Morphological, Anatomical and Palynological Studies on Endemic Matthiola anchoniifolia Hub. -Mor. (Brassicaceae)

In this paper, anatomical, palynological and seed micromorphological properties of an endemic plant Matthiola anchoniifolia Hub.Mor. are recorded for the first time. A description and descriptive illustrations of the species are given based on the collected specimens for morphological study. Seed surface of M. anchoniifolia is examined by scanning electron microscope. The seed of M. anchoniifolia was compressed, brownish in colour and the cells of testa were nearly 60-80 μm in diameter and ranged from isodiametric, tetragonal or pentagonal. The anticlinal walls were straight or weakly curved while the outer periclinal walls were concave to flat with smooth surface. In anatomical study, cross sections of root, stem and stem leaf are examined. The root had secondary structure. Periderm consists of 5-8 layers of cells for phellem. Cortex consists of 9-12 layered parenchymatic tissue under the periderm. Secondary phloem ring-shaped, 6-9 layered and consists of companion cells and grouped sieve tubes. Stem had primary structure when analyzed. It is circular with a few irregular ribs in cross section. Cortex is 8-12 layered and parenchymatous. Stoma cells are present on both epidermis. Leaf is isobilateral. There are unicellular and ramified hairs on both surface. Palisade parenchyma cells are 1-2 layered and spongy parenchyma cells are 5-12 layered. M. anchoniifolia has tricolpate pollen type, prolate pollen shape and reticulate exine ornamentation.


Introduction
Brassicaceae has been represented by 365 genus and 3250 species in the world and it is indicated as a huge family (Simpson, 2006). The major centres of distribution of the family are in the Mediterranean, Irano-Turanian and Saharo-Sindian regions (Hedge, 1976). In terms of the Brassicaceae family, Turkey is one of the richest country with 61 genera and 653 native species (Al-Shehbaz et al., 2007). The genus of Matthiola R.Br. (Brassicaceae) is distributing in Africa, Europe and Asia. This genus represented approximately 50 species in the world and with 10 species in Turkey (Cullen, 1965;Dirmenci et al., 2006;Heywood, 1993 (Dirmenci et al., 2006;Ekim et al., 2000). M. anchoniifolia has been distributed in Sivas province in Turkey (Cullen, 1965). According to Red Data Book of Turkish Plants, treat category of M. anchoniifolia Hub.-Mor. is lower risk/conservation dependent (LR / cd) (Ekim et al., 2000). There is no report on the anatomy, palynology and seed micromorphology of M. anchoniifolia which is an en-demic species in Turkey. In this study, morphological, anatomical and palynological properties of M. anchoniifolia were given in detail for the first time.
In the present study morphology, anatomy, palynology and seed micromorphology of M. anchoniifolia, a narrow deployed endemic species, was examined in detail. Some additions are required for morphological features (Cullen, 1965). Biometric measures belonging to the species is presented in Tab. 1. Cullen (1965) did not report any information about the length of plant, stem leaf, pedicel length at flowering time, inflorescense, calyx, corolla, stamen and pistil features.
Sections were waited in blend waited in blend dye about 5 minutes for staining. The stained sections were mounted in glycerin-gelatin to obtain permanent preparations ( Jensen, 1962). Sections were examined using an Olympus light microscope BX51. Images were taken with an Olympus DP 70. For palynological investigations the pollens supplied from herbarium specimens and were prepared by Wodehouse method (Wodehouse, 1965). For measurements of polar axis, equatorial diameter, exine thickness and lumina size of pollen grains were used 100× objective of Olympus light microscope CX21. Measurements were based on 50 pollen grains. For SEM studies, pollen grains coated with gold in a sputter coater. Morphological observations were made and micrographs were taken with a LEO 440 SEM at magnification 5000×; 8500×; 30000×. For morphological descriptions of pollen, followed the terminology of Punt et al. (1994). For seed micromorphology, seeds were mounted on aluminum stubs. Specimen stubs were fixed to the specimen holder of Scanning Electron Microscope (LEO Supra 35 VP FEG SEM) and maintained at accelerating potential voltage of 1.5 kV and photomicrographs were taken at 150× and 500× magnifications in Sabancı University.
In previous studies, Tantawy et al. (2004) determined seed morphological characters of M. longipetala subsp. bicornis (Sibth) Ball., M. longipetala (Vent) DC. subsp. hitra (Conti) and M. longipetala subsp. incana R.Br. In M. longipetala subsp. bicornis the seed surface sculpture was reticulate with more or less broad and raised anticlinal walls. The periclinal walls were more or less raised with smooth striated lateral sides and central knobs. However in M. longipetala subsp. hitra seeds the seed surface pattern was domate with cracks. The anticlinal walls were shallow raised with smooth irregularly arranged folds running in different directions. The periclinal walls were very narrow and deeply depressed. In addition, in the other examined taxa M. longipetala subsp. incana the seed surface pattern was reticulate. The anticlinal walls were more or less broad, raised with smooth surfaces. The periclinal walls were shallow depressed with smooth-papillate surface in certain areas and smooth-striated in others (Tantawy et al., 2004). Kasem et al. (2011) investigated seed exomorphic characters of 32 taxa of Brassicaceae by LM and SEM. In their study seed morphological characters of M. longipetala (Vent) DC. subsp. incana were determined again and they pointed out that seeds of this taxa were globose and yellowish in colour with glabrous texture. The seed size was 1.5 × 0.2 mm (Kasem et al., 2011). In this study seed of M. anchoniifolia was compressed, brownish in colour and the cells of testa were nearly 60-80 μm in diameter. The anticlinal walls were straight or weakly curved while the outer periclinal walls were concave to flat with smooth surface
Brassicaceae is a stenopalynous family whose pollen grains are usually tricolpate and reticulate (Erdtman, 1972;Reile, 1992) but there are some differences between members of the same genus as in Hesperis L. in terms of some features, such as pollen shape (Pınar et al., 2009). Properties of pollen morphology of M. anchoniifolia were determined with our present palynological measurements and examinations. Our results were compared with those in M. anchoniifolia. It was similar to M. longipetala subsp. incana with this feature.

Anatomical properties
The root had secondary structure when the plants collected. Periderm consists of three different tissues, phellem (5-8 layers of cells are brown due to suberised cell wall), phellogen (1 layer) and phelloderm (1 layer), from exterior to interior. Cortex consists of 9-12 layered parenchymatic tissue under the periderm (Fig. 4).
Parenchyma cells are 40-85 × 45-125 μm in size. Cambium is not distinguishable. Secondary phloem ringshaped, 6-9 layered and consists of companion cells and grouped sieve tubes. Secondary xylem consist of trachea and tracheid. In the secondary xylem parenchyma cells could be observed between xylem vessels (Fig. 4).
Stem had primary structure when analyzed. Stem is circular with a few irregular ribs in cross section. Epidermis cells are 12-30 × 15-40 μm in size, ovoid to ovoid-oblong in shape. There are ramified unicellular eglandular hairs on epidermis. Cortex is 8-12 layered and parenchymatous. Cells of cortex are 20-70 × 25-100 μm in size and ovoidal. There are sclerenchyma fibers between xylem bundles. The pith is large and consists of parenchymatous cells. Pith cells are 35-115 × 45-140 μm in size and circular to circular-ovoid in shape (Fig. 5).
Our results confirm the existence of minor differences in pollen size, shape, apertures and exine ornamentation in pollen grains of the other members of Matthiola genus. In Brassicaceae, pollen shapes vary more or less among genera within tribes but very rarely among species within the same genus (Abdel . The pollen shape of M.

Conclusion
Morphological, anatomical, palynological and seed micromorphological properties of M. anchoniifolia which was an endemic plant in Sivas province were reported in detail for the first time in this study.