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Morphological and Molecular Evidence for a New Species of Bolanthus
(Caryophyllaceae) from Turkey
Authors: Murat Koç, Ergin Hamzaoğlu, and ˙Ilker Büyük
Source: Systematic Botany, 44(1) : 189-196
Published By: American Society of Plant Taxonomists
URL: https://doi.org/10.1600/036364419X698010
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Systematic Botany (2019), 44(1): pp. 189–196
© Copyright 2019 by the American Society of Plant Taxonomists
DOI 10.1600/036364419X698010
Date of publication February 5, 2019
Morphological and Molecular Evidence for a New Species of Bolanthus (Caryophyllaceae)
from Turkey
Murat Koç,
1,4
Ergin Hamzao˘glu,
2
and ˙
Ilker B ¨uy¨uk
3
1
Animal Production High School, Bozok University, TR-66200, Yozgat, Turkey
2
Department of Mathematics and Science Education, Gazi Faculty of Education, Gazi University, TR-06500, Ankara, Turkey
3
Department of Biology, Faculty of Science, Ankara University, TR-06100, Ankara, Turkey
4
Author for correspondence (bozokmuratkoc@yahoo.com)
Communicating Editor: Jocelyn Hall
Abstract—In Turkey, the genus Bolanthus is represented by 10 endemic taxa. Some interesting Bolanthus specimens were collected from
Afyonkarahisar province. After examining the literature and herbarium specimens, it was found that these specimens resemble Bolanthus
thymoides but differ from this species by several characters. The newly collected specimens and B. thymoides were compared with each other in
terms of their general morphology and seed micromorphology. By using DNA sequences of the ITS and trnL–trnF regions, phylogenetic re-
lationships between this collected species and the other species distributed in Turkey were investigated. As a result of the evaluation of the
molecular and morphological data, it was evident that the specimens collected from Afyon karahisar represent a new species, and this new species
has been named B. aziz-sancarii.
Keywords—Afyonkarahisar, ITS, taxonomy, trnL–trnF.
The Caryophyllaceae includes approximately 100 genera
and 3000 species that are almost entirely distributed in the
Northern Hemisphere, and less in the Southern Hemi-
sphere. Specifically, the main distribution of this family is
the Mediterranean phytogeographical region. This family is
split into three subfamilies: Alsinoideae Burnett, Cary-
ophylloideae Arn., and Paronychioideae A.St. (Hern´andez-
Ledesma et al. 2015). The genus Bolanthus, amemberoftribe
Caryophylleae, presents an interesting taxonomic group
containing 11 taxa endemic to Turkey. Despite the fact that
the main distribution area of the genus is the Mediterranean
phytogeographical region, B. minuartioides (Jaub. & Spach)
Hub.-Mor., B. cherlerioides (Bornm.) Bark., and B. turcicus
occur eastward into the Irano-Turanian phytogeographical
region.
The name Bolanthus (Ser.) Rchb. was first used by Seringe
as a section name under Saponaria L. The species Saponaria
hirsuta Labil., S. depressa Div., S. caespitosa DC., S. lutea L., S.
bellidifolia Smith., S. smithii Ser., and S. saxatilis Bory fall within
this section (De Candolle 1824). In their study, Fenzl and
Endlicher kept this section within Saponaria, while making no
other changes (Endlicher 1836). In 1843, Braun moved Sap-
onaria hirsuta to Gypsophila L., presumably because of its re-
semblance to the taxa of this genus (Braun 1843). However,
previously Reichenbach (1841) raised the rank of section
Bolanthus to the genus level (Reichenbach 1841).
Bolanthus (Ser.) Rchb. is placed in the tribe Caryophylleae.
This genus in Turkey was last revised by Huber-Morath in the
Flora of Turkey and the East Aegean Islands, published in 1967. In
this taxonomic treatment, five species and one variety were
reported (Huber-Morath 1967), but Davis et al. (1988) later
added one species to this genus. As a result of more recent
studies, three additional taxa were added to Bolanthus,of
which one is an overlooked species (B. huber-morathii C. Simon)
and three are new species (B. mevlaneae Aytaç, B. turcicus Koç &
Hamzao˘glu, B. sandrasicus Koç & Hamzao ˘glu) ( ¨
Ozhatay et al.
1999; Aytaç and Duman 2004; Koç and Hamzao˘glu 2015;
Hamzao˘glu et al. 2017). Together with these additions, the
number of taxa belonging to the genus Bolanthus that occur in
Turkey increased to 11, all of which are endemic (Barkoudah
1962). The genus is represented by six taxa found in Syria,
Palestine, Israel, and Lebanon (Barkoudah 1962; Zohary 1966;
Barkoudah and Akeroyd 1993). The eight taxa growing in
Europe are known from Greece or the East Aegean Islands
(Phitos 1997). Together with the Turkish taxa, the total number
of taxa in Bolanthus is 25. This study was aimed to define the
taxonomic categories of Bolanthus specimens collected from
Afyonkarahisar province by comparing them with Bolanthus
species distributed in Turkey based on morphological and
molecular data.
Materıals and Methods
Specimen Collection and Analysis—As part of the revision of the genus
Bolanthus in Turkey, a large number of specimens from different pop-
ulations growing in Burdur and Afyonkarahisar provinces were collected.
Among these Bolanthus specimens, the ones growing in the fissures of
tuffaceous volcanic rocks especially attracted the attention of the re-
searchers. These specimens were extensively compared with the re-
lated literature (Barkoudah 1962; Zohary 1966; Barkoudah and
Akeroyd 1993; Phitos 1997) and with specimens at GAZI, HUB, ISTE,
ISTO, EGE, a nd KNYA herbaria; herbarium abbreviations follow Thiers
(2017). The specimens found in K [http://apps.kew.org/herbcat/navigator.
do], G [http://www.villege.ch/musinfo/bd/cjb/chg/? lang5en], and E
[http://elmer.rbge.org.uk/bgbase/vherb/bgbasev herb.php] virtual herbaria
were also examined. The genus Bolanthus contains about 25 taxa worldwide,
and 11 out of 25 are endemic to Turkey, while the remaining 14 species are
Table 1. Diagnostic characters between Bolanthus aziz-sancarii and B. thymoides.
B. aziz-sancarii B. thymoides
Bracts Lanceolate, 5.5–7 mm long Linear-setaceous, 4–5 mm long
Inflorescence 1–3 pedicellate flowers 5–10 sessile or subsessile flowers
Calyx 5.5–7 mm long 4–5 mm long
Petal 7–8.5 mm long, completely white, apex emarginate 5–6 mm long, white with purple veins, apex truncate
Capsule 5–6.5 mm long 3.5–4.5 mm long
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endemic to either Greece or Syria, Palestine, Israel and Lebanon. How-
ever, we were not able to collect plant material of Bolanthus outside of
Turkey.
All of the samples used in the study (Appendix 1) were collected
from the field and turned into herbarium specimens and then these
materials were used in description, seed surface, and molecular
studies. Bolanthus thymoides were collected from five different pop-
ulations of Burdur province. Bolanthus aziz-sancarii were collected from
three different populations of Afyonkarahisar province. Other samples
used in molecular studies were collected from a single population.
Approximately 30 individuals from each population were examined
for plant description studies. At least five mature seeds from each
population were selected and examined for the seed surface studies.
Molecular studies were performed by randomly selecting one indi-
vidual, considered to be a good representative of the taxon, from the
population.
Seeds were placed onto a stamp using double-sided adhesive tape and
then coated with gold palladium. Following this application, the seeds
were monitored using LEO 440 scanning electron microscope (Zeiss, USA)
in the Science and Technology Application and Research Center of Bozok
University. The images were captured with the 5003,1003,and403
objective lens for the details. Thirty measurements were made for
Fig. 1. The maximum likelihood tree showing the genetic relationships between 11 Bolanthus species and outgroup Dianthus sylvestris, based on the
nuclear ITS region. Bootstrap values from 1000 bootstrap replicates are above the branches.
Fig. 2. The maximum likelihood tree showing the genetic relationships between 11 Bolanthus species and outgroup Dianthus sylvestris, based on the
trnL–trnF region of the chloroplast. Bootstrap values from 1000 bootstrap replicates are above the branches.
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quantitative traits and the maximum and minimum values were used for
the plant descriptions. The vegetative parts were measured using a ruler
with 0.5 mm accuracy and floral characteristics were studied using an
ocular micrometer. Photographs were taken with an Olympus C-5060
digital camera.
DNA Isolation—Leaf material of eleven Bolanthus (Caryophyllaceae)
specimens (see Appendix 1) was dried in silica gel and the genomic DNA
was extracted according to the modified CTAB method as described by
Aras et al. (2003). NanoDrop ND-Lite was used to measure DNA con-
centration and purity of each Bolanthus sample.
PCR Amplification and Sequencing—Sequence comparisons of the ITS
and trnL–trnF regions are frequently used in phylogenetic reconstructions
at the infrageneric taxonomic level (Chase et al. 2000; Hao et al. 2009). The
ITS region of the rDNA was amplified using ITS1 and ITS4 primers as
described by White et al. (1990). A primer set (trnL and trnF) described by
Sang et al. (1997) was chosen to amplify the trnL–trnF non-coding
Fig. 3. Bolanthus thymoides (Koç 1848 &Hamzao˘glu). A. Habit. B. Inflorescence. C. Flower. D. Petal. E. Capsule.
KOÇ ET AL.: BOLANTHUS AZIZ-SANCARII IN TURKEY 1912019]
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chloroplast DNA (cpDNA) region. This region comprises two tRNA genes,
denoted as trnL and trnF, and an intergenic non-coding portion located
between these two genes (Sang et al. 1997).
In this study, the nuclear ITS region and trnL–trnF region of the
chloroplast were analyzed in 11 Bolanthus specimens. The PCR reactions
were performed in 50 ml volumes containing 10 mlof103buffer, 1 unit
Taq DNA polymerase (Promega, Madison, Wisconsin), 200 mMdNTPs,
2mMMgCl
2
, and 10 pmol of both primers, ITS1 and ITS4, or trnL and
trnF. PCR amplification was performed in a Biometra TProfessional
Standard thermal cycler (Biometra, GmbH, Germany) with the fol-
lowing thermal cycling conditions: 94°C for 2 min, 35 cycles of 94°C for
30 s, 55°C for 1 min, and 72°C for 1 min, and a final extension step
of 8 min at 72°C. The amplification products were analyzed by elec-
trophoresis in 1.2% agarose gel containing ethidium bromide, and the
product sizes were determined using a nucleotide size marker (100 bp
ladder; Fermentas, Vilnius, Lithuania). The PCR products were
sequenced with a BigDye cycle sequencing kit (Applied Biosystems,
Foster City, California) using an ABI 3130XL genetic analyzer (Applied
Biosystems).
Sequence Analysis—The final matrix included 11 species of Bolanthus
from Turkey and one outgroup (see Appendix 1). Dianthus sylvestris
Wulfen was chosen as the outgroup as the genus Dianthus is closely related
to Bolanthus. The DNA sequences were aligned and assembled using
Multiple Sequence Comparison by Log-Expectation (MUSCLE) by
Geneious R9 (Kearse et al. 2012). Molecular phylogenetic analyses were
performed using the maximum likelihood method based on the Kimura 2-
parameter substitution model via MEGA7 software. This model (Kimura 2-
parameter) was determined as being the best-fit model for the current data
according to the FINDMODEL, which is a web service based on MOD-
ELTEST (Posada and Crandall 1998; Tao 2005; Chen et al. 2009; Tamura
et al. 2011). One thousand bootstrap replicates were performed (Felsenstein
1985). The ITS and cpDNA data were analyzed separately; we decided not
Fig. 4. SEM photographs of the seed coat. A1–A3. Bolanthus thymoides.B1–B3. B. aziz-sancarii.
Fig. 5. Distribution map of Bolanthus aziz-sancarii (triangles) and B. thymoides (squares).
SYSTEMATIC BOTANY [Volume 44192
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to combine them due to the fact that different data sets may reflect different
phylogenetic histories of the genes or genomes (Mason-Gamer and Kellogg
1996).
Results and Dıscussıon
The genus Bolanthus resembles the genus Gypsophila L.,
especially in terms of its stem position, leaf shape, and petal
colours. However, it differs from this genus by having a
tubular calyx that does not contain calcium oxalate crystals.
Additionally, some researchers liken the taxa belonging to
the genus Bolanthus to the genus Acanthophyllum C. A. Mey.
However, Bolanthus differs from this genus by 4–36 seeds
and capsules opening by both teeth and valves (Barkoudah
1962).
The new species that was collected in Afyonkarahisar
province is similar to B. thymoides Hub.-Mor. in having stems
completely covered with eglandular and glandular hairs, lax
cushions, leaves that are 3–6 mm long, and linear-oblong
petals. However, it differs from this taxon by having an im-
bricate leaf arrangement, calyx 5.5–7 mm long, petals 7–8.5 mm
long, entirely white petals, and an inflorescence with 1–3
pedicellate flowers (versus calyx 4–5 mm long, petals 5–6mm
long, petals white with purple veins, and an inflorescence with
4–8 sessile or subsessile flowers in B. thymoides). In addition,
while B. thymoides occurs in serpentinized stony fields of the
Mediterranean phytogeographical region, the new species is
found in fissures in tuffaceous volcanic rocks in the Irano-
Turanian phytogeographical region (Table 1).
This study showed for the first time the molecular phylogenetic/
evolutionary/genetic relationships among the taxa of the genus
Bolanthus distributed in Turkey based on the ITS and trnL–trnF
gene sequences. The phylogenetic trees based on two molecular
markers provided evidence for the divergence of B. aziz sancarii
from the rest of the group (Figs. 1, 2).
The two phylogenetic trees from the ITS and cpDNA ana-
lyses differ in overall relationships amongst the Turkish
Bolanthus species (Figs. 1, 2). Conflicting topologies of trees
Fig. 6. Bolanthus aziz-sancarii (holotype, Koç 1209 &Hamzao˘glu). A. Habit. B. Inflorescence. C. Flower. D. Petal. E. Capsule.
KOÇ ET AL.: BOLANTHUS AZIZ-SANCARII IN TURKEY 1932019]
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derived from nuclear and plastid genome regions might be
explained by the hybrid origin of the new species or one or
more of the others. Despite these differences, analyses suggest
monophyly of B. thymoides,B. minuartioides,B. stenopetalus,andB.
mevlanae. ITS and cpDNA analyses revealed relationships of B.
aziz-sancarii to B. cherlerioides. However, the habitat of these two
species are distinctly different even if they have close geographic
distribution. Bolanthus aziz-sancarii distributes within volcanic
regions at a maximum altitude of 1500 m, whereas B. cherlerioides
prefers alpine or subalpine regions within calcareous areas.
Bolanthus cherlerioides differs from B. aziz-sancarii by having 1–3
pedicellate flowers and 2–2.5 mm calyx teeth.
Although morphologically the new species most closely re-
sembles B. thymoides, these taxa are more distantly related (Figs.
1, 2). As a result, all the obtained morphological and molecular
data showed that the B. aziz-sancarii found in Afyonkarahisar
province is a new species of the genus Bolanthus.
Taxonomıc Treatment
The descriptions and the distribution areas of taxa belonging
to the genus Bolanthus from Turkey were given in the work
Flora of Turkey and the East Aegean Islands published in 1967.
The specimens collected by various researchers within the last
50 yr have revealed new distribution areas and morphological
variations of the taxa. With the aim of determining whether
characters such as bracts, calyces, leaves, and seed surfaces
differ among the taxa, descriptions of the new species and the
similar B. thymoides have been generated after detailed spec-
imen examinations.
1. BOLANTHUS THYMOIDES Hub.-Mor., Bauhinia 2(2): 187 (1963)
TYPE:TURKEY. Burdur: d. Tefenni, Passh ¨ohe s ¨udlich
ob Dirmil, 1600–1660 m, auf Serpentinschutt, 28 Jul
1948, Huber - Morath 8447 (holo. G, G00006008,
photo!).
Perennial herb. Stems forming lax cushions, 2–8cmbroad,
ascendant-erect, 3–8cm,base0.8–1 mm diam, entirely
covered with mixed eglandular and glandular hairs. Stems
3–6-noded, internodes 2–5 mm, glandular-hairy. Leaves
linear-setaceous, 3–630.5–0.7 mm, glandular- hairy, 3-
nerved, greenish, rigid, apex acute; sheath membranous,
0.2–0.3 mm, glandular-hairy. Inflorescence congested in
small sessile or subsessile terminal or subterminal clusters,
Fig. 7. Bolanthus aziz-sancarii (holotype, Koç 1848 &Hamzao˘glu). A. Habit. B. Petals. C. Flower.
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densely glandular-hairy, 4–8-flowered, flowers sessile.
Bracts linear-setaceous, 3–4.5 30.5–0.7 mm, as long as or 1.2 times
longer than calyx; apex acute. Calyx tubular, densely glandular-
hairy, 4–531–1.3 mm, with 5 projecting ribs; teeth lanceolate,
1–1.3 mm long, apex acute. Petals linear-oblong, 5–63
0.7–0.9 mm, 1.2 times longer than calyx, white with purple veins;
apextruncate;basecuneate.Stamens10,filaments3–4 mm. Styles
2, 3–4 mm long. Capsule oblong, 3.5–4.5 31–1.3 mm, 2–4-seeded,
usually as long as calyx. Seeds comma-shaped, with prominent
radicle, 1–1.3 30.6–0.8 mm, brown. Figures 3, 4.
Distribution and Habitat—Bolanthus thymoides occurs in
Bilecik, Antalya, Mu˘gla, and Burdur provinces at altitudes be-
tween 605 and 2000 m (Fig. 5). It usually prefers serpentine locales.
Specimens Examined—Bolanthus thymoides Hub.-Mor. Turkey. —BILECIK:
Boz¨uy ¨uk, 740 m, 07 Jun 1975, R. Çetik (KNYA, No: 1239); —MU˘
GLA:Sandras
mountain, NW of Çiçekbaba hill, Serçegedi˘gi site, 1850 m, 14.07.1979, E. ¨
O
zhatay (ISTO, No: 23928); —BURDUR:Altınyayla, Dirmil pass, Esenli plateau,
1610 m, 06 Jun 1996, N. &E. ¨
Ozhatay, H. Duman (ISTE, No: 72219); Dirmil,
hills above Dirmil, 1750–1800 m, 08 Jul 1993, L. Bekat &E. Leblebici (EGE, No:
18864); Altınyayla, Dirmil pass, 605 m, 20 Jun 2002, A.A. D¨onmez 10922 &E.
D¨onmez (HUB); Çamlıyayla, Dirmil pass,1650 m, 30 Jun 1996, Z. Aytaç 7405
(GAZI); Yes
¸ilova, Salda town, Es
¸eler mountain, 2000 m, 18 Aug 2014, Koç
1848 &Hamzao˘glu (Bozok Univ. Herb.);—ANTALYA: Elmalı,Ça˘glıkara,
1540 m, 28 Jun 1975, R. Çetik (KNYA, No: 1238).
2. Bolanthus aziz-sancarii Koç & Hamzao˘glu, sp. nov. TYPE:
TURKEY. Afyonkarahisar: between Bayat and Iscehisar, 1500 m,
fissures of tuffaceous volcanic rocks, 02 Jul 2010, Koç 1209 &
Hamzao˘glu (holotype:GAZI,isotypes:GAZI,ANK,HUB).
Bolanthus aziz-sancarii is most similar to B. thymoides.It
differs from B. thymoides in having bracts lanceolate, 5.5–7mm
long (vs. linear-setaceous, 3–4.5 mm long); inflorescence with
1–3 pedicellate flowers (vs. with 4–8 sessile or subsessile
flowers); calyx 5.5–7 mm long (vs. 4–5 mm long); petals
7–8.5 mm long (vs. 5–6 mm long), completely white (vs. with
purple veins), apex emarginate (vs. truncate); capsules 5–6.5 mm
long (vs. 3.5–4.5 mm long) (Table 1).
Perennial herb. Stems forming lax cushions, 4.5–15 cm
in diam, ascendant-erect, base 0.8–1.2 mm diam, entirely
covered with eglandular and glandular hairs. Stems short,
8–12-noded, internodes 4–8 mm, glandular-hairy. Leaves
linear-setaceous, 4–5.5 30.5–0.7 mm, glandular-hairy, 3-
nerved, greenish, rigid, apex acute; sheath membranous,
0.2–0.3 mm, glandular-hairy. Inflorescence terminal, densely
glandular-hairy, 1–3-flowered, flowers pedicellate. Bracts
lanceolate, 5.5–730.9–1.2 mm, as long as calyx; edge with
ciliae 0.1–0.15mmlong;apexacute.Pedicels1–2mmlong,
densely glandular-hairy. Calyx tubular, densely glandular-
hairy, 5.5–731.3–1.7 mm, with 5 projecting ribs; teeth
lanceolate, 1–1.5 mm long, apex acute. Petals linear-oblong,
7–8.5 30.8–1.2 mm, 1.5 times longer than calyx, entirely
white, without purple veins; apex emarginate; base cuneate.
Stamens 10, filaments 4–6mm.Styles2,4–6mmlong.
Capsule oblong, 5–6.5 31.3–1.7 mm, 2–4-seeded, usually as
long as calyx. Seeds comma-shaped, with prominent radicle,
0.9–1.1 30.6–0.8 mm, brown. Figures 4, 6, 7.
Distribution and Habitat—Bolanthus aziz-sancarii prefers
fissures of tuffaceous volcanic rocks at an altitude of 1500 m in
Afyonkarahisar province (Fig. 5).
Conservation Status—According to current data, Bolanthus
aziz-sancarii grows in an area of approximately 1000 m
2
cov-
ering the Bayat-Iscehisar districts of Afyonkarahisar province.
The species, which has a discontinuous distribution, has been
reported in only one locality in Afyon. Because of over-
grazing, the habitat of this species is under threat, and de-
struction of the species is leading to the reduction in the
number of plants. For that reason, it is proposed that the
species should be classified as endangered [EN (B1a)]
according to the International Union for Conservation of
Nature (IUCN) categories (IUCN 2013).
Etymology—In honor of our Nobel laureate scientist, Aziz
Sancar (USA), his name was given to this species.
Key to the Species of
Bolanthus
in Turkey
1. Petals entirely white, without purple veins .......................................................................................2
2. Plants prostrate or decumbent; stems long-eglandular-hairy; flowers (5–)10–25 in dense subsessile clusters; calyx 4–5mmlong .............
.............................................................................................................B. minuartioides
2. Plants ascending or erect; stems densely glandular-hairy; flowers 1–3 in pedicellate clusters; calyx 5.5–7mmlong ............B. aziz-sancarii
1. Petals entirely purple or white with purple veins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3. Stems glabrous or puberulent-hairy; leaves setaceous, glabrous; bracts one-half as long as calyx . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
4. Plants prostrate; internodes 5–15 mm long; flowers pedicellate ...................................................B. huber-morathii
4. Plants ascending or erect; internodes 1–3 mm long; flowers sessile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B. mevlanaea
3. Stems glandular-hairy; leaves linear to linear-setaceous; bracts 1–1.2 times longer than calyx . . . ......................................5
5. Cushions very compact; leaves falcate; inflorescence 1–3-flowered; calyx teeth 2–2.5mmlong ..........................B. cherlerioides
5. Cushions lax or prostrate, leaves not falcate; inflorescence 4–15-flowered; calyx teeth 0.5–1.5mmlong .............................6
6. Plants ascending or erect; internodes 2–5 mm long; flowers sessile; petals linear-oblong . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B. thymoides
6. Plants prostrate; internodes 5–15 mm long; flowers pedicellate; petals narrowly oblanceolate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
7. Leaves linear-oblanceolate; calyx 3–3.5 mm; inflorescence of lax clusters . . ...............................................8
8. Leaves long-ciliate; calyx non-viscid, ovary 8–10-ovulate . . . . . . . .........................................B. stenopetalus
8. Leaves not long-ciliate; calyx viscid, ovary 20-ovulate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B. frankenioides
–Pedicels 2–4 mm, as long as calyx . ...............................................................var.frankenioides
–Pedicels 1–2 mm, shorter than calyx .................... ..........................................var.fasciculatus
7. Leaves subulate-setaceous or linear; calyx 3.5–5.5 mm; inflorescence of compact clusters . . . ................................9
9. Leaves linear, 3-nerved; calyx 3.5–4.5 mm long; petals 3.3–4.5 mm long, usually as long as calyx ................B. turcicus
9. Leaves subulate-setaceous, 1-nerved; calyx 4.5–5.5 mm long; petals 6–7 mm long, 1.5 times longer than calyx . . . B. spergulifolius
Acknowledgments
We are indebted to Bozok University for financial support (Project
No:6602CFEF/17-67)andthecuratorsoftheE,K,G,GAZI,HUB,
ISTE, ISTO, EGE, and KNYA for the address information of
specimens.
Author Contributions
The morphological diagnosis and field work were conducted by MK
and EH. DNA extraction, amplification, sequencing, and alignment of
sequences, and phylogenetic analyses were done by IB. All three authors
conceived and contributed to writing the manuscript.
KOÇ ET AL.: BOLANTHUS AZIZ-SANCARII IN TURKEY 1952019]
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APPENDIX 1. Bolanthus specimens sampled in the field (all are kept in
Herbarium of Bozok University). *GenBank accession numbers are ITS and
trnL-trnF, respectively.
Turkey. Bolanthus aziz-sancarii. —AFYONKARAHISAR: Between Bayat and
Iscehisar, volcanic tuff rocky crevices, 1500 m, 05 Jul 2015, Koç 1741 &
Hamzao˘glu, *KY406153, KY421725. Bolanthus minuartioides.—KONYA:be-
tween Bozkır and Ak¨oren, dry hills, 1105 m, 18 Jun 2013, Koç 2006 &
Hamzao˘glu, *KY406151, KY421723. Bolanthus spergulifolius.—K¨
UTAHYA:
Gediz, Murat mountain, road Kaplıcalar, stony places, 1495 m, 25 Jun 2015,
Koç 2043 &Hamzao˘glu, *KY406150, KY421722. Bolanthus cherlerioides.—KONYA:
between Aks
¸ehir and S
¸arkikaraa˘g aç, around Yellibel pass, 1550 m, 14 Jul 2011,
Koç 217 &Hamzao˘glu, * KY406155, KY421727. Bolanthus thymoides. —BURDUR:
Yes
¸ilova,Saldatown,Es
¸eler mountain, 2000 m, 18 Aug 2014, Koç 1848 &
Hamzao˘glu, *KY406152, KY421724. Bolanthus frankenioides var. frank-
enioides.—ANTALYA:Elmalı,Beyda˘gı,K¨uç¨uks ¨o˘gle town, Serkizalan pla-
taeau, Kırkınargedi˘gi site, 2240 m, 28 Jul 2015, Koç 2167 &Hamzao˘glu,
*KY406149, KY421721. Bolanthus frankenioides var. fasciculatus.—MU˘
GLA:
K¨oyce˘giz, Yayla town, above G ¨okçeova lake, Sandras mountain, 2030 m,
05 Aug 2015, Koç 2188 &Hamzao˘glu, *KY406148, KY421720. Bolanthus
stenopetalus.—MU˘
GLA:K¨oyce ˘giz, Yayla town, above G ¨okçeova lake,
Sandras mountain, 2030 m, 05 Aug 2015, Koç 2196 &Hamzao ˘glu,
*KY406147, KY421719. Bolanthus huber-morathii. Bursa: between
So˘gukpınar and Keles, output So ˘gukpınar, 930 m, 19 Jul 2011, Koç 2352 &
Hamzao˘glu, *KY406154, KY421726. Bolanthus mevlanaea.—ANTALYA:be-
tween Akseki and Bozkır, steppee, 10 Jul 2016, Hamzao˘glu 6748 & Koç,
*KY406145, KY421717. Bolanthus turcicus.—AKSARAY:Hasanmountain
above Karkın town, 1950 m, volcanic stony slopes and alpine steppe, 18
Jul 2015, Koç 2226 &Hamzao˘glu, *KY406146, KY421718. Dianthus sylvestris
(GenBank Outgroup), AY594317, EF407923.1.
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