Fig 1.
Cells of the Calypogeia azurea complex and the related taxa with oil bodies.
(a) C. azurea s. str. (European clade; POZW 41390); (b-c) C. azurea—NA (North American clade; POZW 42444); (d-e) C. orientalis (VBGI P-39-4-15, VBGI P-8-1-16); (f-g) C. sinensis (China-56-78-13, China-56-77-13); (h-i) C. granulata (VBGI V-2-73-16, VBGI China 56-19-13); (j) C. peruviana (POZW 42619); (k) C. aeruginosa (VBGI V-8-75-17); (l) C. tosana (VBGI China-56-86-13). Bar = 50 μm.
Table 1.
The length (in bp) of the analyzed DNA regions in units detected within the C. azurea complex and the related taxa.
Fig 2.
Maximum likelihood consensus tree of the studied Calypogeia species based on a combined chloroplast dataset.
The analysis included all accessions, including those with incomplete sequences. Bootstrap values are given above the branches. Species with blue oil bodies are marked in blue.
Fig 3.
Maximum likelihood consensus tree of the Calypogeia azurea complex based on combined chloroplast dataset.
The results of the ABGD analysis are represented by colored stripes on the right side of the diagram. The first bar represents the initial partition, and the second bar represents the recursive partition. Only the accessions containing the sequences of all loci were included in the analysis. The related taxa (C. peruviana, C. granulata, C. tosana) were used for comparison. Calypogeia arguta and C. sullivantii were used as an outgroup. Bootstrap values are given above the branches.
Fig 4.
Maximum likelihood consensus tree of the Calypogeia azurea complex based on nuclear ITS2 sequences.
The results of the ABGD analysis represented by colored stripes on the right side of the diagram. The first bar of the diagram represents the initial partition, and the second bar represents the recursive partition. Only the accessions containing the sequences of all loci were included in the analysis. The related taxa (C. peruviana, C. granulata, C. tosana) were used for comparison. Calypogeia arguta and C. sullivantii were used as an outgroup. Bootstrap values are given above the branches.
Fig 5.
A haplotype network of the Calypogeia azurea complex and the related taxa based on combined plastid (A) and ITS2 (B) data. Colored circles represent the geographic origin of the haplotypes. Diameters denote the number of specimens carrying a particular haplotype, where the smallest circle represents a single individual, and the largest circle represents 16 individuals. Black squares represent median vectors.
Table 2.
Average genetic differences (%) between the units identified in the C. azurea complex and the related taxa, based on the K2P combined plastid loci (below diagonal) and nuclear ITS2 (above diagonal).
Fig 6.
Microscopic images of the leaves and underleaves of the Calypogeia azurea complex: C. azurea s. str. (European clade, 1a, 1b –POZW 41390, 2a, 2b –POZW 41746, 3a, 3b –HRE 57); C. azurea—NA (North American clade, 1a, 1b –MO 6005455, 2a, 2b –MO5241997, 3a, 3b POZW 42444); C. sinensis (1a, 1b –VBGI China-56-78-13, 2a, 2b –VBGI China 56-77-13, 3a, 3b –VBGI V-2-73-16) and C. orientalis (1a, 1b –VBGI P-40-1-12, 2a, 2b –VBGI P-39-4-12, 3a, 3b –VBGI J-7-79-14). The samples are numbered in accordance with S1 Table. Bar = 1000 μm for leaves, bar = 500 μm for underleaves.
Table 3.
Morphological differences between the units detected within the C. azurea complex and the related taxa.
Fig 7.
Calypogeia sinensis Bakalin & Buczkowska: (a) plant segment, dorsal view; (b-c) plant segment, ventral view; (d) lateral leaves; (e) leaf apex; (f) leaf margin (40×); (g-h) underleaves; (i) stem cross sections (10×); (j) oil bodies (40×). Sample VBGI China-56-77-13.
Fig 8.
Calypogeia orientalis Buczkowska & Bakalin: (a) plant segment, dorsal view; (b) plant segment, ventral view; (c) lateral leaves; (d) leaf apex (10×); (e) median leaf cells (10×); (f-i) underleaves; (j-k) stem cross sections (40×). Samples: VBGI P-39-4-12 (parts a, c, d, f, h, i, j, k), VBGI Kor-7-36-11 (parts b, e, g).