First EST-SSRs of Helichrysum italicum (Roth) G. Don (Asteraceae) Revealed Insights into the Genetic Diversity and Population Structure in Corsica

<i>Helichrysum italicum</i> (Roth) G. Don (Asteraceae) is a valuable medicinal and aromatic plant native to a variety of habitats across the Mediterranean region. However, genetic studies of this morphologically diverse species have been limited by the scarcity of species-specific DNA markers. To address this limitation, we generated the first de novo transcriptome assembly comprising 24,806 transcripts from young shoots containing leaves and flowers, developed EST-SSR markers, and evaluated their utility in population genetic analysis. Seventy-eight primer pairs were designed, of which 23 showed successful amplification, polymorphism, and transferability to <i>Helichrysum litoreum</i> Guss. and <i>Helichrysum arenarium</i> (L.) Moench. A subset of 12 EST-SSRs was used to genotype 270 individuals from 12 natural populations of <i>H. italicum</i> in Corsica (France), along with one outgroup population from Croatia. The polymorphic information content ranged from 0.250 to 0.796, and Shannon's information index ranged from 0.588 to 1.843, indicating the markers' suitability for population genetic studies. Analysis of molecular variance revealed that 15% of the total genetic variation was attributable to differences among populations. Discriminant analysis of principal components and Bayesian clustering in STRUCTURE identified distinct population clusters corresponding to geographic locations. Notably, the southernmost coastal populations were clearly differentiated from the others.