Notes on Crocus L. Series Flavi Mathew (Iridaceae) and a New Species with Unique Corm Structure

Phytotaxa 438 (2): 065–079 ISSN 1179-3155 (print edition) https://www.mapress.com/j/pt/ PHYTOTAXA Copyright © 2020 Magnolia Press Article ISSN 1179-3163 (online edition) https://doi.org/10.11646/phytotaxa.438.2.1

Notes on Crocus L. Series Flavi Mathew (Iridaceae) and a new species with unique corm structure

ALMILA ÇIFTÇI1,4, DOERTE HARPKE2,5, RACHEL MOLLMAN1,6, HASAN YILDIRIM3,7 & OSMAN EROL1,8* 1 Botany Division, Department of Biology, Faculty of Science, Istanbul University, 34116 Beyazıt, Istanbul, Turkey. 2 Leibniz Institute of Plant Genetics and Crop Research (IPK), 06466 Gatersleben, Germany. 3 Biology Dept, Botany Section, Science Faculty, Ege Univ., Bornova, Izmir, Turkey. 4 �[email protected]; https://orcid.org/0000-0002-3406-3064 5 �[email protected]; https://orcid.org/0000-0003-1667-2912 6 �[email protected]; https://orcid.org/0000-0002-6873-6212 7 �[email protected]; https://orcid.org/0000-0003-3951-4343 8 �[email protected]; https://orcid.org/0000-0002-6310-1402 *Author for correspondence

Abstract

Crocus asymmetricus (Iridaceae) is described as a new species endemic to the southern part of the Anatolian diagonal in Turkey. It is phylogenetically related to C. vitellinus and morphologically to C. antalyensis, but differs from these species in showing an asymmetric corm and a single point of root emergence. Both C. antalyensis and C. asymmetricus are illustrated and compared in this paper. A Bayesian phylogenetic tree of the nuclear rDNA ITS region confirms the affiliation of C. asymmetricus to C. ser. Flavi, and its close relationship to C. vitellinus. A new identification key to the species of C. ser. Flavi occurring in Turkey is also presented.

Introduction

Crocus Linnaeus (1753: 36) (Iridaceae) is one of the largest genera in the flora of Turkey. This genus is represented by over 135 species according to Rukšāns (2017). Some species are taxonomically doubtful, because of unclear locality data, inadequate type specimens, and lack of clear diagnoses. Despite this, Turkey is the centre of diversity of the genus and scientists still discover new taxa even near heavily trafficked roads and human settlements. Most of the Turkish Crocus taxa belong to C. sect. Nudiscapus Mathew (1982: 61), which is further divided into series. In several cases, a detailed examination of corm tunics allows the placement of a taxon in a series or even identification at the species level (Kerndorff et al. 2015). Therefore, the corm still represents a useful part of the plant for diagnostic purpose. Within C. sect. Nudiscapus, C. ser. Flavi Mathew (1982: 84) is characterized by membranous (sometimes papery) parallel-fibrillated tunics and multifid stigma branches (Mathew 1982), with the exceptions of C. adanensis Baytop & Mathew (1975: 245) and C. paschei Kerndorff (1994: 76), both of which have a three-branched stigma and poorly developed rings at the base of the corm. Phylogenetic studies (Petersen et al. 2008, Harpke et al. 2013) placed the latter two taxa as sister to all the other C. ser. Flavi members. In the current circumscription, C. ser. Flavi includes 16 taxa: C. adanensis, C. antalyensis Mathew (1972: 327) subsp. antalyensis, C. antalyensis subsp. striatus Erol & Koçyiğit (2010: 187), C. antalyensis subsp. gemicii Sik & Erol (2011: 282), C. antalyensioides Rukšans (2015: 6), C. balansae J.Gay ex Baker (1879: 234), C. candidus Clarke (1812: 145), C. mouradi Whittall (1889: 473), C. flavus Weston (1771: 237), C. graveolens Boissier & Reuter (1882:107), C. hyemalis Boissier (1859: 93), C. istanbulensis Mathew (1982: 99) Rukšâns (2017: 271), C. olivieri Gay (1831: 319), C. paschei Kerndorff (1994:76), C. sarichinarensis Rukšans (2010: 157) Rukšans (2017: 442), and C. vitellinus Wahlenberg (1828: 1000). An amateur scientist, Mehmet Çelik, found a Crocus population near the locus classicus of C. adanensis in Maraş province, Turkey. The individuals of this population are characterized by asymmetric corms, a shape unique among all species of the genus, thus suggesting that they represent a new species. Therefore, we carried out a thorough morphological investigation and used the rDNA internal transcribed spacer (ITS) region to confirm its rank as a putative new species of C. ser. Flavi and to identify its closest relatives.

Accepted by Giovanni Astuti: 13 Mar. 2020; published: 6 Apr. 2020 65 FIGURE 1. Crocus asymmetricus from its locus classicus: habit (A, B); peeled corm (C); corms with tunics (D).

The most interesting feature of Crocus asymmetricus sp. nov. is the organization of its corm tunics. Its flower morphology is similar to C. antalyensis, but it differs by its asymmetrically oriented corm and tunics, as well as by its lack of persistent cataphylls (Figs. 1, 2, 3).

Morphological investigations:—Standard deviations (SD) were calculated for each quantitative morphological parameter. The number of individuals measured (indicated by n) for each morphological character is given in the description. Photographs of the plants were taken with a Canon 5Ds digital camera (60 and 100 mm macro-lenses). Macromorphological features were examined under a stereo-binocular microscope. We compared the morphology of the undescribed specimens to the relevant species descriptions available in the taxonomic literature (Maw 1886, Bowles 1952, Mathew 1982, Mathew 1984, Rukšāns 2017) and herbarium material conserved in EGE, ISTE, ISTF, K (acronyms follow Thiers 2019).

FIGURE 2. Illustration of C. asymmetricus: A. Habitus; B. Flower dissection; C. Leaf cross section; D. Corm: D1. General view, D2. Young (above) and mature corm (below); E. Corm tunics (E1 and E2); F. Basal tunic. Drawing made from specimen ISTF 41370.

Notes on Crocus L. Series Flavi Mathew Phytotaxa 438 (2) © 2020 Magnolia Press • 67 FIGURE 3. Illustration of Crocus antalyensis: A. Habitus; B. Flower dissection; C. Leaf cross section; D. Corm: D1. General view, D2. Young (above) and mature (below) corm; E. Tunics (E1, E2, E3). Drawing made from specimen ISTF 41144.

Molecular methods:—The extraction of genomic DNA and amplification of the nuclear ribosomal region ITS of five individuals were performed according to Harpke et al. (2013). Both strands of the PCR products were directly Sanger sequenced with Applied Biosystems BigDye Terminator technology on an ABI3730xl automatic DNA sequencer using the primers from PCR amplifications.

FIGURE 4. Phylogenetic tree obtained by Bayesian inference of the nuclear rDNA ITS. Numbers along branches are posterior probabilities.

sym full prl outer tunics split strap- like at base formed by old cataphylls ab obov obl sac C. paschei C. to p ob ; , with sym full p occasional splits at base short, ab ov sac

; C. antalyensioides C. prl sac at sla ...... continued on the next page spl to to ,

sym full p base into many fibrils formed by old cataphylls ab ov ob C. candidus C. into spl fibrils ob , ; sym full m prl narrow sharp tunic splits ab el

, C. mouradi C. ob a ;

to fibrils ov , distinctly sym full m fibrous with prl formed by old cataphylls ab obl or sac

; at C. flavus C. prl ov spl

or with sac sym full m fibrous points at apex, the base into coarse fibres formed by old cataphylls ab obl to ,

;

C. istanbulensis C. prl obl to sym full c coarsely fibrous, fibrils weakly reticulate at apex bristly ab el sac

C. balansae C. prl sac at base ; sym full m spl into fibrils narrow sharp fibrous splits ab ov

to C. olivieri C. ob to ; fibres or at base sym full m spl into coarse, prl triangular teeth sharp triangular fibrous points ab el, obl obov sac

; C. hyemalis C. ov ob to to strips at length sym full m spl ways into prl base poorly developed, soft fibrous points ab obl sac

ob obl C. vitellinus C. ; ;

and its related species (abbreviations written in bold a list of all abbreviations given at the end table at base obov sac sym full m/c spl into narrow strips or fibrils fibrous points of main tunic ab nd wai to to

prl

ob or C. graveolens C. at to spl obl , ; ac ; sym full m/c base into fibrils fibrous points of main tunic ab wai el ;

sla Crocus asymmetricus Crocus adanensis C. into p sac to spl fibrils to , several sym two – m stripes or prl short, poorly developed obov ob

ob C. antalyensis C. ; fibrils into obl prl spl sac to , sym full p narrow bands or from base formed by old cataphylls ab el to

sac C. asymmetricus C. spl ) to ob ; asym asym one c/m heads towards one side ( into narrow bands, or triangles from base narrow, sharp, triangular teeth ab el Morphological comparison of a b le 1. S pecies/ Characters Corm shape Emergence of root Tunics neck Tunic Basal rings Flower segment shape T alphabetical order).*

gr gla

wh br C. C. -ish -ish gr

br occ usually to deep

with a , , y or creamy, always suffused, speckled or diffusely striped ish to y, dk or speckled, well-defined basal blotch pl y with a distinct zone above throat,

- to sl C. paschei C. gr sil dkr gla to deep , outside buff-colored with a distinct ish speckled basal blotch or only speckled at the base, maybe extending up in three indistinct lines Same as but outer, dkr pl y

antalyensioides

gre - ...... continued on the next page C. C. blotch vio

gr gla or with , uniform bl dkr shading along midrib, ish at base Same as outer y

or y

, C. candidus C. bl y with -ish- spotted or suffused gr pur wh base bright gla

pub y C. mouradi C. -ish or dk gr

to gla to dark , same , pl y color inside and out, sometimes with staining at base pl y ,

, br C. flavus C. y y

gr or o o to to or y y gla

, pl deep sometimes striped or stained vio base pl deep y pub

to C. istanbulensis C. , y o o gla to , uniform to y same colour inside and out, without marking deep o

, C. balansae C. -ish y y or

gre -ish o o br gr y gla to to , y sometimes with translucent striping, very rarely pl y with ish, or basal blotch y pub

C. olivieri C. -ish o or br , rarely or y gla

y ,

usually uniform bright o sometimes pl with striping or staining bright y pub C. hyemalis C.

with , with vio gla -ish , wh pur staining or speckling, sometimes with central band of reaching apex wh pointed gr basal blotch y

- C. vitellinus C. gla y y , , br y y

vio pub o yo to bright to sometimes with ish- stripes or speckles bright to o or

C. graveolens C. vio , pub y or usually -ish- o o

y with 3 major br stripes y deep gla ,

wh C. adanensis C. gla wh - sl y li li

dull pl usually sometimes with center,

C. antalyensis C. vio , or , or with pub or deep staining or deep bl bl

, pl li wh bl on base, sometimes buff-colored with stripes or flecks pl li wh with staining on base y

C. asymmetricus C. gla pur , wh y - li , stripe pur sometimes with one inconspicuous vio bright dull with three distinct stripes and featherings (Continued) S pecies/ Characters Colour of outer segments Colour of inner segments Throat B LE 1. TA

Notes on Crocus L. Series Flavi Mathew Phytotaxa 438 (2) © 2020 Magnolia Press • 71 C. sarichinarensis C. × × primarily 34 16 34 16 17 – – – – – 21 10 21 10 y 10 mul divided into 3 at the apex, divided into secondary branches

C. paschei C. × ×

sl 38 30 , – 15 – 12 10 – – – 28 9 19 6 y 7 tri expanded and fringed at apex

...... continued on the next page C. antalyensioides C. 15 × × – 8 40 20 40 20 20 with – – – – – 35 17 35 17 y 10 mul bra short subdivisions

× C. candidus C. × sle 23 6 23 – – 10 10 10 – – – 20 20 6 6 y 9 mul bra

C. mouradi C. ×

y 30 bra

– 10 15 – – 15 5 15–30 × 5–10 deep 8 mul 6 or more distinct, sle

–) C. flavus C. (4 × ×

35 35 – 12 – 12 15 – – 20 6– 20 4 y 8 mul primarily divided into 3 at the apex, divided into secondary branches

× × C. istanbulensis C. bra

27 27 – 11 – 11 11 sle – – – 20 7 20 7 y 9 mul 6

15 C. balansae C. × × –

o 32 32 15

)12 – 9 – 9 – – – – to 25 7 25 7 y 12 mul (8 bra

C. olivieri C. × × bra

35 35 – 12 – 12 15 sle – – – 15 4 15 4 y 6 mul 6

C. hyemalis C. 15 × y , – m 42 bra -

– 15 13 – – 24 6 23–42 × 6–15 bla rarely 9 mul deeply divided into 8 sle

C. vitellinus C. × × bra 30 30

– 9 – 9 13 – – rarely – 20 6 20 6 y bla 9 mul many 7 –

4 C. graveolens C. × cr 28 – 13 – 18 18–28 × 4–7 y 9 mul many bra 7 7 –

–

4 C. adanensis C. 4 × × 25 25 –

– 9 – 20 20 y 7 tri sometimes with very shallow secondary division, but not widening near the tips

C. antalyensis C. × × sle

35 28 12 y – 11 9 – 12

– – – 20 6 20– 7 pl 10 mul 6 bra

C. asymmetricus C. × × bra

30 28 y – 9 – 10 14

– – – 19 4 20 5 pl 9 mul many (Continued) S pecies/ Characters Outer flower segment size (mm) Inner flower segment size (mm) Colour of anthers Length of anthers (mm) Style B LE 1. TA

eq C. sarichinarensis C. , rarely y sh Mar

red C. paschei C. lon to eq o deep mostly rarely Mar

two = two sides of the basal

C. antalyensioides C. eq y long Mar – -ish to

wh creamy rarely usually rarely Feb C. candidus C. Mar o – or just to y eq exceeding Feb

/ C. mouradi C. sh occ , o -ish lon

to y wh usually sometimes sl Mar

C. flavus C. eq Apr – o sh

y rarely or Mar C. istanbulensis C.

o sh

sl Feb y C. balansae C. lon Mar or o – sh

to sl Jan y

lon C. olivieri C. s , Apr to o – -ish at sh

to sl Feb y wh tip

) C. hyemalis C. Jan eq – – ( or o sh Nov Dec –

( o S;

sh C. vitellinus C. or Jan y Mar

– ) N – eq usually or Nov Feb Apr deep

C. graveolens C. ) sl y Apr – – o or

Jan sh lon ( Feb deep or C. adanensis C. Mar r – to o lon Feb

C. antalyensis C. sl Mar y –

o mostly Feb C. asymmetricus C. Apr = April, Dec = December, Feb = February, Jan = January, Mar = March, N = in Northern part, Nov = November, S = in Southern part Mar = March, N in Northern part, Nov November, Jan = January, Feb = February, April, Dec = December, Apr = o always lon Feb (Continued) col. anth. = colour of anthers, l. length anthers : ac = acute, el = elliptic, gla = glabrous, nd wai = non-distinct waist, ob = obtuse, obl = oblanceolate, ov = obovate, pub = pubescent, sac = subacute, sla = sublanceolate, wai sublanceolate, = sla subacute, = sac pubescent, = pub obovate, = ov oblanceolate, = obl obtuse, = ob waist, non-distinct = wai nd glabrous, = gla elliptic, = el acute, = ac : asym = asymmetric, full = full perimeter of basal node, one = one side of basal node, several = from 1-3 points of basal node, sym = symmetric; bl = blue, bla = blackish, br = brown, dk = dark, dkr= darker, gr = grey, gre = green, li = lilac, lt = light, ltr = lighter, m = maroon, o = orange, occ = occasionally, pl = pale, pur pale, = pl occasionally, = occ orange, = o maroon, = m lighter, = ltr light, = lt lilac, = li green, = gre grey, = gr darker, dkr= dark, = dk brown, = br blackish, = bla blue, = bl ab = absent, c = coriaceous, m = membranous, p = papery, pr = present, prl parallel spl splitting ab = absent, c coriaceous, m membranous, p papery, bra = branches, cr bra = curved branches, d = deeply, eq = equal, lon = longer; mul = multifid, sh = shorter, sl = slightly, sle = slender, tri = trifid sle = slender, sl = slightly, eq = equal, lon longer; mul multifid, sh shorter, bra = branches, cr curved d deeply, Colour of style Length of style according to stamens Flowering time S pecies/ Characters Corm: node Tunic: Flower = waisted Color: vio = violet, wh white, y yellow = purple, r red, sil =silvery, Style: Anthers: Flowering time: * The table was prepared according to previous literature (Bowles, 1952; 1982, Mathew, 1984; 1886; Maw, Ruksans, 2017) and B, specimens E, from ISTE, herbaria ISTF, (GAT, Appendix 2. GB) listed in B LE 1. TA

Notes on Crocus L. Series Flavi Mathew Phytotaxa 438 (2) © 2020 Magnolia Press • 73 FIGURE 5. Mitotic metaphase plate with 2n = 8 chromosomes (a), and idiogram (b) of C. asymmetricus. Scale bar = 10 µm.

Phylogenetic analysis:—Information about the investigated material and accessions numbers are given in the Appendix. For newly obtained sequences, forward and reverse strands were checked and manually edited where necessary, after which they were combined in consensus sequences for each locus and individual. Representatives of other groups of section Nudiscapus where used as outgroups (C. almehensis C.D.Brickell & B.Mathew, C. aleppicus Baker, C. fleischeri J.Gay, C. tournefortii J.Gay). Sequences were submitted to ENA (European Nucleotide Archive) and are accessible through accession number ERZ1300521. Sequences were aligned manually, retaining only one sequence in the case of identical sequences shared by the individuals within a population. The alignment was subjected to phylogenetic analyses using Bayesian phylogenetic inference (BI) with MrBayes 3.2 (Ronquist et al. 2011). For BI, 2 times 4 chains were run for 1 million generations under the appropriate GTR+Γ+I model of sequence evolution, sampling a tree every 100 generations. Converging log-likelihoods, potential scale reduction factors for each parameter and inspection of tabulated model parameters in MrBayes suggested that stationary had been reached in all analyses. The first 25% of trees of each run were discarded as burn-in. Two independent runs of BI analysis were performed to confirm that separate analyses converged on the same result. Each of the two analyses resulted in the same topology and similar posterior probabilities (pp) for nodal support (Fig. 4). Karyological investigations:—Living corms were collected from the type locality on 11 February 2019. The root tips were then fixed with Clarke’s solution (1:3 acetic acid:alcohol) for 3–4 hours, washed in distilled water, hydrolyzed and stained with Carmine. The mitotic metaphase squash preparations (20 slides) were examined under a Zeiss AxioScope A1 microscope, and Kameram Software v3.1.0.0.

Description of the new species

Crocus asymmetricus Erol sp. nov.

Crocus asymmetricus is similar to C. antalyensis. It is easily distinguished from C. antalyensis by its asymmetric corm and lack of long, persistent neck of old cataphylls at the apex. Type:—TURKEY. Maraş: border of Maraş – Osmaniye, Ceyhan Valley 850–920m a.s.l., 6 March 2019, Mehmet Çelik s.n. (holotype ISTF 41370!, isotype: EGE!)

Corm ovoid-globose, asymmetric when mature, 8–18 mm in width (mean 12.28 ± 2.5; n = 17) × 11–19 mm in length (mean 15.52 ± 2.6; n = 17); roots emerge from one side. Corm tunics coriaceous to membranous, split into narrow bands or triangles from base, rings absent; neck narrow sharp triangular teeth, 1.79–4.52 mm long (mean 3.17 ± 0.9; n = 17). Cataphylls 2-3, white, dried cataphylls white. Prophyll absent. Bract and bracteole present, conspicuous, silvery white. Leaves synanthous, 3–8, usually 5 (mean 4.83 ± 1.09; n = 18), green, elongate lanceolate, 0.84–3.06 mm in width at broadest (mean 1.8 ± 0.5; n = 18), white stripe usually 1/2 of leaf diameter, 0.33–1.07 mm (mean 0.76 ± 0.2; n = 18), without ribs, glabrous. Perianth tube creamy white (yellow on herbarium specimens) with purple stripes. Flowers star-shaped, perigone segments elliptic, obtuse to subacute, outer segments 19–30 mm long (mean 25.31 ± 3.4; n = 13), 4–9 mm wide (mean 7.4 ± 1.4; n = 13). Inner segments 20–28 mm long (mean 23.9 ± 2.3; n = 13) and 5–10

74 • Phytotaxa 438 (2) © 2020 Magnolia Press Çiftçi et al. mm wide (mean 7.47 ± 1.9; n = 13). Segments purple on inside, outer segments dull lilac-white on abaxial side with three distinct purple stripes and featherings, inner segments purple, sometimes with one inconspicuous purple stripe and featherings. Throat bright yellow, glabrous. Filaments creamy, 3–6 mm long (mean 4.67 ± 1.05; n = 13), glabrous, anthers 9–14 mm long (mean 10.84 ± 1.5; n = 13), pale yellow, connective white. Styles orange, many branched, with long branches 5–10 mm long (mean 7.66 ± 0.71; n = 13). Styles always longer than stamens (Fig. 3). Mature capsule and seed not observed. Flowering February. Chromosome number 2n = 2x = 8 (Fig. 5). Etymology:—The species epithet is derived from the asymmetric corm of the new species. The Turkish name given to the species is “Karacaoğlan çiğdemi” according to the guidelines laid out by Menemen et al. (2013). Karacaoğlan was a folk poet who lived near the type locality of C. asymmetricus in 1700s. Taxonomic relationships:—The asymmetric corm of Crocus asymmetricus is unique within the genus. Other corm tunic characters and the multifid stigma branches suggest that this species belongs to C. ser. Flavi. Prior to the discovery of this new species, the only species in series C. ser. Flavi showing lilac-blue flowers and multi-branched stigma were C. antalyensis and C. antalyensioides. These two closely related taxa are characterized by long, brown, persistent neck of old cataphylls at the apex (Fig. 3, Table 1), whereas the new species has deep lilac-blue flowers without persistent cataphylls (Fig. 1, 2, Table 1). Besides its slightly asymmetric corms (Figs. 2, 3), C. asymmetricus shows another unique features, i.e. the arrangement of root emergence, where roots emerge from only one point rather than from around the basal node as in other species. Since its original description published in 1975 no one paid enough attention to the corm of C. adanensis. Indeed, after a careful observation of peeled corms (free from tunics) of this species we found that roots only emerge from the base of the corm in two or three bundles (Figs. 6a, b). This feature is shared with C. asymmetricus discovered in the Ceyhan valley. The ITS sequences of individuals of this population are also unique within the genus: eight substitutions and two indels distinguish it from the closest species C. vitellinus. Our phylogenetic analysis places the new species in a clade comprising C. vitellinus, C. graveolens, and C. hyemalis (pp = 1) (Fig. 4). Morphologically, C. asymmetricus can be distinguished from the latter four species by its flowers color in addition to the unique corm shape and root emergence (Table 1, Fig. 1). All species of C. ser. Flavi with the exception of C. hyemalis are part of the Turkish flora. A new identification key for species occurring in Turkey is presented below (the two subspecies of C. antalyensis are excluded, but a key for them was provided by Erol et al. 2011).

Identification key to Crocus ser. Flavi in Turkey

1. Corm asymmetric, roots emerge from one side...... Crocus asymmetricus - Corm symmetric, roots emerge from around basal node...... 2. 2. Cataphylls persistent, old cataphylls form a persistent brown sheath...... 3. - Cataphylls not persistent, white...... 7. 3. Flowers lilac-blue...... 4. - Flowers yellow or pale yellow...... 5. 4. Style yellow-orange, perigone segments narrow, with a length to width ratio of 3.3–3.8...... Crocus antalyensis - Style white-cream, rarely yellow, perigone segments are ovate-sublanceolate, with a length to width ratio of 2.0–2.5...... Crocus antalyensioides 5. Leaves 1.0–1.5 mm wide, outside of outer flower segments always striped...... Crocus sarichinarensis - Leaves 2.5–4 mm wide, outside of outer flower segments usually unstriped...... 6. 6. Style has 3 branches, each splitting into shorter indistinct lobes, and always shorter than anthers...... Crocus flavus - Style has 6–15 branches, reaching the same height as anthers or slightly longer...... Crocus mouradi 7. Style with 3 distinct branches, corm base splits weakly into basal rings...... 8. - Style with more than 3 branches, basal rings absent on base of corm, with parallel fibrils...... 9. 8. Throat white...... Crocus adanensis - Throat yellow with a white zone upper side...... Crocus paschei 9. Corm tunic fibrils coarse, fibrils slightly reticulate at apex...... Crocus istanbulensis - Corm tunic membranous, splitting into narrow triangular strips or fibrils parallel to base...... 10. 10. Style has ca. 6 branches, outside of outer perigone segments usually unmarked, perianth tube may be violet...... 11. - Style with 12–15 branches, outside of outer perigone segments usually with brown-violet stripes or markings...... 12. 11. Leaves 1–4(–5) per corm, width (1.5–)2–5(–7) mm, flowers yellow...... Crocus olivieri - Leaves 1–2(–4) per corm, width 4.5–9 mm, flowers white with yellow base...... Crocus candidus 12. Leaf width 4–6 mm, filaments glabrous rarely pubescent...... Crocus balansae - Leaf width 2–4 mm, filaments pubescent-papillose...... 13. 13. Leaves 2–4, 1.5–3 mm wide...... Crocus vitellinus - Leaves 5–8, 0.5–1.5 mm wide...... Crocus graveolens

Based on our morphological and molecular analyses, C. asymmetricus represents a distinct, new species. Future studies will be aimed to add further details, as other characters, such as those of mature capsules and seeds, are considered of diagnostic importance within the genus (Karaismailoğlu et al. 2018, Kerndorff et al. 2015). Characterization of the karyotypes of this series in Turkey should also be interesting. Indeed, although the variation in chromosome numbers (2n = 6, 8, 14) is comparatively low in this group of crocuses, Karamplianis & Constantinides (2019) reported that karyotypes and number of B chromosomes differ among the four C. ser. Flavi taxa growing in Greece and possessing the same chromosome number. Further studies involving the karyotypes of the other ten C. ser. Flavi taxa could be useful in order to gain detailed insights into the karyotype evolution of this group.

Acknowledgments

We would especially like to thank Mehmet Çelik for collecting and sending us specimens of C. asymmetricus and Kadir Terzioğlu for his kind help in the field trips. This work was supported by the Research Fund of Istanbul University, Istanbul, Turkey (project number 27545 and 24390). We also thank Nezahat Gökyiğit Botanik Bahçesi (ANG Foundation) for funding visits to K, E and GAT herbaria.

References

Baytop, T., Mathew, B. & Brighton, C. (1975) Four new taxa in Turkish Crocus (Iridaceae). Kew Bulletin 30: 241–246. https://doi.org/10.2307/4103158 Boissier, P.E. & Blanche, C.I. (1859) Diagnoses Plantarum Orientalium novarum ser. 2. ser. 2, 4: 93 Lipsiae [Leipzig] pp. 148. Boissier, P.E. (1882) Flora Orientalis sive enumeratio plantarum in Oriente a Graecia et Aegypto ad Indiae fines hucusque observatarum 5 (1). H. Georg, Basileae & Genève, 428 pp. Bowles, E.A. (1952) Handbook of Crocus and Colchicum for gardeners. Edn. 2. pp. 222. Clarke, E.D. (1812) Travels in various countries of Europe, Asia and Africa. London, 145 pp. Erol, O., Koçyiğit, M., Sik, L., Ozhatay, N. & Kucuker, O. (2010) Crocus antalyensis subsp. striatus subsp. nov. (Iridaceae) from southwest Anatolia, Nordic Journal of Botany 28: 186–188. https://doi.org/10.1111/j.1756-1051.2009.00447.x Erol, O., Kaya, H.B., Şik, L., Tuna, M., Can, L. & Tanyolac, M.B. (2014) The genus Crocus, series Crocus (Iridaceae) in Turkey and 2 East Aegean islands: a genetic approach. Turkish Journal of Biology 38: 48–62. https://doi.org/10.3906/biy-1305-14 Erol, O., Şik, L., Kaya, H.B., Tanyolaç, B. & Küçüker, O. (2011) Genetic diversity of Crocus antalyensis B.Mathew (Iridaceae) and a new subspecies from southern Anatolia. Plant Systematics and Evolution 294: 281–287. https://doi.org/10.1007/s00606-011-0465-8 Gay, J. (1879) Notes on New Croci. Gardeners’ Chronicle New series 11: 234. [pp. 828] Gay, J. (1831) Nouvelles Especes de Crocus. Bulletin des Sciences Naturelles et de Géologie 25: 319. [pp. 368] Harpke, D., Carta, A., Tomović, G., Ranšelović, V., Ranšelović, N., Blattner, F.R. & Peruzzi, L. (2015) Phylogeny, karyotype evolution and taxonomy of Crocus series Verni (Iridaceae). Plant Systematics and Evolution 301: 309–325. https://doi.org/10.1007/s00606-014-1074-0 Harpke, D., Meng, S., Rutten, T., Kerndorff, H. & Blattner, F. (2013) Phylogeny of Crocus (Iridaceae) based on one chloroplast and two nuclear loci: ancient hybridization and chromosome number evolution, Molecular Phylogenetics and Evolution 66: 617–627. https://doi.org/10.1016/j.ympev.2012.10.007 Harpke, D., Peruzzi, L., Kerndorff, H., Karamplianis, T., Constantinidis, T., Randelovic, V., Randelovic, N., Juskovıc, M. & Blattner, F.R. (2014) Phylogeny, geographic distribution, and new taxonomic circumscription of the Crocus reticulatus species group (Iridaceae). Turkish Journal of Botany 38: 1182–1198. https://doi.org/10.3906/bot-1405-60 Karaismailoğlu, M.C., Şik, L., Gemicioğlu, A. & Erol, O. (2018) Seed structure of some taxa of the genus Crocus L.(Iridaceae) series Crocus. Turkish Journal of Botany 42: 722–731. https://doi.org/10.3906/bot-1712-17

Notes on Crocus L. Series Flavi Mathew Phytotaxa 438 (2) © 2020 Magnolia Press • 77 Karamplianis, T. & Constantinides, T. (2019) The taxa of Crocus ser. Flavi (Iridaceae) in Greece: a taxonomic and karyomorphometric study. Botanika Chronika 22: 171–194. Kerndorff, H. (1994) Two new taxa in Turkish Crocus. Herbertia 49: 76–86. Kerndorff, H., Pasche, E. & Harpke, D. (2015) The genus Crocus (Liliiflorae, Iridaceae): life-cycle, morphology, phenotypic characteristics, and taxonomical relevant parameters. Stapfia 103: 27–65. Kerndorff, H. (1994) Two New Taxa in Turkish Crocus (Iridaceae), Herbertia 49: 76–78. Mathew, B. (1982) The Crocus, a revision of the genus Crocus. Timber Press, London, 224 pp. Mathew, B. (1984) Crocus L. In: Davis, H. (Ed.) Flora of Turkey and the East Aegean islands 8. Edinburgh University Press, Edinburgh, pp. 413–438. Mathew, B. & Brighton, C.A. (1972) A new Crocus from Turkey, Kew Bulletin 27: 327–329. https://doi.org/10.2307/4109462 Mathew, B., Petersen, G. & Seberg, O. (2009) A reassessment of Crocus based on molecular analysis. Plantsman 8 (1): 50–57. Maw, G. (1886) A Monograph of the Genus Crocus. Dulau & co., London, 54 pp. Menemen, Y., Aytaç, Z. & Kandemir, A. (2013) Türkçe bilimsel bitki adları yönergesi. Bağbahçe Dergisi 47: 28–31. [in Turkish] Rukšâns, J. (2015) Some New Crocus Taxa (Iridaceae) from Western Turkey and East Aegean Islands, International Rock Gardener 64: 1–38. Rukšāns, J. (2010) Crocuses. A complete guide to the genus. Timber Press, Portland, 157 pp. Rukšāns, J. (2017) The world of crocuses. The Latvian Academy of Sciences, Riga, 442 pp. Thiers, B. (2019) Index Herbariorum: A global directory of public herbaria and associated staff. New York Botanical Garden’s Virtual Herbarium. Available from: http://sweetgum.nybg.org/ih/ (accessed 12 September 2019) Wahlenberg, G. (1828) Gewächse im Morgenland gesammelt von J. Berggren, und näher bestimmt von Goran Wahlenberg. Isis (Oken) 21: 972–1006. Weston, R. (1771) Botanicus Universalis et Hortulanus 2: 237. [pp. 384] Whittall, E. (1889) Notes from Smyrna. The garden: an illustrated weekly journal of horticulture in all its branches 35: 473.

78 • Phytotaxa 438 (2) © 2020 Magnolia Press Çiftçi et al. Appendix 1. This appendix lists the source of each sequence used for our phylogenetic analysis and its EMBL/NCBI accession number (taxon, herbarium vouchers, collection number, origin, EMBL accession nrITS region. Missing data is indicated with “-“.

Crocus adanensis T.Baytop & B.Mathew, GAT7148, -, Turkey,HE663988, Crocus aleppicus Baker, IABH18357, -, Jordan, HE801175; Crocus almehensis C.D.Brickell & B.Mathew, TARI69170, -, Iran, HE801162; Crocus antalyensis B.Mathew, HKEP9511, GAT7237, Turkey, HE664015; Crocus antalyensis, HKEP1006a, GAT7186, Turkey, HE663990; Crocus asymmetricus Erol, ISTF 41370, Crocus sp. DH-2020, Turkey, LR761644; Crocus candidus E.D.Clarke, GAT7137, HKEP 0907, Turkey, HE66398; C. flavus Weston, GAT7140, HKEP0915, Turkey, HE663984; Crocus fleischeri Baker, GAT7139, HKEP0914, Turkey, HE663983; Crocus graveolens Boiss. & Reut., GAT7457, HKEP1111, Turkey, HE664010; Crocus hyemalis Boiss. & C.I.Blanche, GAT7118, -, Jordan ,HE801057; Crocus olivieri J.Gay, GAT23020, HKEP1342a, Serbia, LR778179, Crocus vitellinus Wahlenb., -, -,Turkey, LR778178; Crocus paschei Kerndorff,-,-, Turkey, LR778177; Crocus tournefortii J.Gay, GAT7202, -, Greece, HE801123;

Appendix 2. Voucher numbers of herbarium specimens used for the morphological investigations.

C. adanensis K802478, ISTE23969, ISTE12468, ISTE95577, GAT7148; C. antalyensioides ISTF41127, GAT56933, GB0152368; C. antalyensis ISTF40145, ISTF40143, ISTF40142, ISTF40168, ISTF40079, ISTF41127, ISTF41126, ISTF41117, ISTF41116, ISTF41113, ISTF41109, ISTF41105, ISTF41103, ISTF41144, ISTF41149, ISTF41141, K802490, E333389, E333390, ISTE36684, ISTE31337, ISTE31340, ISTE9825, ISTE9826, ISTE60001, ISTE12482, ISTE23980, ISTE21323, ISTE93252, ISTE93289, ISTE99292, GAT7186, GAT7237, GAT7458; C. balansae K802441, K802440, K802442, ISTF41122, E333325, E333326, E333327, E333328, E333329, E333330, ISTE21342, ISTE95469, ISTE93253, ISTE19651, ISTE36513, ISTE38935, ISTE23806; C. candidus ISTF41161, E346204, E346205, E333324, ISTE79127, ISTE65012, ISTE62669, ISTE64999, ISTE65002, ISTE64993, ISTE106596, ISTE23812, ISTE79119, ISTE65009, ISTE65015, ISTE99290, ISTE96004, GAT7137; C. flavus ISTF41130, ISTF41128, ISTF41151, ISTF41150, K802444, K802400, B100151920, B100151921, B100151922, B100151923, B100210241, B100510434, GAT23031, GAT23032, GAT23034, GAT31633, GAT33768, GAT33769, GAT33775, GAT33780, GAT33781, ISTE27221, ISTE27225, ISTE27239, ISTE27281, ISTE27286, ISTE27294, ISTE27296, ISTE27212, ISTE27214, ISTE27217, ISTE95468, ISTE99247, ISTE21333, ISTE108652, ISTE108651, ISTE10804, ISTE31535, ISTE23990, ISTE10667, ISTE10672, ISTE95467, ISTE31341, ISTE31346, ISTE31352, ISTE31306, ISTE31316, ISTE23750, ISTE23807, ISTE23808, ISTE23815, ISTE23816, ISTE23817, ISTE27257, ISTE31320, ISTE96668, ISTE99289, ISTE60018, ISTE66649, ISTE65008, ISTE10790, ISTE12446, ISTE21283, ISTE21289, ISTE21321, ISTE34806, ISTE3482, ISTE3498, ISTE66248, ISTE80850, ISTE82998, ISTE83516, ISTE96168, ISTE99266; C. graveolens E333392, E333393, E333394, E333395, E333396, E333397, E333398, E333399, E333400, E333401, E333402, E333403, E333404, E333405, E333406, E333407, E333408, ISTE96002, ISTE99287, ISTE99237, ISTE99240, ISTE99250, ISTE99248, ISTE96031, ISTE78351, ISTE12460, ISTE12469, ISTE4173, ISTE65127, ISTE93255, GAT7144, GAT7256, GAT7457, GAT30297, GAT56985; C. hyemalis K318284, K802431, K802425, K802428, K318285, K802429, K802426, K802430, K802427, E333294, E333295, E333297, E333298, E333299, E333300, E333301, E333302, E333303, GAT7218; C. istanbulensis K802399, ISTE46030, ISTE69709, ISTE99254, ISTE96169, GAT7399; C. mouradii ISTF36901, K400300, ISTE23390; C. olivieri ISTF34678, ISTF41119, ISTF41120, ISTF41287, K802443, K341684, K99742, E333311, E333310, E333312, E333313, E333314, E333315, E333316, E333317, E333318, E333319, E333320, E333321, E333322, E333323, ISTE69768, ISTE1162, ISTE1365, ISTE10247, ISTE21255, ISTE21313, ISTE21326, ISTE99249, ISTE99291, ISTE99279, ISTE99246, ISTE96024, ISTE95484, ISTE95479, ISTE96669, ISTE95478, ISTE21364, ISTE21370, ISTE27231, ISTE24197, ISTE25002, ISTE25979, ISTE27251, ISTE31556, ISTE34411, ISTE34416, ISTE34560, ISTE34573, ISTE48464, ISTE50018, ISTE52903, ISTE66244, ISTE66255, ISTE7919, ISTE80846, ISTE88431, ISTE95817, GAT7135, GAT23020, GAT33772, GAT47240; C. paschei ISTF41083, ISTF41085, ISTF41086; C. sarichinarensis ISTF41130; C. vitellinus ISTF13965, ISTF13974, ISTF41094, ISTF1443, ISTF1444, ISTF1469, ISTF1470, ISTF12118, ISTF820, K802437, K802439, K802436, K802438, E333309, B100355309, B100355310, B100355311, ISTE99282, ISTE99283, ISTE77725, ISTE77731, ISTE5129, ISTE10243, ISTE23824, ISTE23825, ISTE23828, ISTE23967, ISTE23972, ISTE23981, ISTE23982, ISTE23989, ISTE31390, ISTE31400, ISTE5788, ISTE23983;