Crocus novicii V.Randjel. & Miljković, 2016

Miljković, Milica, Ranđelović, Vladimir & Harpke, Dörte, 2016, A new species of Crocus (Iridaceae) from southern Albania (SW Balkan Peninsula), Phytotaxa 265 (1), pp. 39-49 : 41-46

Crocus novicii V.Randjel. & Miljković sp. nov.

Crocus novicii differs from other species of the C. sieberi aggregate by its color of tepals (white with lilac coloring at the base), white stylus and white filaments ( Table 1, Figure 1 and Figure 2 ).

Type: — Albania, Nemërçkë Mt., alpine grasslands around snowmelts, limestone, 1860 m, 30 May 2014, V.Randjel. & Miljković 17112 (holotype BEOU!)

Corm about 0.5–0.9 cm in diameter, flattened-subglobose; corm tunics brown and papyraceous with thin fibres indistinctly reticulated. Cataphylls 3–4 (–5), white, membranous-papyraceous. Leaves 3–5–6 (–7), synanthous, mainly shorter than flower at anthesis, 2.7–5.7 mm wide, erect, glabrous, green, with revolute margins and with a prominent white median stripe on the adaxial surface with width of 1/ 3 in relation to the leaf diameter. Flowers vernal, solitary, slightly scented. Throat white, glabrous. Prophyll absent. Bract and bracteole present, subequal, membranous, both 3.2–4 cm long. Perigone tube 2.7–4.5 cm long, lilac in the upper part. Perigone segments obovoid, obtuse or subacute, white and at the base of the outer side lilac almost to the edge, rarely segments completely pale lilac with a darker lilac coloring almost to the edge at the base of outer side; outer segments 2.2–4.9 cm long, 0.6–1.2 cm wide; inner segments 2.2–4.6 cm long, 0.6–0.7 cm wide. Filaments 1–2.3 cm long, white, glabrous; anthers 0.8–1.3 cm long, yellow. Style white, up to 0.3 cm longer than anthers (rarely shorter than them); stigma divided into 3 white branches 0.3–0.8 cm long; branches with extended and frilled tips. Capsule ellipsoid, 0.8–1–1.2 long. Seeds reddish-brown to dark brown, 0.21–0.27 cm long, 0.15–0.2 cm wide.

Distribution and habitat:— Crocus novicii is endemic to the Nemërçkë mountain in the southeastern part of Albania (southwestern Balkan peninsula) and grows on limestone substrate at an elevation of 1700–1900 m. This species is associated with dominant Plantago atrata Hoppe (1799: 85) , Armeria canescens Boissier (1848: 686) , Myosotis alpestris Schmidt (1794: 26) and Ranunculus montanus Willdenow (1799: 1321) , building alpine association from the class Salicetea herbaceae Br.-Bl. 1947, order Salicetalia herbaceae Br.-Bl. 1926 and the alliance Salicion herbaceae Br.-Bl. 1926. Besides these species also Poa alpina Linnaeus (1753: 67) , Astragalus depressus Linnaeus (1756: 29) , and Scilla bifolia Linnaeus (1753: 309) are notable.

Etymology:—Novica Ranđelović (1937–) is one of the most prominent Serbian botanists. He has collected and studied a lot of specimens from the Balkan Peninsula. The results of his investigations were published in numerous papers. However, crocuses were always one of his favorite study objects. He described two new species, C. rujanensis and C. jablanicensis , published a monograph about Serbian crocuses ( Ranđelović et al. 1990) and wrote a publication about the genus Crocus in Macedonia ( Ranđelović et al. 2007). He was also a great teacher, who influenced and inspired many young scientists.

Phenology:—Flowering period from late May to June.

Morphological comparison:—Morphologically, Crocus novicii sp. nov. belongs to the C. sieberi aggregate, with whom it shares the absence of a prophyll and the reticulated corm tunic. It can be distinguished from most of the species of the C. sieberi aggregate by the white color of style and stigma (others yellow). The only other species also having a white style and stigma is C. jablanicensis . Both taxa share corm shape, corm-tunic form, similar plant size, length of bract, bracteole, perigone tube, perigone segments, filaments, anthers, the same white, glabrous perigone throat, reddish brown to dark brown seed color (see Table 1). However, C. novicii differs from C. jablanicensis by its higher number of cataphylls and leaves and by the presence of wide lilac coloring at the base of perigone segments and perigone tube, which is lilac in the upper part ( Table 1).

Karyological analysis: — Crocus novicii proved to be diploid with 2n = 2x = 20 chromosomes ( Fig. 6).

Chloroplast DNA analysis: — The sequence lengths of the two chloroplast regions analyzed for this study are 640–642 base pairs (bp) for rps 16– trn Q (4 variable positions), and 620–630 bp for mat K– trn K (12 variable positions) within the C. sieberi aggregate. Concatenation of the two loci for the C. sieberi aggregate resulted in an alignment length of 1272 bp. Shortening of gaps and excluding a mono-nucleotide repeat (T stretch) for the statistical parsimony analysis in TCS resulted in an alignment of 1264 bp. TCS calculated a network with a connection limit of 8 steps for this alignment, and inferred 6 chloroplast haplotypes and 7 missing intermediates not present in the data. Crocus novicii shares the same haplotype as the investigated C. veluchensis from a geographically close population in Greece ( Fig. 3 ).

Nuclear DNA analysis:—The length of the ETS region varies from 523–526 bp for the C. sieberi aggregate species, and has a size of up to 522 bp in the outgroup taxa. In some individuals direct sequencing showed a few ambiguous nucleotide positions, which were coded using the nucleotide ambiguity code (IUPAC). In contrast to ETS, length differences in the ITS data set are more than six times higher (671–696 bp in the C. sieberi aggregate, 634–696 with outgroups). Intra-individual length polymorphism resulted in unreadable sequences in C. novicii and C. veluchensis . Hence, cloning was necessary. In the combined ETS and ITS Bayesian analysis C. novicii and C. veluchensis form a monophyletic group within the highly supported C. sieberi aggregate ( Fig. 4 ).

In the NN of the ITS data set C. novicii and C. veluchensis sequences, derived from cloning, grouped (i) either together close to C. dalmaticus and C. sieberi subsp. atticus or (ii) without any other close species, and (iii) two copies of the ITS region of C. veluchensis grouped together close to C. rujanensis ( Fig. 5 ). Different ITS copies obtained from cloning were unique to the species to which the sequenced individual belongs.

The length of the investigated part of TOPO 6 ranged from 677 to 687 bp. It was not found to be single-copy in C. jablanicensis , C. rujanensis and C. veluchensis . Crocus jablanicensis and C. rujanensis copies differed in a highly variable region (a up to 16 bp long T stretch sometimes including one or two C in the C. sieberi aggregate). In C. veluchensis , one type of the copies found had an insertion, which was unique in the data set while the other type of copy was highly similar to C. novicii (differing only by 3 bp; see Fig. 5B ). Both copies of C. veluchensis clustered together with C. novicii . Crocus jablanicensis was found to be sister to C. cvijicii in the analysis of all nuclear markers ( Figs. 4–5 ).

Figures

FIGURE 1. Photos of plants from the type population of Crocus novicii (a), its typical flower (b), the anthers and filaments (c), the corm (d), and the stylus (e) photographed by V. Ranđelović.

FIGURE 2. Photos of C. novicii (a), C. jablanicensis (b), C. cvijicii (c), and C. veluchensis (d) photographed by V. Ranđelović.

FIGURE 3. Chloroplast haplotype genealogy for the C. sieberi aggregate derived from concatenated sequences of the rps16–trnQ, and matK–trnK regions of the chloroplast genome. Black dashes depict missing haplotypes, i.e. alleles not found in the analyzed individuals. Crocus novicii is marked bold.

FIGURE 4. Phylogenetic tree obtained by Bayesian phylogenetic inference of the combined sequences of the nuclear rDNA ETS and ITS regions. Numbers along branches give posterior probabilities. Species names followed by a small letter refer to cloned sequences of the ITS region. Crocus novicii is marked bold.

FIGURE 5. Neighbor-net splits graph of (A) the ITS region and (B) the Topo6 region. Species names followed by a small letter refer to cloned sequences. Crocus novicii is marked bold.