Abstract
The Balkans endemic species Alkanna primuliflora Griseb., A. stribrnyi Velen., and A. graeca Boiss. & Spruner have limited distribution in the Balkan Peninsula and a large variation in the morphological characteristics. The populations of the three Alkanna species in the Bulgarian flora are small and fragmented. There are no previous reports on the chemical profile or on the embryology of these species. The hypothesis was that the limited distribution of A. primuliflora, A. stribrnyi, and A. graeca was due to their reproductive capacity and genetic diversity. Furthermore, we hypothesized that the three species will contain pyrrolizidine alkaloids (PAs), as other species of the genus Alkanna (Boraginaceae), but they would have differential alkaloids composition. The population genetic structure and differentiation showed a clear distinction between species and revealed average levels of genetic diversity among the natural populations of the three Alkanna species. The embryological investigation observed stability of the processes in the male and female generative spheres and high viability of mature pollen and embryo of the three species. The normal formation of male and female gametophytes without deviations or degenerative processes, and observed levels of genetic diversity between Alkanna individuals are important in maintaining the size and resilience of the Alkanna populations. Eight alkaloids were identified by GC-MS in A. primuliflora and A. graeca and six alkaloids in A. stribrnyi. The main pyrrolizidine alkaloids (PAs) in all investigated species was triangularine. A. primuliflora and A. graeca showed similar chemical composition that comprised 9-angeloylretronecine, 7-tigloylretronecine, 9-tigloylretronecine, triangularicine, dihydroxytriangularine, dihydroxytriangularicine, whereas, in A. stribrnyi 9-tigloylretronecine, triangularicine and dihydroxytriangularicine were not found. This is the first report on the presence of PAs in A. primuliflora, A. stribrnyi and A. graeca. Besides, this is the first report on the embryology of these endemic species. The results contribute to the knowledge of the three endemic Alkanna species and will facilitate policy-making and defining new strategies for their conservation.
Highlights
The genus Alkanna belongs to family Boraginaceae, subfamily Boraginoidae, tribe Lithospermeae [1]
The hypothesis was that the limited distribution of A. primuliflora, A. stribrnyi, and A. graeca was due to their reproductive capacity and genetic diversity
The investigated Alkanna species had a total of 20 private alleles, nine of A. primuliflora and A. graeca and two of A. stribrnyi (Table 3)
Summary
The genus Alkanna belongs to family Boraginaceae, subfamily Boraginoidae, tribe Lithospermeae [1]. Genus Alkanna is represented by a large number of local and regional endemic species [3]. Six Alkanna species are found in the Bulgarian flora and five of them are either Balkan or Bulgarian endemics. The three studied species have very limited distribution in Bulgaria as follow: A. graeca—in two floristic regions, Pirin Mt (South), Thracian Lowland; A. stribrnyi–in three floristic regions, Slavyanka Mt, Thracian Lowland, Pirin Mt (South); and A. primuliflora—mainly in four floristic regions, Thracian Lowland, Pirin Mt (South), Valley of Struma river, and Slavyanka Mt. The populations of A. primuliflora, A. stribrnyi, and A. graeca are included in the European ecological network EU NATURA 2000 under Directive 92/43 / EEC on Habitats. The populations of three Alkanna species are small, fragmented and represented by a very limited number of individual plants
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