Research Article |
Corresponding author: Necmettin Güler ( nguler@trakya.edu.tr ) Academic editor: Murielle Simo-Droissart
© 2016 Necmettin Güler.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Güler N (2016) Seed micromorphology of Orchis Tourn. ex L. (Orchidaceae) and allied genera growing in Edirne province, Turkey. PhytoKeys 68: 9-25. https://doi.org/10.3897/phytokeys.68.8746
|
In this study, the seed micromorphologies of eight taxa of Anacamptis, Neotinea and Orchis growing around Edirne province (Turkey) were investigated using light microscopy and scanning electron microscopy (SEM). Slides prepared with glycerin jelly were used for measurements under the light microscope and fine details of seed testae characteristics were observed with SEM. Seeds of the investigated orchid taxa are fusiform shaped and of different shades of brown. Their lengths and widths are different among the taxa and range between 0.263–0.640 mm and 0.118–0.208 mm, respectively. Testa surfaces of Orchis mascula subsp. mascula, O. purpurea subsp. purpurea and O. simia subsp. simia, are smooth while those of Anacamptis coriophora, A. laxiflora subsp. laxiflora, A. morio subsp. morio, A. papilionacea and Neotinea tridentata subsp. tridentata are reticulate. An identification key based on seed morphologies and sizes is suggested for the first time, including testae structures of orchids growing in Edirne province. The overall results of the study showed that morphological structures of orchid’s seeds could be used as diagnostic characters in identification.
Anacamptis , Neotinea , Orchis , Orchidaceae , seed micromorphology
Orchidaceae are one of the most diversified and evolved families in the flowering plants (
According to
The distribution area of the genus Anacamptis reaches to the Atlantic in the west and to the Hebrides and southern Scandinavia in the north. It includes the North African mountains in its southwest border, whereas other parts of North Africa and the Canaries remain blank, although it penetrates along the Levant considerably further to the south. The genus in the east reaches to Lake Balchaš in central Asia and its representatives are also found on all the larger islands of the Mediterranean. The ecological demands of the different species are various, but all commonly prefer to settle within biotopes that have seasonal changes, really humid winters, which temporarily become very dry in summer (
The genus Orchis (Orchidaceae, Orchidinae) is limited in its distribution exclusively to the northern hemisphere. Its mainly distribution area is Mediterranean Basin where the maximum density of species is reached; however, other part of Europe are also settled to great extent. In addition the genus with some species, divert out of its main range and reaches northwards to Scandinavia, whilst in an easterly direction to Mongolia and reaches last Lake Baikal. On the north coast of Africa the eastern part is blank to great extent due to the absence of suitable biotopes; however, areas of Asia Minor and further on to Iraq and Iran are included. The ecological demands of the different species are various (
The genus Neotinea is limited to Europe, Asia Minor, the Caucasus and the north-west coastal regions of North Africa. The ecological demands of the different species are various (
Seed morphology is one of the important taxonomic characters of orchids.
The rather small sizes of seeds make them difficult to study their details and to compare some features with only light microscopy. Therefore, making comparisons and determining details that could be used as taxonomical characters without SEM techniques appear to be a challenging task (
Most of the studies performed on orchid seeds were based on tropical orchids whereas the non-tropical species were generally neglected (
Several authors published different papers about seed morphology in the genera of Orchis, Anacamptis and Neotinea.
Few studies exist on seed morphology of Turkish orchids. One of them was performed by
We analyzed seeds of eight taxa belonging to the genera Orchis, Anacamptis and Neotinea. The study material consisting of specimens of eight orchid taxa were collected from the region within Edirne provincial borders in 1995 and 1996 and are kept in EDTU Herbarium. A list of voucher specimens and localities is given in the Table
Species | EDTU | Source Locality | Collectors | Date Received |
---|---|---|---|---|
A. coriophora | 6075 | Kesan, Yayla village | N. Güler | 02.06.1995 |
A. laxiflora subsp. laxiflora | 6074 | Kesan, Mecidiye village | N.Güler & M.Aybeke | 06.05.1995 |
A. morio subsp. morio | 6056 | Kesan, Yerlisu village | N.Güler & M.Aybeke | 22.04.1995 |
A. morio subsp. morio | 6058 | Kesan, Camlica village | N.Güler & M.Aybeke | 06.05.1995 |
A. morio subsp. morio | 6059 | Kesan, Camlica-Gökcetepe villages | N.Güler & M.Aybeke | 06.05.1995 |
A. morio subsp. morio | 6062 | Kesan, Mecidiye village | N.Güler & M.Aybeke | 06.05.1995 |
A. morio subsp. morio | 6063 | Kesan, Yayla village | N.Güler & M.Aybeke | 07.05.1995 |
A. morio subsp. morio | 6065 | Enez, Haskoy village | N.Güler | 09.05.1995 |
A. morio subsp. morio | 6067 | Lalapasa, Hanliyenice village | N.Güler | 16.05.1995 |
A. morio subsp. morio | 6265 | Enez, Abdürrahim village | N.Güler & M.Kirec | 02.05.1996 |
A. morio subsp. morio | 6267 | Kesan, Kizkapan village | N.Güler & M.Aybeke | 11.05.1996 |
A. papilionacea | 6079 | Kesan, Yayla village | N.Güler | 02.06.1995 |
O. mascula subsp. mascula | 6132 | Enez, Candir village | N.Güler & M.Kirec | 02.05.1996 |
O. purpurea subsp. purpurea | 6119 | Uzunköprü, Turnaci village | N.Güler & M.Aybeke | 27.05.1995 |
O. purpurea subsp. purpurea | 6103 | Hasanağa village | N.Güler | 25.04.1995 |
O. purpurea subsp. purpurea | 6110 | Kesan, Suluca village | N.Güler & M.Aybeke | 09.05.1995 |
O. purpurea subsp. purpurea | 6116 | Lalapasa, Dogankoy village | N.Güler | 19.05.1995 |
O. simia subsp. simia | 6080 | Kesan, Yerlisu village | N.Güler & M.Aybeke | 15.04.1995 |
N. tridentata subsp. tridentata | 6136 | Kesan, Yayla village | N.Güler & M.Aybeke | 11.05.1996 |
N. tridentata subsp. tridentata | 6120 | B.Ismailce village | N.Güler | 19.05.1995 |
N. tridentata subsp. tridentata | 6092 | Kesan, Kizkapan village | N.Güler & M.Aybeke | 07.05.1995 |
The terminology and methods were adopted from those of
Ve = 4/3pab2
where a is ½ of embryo length, b is ½ of embryo width, and 4/3p is equal to 4.188. Percentage air space was calculated by using the formula: [(Vt-Ve)/Vt] .100.
All investigated orchid seeds were fusiform in shape and had transparent and elliptical embryos (Figures
Species | EDTU | Figure | Embryos | Seeds | Vs/Ve (mm3 x10-3) | Percent Air Space | ||||
---|---|---|---|---|---|---|---|---|---|---|
L/S.D. (mm) |
W/S.D. (mm) |
L/W | L/S.D. (mm) | W/S.D. (mm) | L/W | |||||
A. coriophora | 6075 | 1A−1C | 0.177/0.015 | 0.116/0.014 | 1.532 | 0.397/0.040 | 0.186/0.021 | 2.137 | 3.59/1.24 | 65.60 |
A. laxiflora | 6074 | 1D−1F | 0.225/0.039 | 0.140/0.015 | 1.606 | 0.599/0.097 | 0.208/0.022 | 2.880 | 6.78/2.31 | 65.98 |
A. morio subsp. morio | 6056 | 0.124/0.011 | 0.092/0.011 | 1.351 | 0.400/0.051 | 0.142/0.007 | 2.824 | 2.10/0.54 | 74.19 | |
A. morio subsp. morio | 6058 | 0.137/0.015 | 0.097/0.007 | 1.413 | 0.420/0.053 | 0.143/0.089 | 2.927 | 2.26/0.68 | 70.11 | |
A. morio subsp. morio | 6059 | 0.158/0.019 | 0.118/0.018 | 1.339 | 0.376/0.045 | 0.171/0.024 | 2.191 | 2.89/1.15 | 60.34 | |
A. morio subsp. morio | 6062 | 0.173/0.020 | 0.129/0.014 | 1.344 | 0.513/0.037 | 0.161/0.018 | 3.196 | 3.46/1.50 | 56.82 | |
A. morio subsp. morio | 6063 | 0.156/0.026 | 0.097/0.017 | 1.609 | 0.640/0.067 | 0.152/0.017 | 4.197 | 3.89/0.76 | 80.41 | |
A. morio subsp. morio | 6065 | 2A−2C | 0.152/0.012 | 0.115/0.011 | 1.319 | 0.506/0.068 | 0.147/0.019 | 3.435 | 2.87/1.05 | 63.38 |
A. morio subsp. morio | 6067 | 0.142/0.019 | 0.106/0.017 | 1.335 | 0.452/0.031 | 0.141/0.023 | 3.209 | 2.34/0.83 | 64.50 | |
A. morio subsp. morio | 6265 | 0.183/0.014 | 0.128/0.011 | 1.425 | 0.573/0.095 | 0.177/0.023 | 3.247 | 4.67/1.57 | 66.43 | |
A. morio subsp. morio | 6267 | 0.160/0.026 | 0.106/0.012 | 1.517 | 0.503/0.077 | 0.148/0.018 | 3.398 | 2.88/0.93 | 67.60 | |
Average for A. morio | 0.157 | 0.103 | 1.526 | 0.482 | 0.156 | 3.096 | 3.08/1.07 | 65.14 | ||
A. papilionacea | 6079 | 2D−2F | 0.138/0.027 | 0.104/0.022 | 1.327 | 0.451/0.076 | 0.162/0.027 | 2.778 | 3.11/0.78 | 74.84 |
O. mascula subsp. mascula | 6132 | 3A−3C | 0.124/0.016 | 0.104/0.016 | 1.191 | 0.326/0.035 | 0.195/0.032 | 1.674 | 3.24/0.70 | 78.32 |
O. purpurea subsp. purpurea | 6103 | 3D−3F | 0.138/0.022 | 0.086/0.012 | 1.602 | 0.450/0.030 | 0.144/0.017 | 3.119 | 2.45/0.53 | 78.21 |
O. purpurea subsp. purpurea | 6110 | 0.119/0.016 | 0.086/0.009 | 1.381 | 0.356/0.082 | 0.142/0.018 | 2.514 | 1.87/0.46 | 75.30 | |
O. purpurea subsp. purpurea | 6116 | 0.118/0.014 | 0.079/0.008 | 1.484 | 0.263/0.026 | 0.118/0.012 | 2.221 | 0.96/0.39 | 59.50 | |
O. purpurea subsp. purpurea | 6119 | 0.143/0.016 | 0.098/0.008 | 1.461 | 0.480/0.042 | 0.166/0.014 | 2.902 | 3.44/0.72 | 79.16 | |
Average for O. purpurea | 0.129 | 0.111 | 1.169 | 0.387 | 0.142 | 2.719 | 2.18/0.53 | 75.90 | ||
O. simia subsp. simia | 6080 | 4A−4C | 0.148/0.017 | 0.093/0.015 | 1.593 | 0.357/0.029 | 0.166/0.022 | 2.147 | 2.58/0.67 | 73.92 |
N. tridentata subsp. tridentata | 6092 | 0.158/0.024 | 0.125/0.024 | 1.260 | 0.578/0.075 | 0.185/0.025 | 3.117 | 5.19/1.29 | 75.12 | |
N. tridentata subsp. tridentata | 6120 | 4D−4F | 0.145/0.016 | 0.101/0.012 | 1.428 | 0.448/0.050 | 0.157/0.029 | 2.865 | 2.87/0.78 | 72.96 |
N. tridentata subsp. tridentata | 6136 | 0.157/0.013 | 0.104/0.013 | 1.511 | 0.449/0.044 | 0.153/0.017 | 2.942 | 2.73/0.89 | 67.24 | |
Average for N. tridentata | 0.153 | 0.110 | 1.391 | 0.492 | 0.165 | 2.983 | 3.60/0.99 | 72.55 | ||
Average for orchids studied | 0.151 | 0.106 | 1.430 | 0.454 | 0.160 | 2.853 | 3.152/0.942 | 70.00 |
When testae and embryos were investigated for their colors, the following patterns were obtained: Orchis mascula subsp. mascula and Anacamptis laxiflora subsp. laxiflora were light brown, A. coriophora, A. morio subsp. morio and A. papilionacea were brown, O. purpurea subsp. purpurea and Neotinea tridentata subsp. tridentata were dark brown and O. simia subsp. simia was darker brown than the rest.
It is possible to divide the orchid species found in Edirne into two groups according to their testa morphologies. The first group includes Anacamptis coriophora (Fig.
When the reticulations were analyzed, it appeared that they showed minute anastomosis. Some orchids, especially the tropical ones, have conspicuous reticulations such as Calypso bulbosa (L.) Oakes (
Seed lengths and widths ranged between 0.263–0.640 mm and 0.118–0.208 mm, respectively. The length and width measurements for embryos were 0.118–0.225 mm and 0.079–0.140 mm, respectively. All species are listed in Table
When the mean values of orchid seed morphometry obtained in the present study were compared to those reported in
L/W ratios provide data on the relative degree of truncation (Arditti 1979). The lowest L/W of 1.674 in Orchis mascula subsp. mascula showed that seeds of this species were the most truncate seeds. This species is followed by O. simia subsp. simia, Anacamptis coriophora and O. purpurea subsp. purpurea with their low L/W ratios implying a high truncate nature. On the other hand, higher L/W values were obtained for A. papilionacea, A. laxiflora subsp. laxiflora and Neotinea tridentata subsp. tridentata indicating that they have more elongate seeds. The highest L/W ratio of A. morio subsp. morio seeds (4.197) shows that the seeds of this species are elongate.
The mean lengths and widths of the embryos of the investigated eight taxa were 0.151 mm and 0.106 mm, respectively. The embryos were found to be elliptical with an average L/W value of 1.43. The lowest L/W value of O. mascula subsp. mascula led us to conclude that the embryos of this species were sphere-like. This species is followed by Anacamptis papilionacea. The high L/W values of the other species is an indication that their embryos are elliptical rather than spherical.
Percentage air space affects the length of time the orchid seeds are in air. Specimens with high percentage air space values are known to spread over longer distances via wind (
As shown in previous studies on orchids, there are a number of diagnostic and phylogenetically informative characters present in orchid seeds. In this study, seed morphologies of eight orchids taxa growing in Edirne province were investigated and criteria that could be used to differentiate the seeds are presented. Also, a key is constructed below, based on seed morphology.
1 | Testa walls reticulate | 2 |
– | Testa walls smooth | 6 |
2 | Reticulations occurring in periclinal walls more or less conspicuous or not at all | 3 |
– | Reticulations occurring in periclinal walls conspicuous | 4 |
3 | Thickenings in anticlinal walls rather conspicuous | A. laxiflora subsp. laxiflora |
– | Thickenings in anticlinal walls inconspicuous | A. morio subsp. morio |
4 | Reticulations in testa cells transversely | N. tridentata subsp. tridentata |
– | Reticulations different | 5 |
5 | Reticulations in testa cells transversely diagonal | A. coriophora |
– | Reticulations in testa cells longitudinally diagonal | A. papilionacea |
6 | Seed fusiform | O. purpurea subsp. purpurea |
– | Seed fusiform-oblong | 7 |
7 | Seed light brown | O. mascula subsp. mascula |
– | Seed dark brown | O. simia subsp. simia |
The author would like to thank Scientific Research Projects Unit of Trakya University for supporting this research with the project N° TUBAP- 2016/110. The author is indebted to Prof. Dr. Ekrem Sezik, Prof. Dr. Göksel Olgun and Dr. Nesibe Başak for their comments and advices. He also wish to thank Dr. Volkan Aksoy for translation and TUTAGEM (Trakya University Technology Research, Development and Application Center) staffs for SEM photographs.