensive fi res protocols have not been established. Within and soil erosion (Demmig-Adams & Adams the Oleaceae family, the genus Olea is char- 1996, Chaves et al. 2002, Dimitrakopoulos acterized by physiological dormancy and a et al. 2013). Climate change models project a woody endocarp that prevents the expansion reduction in total precipitation and drier sum- of the embryo (Morales-Sillero et al. 2012), mers (Christensen et al. 2007) and associated and even after total removal of the endocarp, increase in fi re hazard in Mediterranean re- seed germination is slow and erratic (Mitrakos gions (Piñol et al. 1998, Pausas et al. 2008). & Diamantoglou 1984). Seed germination of Therefore, in the Mediterranean areas, plant Phillyrea species was showed to be poor and regeneration strategies and environmental fac- unreliable in nurseries (Catalán 1991). This er- tors are closely interlinked, and knowledge on ratic germination of Phillyrea could have an seed germination of Mediterranean species effect on the presence of plants at different would be fundamental for the ecosystem con- physiological phases, complicating their man- servation and for directing regeneration efforts agement and commercialization. (Varela 2000, Pardos et al. 2005). The general aim of this work was to in- Phillyrea L. is a genus of 2 species in the Ole- vestigate the seed germination behavior of aceae family: P. angustifolia L. and P. latifolia Phillyrea species in order to develop an opti- L. (Andrés 2011). They are evergreen shrubs mized propagation protocol. The specifi c ob- very representative of the Mediterranean for- jectives of the present study were to determine est and with high importance in the ecological (i) the optimal temperature requirement for dynamics (Kutbay & Kilinç, 1994, Herrera et germination of P. angustifolia and P. latifolia al. 1994). Many of the natural and semi-natu- seeds, (ii) the effect of treatments before sow- ral forests containing P. angustifolia are under ing on germination for both species, (iii) the decline due to the impact of anthropogenic water uptake capacity of P. latifolia seeds with disturbance, and restoration efforts to address and without endocarp and (iv) the effect of dif- this are underway in many countries. Phillyrea ferent scarifi cation treatments on germination species are thermophilic and with low water- of P. angustifolia seeds. ing requirements, and have been recently con- sidered suitable for landscaping purposes for their little maintenance needs (De Marco et Materials and methods al. 2005). Phillyrea species are, therefore, of growing economical importance. The species Plant material can be propagated by seed germination and vegetatively (Piotto & Di Noi 2003), but plant Ripe fruits of P. angustifolia and P. latifolia rooting is diffi cult (Catalán 1991). Phillyrea were collected in 2011 from wild plants grow- species produce fl eshy-fruits (drupes), usually ing in Cáceres province (CW Spain) and Anda- containing a single seed enclosed by a lignifi ed lucía (S Spain), respectively. To carry out the endocarp. Typically, a lignifi ed endocarp may trials in which seeds without endocarp were be water-impermeable, thereby preventing im- used, the endocarp of the fruit was completely 2 Mira et al. Seed germination characteristics of ... removed using pliers and a scalpel. Seeds were P. latifolia assays were also performed with manually cleaned, kept in paper bags, and then seeds having intact endocarp. P. angustifolia stored dry under laboratory conditions (at ~23 seeds with endocarp where not included on the ºC under darkness, 35% relative humidity) assay due to the scarcity of seed material. In until the start of trials. Visible damaged seeds all trials, seeds that had not germinated at the were excluded from experiments. end of the incubation period were opened to determine whether the seed was empty. If so, Effect of temperature regimes on seed ger- they were excluded from calculation of fi nal mination germination percentages (Baskin & Baskin 1998). The number of empty seeds was always Seeds without endocarp of P. angustifolia and equal or less than 5% of the total seeds. P. latifolia were tested for germination at dif- ferent constant temperatures (5 ºC, 10 ºC, 15 Effect of treatments before sowing on seed ºC, 20 ºC, 25 ºC), with a 16-h light photope- germination riod (provided by cool white fl uorescent tubes with an irradiance of 35 μmol m2 s-1), and the 3 treatments before sowing were applied by alternate temperature regimes of 20/7 ºC and using seeds of P. angustifolia and P. latifolia. 4 25/15 ºC (the highest temperature for 16 h in replicates of 25 seeds each were tested for each light and the lowest one for 8 h in dark). In treatment: (i) soaking in distilled water. Seeds each trial, four replicates of 25 seeds each for without endocarp were soaked in distilled wa- P. angustifolia and two replicates of 30 seeds ter at room temperature (~ 23 ºC) for 24 or 72 each for P. latifolia were tested for germination h, (ii) soaking in gibberellic acid (GA3). Seeds on top of two sheets of fi lter paper (previously without endocarp were soaked in a GA3 solu- moistened with 3.5 mL of distilled water) in tion (1000 mg L-1) at room temperature (~ 23 7-cm-diameter glass Petri dishes. Filter papers ºC) for 24 h, (iii) cold stratifi cation. Seeds with were rewetted regularly with distilled water, as endocarp were stored in moist vermiculite un- required. Dishes were checked three times a der darkness at 5 ºC for 30 and 90 days. After week and germinated seeds were counted and the cold stratifi cation period, the endocarp was removed. Seeds were considered germinated completely removed from the seeds before on emergence of the radicle from the seed they were sowed. coat. The initial incubation period was 35 days After these three treatments, seeds were for P. angustifolia and 55 days for P. latifolia, tested to germinate at 15 ºC under a 16-h light which showed a slower germination. To study photoperiod for 65 days. Untreated seeds with- the effect of high and low temperatures of in- out endocarp were sown in the same conditions cubation on seed germination at the optimum and they were used as control. conditions, after incubation at different tem- peratures, non-germinated seeds were trans- Water uptake during seed soaking ferred to 15 ºC (which was the temperature that induced the highest germination percentages). To determine water uptake capacity during Thus, non-germinated seeds of P. angustifolia seed soaking, 2 replicates of 25 P. latifolia after 35 days at 5 ºC, 25 ºC and 25/15 ºC were seeds (with and without endocarp) each were transferred to 15 ºC and then incubated for weighted and then placed in Petri dishes on fi l- another 35-day period.
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