Crocus adamioides (Iridaceae) in the Bulgarian flora

Crocus adamioides Kernd. et Pasche, as it is currently known, was originally treated as C. biflorus Mill. subsp. adamii B. Mathew in the flora of Bulgaria by Mathew (1982) and verified by Rukšāns (2017). The taxon was afterwards described as a separate species (Kerndorff et al. 2012), the holotype being collected in Kırklareli Province, European Turkey. The species was for the first time mapped in two floristic regions of Bulgaria. The diagnostic characters based on the general morphology and leaf anatomy were defined from the natural populations of the species and compared to the type specimen and relevant data from the literature. The phylogenetic position of the species was clarified by sequencing of the internal transcribed spacer region (ITS: ITS1 + 5.SsrDNA + ITS2) and comparison of the obtained sequence with those annotated in NCBI. A phylogenetic tree was built using Bayesian phylogeny. Results have shown the highest degree of phylogenetic similarity with C. adamioides from Turkey. The closest relative C. ranjeloviciorum Kernd., Pasche, Harpke et Raca remains in the proximity. Our morphological, anatomical and molecular analyses have revealed that the Bulgarian population shows a peculiar combination of characters specific to C. adamioides

EVALUATION OF THREE SPECIES FROM PYROLACEAE FAMILY USING ISSR MARKERS

The family Pyrolaceae is represented in Bulgaria by six species, belonging to three genera. The viewpoints for the taxonomical position of the species in this family are not clearly unilateral. The aim of this study is to find molecular markers, allowing the objectively differentiation of the genera in this family, without use of morphological characters. Three species of the family were examined, representing the tree genera distributed in Bulgaria: Moneses uniflora (L.) A.Gray, Orthilia secunda (L.) House and Pyrola minor L. DNA was isolated from leafs and amplified via PCR reactions using 4 primers. The results were processed using cluster analysis and non-metric multidimensional scalling. The consequent cladogram, obtained by the mean euclidean distances, displayed grouping of the samples by genera, independently of their locality. The independent multitudes confirm the accuracy of the method for determination of the species in Pyrolaceae

<i>Crocus speciosus</i> (Iridaceae)—A New Species for the Bulgarian Flora

This is the first report on the autumn-flowering species Crocus speciosus, belonging to C. ser. Speciosi from the Bulgarian flora. The species was found in Southeastern Bulgaria, in the area between Ahtopol and Rezovo. Re-examining the Bulgarian collections, the earliest specimen was collected in 1975, was probably overlooked, and most likely determined as C. pulchellus. The nearest known localities of the species are on the territory of Türkiye. In this study, we compared C. pulchellus and C. ibrahimii using DNA sequence data from the nuclear ITS1/2 region and morphological features. Our study showed a close relationship between the specimens from Bulgaria and the recently deposited data of C. speciosus and their separation from the closely related C. pulchellus. Together with the previously cited white anthers as a key feature for determination, the molecular data confirmed a clear distinction between the samples with white anthers in the two species. The morphological data of our taxon overlapped with the description of C. ibrahimii. The molecular data strongly supported the affiliation of C. speciosus s.l., but did not support the recognition of C. ibrahimii as a separate species and it should be referred to as a subspecies of C. speciosus subsp. ibrahimii Rukšāns

Crocus pallidus (Iridaceae)&mdash;A Neglected Species for the Bulgarian Flora and Critical Taxon in the Balkans

For a long time, the Balkan endemic species Crocus pallidus has been unconfirmed and neglected for the flora of Bulgaria. It has remained an uncertain species from the Balkans, often listed as a synonym of C. weldenii. The morphological resemblance to the albinistic forms of C. chrysanthus has led to incorrect identification in the past, resulting in uncertainty regarding the distribution of this species in Bulgaria. In this regard, a detailed morphological and anatomical study of Bulgarian natural populations was carried out. A phylogenetic comparison in the ITS region of two Bulgarian populations of C. pallidus with other related species was conducted, revealing the distinction of C. pallidus populations from the closely related C. weldenii. The recently described new endemic species from Turkey, C. thracicus, shows very similar morphological and anatomical characteristics to C. pallidus and logically continues the distribution range in Eastern Thrace, along the Black Sea coast. We have a basis for suggesting that it should be treated as a synonym of C. pallidus

GENETIC DIFFERENTIATION BETWEEN SPECIES OF THE GENUS CROCUS BASED ON THE ISOENZYME ANALYSIS

This preliminary study represents data for the electrophoretic variations in the expression of the enzyme superoxide dismutase in six species of genus Crocus. Polymorphism at two superoxide dismutase loci was reported, with two alleles at the locus Sod-1 (Sod-1100 и Sod-189) and three alleles at the locus Sod- 4 (Sod-4100, Sod-490 и Sod-478). A comparative analysis of allelic frequencies was made and the degree of genetic differentiation between the studied species was characterized. The genetic distance according to Nei was determined and the phylogenetic dependences at the interspecific level were analyzed