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Dracocephalum (dragonhead) with about 60 to 70 species is mostly annual or perennial herbs or subshrubs. These species have medicinal values including anticancer, antioxidant, ant hypoxic and immune modulator activities. Dracocephalum thymiflorum grows in limited areas in Iran and forms few local populations and is extensively used by locals. We have no information on anatomical and morphological features of these medicinal species in the country. Clustering and PCA ordination of the studied populations based on morphological and anatomical data separated some of these populations from the others suggesting the existence 2 varieties within this species.

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The genus Dracocephalum L. (Lamiaceae) with about 60 to 70 species is a genus in the sub-tribe Nepetinae, tribe Mentheae of Lamiaceae family, native to temperate regions of the Northern Hemisphere. They are mostly perennial herbs, and rarely annual. Flora Iranica reports 8 Dracocephalum species and the Flora of Iran reports 10 Dracocephalum species in Iran out of which, 4 species are endemic. We collected 7 Dracocephalum species and studied species delimitation and species relationship by morphometric and anatomic results. The species were efficiently delimited by morphological and anatomical characters. Morphological and anatomical characters revealed closer affinity between D. moldavica and D. subcapitatum and D. thymiflorum were placed with distance from these species.

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Tamarix L. (Tamaricaceae) is a halophytic shrub in different parts of Asia and North Africa. Taxonomy and species limitation of Tamarix is very complex. This genus has three sections as Tamarix, Oligadenia, and Polyadenia, which are mainly separated by petal length, the number of stamens, the shape of androecial disk and attachment of filament on the androecial disk. As there was no palynological data on pollen features of Tamarix species of Iran, in the present study 12 qualitative and quantitative pollen features were evaluated to find diagnostic ones. Pollen grains of 8 Tamarix species were collected from nature. Pollen grains were studied without any treatment. Measurements were based on at least 50 pollen grains per specimen. Light and scanning electron microscopes were used. Multivariate statistical methods were applied to clarify the species relationships based on pollen data. All species studied showed monad and tricolpate (except some individuals of T. androssowii). Some Tamarix species show a high level of variability, in response to ecological niches and phenotypic plasticity, which make Tamarix species separation much more difficult. Based on the results of the present study, pollen grains features are not in agreement with previous morphological and molecular genetics about the sectional distinction.

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Acta Botanica Hungarica
Authors: M. Sheidai, S. Darini, S. M. Talebi, F. Koohdar, and S. Ghasemzadeh-Baraki

The genus Linum L. is an important plant genus as it contains the species with economic values and particularly Linum usitatissimum L. that is source of fibre and linseed oil. This genus contains 230 species throughout the world and has about 22 species in Iran. Little is known about Linum species relationship and phylogeny. Therefore, the aim of present study was molecular phylogenetic investigation of the Linum species growing in Iran and to present data on their biogeography. We used both ITS and chloroplast DNA sequences (psbA-trnHGUG region) for inferring the species phylogeny and relationship. We also used cpDNA for inferring the species time of divergence and with ISSR markers to identify the path of species distribution in the country. The phylogenetic trees obtained for both ITS and cpDNA sequences were almost congruent. NeighborNet diagram and BEAST tree based on Bayesian method separated the outgroup species Hugonia and Anisadenia from the other species studied. The subspecies studied in Linum macronicum were placed close to each other and along with L. corymbulosum comprised a separate clade. The clades obtained showed divergence time between 5–20 mya. The present study revealed that the species of the sect. Linum are monophyletic, while members of the sections Linastrum and Syllinum are intermixed and seem to be paraphyletic.

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Acta Botanica Hungarica
Authors: H. Azizi, M. Sheidai, V. Mozaffarian, and Z. Noormohammadi

Tragopogon L. (Cichorioideae, Lactuceae, Scorzonerinae) is an Old World genus with 150 species. Pollen morphology has proved useful in the systematics of some genera and species of Asteraceae as well as in that of some of its genera and species. The pollen morphology of 24 taxa of the genus Tragopogon was investigated in detail by scanning electron microscopy (SEM). The pollen grain type ranged from suboblate, oblate-spheroidal to prolate-spheroidal in equatorial view and hexagonal, obtuse-hexagonal to hexagonal-angular in polar view. In this study separation of the species of the sections Majores, Profundisulcati, Sosnovsky, Chromopappus, Rubriflori according to Flora Iranica is presented from the other species of Tragopogon. T. jezdianus, T. porphyrocephalus, T. rezaiyensis are suggested to belong to Rubriflori section. The results indicate that the palynological characters of the genus Tragopogon are valuable for taxonomic applications and are useful for classification.

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The genus Geranium (Geraniaceae); with about 320 species throughout the temperate regions, is chemically characterised by the presence of tannins, flavonoids, anthocyanins and essential oils which interfere with the extraction of pure genomic DNA. It is necessary to optimise the extraction protocols to reduce the effects of the presence of these compounds to the lowest level.

The present study compares the plant genomic DNA extraction Kit (DNP™ Kit), CTAB DNA extraction method by Murray and Thompson and Sahu et al., from the extracting DNA point of view Geranium species. The results showed significant differences in DNA contents between the three methods. Quantity and quality of extracted genomic DNAs were compared by employing the spectrophotometer, Nano-Drop, agarose gel electrophoresis, and polymerase chain reaction (PCR) methods and molecular marker such as (ITS and trnL-F) and ISSR. The method of Sahu et al., provided the best results (200 ng/µL) in terms of quantity and quality of DNA, therefore, this method was taken and optimised for DNA extraction. Our results proposed that this method could be effective for plants with same polysaccharides, proteins and polyphenols components. The advantage of this method is that it omits the use of liquid nitrogen and toxic phenols which are expensive. The success of this method in obtaining high-quality genomic DNA has been demonstrated in the Geranium species group and the reliability of this method has been discussed.

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Persicaria with 70–100 species in the world is distributed in temperate regions of both hemispheres. It has 11 species in Iran growing in moist areas and margins of rivers. Through hybridisation, plasticity and existence of overlapping habitats, species identification shows difficulty. In this study, we aimed to investigate karyotype characters and morphological features, evaluate genetic variability within and among species studied and examine species relationship using ISSR data. Nine annual taxa of Persicaria were gathered from different localities in Iran and used for studies. Our studies showed that combination of karyological, morphological and molecular data can delimit species studied. Based on karyological results, three chromosome counts (P. hydropiper (2n = 2x = 20), P. maculosa (2n = 2x = 22), P. orientalis (2n = 2x = 22)) were the first reports for the Flora of Iran. Analyses of morphological characters showed diagnostic features among taxa. STRUCTURE and AMOVA analyses showed high intraspecific genetic diversity. Our results suggested that phenotypic plasticity and hybridisation may cause genetic diversity within Persicaria species.

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A cogent medicinal and aromatic plant, Ziziphora clinopodioides (Lamiaceae) is a perennial herb, its aerial parts is used as a wild vegetable or additive in foods to proffer pleasant aroma and flavour. There are many discussions about the intraspecific classification of this species and several subspecies have been introduced for it in different flora. These subspecies are morphologically very similar and identification of them is very difficult and in some cases, impossible. Therefore, in the present study, the pollen grains morphology of nine subspecies (32 specimens) of Z. clinopodioides were probed and documented in details utilising the scanning electron microscopy (SEM). In total, eleven pollen morphological characteristics were investigated and analysed by PAST software. The obtained results showed that pollen grains were hexacolpate and their equatorial views were sub-oblate to prolate. The exine ornamentation types of the pollen grains are bireticulate, microreticulate, reticulate and bireticulate-reticulate. The ANOVA test did not show significant difference for the studied quantitative traits. Although, the results of the multivariate analysis revealed a high diversity amongst the specimens even in the specimens of a single subspecies; it did not confirm the separation of subspecies in Z. clinopodioides.

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Species delimitation is essential since species is regarded as the basic unit of analysis in nearly all biological disciplines, such as ecology, biogeography, conservation biology, and macroevolution. The genus Geranium (Geraniaceae) comprises about 350 species distributed throughout most parts of the world. The subg. Robertium comprises 30 species which are arranged in 8 sections. This subgenus is represented in Iran by 10 species. These species are grouped into 5 sections. In spite vast distribution of many Geranium species that grow in Iran, there are not any available report on their genetic diversity, mode of divergence and patterns of dispersal. Therefore molecular (ISSR markers) and morphological studies of 147 accessions from 10 species of Geranium (subg. Robertium), that were collected from different habitats in Iran were performed. The aims of the present study are: 1) to find the diagnostic value of ISSR markers in delimitation of Geranium species, 2) to find the genetic structure of these taxa in Iran, and 3) to investigate the species inter-relationship. The present study revealed that combination of morphological and ISSR data can delimit the species. AMOVA and STRUCTURE analysis revealed that the species of subg. Robertium are genetically differentiated but have some degree of shared common alleles.

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Geranium subg. Robertium (Geraniaceae) comprises eight sections, of which sect. Batrachioidea contains four species centred in Eurasia, Mediterranean region and the Himalaya Mountains. Three species of Geranium pusillum, G. molle and G. pyrenaicum occur in Iran show some degree of morphological overlaps that make the species delimitation difficult. Moreover, hybrids are known to be formed between these species elsewhere. Till present time, there has been no detailed information available on molecular phylogeny and genetic structure of these species in the country. Therefore, the present study was conducted with the aim to investigate species delimitation by both morphological and molecular data and to reveal genetic diversity and population structure in these three Geranium species. For this study, 216 randomly collected plants from 30 geographical populations in three Geranium species were used. We encountered extensive within species genetic and morphological diversity. ISSR molecular markers could not delimit the studied species. STRUCTURE analysis revealed the occurrence gene flow between these species. The Mantel test showed no correlation between genetic distance and geographical distance of the populations studied. Statistical analysis showed no significant difference between ITS and rbcL sequences and phylogenetic tree was constructed based on combined data set which separated outgroups from the studied species. Genetic affinity of the studied species has been discussed.

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