Noccaea birolmutlui, a New Crucifer Species from South West Anatolia, Turkey

Abstract

Noccaea birolmutlui Özgişi & Özüdoğru is described as a new species from Kızıldağ National Park, Isparta, in SW Anatolia. It resembles N. ochroleuca but can be readily distinguished by several morphological characters including shape and size of basal/stem leaves, fruit shape, style length, ovule number etc. Nuclear-encoded ribosomal internal transcribed spacer (ITS) region based phylogeny proved that this species clearly differs than N. ochroleuca and these two species belong to different clades in Noccaea s.str. Diagnostic characters, a description, detailed photos and taxonomic comments on the species are given. Morphological affinities and ITS based phylogenetic relationships of new species are discussed. Furthermore, the IUCN Red List category of N. birolmutlui is proposed as critically endangered (CR) due to its limited distribution.

Full text

Phytotaxa 345 (1): 059–067
http://www.mapress.com/j/pt/
Copyright © 2018 Magnolia Press Article PHYTOTAXA
ISSN 1179-3155 (print edition)
ISSN 1179-3163 (online edition)
Accepted by Karol Marhold:22 Feb. 2018; published: 20 Mar. 2018
https://doi.org/10.11646/phytotaxa.345.1.7
59
Noccaea birolmutlui, a New Crucifer Species from South West Anatolia, Turkey
KURTULUŞ ÖZGİŞİ1, ATILLA OCAK1 & BARIŞ ÖZÜDOĞRU2*
1
2

Abstract
Noccaea birolmutlui Özgişi & Özüdoğru is described as a new species from Kızıldağ National Park, Isparta, in SW Anatolia.
It resembles N. ochroleuca but can be readily distinguished by several morphological characters including shape and size of
basal/stem leaves, fruit shape, style length, ovule number etc. Nuclear-encoded ribosomal internal transcribed spacer (ITS)
region based phylogeny proved that this species clearly differs than N. ochroleuca and these two species belong to different
clades in Noccaea s.str. Diagnostic characters, a description, detailed photos and taxonomic comments on the species are
given. Morphological affinities and ITS based phylogenetic relationships of new species are discussed. Furthermore, the
IUCN Red List category of N. birolmutlui is proposed as critically endangered (CR) due to its limited distribution.
Keywords: Brassicaceae, Coluteocarpeae, Cruciferae, Noccaea, Turkey
Introduction
The genus Noccaea Moench (1802: 89) is one of the most taxonomically problematic genera among crucifers and
its generic circumscription varies according to different concepts (Al-Shehbaz 2014; https://brassibase.cos.uni-
heidelberg.de/; accessed 01 December 2017). Although Noccaea was described for the first time by Moench (1802),
species of within the genus were subsumed under  Linnaeus (1753: 645) (Prantl 1891; Schulz 1936) until
Meyer (1973; 1979) divided  into 12 segregate genera. Meyer’s concept was generally supported by early
molecular studies (Mummenhoff & Zunk 1991, Mummenhoff & Koch 1994, Mummenhoff et al. 1997a,b). However,
subsequent studies (Koch & Mummenhoff 2001, Koch & Al-Shehbaz 2004) showed that this concept was artificial.
Because Meyer’s concept was impractical for taxonomist, his treatment was not followed by some authors and generic
limits of segregates are still controversial (Greuter & Raus 1983, Greuter et al. 1986, Al-Shehbaz, 1986, 2014, Ali et
al. 2016). Based on both morphological and molecular studies, Al-Shehbaz (2014) united all of Meyer’s (1973; 1979)
segregates in a more inclusive Noccaea in the tribe Coluteocarpeae whereas database for Brassicaceae (BrassiBase,
https://brassibase.cos.uni-heidelberg.de/) presented an alternative view including 14 genera.
In the first volume of  Hedge (1965) recognized 25  species
and two doubtful records. After this account, 26 species were added to  or to others of Meyer’s segregates
including Callothlaspi Meyer (1973: 457), Microthlaspi Meyer (1973: 452), Noccaea, etc. (Davis et al. 1988, Gemici
& Leblebici 1995, Yıldırımlı 2000, Aytaç et al. 2006, Fırat et al. 2014). Based on both morphological and molecular
studies, Al-Shehbaz (2014) subsumed some Eunomia Candolle (1821:555) species and all of Meyer’s segregates under
the genus Noccaea s.l. in the tribe Coluteocarpeae except  s.str. and Noccidium Meyer (1973: 456). Following
this treatment the current number of Noccaea species in Turkey is 50.
During the herbarium studies for “The Revision of Turkish Noccaea Species” project an interesting voucher
was found in the Herbarium of Biology Department, Hacettepe University (HUB), which was collected by Dr. Birol
Mutlu at Kızıldağ National Park, Isparta. This Noccaea specimen could not be identified with the Flora of Turkey and
related references (Hedge 1965; Meyer 2006; Al-Shehbaz 2014). Later, specimens in flowering and fruiting stage were
collected from the original location by the authors and were used in the current study.

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Material and Methods
Flowering and fruiting specimens of Noccaea ochroleuca (Boissier & Heldreich 1849:39) Meyer (1973:461) and
new species were collected in the field. All morphological analyses were undertaken on both living and herbarium
specimens. Herbarium specimens of the new species were compared with its morphologically closely related species N.
ochroleuca based on relevant literature (Hedge 1965, Meyer 2006, Al-Shehbaz 2014). There are some taxonomically
ambiguous populations around Isparta, Konya and Antalya border which may or may not belong to N. ochroleuca.
Therefore, we only used N. ochroleuca samples from Davraz Mountains following Meyer’s treatment (Meyer 2006).
For scanning electron microscopy (SEM) of seeds, dry seeds were mounted directly on stubs and coated with gold in
a sputter coater. SEM examination was carried out using a Jeol 5600 LV-SEM microscope at Eskişehir Osmangazi
University. Photographs of living material were taken with a Nikon D90 digital camera.
Internal transcribed spacer regions (including ITS1 and ITS2) and the 5.8S gene of nuclear ribosomal DNA
(hereafter ITS) were sequenced for the new species, N. ochroleuca and N. papillosa (Boissier 1867:330) Meyer
(1973:460). Additionally, ITS sequences of 32 available Coluteocarpeae members representing all Meyer’s segregate
plus Linnaeus (1753: 646) as an outgroup were taken from Genbank. (http://www.ncbi.nlm.nih.gov/
genbank).
DNA extraction, amplification, and sequencing:—Total genomic DNA was isolated using DNeasy Plant Mini
Kit (Qiagen, Hilden, Germany) following the manufacturer’s instructions. ITS region was amplified using primers
ITS1 and ITS 4 (White et al. 1990). Amplification of ITS followed the protocol in Warwick et al. (2004). Purification
and sequencing were performed by MedSanTek (İstanbul, Turkey).
Phylogenetic analysis:—Sequences were edited with Codon Code Aligner (CodonCode Corporation) and aligned
with MUSCLE v.3.6 (Edgar 2004). Alignment file was provided as supplementary data. The best fitting nucleotide
substutition model was selected by the Akaike information criterion (AIC) implemented in MEGA v.6 (Tamura et
al. 2013). The GTR + G model was selected for Bayesian analysis and a Yule Model process of speciation was used
as the tree prior. Bayesian analyses were implemented in BEAST v.1.8 (Drummond et al. 2012). Two independent
MCMC runs were conducted with chain lengths of 10 million and sampled every 1000 generations. Each run was
checked using Tracer v.1.5 (Rambaut & Drummond, 2009), with the first 10% of samples discarded as burn-in, and
then combined in LogCombiner (Drummond et al. 2012).
Results
The aligned ITS data set included 38 sequences that belong to 34 species and 648 bp positions were available for
phylogenetic analysis long, of which 228 were variable and 123 were potentially parsimony informative.
Noccaea birolmutlui was placed in a clade with some representatives of Noccaea s.str including N. jankae (Kerner
1867:35) Meyer (1973:462), N. montana (Linnaeus 1753: 647) Meyer (1973:462), N. densiflora (Boissier & Kotschy
(Boissier 1867:328) Meyer (1973:462), N. macrantha (Lipsky 1894:271) Meyer (1973:464) and Raparia bulbosa
(Spruner 1843:74) Meyer (1973:458), whereas its morphologically closely relative N. ochroleuca is found in different
clade along with N. papillosa (Fig. 1).
Taxonomy
Noccaea birolmutlui
Type:—TURKEY. B3 Isparta: Şarkikaraağaç; Kızıldağ National Park, 1400–1500 m, clearings in Cedrus libanii forest, 09.06.2017,
 5017 &  (holotype: HUB!, isotypes: OUFE!, GAZI!, EGE!, INU!). Paratypes: B3 Isparta: Şarkikaraağaç; Kızıldağ
National Park, ormanevleri – Hastane inşaatı arası, Cedrus libani-Quercus karışık ormanı, 1150–1250 m, 26.05.1994, Mutlu 443
(HUB!); Isparta, Şarkikaraağaç, Kızıldağ Milli Parkı, Piknik Alanı Çevresi, Cedrus libani orman açıklığı 38° 02’ 25”K, 31° 21’
54”K, 1400 m, 26.06.2016,  1211 (ANK!, HUB!, OUFE!, NGBB!)

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 Phylogenetic tree of tribe Coluteocarpae based on the Bayesian analysis of the ITS data. Posterior probability
values > 0.5 are shown at the nodes. Taxon names follow concept presented in BrassiBase (https://brassibase.cos.uni-
heidelberg.de/).
Diagnosis
Noccaea birolmutlui differs from the related N. ochroleuca by having long petiolate (up to 28 mm) and entire (vs.
short petiolate (up to 14 mm) and dentate) basal leaves, oblong to ovate (vs. lanceolate) cauline leaves, obdeltoid (vs.
obcordate) siliculae, 5-6 ovules (vs. 4) per locule.
Description
Glabrous, glaucescent perennial herbs 15–40 cm tall; stems single or rarely several from the base, terete. Basal leaves
rosette forming, green, spatulate, tapering into a long petiole, 10–72 × 4–14 (16) mm (inc. petiole), apex rounded,
margins entire; cauline leaves subcoriaceous, glaucus, oblong to ovate, ca. 6–17 (19) × 5–13 (15) mm, apex obtuse
to acute, margins entire, amplexicaul or auriculate, sometimes overlapping, auricles obtuse. Inflorescence simple
or compound raceme (up to 6 racemes), racemes 25–107-flowered, congested in flower, elongating in fruit, lateral
racemes shorter than terminal in fruiting time. Pedicel 1.5–4.5 mm in flower; horizontally spreading up to 15 mm in
fruit. Sepals 2.1–2.2 × 0.9–1 mm. Petals 5.2–5.5 × 1.5–2 mm, white. Siliculae glaucus, pruinose, obdeltoid, 2.5–8.5 ×
1.8 (2.1) –6 (6.2) mm with a apical notch. Tips of notch retuse (Notch sometimes absent). Valves of siliculae winged up
to 1.8 mm at apex, narrower at base. Style 0.8–2.1 mm long in fruit, always exceeding sinus. Stigma capitate. Ovules
(5)6 per locule. Seeds orange to orange-brown, tuberculate, 2(–5) per locule, ca. 1.3 (1.5) × 0.7 mm.

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 General view of Noccaea birolmutlui and N. ochroleuca. N. birolmutlui  N. ochroleuca

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 Noccaea birolmutlui: Basal leaves;  Cauline leaves;  Fruits. N. ochroleuca:  Basal leaves;  Cauline
leaves;  Fruits.
Habitat: Cedar forest.
 : May–June.
 The specific epithet is named after Dr. Birol Mutlu, who is the first collector of new species. The
Turkish name of this species is given as “Kızıldağ Dağarcığı” according to the guidelines of Menemen et al. (2013).
Seed morphology: Seeds of N. birolmutlui are orange to orange-brown, tuberculate, ca. 1.3 (1.5) × 0.7 mm and
2(–5) per locule (Fig. 4). Seed coat is colliculate, lacking the mucilage and has epicuticular wax only. Epidermal
cells are irregularly pentagonal to elliptic. Cell walls are evenly thickened, the boundaries are below the cell centers.
Anticlinal walls are flat, periclinal walls are obviously convex. Surface ornamentation is ruminate.

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 Seed of Noccaea birolmutlui  General appearance,  Seed surface
  N.biromutlui is a local endemic restricted to the part of Cedar forest in Kızıldağ
National Park (Isparta, Turkey) (Fig. 4). The occupancy area (AOO) of new species was calculated as ca. 1 km2 in
which about <1000 individuals were estimated to occur. Although N. birolmutlui occurs in a protected area (National
Park), it has very small distribution area with low population size and therefore, in accordance with the criteria of the
IUCN (2017), it is here assessed as “Critically Endangered”
Examined specimens:—N. ochroleuca: —TURKEY. C3 Isparta: Davraz mountain, 1800–1900 m, 09.06.2017,
 5009 & (HUB!), Mons Davros dagh Pisidiae, ad nives, 05.1845, Heldreich (isolectotype B photo!
(http://herbarium.bgbm.org/object/B100249985).

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 Distribution of Noccaea birolmutlui ( ) and N. ochroleuca ( ).
Discussion
Noccaea birolmutlui is morphologically similar to N. ochroleuca but it is distinguished by several morphological
characters including basal and cauline leaves, fruit shape and ovule numbers.
These characters are typically used in species delimitation of Noccaea (Meyer, 2006; Al-Shehbaz, 2014 etc.).
Of these, fruit shape can be extremely variable for Noccaea species in the different stages of plant development
as indicated by Özgişi et al. (unpublished data). But as our collections represent different ontogenetic stages of N.
birolmutlui, fruit shape can be used as a diagnostic character.
Bayesian based molecular phylogenetic analysis of ITS data clearly show that N. birolmutlui and N. ochroleuca
are distinct species placed in different clades (Fig. 1). Raparia bulbosa is the phylogenetically closest species to N.
birolmutlui, but morphologically it is distinguished from latter by having napiform root and large rose-lilac petals.
However, to reveal the true phylogenetic position of N. birolmutlui, more representatives of Noccaea (128 species
according to Al-Shehbaz 2014) and additional molecular markers should be included in a phylogenetic analysis.
In his  account for Flora of Turkey, Hedge (1965) used broad species delimitation for N. ochroleuca and
apart from type specimen from Davraz Mountain (Isparta, Turkey), he included some additional accessions which are
treated as N. phyrgia (Bornmueller 1929: 57) Meyer (1973: 465), N. edinensium Meyer (1973: 460) and 
(Stojanov & Kitanov 1943: 97) Meyer (2006: 116) at the present time. Of them, N. phyrgia has a spotted distribution
in Anatolia and reported from Ankara, Bolu, Bursa and Konya provinces, whereas N. edinensium and  are
restricted to Murat Mountain (Kütahya, north-east Aegen Part of Turkey) and Uludağ (Çanakkale, north-west Turkey).
This would indicate that these species are allopatric. Finally, taxonomically ambiguous populations that outlined in
Material and Methods chapter, needs further evaluations using comprehensive morphometric and also genetic analyses.
Thus, species boundary of N.ochroleuca can be outlined correctly.
 Morphological comparison of Noccaea birolmutlui and its morphologically closest relative N. ochroleuca.
Characters Noccaea birolmutlui Noccaea ochroleuca
Basal leaves
Petiole (mm) 4.2–28 × 0.5–2.3 4–14 × 0.5–3.1
Size (mm) 10–72 × 4–14 (16) 8–52 × 2–9
Margin Entire Somewhat dentate
Cauline leaves
Shape Oblong to ovate Lanceolate
Size (mm) 6–17 (19) × 5–13 (15) 3–7 × 6–31
Apex Obtuse to acute Acute
Inflorescence A simple raceme or a compound raceme (up to 6
racemes)
A simple raceme or compound raceme (up to 4
racemes)
Racemes 25–107 flowered 17–47 (50) flowered
Fruits
Shape Obdeltoid Obcordate
Size (mm) 2.5– 8.5 × 1.8 (2.1)–6 (6,2) 5–10 (12) × 2–7
Margins Winged up to 1.8 mm at apex, green Winged up to 1.2 mm at apex, often purplish
Style 0.8–2.1 mm Up to 4.7 (5.0) mm
Ovules per locule (5) 6 4

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66 • Phytotaxa 345 (1) © 2018 Magnolia Press
Acknowledgements
This study was supported by Eskişehir Osmangazi University BAP Coordinatorship (project number: 2015/681) and
Hacettepe University Research Project Coordination Unit (project number: FBA-2017-12879). We thank Yelda Güzel
for helping us to describe seed surface of new species and Dmitry German for providing a reference.
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