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tinaei and its diploid (2n = 2x = 60) parents: C. hispanica and C. maderensis. Cheilanthes tinaei and C. maderensis are both located in the western Mediterranean basin and in some Macaronesian archipelagos whereas C. hispanica is subendemic to the Iberian Peninsula, with some populations also in Algeria and southern France. All three species grow in crevices of exposed siliceous outcrops from 0 to 1350 m above sea level (Garmendia 1986) and can be occasionally found in mixed populations. However, they tend to occupy different microhabitats, with an increase of water requirements in the order: C. hispanica (most xerophytic) GDC-0941 solubility dmso hemicryptophytes and have rhizomes that may branch but do not display extensive clonal growth. The leaves, which form dense apical rosettes, are bigger in C. tinaei (up to 30 cm long) than in C. hispanica (diglyceride province, 38��40��N, 2��38��W), C. maderensis in La Fregeneda (Salamanca province, 41��01��N, 6��55��W) and C. tinaei in Torrelodones (Madrid province, 40��35��N, 3��56��W). These localities followed the same relative position along the moisture gradient described above for the three species. Fragments of leaves with mature sporangia were collected between April and May 2005. Spore release was promoted by drying the leaf fragments on sheets of smooth paper for 1 week in the laboratory. Spores from the 15 individuals of each species were pooled prior to beginning the culture experiment. Experimental treatments Spores were sown on sterilized mineral agar (see Dyer 1979, p. 282) in 5.5 cm-diameter Petri dishes. The dishes were sealed with Parafilm (American National Can, Chicago, IL, USA) and incubated in a growth chamber (20 ��C, PAR 50 ?mol m?2 s?1, 16/8 h photoperiod). Five weeks after Selleckchem AZD0530 sowing, the resulting gametophytes, still asexual at that time, were transplanted to square Petri dishes divided into 25 isolated square cells of 2 �� 2 cm each (Bibby Sterilin; Barloworld Scientific, Stone, Staffordshire, UK). Into each cell, one gametophyte was transplanted on 3 cm3 of commercial potting soil (��Blumenerde��, Floragard, Oldenburg, Germany). Three moisture levels were established by watering weekly with 100, 200 or 500 ?L of distilled water per cell (hereafter low, medium and high moisture, respectively).