Limonium species endemic to Zakynthos Island 595
private allele in subpopulation Lp7.Ld423 was highly poly- morphic, with 22 different alleles across all subpopulations. Of these, seven were present in both species, and the others were species-specific. Limonium phitosianum had 11 private alleles, some of which were subpopulation-specific (three in subpopulation Lp7, six in subpopulation Lp10), and two occurred in more than one subpopulation (Lp1 and Lp7; Lp7 and Lp10). Limonium zacynthium had four private alleles, one in subpopulation Lz1 and three in Lz2. Mean number of alleles (Na) ranged from 1.000 to 3.200
(for subpopulationa Lp1 and Lp10, respectively), and the mean effective number of alleles (Ne) ranged from 1.000 to 2.706 (for the same subpopulations). Shannon’s informa- tion index (I) ranged from 0.000 to 0.474 (for Lp1 and Lp10, respectively). Observed heterozygosity (Ho) was lowest in Lz3 and Lp1 (0.000) and highest for Lp7 (0.220), and ex- pected heterozygosity (He) had the lowest and highest va- lues in Lp1 and Lp7, respectively (Supplementary Table 3). The inbreeding coefficient (Fis) values were positive for all subpopulations, except for Lp1 where no value was obtained by GENEPOP. Fis was 1.000 for Lz3, and for the other sub- populations the following values were estimated: 0.0913 (Lz2), 0.1091 (Lp7), 0.1884 (Lp10), 0.3000 (Lz1). The mean fixation index (FST) value was 0.172 when calculated only among L. phitosianum subpopulations and 0.151 among L. zacynthium subpopulations. The overall FST calculated using the data available for both species was 0.159, and the value of gene flow (Nm) was 0.424.
FIG. 3 Flowering and fruiting periods of the three species of Limonium during 2014–2018.
and C2a(ii), with ,250 mature individuals (C) and a pro- jected continuing decline in number of mature individuals (2), and all individuals in a single subpopulation (a(ii)). We assess L. phitosianum as Near Threatened, as it is close to meeting the criteria for Vulnerable based on its EOO and AOO, with the exception of the number of locations, which is.10.We assess L. zacynthiumas Endangered based on cri- teria B1ab(iv,v) + B2ab(iv,v), with EOO,100 km2 (B1)and AOO,10 km2 (B2), and #5 locations (a), and a projected continuing decline in number of mature individuals (b(v)) and number of locations or subpopulations (b(iv)).
Genetic analyses
Analysis of the five microsatellite loci revealed 28 alleles (1–22 alleles/locus) in total. Ln115 and Ln152 loci were monomorphic in both species, and Ln39 is monomorphic in L. phitosianum but polymorphic in L. zacynthium, with a private allele in subpopulation Lz2.Ln68wasmonomorphic in L. zacynthium and polymorphic in L. phitosianum,witha
Discussion Population size and reproductive biology
In this study, we monitored all extant subpopulations of three rare Limonium species endemic to Zakynthos Island for 5 years. The EOO and AOO remained stable throughout the study for all three species. Therewas aprevalenceofmatureindividuals of L. korako-
nisicum, with low seedling abundance. The rocky calcareous substrate and potential competition with seedlings of the sym- patric L. phitosianum could impede successful establishment and growth of L. korakonisicum seedlings, although relative reproductive success was consistently high, indicating this is not a limiting factor for the species’
survival.Given a seed ger- mination rate of up to 86%(Valli&Artelari, 2015),we suggest that seedling establishment and survival could be a bottleneck in the species’ reproductive cycle. The population viability analysis indicated a 3.9% extinction risk over the next 50 years. However, deterministic models may underestimate quasi-extinction probability by not considering other factors influencing population viability (Akçakaya et al., 1999; Morris & Doak, 2002). There were fluctuations in the seedling survival rate of L. phitosianum. The lowest spring rainfall duringmonitoring,
Oryx, 2024, 58(5), 587–599 © The Author(s), 2024. Published by Cambridge University Press on behalf of Fauna & Flora International doi:10.1017/S0030605324000140
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