Haplophyllumermenekense (Rutaceae), a new species from Turkey

Abstract A new species of Haplophyllum, Haplophyllumermenekense (Rutaceae) is described and illustrated in line drawing. It grows on stony slopes of Ermenek town, Karaman province, in southern Turkey. It is compared with the closely related species H.myrtifolium. H.ermenekense is distinguished from the morphologically similar H myrtifolium chiefly by sepal shape, petal size, capsule size, presence of capsule hair and appendage form. On the other hand, the seed coat and pollen grains surface of H.ermenekense and H.myrtifolium are demonstrated in SEM photographs. In addition to the detailed description, the illustration, distribution map, conservation status and ecology of the new species are also provided.


Introduction
With its 69 species, Haplophyllum Jussieu is one of the richest genera in the Rutaceae family (Townsend 1986, Navarro et al. 2004, Soltani and Khosravi 2005, Tugay and Ulukuş 2017. The genus is widely distributed in subtropical and tropical regions of the northern hemisphere of the Old World, notably in Iran, Turkey and Central Asia (Townsend 1986).
Some authors have endeavoured to subdivide Haplophyllum into different sections by using morphological characters (Spach 1849, Boissier 1867, Engler 1896, Vvedensky 1949, Townsend 1986. The most comprehensive studies conducted with re-gard to Haplophyllum were published by Vvedensky (1949) and Townsend (1986). In these studies, the genus was divided into four sections by Vvedensky (1949), based on capsule opening, ovule and carpel number. In the last monographic study, Townsend (1986) divided the genus into three sections according to carpel number, capsule opening, petal colour, plant architecture, stamen form and ovary shape.
Haplophyllum species are perennial herbs, growing mainly on sandy soil, rocky hills, slopes, stony landscapes or steppes (Townsend 1986). Morphologically, the genus is characterised by the presence of exstipulate, cymose inflorescences with, bracts, lax to dense. Flowers have five petals and five sepals, creamy-white to bright yellow petals, ten stamens with free filaments expanded below and pubescent on the inner surface. Fruits have three to five connate carpels, five-lobed capsules which are dehiscent or indehiscent (Townsend 1986). Pollen grains are tricolporate, radially symmetrical and isopolar (Ulukuş et al. 2016). The equatorial view of pollen of Haplophyllum is distinctly rhomboid (Townsend 1986). Tectum ornamentation is commonly striate or striate perforate (Townsend 1986, Ulukuş et al. 2016, Tugay and Ulukuş 2017. Turkey is one of the most important centres for Haplophyllum diversity with three phytogeographical regions; Euro-Siberian, Irano-Turanian and Mediterranean (Ulukuş et al. 2016). Boissier (1867), in his Flora Orientalis, recognised 15 species in Turkey. At a later date, Townsend (1967) recognised 17 taxa in the Flora of Turkey. Recently, one Haplophyllum species was published by Tugay and Ulukuş (2017), bringing the genus to 18 taxa. With the new species described in this paper, Turkey harbours 19 Haplophyllum taxa, 11 (58%) of which are endemic.
The Irano-Turanian region in SW Asia is one of the richest floristic areas of the Holarctic Kingdom. Most of its species diversity is concentrated in the Anatolian plateau, Iranian plateau and Central Asia (Zohary 1973, Manafzadeh et al. 2014.
Haplophyllum has mostly been studied from a morphological point of view by several authors (Jussieu 1825, Spach 1849, Boissier 1867, Engler 1896, Vvedensky 1949, Townsend 1986, Salvo et al. 2011, Ulukuş et al. 2016, Tugay and Ulukuş 2017. Only a few palynological studies have been conducted in the genus Haplophyllum (Townsend 1986, Perveen and Qaiser 2005, Akyol et al. 2012, Al-Eisawi and Al-Khatib 2015, Ulukuş et al. 2016, Tugay and Ulukuş 2017. There are several studies about the seeds of some species belonging to the Haplophyllum genus (Townsend 1986, Navarro et al. 2004, Tugay and Ulukuş 2017. From a biogeographical standpoint, Manafzadeh et al. (2014) showed that the clade, formed by the Mediterranean species of Haplophyllum and Anatolian H. telephioides, diverged from its geographically diverse sister clade in the middle Miocene probably in the Irano-Turanian region and, from there, it quickly invaded the eastern Mediterranean region.
Ermenek, located within the boundaries of Karaman province, in the Mediterranean region of Turkey, is one of the most interesting plant diversity centres in Turkey.
The aim of this study is to describe the new species, H. ermenekense, found in Ermenek and to compare it with similar species, especially H. myrtifolium Boiss., based on evidence from (micro) morphology and palynology.

Material and methods
Between 2011 and 2015, during the process of writing a revision of the Haplophyllum genus in Turkey, the authors carried out fieldwork around Ermenek and collected samples. All available specimens of Haplophyllum harboured in Turkish herbaria (ANK, EGE, GAZI, HUB, ISTE, KNYA), relevant Turkish collections from herbaria out of Turkey (E, K), as well as all specimens collected during recent fieldwork, were examined under dissecting microscopes. Examined specimens were checked and evaluated comprehensively by relevant literature (Boissier 1867, Vvedensky 1949, Townsend 1966, 1967, 1968, 1985, 1986. The Townsend (1967Townsend ( , 1986 terminology was used to describe the new species. For palynological investigations, the pollen slides were prepared according to Wodehouse's (1935) technique. The pollen micromorphology of H. ermenekense and H. myrtifolium were examined by using scanning electron microscopy (SEM) techniques. For SEM, pollen grains were first mounted on double-sided carbon tape affixed to aluminium stubs, covered with gold with a Hummle VII sputter coater and photographed at a magnification of 2000× to 7000× with a JEOL-5600. SEM micrographs were used to determine exine sculpturing of the pollen. For pollen morphology, Punt et al. (2007) terminology was used.
Morphometric measurements of seeds were made under a stereomicroscope (Leica S8AP0) coupled to a Leica DFC 295 digital camera. The seed length and width of (10-) 30-35 seeds per species were measured. Measurements were made using the Image Tool software. Minimum-maximum ranges, mean, standard deviations of seed length and width, as well as length/width ratio, were calculated. SEM micrographs were used to determine seed coat sculpturing of the seeds. The terminology of Stearn (1983) was adopted to describe the SEM aspects of the seed coat.
Etymology. The name of Ermenek town where new species found is given to the species epithet.
Proposed Turkish name for the new species. Ermenek sedosu. Distribution and conservation status. H. ermenekense is endemic to Karaman province. It is an element belonging to the east Mediterranean phytogeographic region (Fig. 1). The range of this new species is limited to a single locality and its area of occupancy is estimated to be less than 5 km or 5 km 2 . The number of mature individual plants is estimated to be less than 250. As it is perennial, this new species has a crucial advantage for its future as destruction of the bushes by local people, road construction and deterioration of habitats may cause some threats. Thus, according to criterion D, it can be included in the EN (Endangered) category (IUCN 2001;.

Seed morphology
The seed features of H. ermenekense and its immediate relative H. myrtifolium were investigated. It was seen that H. ermenekense has a reniform seed type. Seeds are 1.40-1.83 mm × 1.18-1.39 mm and the range of the L/W ratio is 1.28 ± 0.09. The seeds of H. ermenekense have widely spaced transverse ridges and micromorphologically, the sculpturing of the seed coat is not clearly striate. On the other hand, H. myrtifolium has narrowly spaced longitudinal ridges and a clear striate sculpturing pattern on the seed coat surface in the detailed view (Fig. 5).

Discussion
Haplophyllum ermenekense is morphologically similar to H. myrtifolium, but it differs from H. myrtifolium by significant vegetative and reproductive characters (Table 1). According to Townsend (1986), H. molle, H. amoenum and H. viridulum have white petals yet H. canaliculatum has creamy-white petals. However, these species are basically separated from the other white or creamy-white flowering group without appendage. According to Townsend (1967), H. vulcanicum differs from H. myrtifolium and H. megalanthum by having stipitate glands on stems and inflorescence and H. megalanthum differs from H. myrtifolium by having larger corollas and rhomboidlanceolate upper leaves. In addition, Townsend (1967) reported that H. megalanthum could be a large flowered variety of the western variant of H. myrtifolium. Ulukuş et al (2016) reported that H. myrtifolium differs from H. vulcanicum by its white to sulphur-yellow petals (versus cream-white), conical and rod-like appendages on the ovary (versus smooth line and acute), patent and crisped hairy indumentum (versus stipitate glands) and it differs from H. megalanthum by its lanceolate, lanceolate-elliptic and rarely ovate upper leaves (versus rhomboid-lanceolate), crisped or patent hairy leaf indumentum (versus silky), oblong-ovate (versus lanceolate-elliptic), white to sulphuryellow (versus creamy-white) and larger petals (in H. myrtifolium 6−8.5 × 3−4.5 mm, Haplophyllum ermenekense is related to H. canaliculatum (not distributed in Turkey), differing in its apical appendage on ovary, tuberculate glands on the ovaries (versus non-tuberculate glands) and in its linear bracts (versus broad bracts). Townsend (1986) reported that seeds of Haplophyllum commonly have transverse and longitudinal ridges. Tugay and Ulukuş (2017) showed that seeds are significant characters for differentiation between related species. In addition, in this study, the micromorphological study of the seeds showed that there are clear differences between the studied species. However H. ermenekense does not have a striate sculpturing pattern on the seed surface and H. myrtifolium has a distinctly striate sculpturing pattern in detailed view. On the other hand, H. ermenekense has transverse ridges while H. myrtifolium has longitudinally ridges (Fig. 5). According to Townsend's (1986) palynologic study on some genera of the Rutaceae, including 14 species of Haplophyllum, it was shown that H. myrtifolium has striate pollen exine sculpturing. In this study, palynological results showed that there are no clear differences between the studied species. Both species have often striate-perforate and striate-microreticulate exine sculpturing patterns (Fig. 6).

Conclusion
With the description of this new species, the number of species within Haplophyllum has risen to 70. This study provides material and data to aid further research on this important genus of the Rutaceae.