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Genetic diversity, population structure and chromosome numbers in medicinal plant species Stellaria media L. VILL.

Shahram Mehria, Department of Agriculture, Pars Abad Moghan Branch, Islamic Azad University, Pars Abad Moghan, Iran.

Accepted: 2019-10-31 | Published Online: 2020-01-20 | DOI: 10.13128/caryologia-680


Stellaria media L. VILL., is known under the name of chickweed, it is an annual plant in the family Caryophyllaceae. Stellaria media is distributed in the all regions of Iran and has been introduced to many habitats of the world. S. pallida is very similar to S. media. This plant is considered to be as a herbal remedy and is used in folk medicine. Stellaria media is edible and nutritious. In the present study, we used morphological and ISSR data for this species. For this, 43 morphological characteristics, including 16 qualitative and 26 quantitative. AMOVA and Gst analyses showed that the populations of this species are genetically differentiated. Nm analysis revealed very low value of genetic diversity among the studied population and mantel test indicated isolation by distance occurred among them. The present study showed that the studied populations of S. media are differentiated in morphological characteristics and genetic content. In general, species relationships obtained from morphological and molecular data were largely congruent.

A new diploid cytotype of Agrimonia pilosa (Rosaceae)

Andrey Erst, Laboratory of Herbarium, National Research Tomsk State University, Tomsk, Russia.

Accepted: 2020-02-23 | Published Online: 2020-02-28 | DOI: 10.13128/caryologia-170


A new diploid cytotype of Agrimonia pilosa Ledebour (Rosaceae) collected in China has been revealed. Karyotype formula is 2n = 2x = 16 = 14m + 2sm. Previously, chromosome numbers in A. pilosa established by other researchers were 2n = 28; 56; 70 with basic chromosome number x = 7. All the other members of genus Agrimonia Linnaeus have the same basic chromosome number. In the meanwhile, some members of fam. Rosaceae have different basic chromosome numbers: x = 8 (e.g., in genera Amygdalus L., Aphanes L., Cerasus Mill., etc.), x = 9 (e.g., in genera Adenostoma Hook. & Arn., Chamaebatia Benth., etc.), x = 17 (e.g., in genera Amelanchier Medik., Chaenomeles Lindl., etc.). We suppose that the new basic chromosome number x = 8 was revealed in Agrimonia pilosa collected in China.

Clethodim induced pollen sterility and meiotic abnormalties in vegetable crop Pisum sativum L

Sazada Siddiqui, Department of Biology, College of Science, King Khalid University, Abha 61413, Saudi Arabia.

Accepted: 2020-02-23 | Published Online: 2020-02-28 | DOI: 10.13128/caryologia-269


Pesticides are highly noxious materials.Their poisonousness might not be unequivocally precise to the target entities but can unfavorably disturb various procedures in the non-target host plants. In the present study, the effect of application of clethodim on pollen sterility and meiotic anomalies of Pisum sativum L are studied. Pisum sativum L seeds are treated with different concentrations of clethodim varying from 0.01%, 0.02%, 0.03%, and 0.04% for exposure time of 1 hour and their effect on pollen sterility and chromosomal anomalies were investigated. The outcomes reveal that treatment of clethodim on Pisum sativum L seeds induces pollen sterility (PS) and chromosomal anomalies (CA) in a dose-dependent manner. Also in clethodim treated seeds, an elevation in the proportion of abnormal meiotic phases were observed which was time and concentration dependent. Secondary association (SeA), precociuos separation (PS), clumped nuclei (CNu) were reported in metaphase I & II, stickiness (Stc), bridges (Br) and laggards (Lg) in anaphase I & II. The results of the present study reveal that frequently used herbicide clethodim has a substantial cytotoxic effect on meiotic cells of Pisum sativum L.

Karyological studies in thirteen species of Zingiberacaeae from Tripura, North East India

Kishan Saha, Cytogenetics and Plant Biotechnology Laboratory, Department of Botany, Tripura University, Suryamaninagar-799022, Tripura, India.

Accepted: 2020-02-23 | Published Online: 2020-02-28 | DOI: 10.13128/caryologia-186


Tripura being a state in North East India belongs to Indo-Burma bio-diversity hotspot and is considered as a centre of origin of many species of Zingiberaceae.  Alpinia calcarata, Alpinia malaccensis, Alpinia nigra, Amomum aromaticum, Amomum koenigii, Amomum maximum, Curcuma amada, Curcuma caesia, Curcuma longa, Curcuma picta, Hedychium coccineum, Hedychium coronarium and Hedychium thyrsiforme are found in wild state in different geographical locations of Tripura. Their karyotypes were analyzed both at interspecific and intraspecific levels. The somatic chromosome number of Alpinia spp. and Amomum spp. was found to be 2n = 4X = 48. The Curcuma spp. represented by C. amada had 2n = 42 chromosomes and three other species viz., C. caesia, C. longa and C. picta had 2n = 63 chromosomes having X=21, indicating that polyploidy is a common feature in this genus. The somatic chromosome number of Hedychium spp. was found to be 2n = 34 chromosomes having a basic no. X = 17. Chromosomal data based clustering pattern and their sub-grouping at intra-specific level validates the taxonomic status of these species. Gower’s similarity matrix is an indicator of cryptic changes leading genus specific karyotype conservatism.

Phagocytic events, associated lipid peroxidation and peroxidase activity in hemocytes of silkworm, Bombyx mori induced by microsporidian infection.

Appukuttan Nair R. Pradeepa, Proteomics Division, Seribiotech Research Laboratory, CSB-Kodathi Campus, Carmelaram P.O., Bangalore-560035, Karnataka, INDIA.

Accepted: 2020-02-22 | Published Online: 2020-03-10 | DOI: 10.13128/caryologia-112


Microbial infections induced humoral and cell-mediated immune events in hemocytes. After infection by the microsporidian Nosema bombycis in the commercially important silkworm, Bombyx mori, hemocytes exhibited deformed nucleus and degranulation of structural granules by exocytosis. Granulocytes showed signs of phagocytosis included the formation of microvilli, pseudopodia, engulfment of spores, phagosome formation and membrane porosity. Association of membrane disintegration with infection-induced lipid peroxidation (LPO) was revealed by testing the level of malondialdehyde, a byproduct of LPO. LPO activity enhanced significantly (P<0.0002) throughout infection with peak activity in later stages of infection from day 11 accompanied by hemocyte plasma membrane disintegration. Partial increase in LPO activity coupled with increased peroxidase activity recorded in early and mid-stages of infection. In later stages, peroxidase activity decreased however PO increased accompanied by phagocytosis events. In hemocytes, phagocytic events are initiated by activation of genes encoding recognition proteins, aggregation factors, and immune-associated proteins. ß-GRP expression was down-regulated after the infection whereas CTL-11 enhanced expression on day 10. Humoral lectin enhanced expression on day 6 whereas apolipophorin showed 2.59 fold increase on day 10 after infection. Gene encoding cytoskeletal protein, ß- Actin showed stable enhanced expression throughout infection showing positive correlation (R2 = 0.65) with age after infection. Phagocytosis- associated gene Eater from Drosophila showed enhanced heterologous expression. Altogether phagocytic events induced by microsporidian infection are accompanied by increased LPO, decreased peroxidase activity and modulated gene activity in hemocytes of B. mori.

Chemical composition, antioxidant and cytogenotoxic effects of Ligularia sibirica (L.) Cass. roots and rhizomes extracts

Nicoleta Anca Şuţan, University of Piteşti, Faculty of Sciences, Physical Education and Informatics, Department of Natural Sciences, 1 Targu din Vale Str., 110040 Pitesti, Romania.

Accepted: 2020-02-22 | Published Online: 2020-03-11 | DOI: 10.13128/caryologia-116


Through time, in the traditional medicine Ligularia genus has been used as a remedy to cure several diseases and affections. The paper represents an essential step in offering more information about the antioxidant activity, chemical composition and cytogenetic activity of L. sibirica (L.) Cass. rhizomes and roots extracts. The antioxidant activity of the extracts has been achieved by analyzing the total phenolic content, total flavonoids, the organic chemical compound and trolox equivalent antioxidant capacity and their major polyphenolic constituents were quantified by liquid chromatography electrospray ionization-tandem mass spectrometry. The extracts were obtained by the Supercritical fluid extraction (SFE-CO2) technique, SFE-CO2 extraction with co-solvent and absolute ethanol extraction. The best results for the antioxidant activity have been fulfilled through the last two techniques. High-performance liquid chromatography (HPLC) has been applied in order to identify and quantify selected phenolic acids and flavonoids in the ethanolic extracts of L. sibirica (L.) Cass. The cytogenotoxic effects of the extracts completed the present study, with a furtherance of antiproliferative potential highlighted by the statmokinetic effect and an additional genoprotective effect.

Electrophoretic study of seed storage proteins in the genus Hypericum L. in North of Iran

Arman Mahmoudi Otaghvari, Department of Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Iran.

Accepted: 2020-02-23 | Published Online: 2020-03-11 | DOI: 10.13128/caryologia-122


In this research, we studied the electrophoretic of seed storage proteins in the genus Hypericum L. from Iran. The plant samples were collected from various phytogeographical regions of Iran to study the seed storage proteins. The study was performed to determine the boundary among different species of genus Hypericum using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). All samples belong to three species of H. perforatum, H. tetrapterum and H. androsaemum. A total of 22 protein bands were observed in the studied species. The results show that H. perforatum, H. tetrapterum are closely related based on seed storage proteins. A closely relationship and high protein similarity (J=0.66) were found between H. perforatum, H. tetrapterum. Electophoretic results compared with earlier molecular and morphological studies. The highest number of bands was observed in Kordkoy1 population (Pop12) and Gardane heyran population (Pop20) of H. perforatum and the lowest in Gorgan/Naharkhoran population (Pop 25) of H. androsaemum. Our results showed the species of Hypericum were placed intermixed. The aim of this study to delimit species in the genus Hypericum and used these seeds storage protein for the correct identification.

Melissa officinalis: A potent herb against EMS induced mutagenicity in mice

Hilal Ahmad Ganaie, Cytogenetics and Molecular Biology Research Laboratory, Centre of Research for Development (CORD), University of Kashmir, Srinagar-190 006, J & K, India.

Phytochemistry Research Laboratory, Centre of Research for Development (CORD), University of Kashmir, Srinagar-190 006, J & K, India.

Accepted: 2020-02-23 | Published Online: 2020-03-11 | DOI: 10.13128/caryologia-136


Melissa officinalis (L) is used traditionally for different medical purposes such as tonic, antispasmodic, carminative, diaphoretic, surgical dressing for wounds, sedative hypnotic, strengthening the memory and relief of stress induced headache. The methanolic extract of Melissa officinalis (Mo-ME) was investigated antimutagenic activity. The extraction was done by Soxhlet extraction method and the extract was evaluated for antimutagenic assay against EMS induced mice by micronucleus and chromosomal aberration assay. Briefly, mice were treated with methanolic extract of Melissa officinalis (Mo-ME) (100, 200 300 & 400 mg/kgbw) for 15 days. Without the doses of EMS, no and mutagenic effects were observed in blood and bone marrow samples of the mice. Micronucleus and chromosomal aberration test revealed the protective effects of Mo-ME when administered at high doses. The reduction profiles in the MN induction of methanolic extract of Melissa officinalis at the concentration (100, 200, 300 and 400 mg/kgbw) with EMS were estimated as 14.5%, 28.0%, 47.7%, and 81.5% respectively. The methanolic extract of Melissa officinalis exhibited no cytotoxic and mutagenic effects but only have antimutagenic effects, an effect that can be attributed to the presence of majority compounds, and the antimutagenic property of Mo-ME is an indication of its medicinal relevance.

Study regarding the cytotoxic potential of cadmium and zinc in meristematic tissues of basil (Ocimum basilicum L.)

Aurelian Rosculete, University of Craiova, Faculty of Agronomy, Craiova, Romania

Accepted: 2020-02-23 | Published Online: 2020-03-11 | DOI: 10.13128/caryologia-138


The cytogenetic study on the meristematic tissues of basil (Ocimum basilicum L.) aimed to evaluate some cytotoxic effects induced by two heavy metals (cadmium - Cd and zinc - Zn) applied in three different concentrations: 10, 50 and 100 ppm. Cytogenetic tests reveal a decrease of the mitotic index and the occurrence of various chromosomal aberrations following heavy metal treatments. The cell division was significantly affected, especially in the case of Cd treatment, which showed the highest degree of toxicity in all variants compared to the control variant. Instead, Zn has a lower degree of toxicity but only at concentrations of 50 ppm and 100 ppm. Types of chromosomal aberrations were relatively varied, being randomly distributed and concentration dependent, for both Cd and Zn. Were observed cells with large nucleus and disorganized-looking; interphases with pyknotic nucleus; cells with laggard chromosomes, pyknotic and sticky chromosomes, as well as cells with telophase bridge. The results reveal that Cd (at all tested concentrations) and Zn in concentrations higher than 10 ppm exhibit the significant cytotoxic potential to Ocimum basilicum L. as a result of the effects reported in cell divisions of the meristematic tissues. We can also appreciate that the Ocimum basilicum L. species could be used as a test plant to determine the degree of soil pollution with heavy metals.

Population genetic studies in wild olive (Olea cuspidata) by molecular barcodes and SRAP molecular markers

Alireza Iaranbakhsh, Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.

Accepted: 2020-02-23 | Published Online: 2020-03-11 | DOI: 10.13128/caryologia-147


Olive is an important horticultural plant having both cultivated and wild forms. The aim of the present study was investigating the genetic diversity of 13 wild olive trees belonging to four geographical populations in IRAN using SRAP neutral molecular markers as well as cp-DNA rpl intergenic sequences and ITS region. Genetic diversity parameters determined for 76 SRAP loci within the studied olive populations identified the most variable loci. Population differentiation parameters determined for SRAP loci, identified 13 SRAP loci with Gst value of 1, that means they differentiate the studied trees. PCoA analysis based on SRAP data separated olive trees from each other due to genetic difference. Distribution of the samples in PCoA plot indicated that the population 1 is more spread due to population genetic variability. However, the SRAP result reveals that these molecular markers can be used in population genetic investigations and germ plasm analysis. AMOVA showed significant genetic difference among the studied olive populations. Cp-DNA analysis produced 366 bp long sequences, out of which 224 sites were segregating among the studied plants. The mean nucleotide diversity was 0.32. TCS network based on cp-DNA separated most of the studied populations. Therefore, it seems that cp-DNA rpl sequences are a suitable barcode molecular marker for population genetic studies. Phylogenetic tree of ITS data could partially differentiate the wild olive population. In conclusion, a combined use of SRAPs and cp-DNA sequences are suggested for wild olive population genetic investigation.

In Vitro Polyploidy Induction in Persian Poppy (Papaver bracteatum Lindl.)

Saeed Tarkesh Esfahani, Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Tarbiat Modares University (TMU), P. O. Box. 14115-336, Tehran, Iran.

Accepted: 2020-02-23 | Published Online: 2020-03-11 | DOI: 10.13128/caryologia-169


Papaver bracteatum Lindl. grows as a wild perennial medicinal plant in Northern Iran and is known mainly for its high amounts of the pharmaceutically valuable alkaloid of thebaine. In vitro production of tetraploid P. bracteatum through colchicine treatment of imbibed seeds is reported. Resulted tetraploid and mixoploid plants were effectively identified by chromosome counting and flow cytometry technique. The chromosome number in diploid and successfully induced tetraploids were confirmed to be 2n=2x=14 and 2n=4x=28, where their calculated 2C DNA values were 6.15±0.03 and 11.95±0.07 pg, respectively. The highest induction efficiency was obtained by colchicine concentration of 0.05% and the treatment duration of 24 h. The effects of colchicine toxicity on plant survival and growth were proportional mainly to its concentration rather than the duration of exposure to colchicine. Tetraploid plants possessed significantly larger and less frequent leaf stomata as well as larger cell size. These attributes may serve as criteria for preliminary screening of P. bracteatum populations for the ploidy level.

Long-term Effect Different Concentrations of Zn (NO3)2 on the Development of Male and Female Gametophytes of Capsicum annuum L. var California Wonder

Sedigheh Arbabian, Department of Biology, Faculty of Sciences, Islamic Azad University, North-Tehran Branch, Tehran, Iran.

Accepted: 2020-02-23 | Published Online: 2020-03-11 | DOI: 10.13128/caryologia-174


Pepper is one of the most important crop plants. Recently, the global need for this plant has been widely increased due to its use in the food and pharmaceutical industry. we invested the effects of different concentrations of zinc on the development of male and female gametophytes of bell pepper (Capsicum annuum L. var California Wonder). The plants were cultivated with different concentrations of zinc nitrate (0 (control), 2.5, 5, 7.5, 10 and 15 mM) in a greenhouse under experimental conditions. Buds and Flowers are harvested at different stages of development (in 6 sizes) from May to Jun. They were fixed in FAA and maintained in 70% alcohol, then embedded in paraffin, sliced with a microtome and analyzed using a light microscope. Microscopic studies showed that the developmental process of ovule, gynoecium and pollen grain in bell pepper plants was taking to ordinary process in dicotyledonous plants. According to the results, increased zinc concentration resulted in a disorder in the reproductive phase, which caused the treatment 1 to enter the reproductive phase with a 4-week delay. In addition, the other of the treatments did not enter the reproductive phase and were wilt during the growth period. The developmental stages of gynoecium and anther in treatment 1 were similar to the control. However, a number of abnormalities and irregularities were observed including signs of nuclei disintegration, deformation of embryo sac, accumulation of dark materials and deformation of pollen grains.

A karyological study of some endemic Trigonella species (Fabaceae) in Iran

Majid Azizi,Department of Horticultural Sciences, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.

Accepted: 2020-02-23 | Published Online: 2020-03-11 | DOI: 10.13128/caryologia-184


Karyotypes of nine populations belonging to six endemic species of the genus Trigonella (Trifolieae/Fabaceae) studied in this survey. All studied species are perennial and recorded only from the Northeast of Iran. Excluding the karyotype of Trigonella subenervis, which was previously reported, all of the other species studied here (six species) for the first time. Our results present that all assessed genotypes are diploid with 2n=2x=16 and the chromosomal basis of x=8. In addition to the chromosome counts, length of long and short arms of the chromosome and their ratios analyzed and presented in this study.

Temporal Analysis of Al-Induced Programmed Cell Death in Barley (Hordeum vulgare L.) Roots

Filiz Vardar, Marmara University, Faculty of Arts and Sciences, Department of Biology, Göztepe, 34722, İstanbul, Turkey.

Accepted: 2019-11-02 | Published Online: 2020-03-11 | DOI: 10.13128/caryologia-185


Aluminum (Al) is the third most element found in the earth's crust and Al toxicity is one of the most dangerous toxicants in terms of plants. As soil acidity increases due to a number of environmental factors, Al becomes soluble and transforms into toxic forms. In the present study, barley (Hordeum vulgare L.) roots were exposed to 100 µM AlCl3 solution for short (1/2, 1, 2, 3, 4, 5, 6 and 7 h) and long (24, 48, 72 and 96 h) term to reveal time-dependent programmed cell death evidence. At the end of time periods, Al+3 accumulations, loss of plasma membrane integrity and lipid peroxidation increased time-dependently. On the other hand, the increase in caspase-1 like enzyme activities was observed in Al toxicity beginning from ½ h. Similar to apoptosis seen in animals, cytochrome c release from mitochondria to the cytoplasm was also determined quantitatively. As a result of our research, the increase of cytochrome c release from mitochondria to the cytoplasm was time-dependent which is one of the indicators of programmed cell death. Finally, under Al stress, genomic DNA fragmentation was measured by Flow Cytometry, and it was determined that DNA fragmentation was visible at first hours, but it was more significant after long term application in barley roots. In conclusion; the presented study highlights the adverse effects of Al on barley roots and the importance of clarifying the relationship between Al toxicity and time-dependent programmed cell death mechanism.

Cytogenetic effects of C6H4 (CH3)2 (xylene) on meristematic cells of root tips of Vicia faba L. and mathematical analysis

Canan Özdemir, Celal Bayar University, Science and Art Faculty, Biology Department, Manisa, Turkey.

Accepted: 2020-02-26 | Published Online: 2020-03-11 | DOI: 10.13128/caryologia-279


Xylene is a readily flammable and poisonous liquid with a chemical formula of C6H4 (CH3)2. It is used as raw material or auxiliary raw material in many industrial products such as dye, pencil, agricultural chemicals, rubber, fiber, glue, and diaper. In this study, cytogenetic effects of xylene, on the meristematic cells of root tips of V. faba L. used as food have been investigated. For this purpose, the seeds of the plant have been treated with xylene solutions prepared in different concentrations for different time periods. Chromosomes at the root tips have been looked and the effect of xylene has been determined. The abnormalities as chromosome breaking, chromosome dispersion, bridge chromosome, chromosome adherence, ring chromosome have been observed. Abnormalities have been seen at each treatment depended on the time periods. In addition to these visible damages of xylene in the study, possible damages on chromosomes carrying genetic codes of living beings to future generations have been investigated and mathematical analyzes have been made. The results obtained have been evaluated statistically.

High genetic diversity and presence of genetic structure characterise the Corso-Sardinian endemics Ruta corsica and Ruta lamarmorae (Rutaceae)

Gianluigi Bacchetta, Centro Conservazione Biodiversità (CCB), Dipartimento di Scienze della Vita e dell’Ambiente - Università degli Studi di Cagliari. Viale S. Ignazio da Laconi, 13 - IT-09123 Cagliari, Italy.

Banca del Germoplasma della Sardegna (BG-SAR), Hortus Botanicus Karalitanus (HBK), Università degli Studi di Cagliari. Viale Sant’Ignazio da Laconi, 9-11, IT-09123, Cagliari, Italy.

Accepted: 2020-02-25 | Published Online: 2020-03-12 | DOI: 10.13128/caryologia-510


Corsica and Sardinia form one of the ten areas with highest biodiversity in the Mediterranean and are considered one of the priority regions for conservation in Europe. In order to preserve the high levels of endemism and biological diversity at different hierarchical levels, knowledge of the evolutionary history and current genetic structure of Corso-Sardinian endemics is instrumental. Microsatellite markers were newly developed and used to study the genetic structure and taxonomic status of Ruta corsica and Ruta lamarmorae, rare endemics of Corsica and Sardinia, respectively, and previously considered a single species. Our analyses identified high levels of genetic variation within each species (P=0.883, He=0.543 for R. corsica; P=0.972, He=0.627 for R. lamarmorae). Intrinsic traits of the species (hermaphroditism, proterandry and polyploidy) and island-dependent factors (i.e. age, origin and history of the islands) might explain the detected high levels of genetic variation. We discovered differentiation between R. corsica and R. lamarmorae, and genetic structure within each species, which are consistent with the observation of low dispersal ability for both species. Our genetic results support the recent taxonomic classification of R. corsica and R. lamarmorae as separate species and suggest that they diverge at only few loci. One R. corsica population (SA) strongly differed from all other studied populations and appeared to be the product of hybridization between the two species in STRUCTURE analyses. Our results provide important insights for the conservation of the two rare endemics. Further genetic analyses are recommended for R. lamarmorae and for population SA (R. corsica).

Karyotypic investigation concerning five Bromus Species from several populations in Iran

Sara Sadeghian, B.Sc. of Research Division of Natural Resources Department, Fars Agricultural and Natural Resources Research and Education Center, AREEO, Shiraz, Iran.

Accepted: 2020-02-29 | Published Online: 2020-03-12 | DOI: 10.13128/caryologia-863


Karyotypes of five taxa (fourteen populations) of the genus Bromus from different geographic origins is presented: B. scoparius, B. japonicus, B. madritensis, B. rubens and B. tomentellus. The ploidy levels were different. B. scoparius and B. japonicus were found 2n=2x=14, B. madritensis and B. rubens were found 2n=4x=28 and B. tomentellus were found 2n=6x=42. Detailed karyotype analysis allows us to group the different populations and to postulate relationships among them.