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Insights on cytogenetic of the only strict African representative of genus Prunus (P.
africana): first genome size assessment, heterochromatin and rDNA chromosome pattern

Sonja Siljak-YakovlevEcologie, Systématique et Evolution, Univ. Paris-Sud, CNRS, AgroParisTech, Université
Paris-Saclay, Bât 360, 91405 Orsay, France

Accepted: 2019-10-24 | Published Online: 2019-10-25 | DOI: 10.13128/cayologia-253


Prunus africana is a multipurpose evergreen species endemic to Africa and an endangered species because of overexploitation. The great importance of this species resides particularly in the use of its bark against benign prostatic hyperplasia. As for most tropical trees and generally woody species, cytogenetic studies are scarce. Standard and molecular cytogenetic approaches have been implemented for the first time to study P. africana from Cameroon. This is the tetraploid species with a chromosome number of 2n=4x=32. Genome size estimated by flow cytometry was 2C=1.44 pg. Five loci (ten signals) of 35S rRNA genes were observed after fluorescence in situ hybridization. Ten G-C rich DNA regions were detected by chromomycin A3 fluorochrome banding. All chromomycin positive bands were co-localizedwith 35 S rDNA signals. Prunus africana, the only strict African representative of genus Prunus, is in need of the conservation strategy and in situ management that we are also
discussing in this work.

Cytogenetic effects of Fulvic acid on Allium cepa L. root tip meristem cells

Özlem Sultan ASLANTÜRKAdnan Menderes University, Faculty of Art and Science, Department of Biology
09010 Aydın – TURKEY

Accepted: 2019-10-24 | Published Online: 2019-10-25 | DOI: 10.13128/cayologia-257


Fulvic acid is a class of compounds of humic substances and is found in a significant proportion of the substances in the environment. It has been used for many years in industry, agriculture, and complementary medicine.

In this study, cytogenetic effects of fulvic acid purified from Muğla Milas Hüsamlar leonardite (TURKEY) on Allium cepa root tip meristem cells were investigated using the Allium test. For this purpose, 10 mg/ml stock solution of fulvic acid was prepared by dissolving in citric acid and it was diluted with distilled water to 10, 20, 40, 80 and 100 µg/mL concentrations.

Onion bulbs were exposed to these concentrations of the fulvic acid for macroscopic and microscopic analysis. Tap water was used as a negative control, 40 µg/mL citric acid was used as solvent control (fulvic acid solvent), and 0.02M Ethyl methane sulfonate (EMS) (a mutagenic, teratogenic, and possibly carcinogenic organic compound) was used as a positive control. There has been statistically significant stimulation of root growth depending on fulvic acid concentration in comparison with the control groups (p<0.05). Furthermore, in fulvic acid treatment groups, breaks, stickiness and polar deviations appeared at very low rates, and total chromosome aberration ratios were insignificant compared to the control groups. These results suggest that fulvic acid does not have cytotoxic and genotoxic effects on A. cepa.

Evaluation of the cytotoxic and genotoxic potential of some heavy metals by use of Allium test

Elena Bonciu, Department of Agricultural and Forestry Technology, University of Craiova, Craiova,

Accepted: 2019-10-24 | Published Online: 2019-10-25 | DOI: 10.13128/cayologia-256


The present study aimed to evaluate the cytogenetic effects induced by heavy metals nickel (Ni) and lead (Pb) to crop plants, using the Allium sativum (garlic) as a test plant. For this purpose, were used solutions of nickel nitrate - Ni(NO3)2 - and lead nitrate -Pb(NO3)2 - at concentrations of 50, 150 and 450 ppm for 72 hours, along with an untreated control variant immersed in plain water. The biological material was immersed
from the beginning in the tested solutions. The results obtained showed a strong inhibitory effect of these heavy metals on the process of rhizogenesis, as well as a significant mitodepresive effect in the meristematic
cells, both phenomena being correlated with increasing concentration of the tested solutions. At the same time, several types of chromosomal aberrations (c-mitosis, vagrants, star-anaphase, star-telophase, fragments, clumping, stickiness, bridges) have been recorded in all treatment variants. The presence of these chromosomal aberrations in all treatment variants indicates the aneugenic effects of nickel nitrate and lead nitrate in the meristematic cells of A. sativum. The results suggest the ecotoxicity potential of nickel and lead on plants even at low concentrations and confirm the suitability of A. sativum as a test plant for assessing the
cytotoxicity and genotoxicity of heavy metals to plants.

Fluorescence In Situ Hybridisation Study of Micronuclei in C3A Cells Following Exposure to ELF-Magnetic Fields

Luc VerschaeveSciensano, Risk and Health Impact Assessment Service, Brussels, Belgium. Department of Biomedical Sciences, University of Antwerp, Antwerp, Belgium.

Accepted: 2019-10-24 | Published Online: 2019-10-25 | DOI: 10.13128/cayologia-254


Human C3A cells were exposed to extremely low frequency (50 Hz) magnetic fields (ELF-MF’s) up to 500 μT. They were subjected to the micronucleus assay using a Fluorescence In Situ Hybridization (FISH) technique with an in-house pan-centromere probe. We found no increased frequency in micronucleated cells and no change in the proportion of centromere positive over centromere negative micronuclei compared to the unexposed control cells. These results are in accordance with some, but in contradiction with other previously published
investigations underlining that effects of environmental ELF-EMF’s on cellular DNA may be very subtle and that small changes or environmental influences may determine the outcome of a (geno)toxicity study. Interestingly, a low-level (5μT) exposure resulted in less than the background micronucleus frequency.

Phytochemical analysis and in vitro assessment of Polystichum setiferum extracts for their cytotoxic and antimicrobial activities

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: 2019-10-24 | Published Online: 2019-10-25 | DOI: 10.13128/cayologia-255


Ferns are traditionally used by some nations to treat rheumatism, lungs, gynecology, blood and digestion dysfunctions, and several others illnesses. The present study evaluates the bioactivity of methanol and ethanol extracts from Polystichum setiferum (Forssk.) Moore ex Woyn. in an Allium cepa test and disk diffusion test. In the Allium cepa test the methanol and ethanol extracts induced a significant time-related increase in the mitotic index. The tested extracts were non-mutagenic by used assay, with no occurrence either the structural or numerical aberrations detected. The extracts were also evaluated in terms of trace elements (by EDXRF) and qualitative composition (by UV-VIS, FTIR and total phenolic content). In the disk diffusion test, methanol extracts from leaves determined a small inhibition of bacterial growth for Enterobacter cloacae and Citrobacter freundii strains relatively to control sample (methanol). The ethanol extracts were more efficient, the diameter of inhibition growth zones measured from 7 to 10 mm, the most affected strain was Chryseobacterium meningosepticum.

Telomeric heterochromatin and meiotic recombination in three species of Coleoptera (Dorcadion olympicum Ganglebauer, Stephanorrhina princeps Oberthür and Macraspis tristis Laporte)

Bernard DutrillauxSystématique, Évolution, Biodiversité, ISYEB - UMR 7205 – CNRS MNHN UPMC EPHE, Muséum National d’Histoire Naturelle, Sorbonne Universités, 57 rue Cuvier CP50 F-75005, Paris, France.

Accepted: 2019-10-24 | Published Online: 2019-10-28 | DOI: 10.13128/cayologia-194


Centromeres are generally embedded in heterochromatin, which is assumed to have a negative impact on meiotic recombination in adjacent regions, a condition required for the correct segregation of chromosomes at anaphase I. At difference, telomeric and interstitial regions rarely harbour large heterochromatic fragments. We observed the presence at the heterozygote status of heterochromatin in telomere region of some chromosomes in 3 species of Coleoptera: Dorcadion olympicum; Stephanorrhina princeps and Macraspis tristis.
This provided us with the opportunity to study the relationship between heterochromatin, chiasma location and meiotic recombination independently from the proximity of centromeres in this order of insects. In acrocentric chromosomes, the presence of heterochromatin in telomere region of the long arm displaces recombination near the centromere. In sub-metacentrics, recombination is almost always restricted to the other arm. This at distance effect of heterochromatin may deeply influence genetic drift.

Differences in C-band patterns between the Japanese house mice (Mus musculus) in Hokkaido and eastern Honshu

Masahiro A. IwasaCourse in Natural Environment Studies, Graduate School of Bioresource Sciences,
Nihon University, Kameino 1866, Fujisawa, Kanagawa 252-0880, Japan.

Accepted: 2019-10-24 | Published Online: 2019-10-28 | DOI: 10.13128/cayologia-237


We characterized and categorized the C-band patterns of the house mouse Mus musculus from four areas in Hokkaido and Honshu of the Japanese Islands as a biparental marker. The C-band patterns are categorized as
polymorphic, monomorphic, or intermediate, corresponding to those of the Korean mice, C57BL/6, and F1 hybrid mice bred from the Japanese mice and a laboratory mouse, respectively. The C-band patterns mainly differ between mice from Hokkaido and Honshu. The polymorphic patterns are shown in mice in Honshu, while the intermediate patterns are shown in mice in Hokkaido, with an exceptional case of a monomorphic pattern found in one locality. In the other localities of Hokkaido and northeastern Honshu, the C-band patterns are not
congruent with an estimation by maternal element in our previous study, whereas the congruence is observed in other localities. It is suggested that the characteristics of the Japanese house mice have been formed through complicated processes by different expansions between biparental and maternal elements.

A whole genome analysis of long-terminal-repeat retrotransposon transcription in leaves of Populus trichocarpa L. subjected to different stresses

Tommaso GiordaniDepartment of Agriculture, Food and Environment, University of Pisa, Via del Borghetto 80, I-56124 Pisa, Italy

Accepted: 2019-10-24 | Published Online: 2019-10-28 | DOI: 10.13128/cayologia-232


Long terminal repeat retrotransposons have a main role in shaping the structure of plant genomes. We used available genomic resources to study as several factors affect the expression of long terminal repeat retrotransposons in Populus trichocarpa. Such factors included redundancy of a retrotransposon in the genome, chromosomal localization, "genotype" of the retrotransposon, and changes in the environment. Overall, we identified and annotated 828 full-length retrotransposons, and analyzed their abundance in the genome. Then, we measured their expression in leaves of plants subjected to several stresses (drought, cold, heat, and salt) as well as in control plants. Our analyses showed that the expression of retrotransposons was generally low, especially that of abundant elements. The transcription of an element was found to be only slightly dependent on its chromosomal localization, rather it depended on the superfamily and the lineage to which the retrotransposon belonged. Finally, some retrotransposons were specifically activated by different environmental stresses.

Structure and development of male gametophyte in Carya illinoensis (Wangenh.) K. Koch

Fangren PengCo-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry, University, Nanjing 210037, P. R. China

Accepted: 2019-10-24 | Published Online: 2019-10-31 | DOI: 10.13128/cayologia-152


In order to understand the differentiation of staminate flowers of pecans (Carya illinoensis (Wangenh.) K. Koch), we carried out an integrated study of staminate flower development in a protogynous cultivar, Mahan, by assessing changes in external morphology and microstructure at multiple levels. Results showed that the staminate inflorescence differentiation cycle for pecans was 1 year. Staminate inflorescence development was acropetal. When inflorescences developed to 5–8 cm, the microspore mother cells in the base florets of the inflorescences entered into meiosis prophase and the middle layer started to degrade. When inflorescences grew to 8–10 cm, the microspore mother cells in the based florets of the inflorescences were at the peak of meiosis and cytokinesis was synchronous. When bracts have opened to 15°, the microspore mother cells of the basal florets had undergone two divisions to form tetrads. When bracts have opened to 45°, the basal florets entered the mid-late uninucleate stage and the tapetum underwent degradation and autolysis. When bracts opened to >90°, mature pollen grains were 2-celled, with three germ pores and the middle layer tapetum completely degraded. Anther wall development followed the basic type, which was composed of an epidermal layer, an endothecial layer, middle layer (1–3 layers, fibrous thickening absent) and the tapetal layer (cell division was from uninucleate to an octonucleate cell). In summary, external morphology and gametophyte development in pecan staminate flowers were consistent to related; thus, the internal gamete development status can be determined from external morphological characteristics of the flower. This provided a sampling basis and theoretical foundation for in vitro culture of pollen grains and elucidation of flowering mechanisms.

Karyotypic description and repetitive DNA chromosome mapping of Melipona interrupta Latreille, 1811 (Hymenoptera: Meliponini)

Denilce Meneses Lopes, Laboratório de Citogenética de Insetos, Departamento de Biologia Geral, Universidade Federal de Viçosa, CEP 36570-900, Viçosa, Minas Gerais, Brazil.

Accepted: 2019-10-24 | Published Online: 2019-10-31 | DOI:10.13128/cayologia-239


Heterochromatic patterns in the genus Melipona vary among subgenera species. Melikerria is the only subgenus that presents species with different content of heterochromatin. Thus, the cytogenetic knowledge of other species of this subgenus is important for the understanding of karyotype evolution in Melipona. Here, we describe the karyotype of Melipona (Melikerria) interrupta based on the chromosomal heterochromatic patterns, AT and GC richness, mapping sequences of rDNA, microsatellites and telomeric regions. We observed 2n=18 chromosomes, with a high heterochromatin content rich in AT and euchromatic regions rich in GC base pairs. The high GC content was observed at interstitial region near the junction of the stained euchromatin and heterochromatin of the first chromosomal pair, the same region marked for the rDNA 18S locus. Microsatellites hybridized only on euchromatin regions and the telomeric probe on terminal regions of all chromosomes. Melipona (Melikerria) interrupta belongs to previous described heterochromatic 48 Group II, suggesting there has been an increase in heterochromatin content in Melikerria. The M. quinquefasciata, belonging to the same subgenus as Melipona (Melikerria) interrupta, has low content of heterochromatin and appears to be evolving independently. So, the differences in the content heterochromatin, in the marker regions of CMA3 and the rDNA 18S locus in species of Melikerria is an important feature to be investigated further.

Genome size in ants: retrospect and prospect

Danon Clemes Cardoso, Maykon Passos Cristiano,Departamento de Biodiversidade, Evolução e Meio Ambiente/ICEB, Universidade Federal de Ouro Preto, Campus Morro do Cruzeiro, Ouro Preto, Minas Gerais
35400-000, Brazil

Accepted: 2019-10-24 | Published Online: 2019-10-31 | DOI:10.13128/cayologia-172


Genome size is very useful in studies regarding taxonomy, evolution, and reproductive biology in many animal groups, including insects. Herein, we assembled the information about genome size in ants, compiling the DNA content estimated so far, in order to evaluate the methods, the tissues and the internal standard applied to estimate the genomes size. All values were placed in a phylogenetic tree to put it in an evolutionary context and the means of the subfamilies were further compared statistically to investigate changes and trends in the variation across taxa. The compiled data resulted in 86 specimens of ants, comprising 69 different species. This
number represents 0.52% of the total number of 13,369 ant species described, covering only 40 from 333 valid extant genera. The average Formicidae genome size was 0.36 pg (± 0.13). Most of the estimates were obtained through flow cytometry (83.5%), commonly using brain tissues, with Drosophila melanogaster as internal standard (76%). Differences in DNA content of ant species may be related to differences in the amount of heterochromatin and is not related with chromosome number. The evaluation of the genome size estimations currently available for ants has highlighted their scarcity. Such information would be valuable as independent data for the study of ant diversity and evolutionary biology. Further, we conclude that the standardization of the techniques used and a large–scale study on ant genome size are urgently required, given the importance of this insect group and the needs for the improvement in our knowledge on ant genome.

Role of humic acid against salt-induced cytotoxicity in Hordeum volgare L.

Selma Tabur,Süleyman Demirel University, Arts and Sciences Faculty, Department of Biology, 32260 Isparta, Turkey.

Accepted: 2019-10-30 | Published Online: 2019-11-06 | DOI:10.13128/cayologia-317


The effect of humic acid, which is an replace by a biostimulant on mitotic activity and chromosome behaviors in meristem cells of Hordeum vulgare L. germinated under different salt concentrations were investigated. In the parallel to increasing salt concentrations, mitotic index partly decreased and observed the higher number of chromosomal abnormalities as compared to control. Also, it was determined that the mitotic index of seeds pretreated with only humic acid increased by 30% according to control and by 42% of mitotic aberrations. Whereas, humic acid along with salt significantly inhibited to mitotic index with parallel to increasing salt concentrations. Moreover, the frequency of chromosomal aberrations in seeds germinated in humic acid and salty medium significantly decreased according to its own control. Humic acid revealed to a successful performance in ameliorating of the detrimental effect of salinity in the all concentrations studied. Humic acid application at 0.35 M salinity displayed perfectly successful by reaching to the same abnormality percentage of control.

Assessment of cytotoxicity and mutagenicity of insecticide Demond EC25 in Allium cepa and Ames Test

Arzu Öskara, Department of Molecular Biology and Genetic, Faculty of Arts and Sciences, Afyon Kocatepe University, Afyonkarahisar/TURKEY

Accepted: 2019-10-30 | Published Online: 2019-11-14 | DOI:10.13128/cayologia-698


The mutagenicity and cytotoxicity of Demond EC25, a synthetic pyrethroid insecticide, was assessed using two standard genotoxicity assays of the Salmonella typhimurium mutagenicity assay (Ames test) and Allium cepa test. Cytogenetic effects of Demond EC25 were evaluated in the root meristem cells of Allium cepa. The test concentrations of compounds were selected by determining EC50 of the Allium root growth and onion seeds were exposed to Demond EC25 (50, 100, and 200 ppm) for 24, 48, and 72 hours. The concentrations Demond EC25 was compared with the value for the negative control using Dunnet-t test, 2 sided. The results indicated that mitotic index was clearly decreased with increasing the concentration of Demond EC25 in each treatment group as compared to the controls. Demond EC25 was tested for mutagenicity in bacterial reversion assay systems with two strains (TA98 and TA100) of Salmonella typhimurium absence and presence of S9 fraction. The doses of Demond EC25 were 50, 100, 200, 400, 800 µg/plate and test materials were dissolved in DMSO. Our results show that Demond EC25 was found to be mutagenic in 800 and 400 μg/plate doses of TA98 in the without S9 mix and 800 μg/plate in the with S9 mix. In TA100, Demond EC25 was found to be mutagenic only 800 μg/plate doses without S9 mix. The other doses of this insecticide was not found to be mutagenic in both test strains.

Karyotype analysis of a natural Lycoris doubleflowered hybrid

Kun LiuAnhui Provincial Key Laboratory of the Conservation and Exploitation of Biological
Resources, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, China

Accepted: 2019-11-18 | Published Online: 2019-11-19 | DOI:10.13128/cayologia-720


A putative natural double-flowered hybrid in Lycoris Herb. was found on Mt. Zhangjiajie in Hunan, China. The putative natural hybrid had a chromosome number of 2n = 18 and was karyotypically formulated as 2n = 4m + 6st + 5t + 3T. The karyotype of the putative natural hybrid was classified as 2B type according to the degree of
asymmetry and Stebbins’ criteria. According to the gross morphology, phenology and karyotype of the putative natural hybrid, it was suggested that this taxon was probably from the natural hybridization between L. aurea and L. radiata.

Meiotic irregularities associated to cytomixis in Buddleja iresinoides (Griseb.) Hosseus. (Buddlejaceae) and Castilleja arvensis Schltdl. & Cham. (Orobanchaceae)

Aldo Ruben AndradaFundación Miguel Lillo. Miguel Lillo 251, San Miguel de Tucumán, Tucumán, Argentina.

Accepted: 2019-11-18 | Published Online: 2019-11-19 | DOI: 10.13128/cayologia-298


The current paper analyzes the male meiotic behavior in wild populations of Buddleja iresinoides and Castilleja arvensis from Piedmont areas in Northwest Region, Argentina. C. arvensis showed tetraploid number of chromosome of 2n = 24. Interestingly, on the other hand our results are not in agreement with the basic number reported for Buddleja and the chromosome number n = 28 found for B. iresinoides is atypical in the genus. Around 7 % pollen mother cells were aneuploid as they showed meiotic chromosome count of n = 20-21 bivalents. Possible origin for such atypical chromosome number has been discussed in this paper. During the cytological studies we also came across pollen mother cells showing meiotic abnormalities such as cytomixis, chromatin stickiness and chromatin bridges. Consequently microsporogenesis was also irregular showing dyads and triads. However, the percentage of these irregularities during meiosis and microsporogenesis was not higher, and pollen fertility was not affected to a great extent. Cytomixis and other meiotic abnormalities have been reported here first time in these species.

Population Genetic Studies in Ziziphus Jujuba Mill.: Multiple Molecular Markers (ISSR, SRAP, ITS, Cp-DNA)

Farah Farahani, Department of Microbiology, Qom Branch, Islamic Azad University, Qom, Iran.

Accepted: 2019-11-18 | Published Online: 2019-11-19 | DOI:13128/cayologia-390


Ziziphus jujuba (jujube) is an important horticultural crop with medicinal value. It is under cultivation in many areas of Iran and also grows as wild in several geographical populations throughout the country. We have no information on genetic variability and population structure of this important plant species in our country. Therefore, the aim of the present study was to perform genetic fingerprinting of 13 geographical populations of jujuba for the first time and provide data on population genetic structure, admixture versus genetic fragmentation of this important crop. We used multilocus molecular markers (ISSRs and SRAPs) for genetic fingerprinting and also compared the results with bioinformatics investigation results we did on jujuba cultivars by using nuclear r-DNA and chloroplast inter-genetic cp-DNA sequences. Genetic diversity parameters and AMOVA test as well as Ivanno test support some kind of genetic distinctness of the jujuba populations studied. We found that cp-DNA inter-genic sequences can also discriminate jujuba cultivars as efficient as multilocus molecular markers and therefore, a multiple molecular approaches may be used for genetic fingerprinting of
jujuba. The present study revealed good level of genetic diversity among wild/ uncultivated populations of jujuba which can be used in conservation and breeding of this important horticultural crop plant within the country. As this crop has several wild geographical populations throughout the country, we plan to continue our quest to investigate many more populations in nearby future and try to utilize cp-DNA inter-genic sequences along with multilocus molecular markers for genetic discrimination of wild populations.

Karyotypes of Danubian lineage brown trout and their hybrids

Ilhan AltinokDepartment of Fisheries Technology Engineering, Surmene Faculty of Marine Sciences, Karadeniz Technical University, 61530 Surmene, Trabzon, Turkey.

Accepted: 2019-11-18 | Published Online: 2019-11-19 | DOI: 10.13128/cayologia-160


Cytogenetic analysis of brown trout, Salmo trutta, have been described for different populations and morphs; however, cytogenetic analysis of interspecific brown trout hybrids is unknown. Cultured kidney cells from four brown trout subspecies (Salmo trutta abanticus, S.t. caspius, S.t. fario and S.t. labrax) and their reciprocal hybrids were karyotyped using conventional staining, C-banding and Ag-NOR staining techniques. Chromosome number (2N) and chromosome arm number (NF) ranged from76 to 80 and 98 to 102, respectively. Silver staining revealed the presence of NOR sites on the short arm of the submetacentric chromosome. The size and number of NOR sites showed uniformity. The presence of heterochromatin on different chromosome arms was confirmed by C-banding. The presence and position of constitutive heterochromatin showed variability among individuals. Chromosome structures of purebred brown trout subspecies belonging to the Danubian linage and their hybrids were similar, and no distinctive characteristics were observed in any of the species. The results of this study are applicable to the development of improved conservation and management strategies for brown trout.

Chromosome counts and karyotype analysis of species of family Apocynaceae from Egypt

Samia HENEIDAKDepartment of Botany, Faculty of Science, Suez University, Suez, Egypt.

Accepted: 2019-11-18 | Published Online: 2019-11-20 | DOI: 10.13128/cayologia-192


The chromosome counts of 13 species of family Apocynaceae in the flora of Egypt have been reported; one species from subfamily Periplocoideae and the other 12 species from subfamily Asclepiadoideae. The chromosome numbers are 2n = 22 for Periploca angustifolia, Glossonema boveanum, Pentatropis nivalis, Cynanchum acutum, Calotropis procera, Gomphocarpus sinaicus, Pergularia daemia and Pergularia tomentosa; 2n = 24 for Leptadenia arborea and Solenostemma arghel; 2n = 22, 44 for Caudanthera edulis, Caudanthera sinaica and Desmidorchis acutangulus. The chromosome numbers and karyotype analyses were firstly reported in Leptadenia arborea (2n = 24). The polyploid nature was demonstrated by the prevalence of cells with 2n = 4x = 44 chromosomes in Caudanthera edulis, Caudanthera sinaica and Desmidorchis acutangulus. The chromosomes are median and submedian as most species in the Apocynaceae. The intrachromosomal asymmetry and interchromosomal asymmetry were estimated with MCA and CVCL values. In intrachromosomal asymmetry, Desmidorchis acutangulus is the most symmetrical karyotype, while Pergularia tomentosa is the most asymmetrical karyotype. In interchromosomal asymmetry, Glossonema boveanum is the most symmetrical karyotype, while Cynanchum acutum is the most asymmetrical karyotype.

Insights on cytogenetic of the only strict African representative of genus Prunus (P. africana): first genome size assessment, heterochromatin and rDNA chromosome pattern

Sonja Siljak-YakovlevEcologie, Systématique et Evolution, Univ. Paris-Sud, CNRS, AgroParisTech, Université
Paris-Saclay, Bât 360, 91405 Orsay, France.

Accepted: 2019-11-18 | Published Online: 2019-11-20 | DOI: 10.13128/cayologia-253


Prunus africana is a multipurpose evergreen species endemic to Africa and an endangered species because of overexploitation. The great importance of this species resides particularly in the use of its bark against benign prostatic hyperplasia. As for most tropical trees and generally woody species, cytogenetic studies are scarce. Standard and molecular cytogenetic approaches have been implemented for the first time to study P. africana from Cameroon. This is the tetraploid species with a chromosome number of 2n=4x=32. Genome size estimated by flow cytometry was 2C=1.44 pg. Five loci (ten signals) of 35S rRNA genes were observed after fluorescence in situ hybridization. Ten G-C rich DNA regions were detected by chromomycin A3 fluorochrome banding. All chromomycin positive bands were co-localized with 35 S rDNA signals. Prunus africana, the only strict African representative of genus Prunus, is in need of the conservation strategy and in situ management that we are also discussing in this work.

Analysis of the chromosome variation within some natural populations of subterranean clover (Trifolium subterraneum L., Fabaceae) in Algeria

Rachida IssolahINRAA, CRP Mehdi Boualem, Division de Recherche sur les ressources phytogénétiques, BP 37. Baraki. Alger. Algérie.

Accepted: 2019-11-18 | Published Online: 2019-11-20 | DOI: 10.13128/cayologia-164


Nine natural populations of subterranean clover (Trifolium subterraneum L.) coming from different eco-geographical sites of the North-East Algeria, have been studied for their chromosome number and karyotype features. The study is part of the evaluation and valorization of plant genetic resources of fodder and pastoral interest in Algeria. The results of mitosis detect two groups of populations, and reveal diversity in the number among and within populations. The Algerian populations of T. subterraneum are characterized by two chromosomic formulas. The first formula (2n=2x=16m) (median), more common in most of the studied populations, is in conformity with previous reports in this species. The karyotype of these populations is symmetrical for size and form. The second (2n=2x=18m), is detected for the first time and described as a new chromosomal formula in T. subterraneum. The latter is relatively more frequent than the first one and characterizes the populations coming from high altitude areas. The karyotype (2n=2x=18m) is relatively symmetrical. At the level of the two established Karyotypes, satellites are highlighted at the first pair. A variation in the size and frequency of these satellites is observed. The species exhibits regular meiotic behaviour, confirming the presence of two basic chromosome numbers (x=8 and 9). The study also highlights the role of ecological factors (Altitude and Rainfall) of the originating environment of Algerian populations in the variation and evolution of chromosome numbers in T. subterraneum. The new cytogenetic data can be exploited in the taxonomy of the species in Algeria in order to select and develop this plant genetic resource in the agricultural field.

Geographical distribution and karyotype of Nannospalax ehrenbergi (NEHRING 1898) (Rodentia, Spalacidae) in Iraq

Yüksel CoșkunDicle University Science Faculty, Department of Biology, Diyarbakır/Turkey

Accepted: 2019-11-18 | Published Online: 2019-12-03 | DOI: 10.13128/caryologia-318


This paper concerns the karyological analysis of fourteen mole rats collected in four different localities of North-Iraq (Kurdistan Region). The result showed that they belong to the following cytotypes of Nannospalax ehrenbergi: «Duhok-Bardarash Population» 2n = 52, NF = 76, and NFa = 72 and «Arbil-Sulaimania-Kirkuk populations» 2n = 52, NF = 80 and NFa = 76. The karyotypes of the Duhok population are similar to those from Mosul, but the Arbil-Sulaimania-Kirkuk populations’ karyotype represents a new chromosomal form. Their distribution extends from North Iraq to Sulaimania.

Population genetic study of Ziziphus  jujuba Mill. : Insight in to wild and cultivated plants genetic structure

Seyyedeh Tahereh NabaviDepartment of Horticulture Science, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran.

Accepted: 2019-11-18 | Published Online: 2019-12-03 | DOI: 10.13128/caryologia-405


Ziziphus jujuba (jujube) of buckthorn family (Rhamnaceae) is an important medicinal crop plant cultivated in different provinces of Iran. It has also wild populations in some geographical areas. We carried out population genetic study on 8 populations of cultivated versus wild jujuba by using ISSR molecular markers to produce data on population genetic structure, gene flow, and genetic variability in the studied populations. We also aimed to investigate genetic differentiation between wild and cultivated plants and identify the potential gene pools of this medicinal plant species. The studied populations had a moderate genetic variability and were grouped in two major groups by PCoA plot. AMOVA revealed significant genetic difference among these cultivars. Mantel test showed significant correlation between genetic distance and geographical distance in the studied populations. PCoA analysis showed genetic differentiation between wild and cultivated plants within each province. STRUCTURE analysis identified two potential gene pools for jujube cultivars. Data obtained may be used in genetic conservation and future breeding programs of this medicinal plant species in the country.

Effects of zinc on pollen gamete penetration to pistils in some apple crosses assessed by fluorescence microscopy

Yavar SharafiDepartment of Horticultural sciences, Faculty of Agriculture, Shahed University, Tehran, Iran

Accepted: 2019-11-18 | Published Online: 2019-12-05 | DOI: 10.13128/cayologia-258


Zinc is classically the second abundant moveable metal in plants after iron and represented in all enzyme classes. Zinc generally contributes in the biosynthesis of auxin IAA and GA3 phytohormones which play the major role in fertilization and fruit set. Zinc deficiency leads to reduction in leaf and shoot size, photosynthesis and finally decreases the apple fruit set. Foliar Zinc spray was shown to be efficient and fast for improving Zinc deficiency in fruit trees. In this research the effects of Zinc solution by (0, 3000 and 5000 mg. L-1) were studied on pollen penetration to the pistil and ovary in the four apple cultivars crosses which included "Golden Delicious", "Red Delicious", "Gala" and "Fuji". Spraying was done on the shoots two weeks before blooming in the spring. Pollen penetration was studied using fluorescent microscopy technique 72 and 120 hours after field pollination. Results revealed that the effects of Zinc, crosses and their interaction were significant on pollen germination on the stigma and tube penetration into the primary, middle and beginning of the ovaries and the highest pollen germination on the stigma (43.5%) was observed in the cross (♀Golden Delicious × Gala♂), in 3000 mg. L-1 of Zinc 120 hours after pollination and highest pollen tube penetration into the ovary (12/88%) was observed in this cross, respectively. Finally, it was shown that fluorescence microscopy is an accurate technique for nutrition assay in pollination and fruit set. The foliar application of Zinc increased pollen germination and pollen tube growth in all of the crosses.

Characterization of intraspecific hybrid in Clitoria ternatea (L.) using morpho-physiological, cytogenetic, metabolic and molecular markers

Jogeswar Panigrahi, Department of Biotechnology, Central University of Rajasthan, NH-8, Bandarsindri, Kishangarh, Rajasthan

Accepted: 2019-11-18 | Published Online: 2019-12-06 | DOI: 10.13128/caryologia-754


Clitoria ternatea (L.) is a medicinal plant possessed with bioactive molecules such as taraxerol, delphinidin, kaempoferol and quercetin etc. For the development genotype with higher content of these bioactive molecules, marker-assisted breeding is one of the best strategies and it initiates with the development of F1 hybrids. Thus, an intraspecific F1 hybrid was raised involving two contrasting genotypes of C. ternatea acc. CtB3-SL1 (Blue flowered) and C. ternatea acc. CtW2-BL1 (white flowered). The hybridity of the F1 plant was confirmed by assessing the phenotypic traits, such as colour of the petal, pod shape and seed coat colour, 100 seed weight, and the content of taraxerol and delphinidin. The pollen mother cells in the F1 hybrid showed eight bivalents with preponderance of ring bivalents and 8I:8I segregation at metaphase-I and Anaphase-I, respectively. SDS-PAGE seed albumin and globulin detected three pollen parent-specific polypeptides (Mw 31.62, 22.38 and 18.81KDa), and were inherited to F1 hybrids, which evidenced the hybridity of putative F1 plants. Further, DNA marker analysis also showed the inheritance of 11 RAPD, six SCoT and one ISSR markers to putative F1 plant, which affirmed the hybrid nature of the F1 plant. This study also evidenced that combined use of morphophysiological, cytogenetic, protein and DNA marker analyses could be effective for precise characterization of intra-specific hybrids in C. ternatea. These F1 hybrid and its derived future progenies could also be used for mapping of QTLs or genes contributing higher accumulation of taraxerol and delphinidin in different plant parts.

Floral architecture, breeding system, seed biology and chromosomal studies in endangered Himalayan Edgew. (Apiaceae)

Kamini Gautam, YSP University of Horticulture and Forestry, Solan, Himachal Pradesh, India; Grassland and Silvipasture Management Division, ICAR-Indian Grassland and Fodder Research Institute, Jhansi, Uttar Pradesh, India

Accepted: 2019-11-18 | Published Online: 2019-12-06 | DOI: 10.13128/caryologia-755


Endangered Angelica glauca, an important medicinal plant of temperate Himalaya is valued for its roots which are used to treat several diseases besides food flavouring. Reproductive biology studies conducted in this species for the first time have revealed i). presence of umbels of different orders with only bisexual flowers ii). occurrence of sterile seeds (without embryo) apart from fertile ones iii). seed set in only early blooming umbels (primary and lateral-I) iv). 2n=22 chromosomes besides presence of chromosomes in a group at metaphase and anaphase-I and cytomixis in some pollen mother cells and v). extreme protoandry and cross pollination behavior (upto 95%) of the species. These observations have implications for developing any conservation plan for the species.

Cytotoxic and genotoxic activity of Plantago major L. extracts

Adisa ParićDepartment of Biology, Faculty of Science, University of Sarajevo, Zmaja od Bosne 33-35,
71000 Sarajevo, Bosnia and Herzegovina

Accepted: 2019-11-18 | Published Online: 2019-12-10 | DOI: 10.13128/caryologia-759


Plantago major L. is a perennial, wild plant that belongs to the Plantaginaceae family, and is used as a good indicator in the assessment of destructive anthropogenic impact on the environment. The aim of the present study was to evaluate cyto/ genotoxic effects of methanol extracts of Plantago major, collected from two locations (Tetovo and Smetovi), using Allium cepa test. We demonstrated that the highest concentration of P. major extracts from both locations reduced the mitotic index, while the lowest increased mitotic index value comparing to the positive control. As for genotoxic effects of extract from Tetovo, all concentrations increased the frequency of sticky chromosomes and chromosome missegregations in comparison with both controls, and frequency of multipolar anaphases when compared to the negative control. Higher number of cells with vagrants in comparison with positive control was detected after the treatment with 0.005 and 0.02 mg/ml concentrations. P. major extract from Smetovi (0.005 and 0.01 mg/ml) induced an increase in the number of vagrants as compared to the positive control, and frequency of sticky chromosomes when compared to both controls (0.01 mg/ml). Exposure to extract (0.005 and 0.02 mg/ml) caused increased number of multipolar anaphases in comparison with negative control. Apoptosis were not detected for P. major extract from Smetovi, while its highest concentration (0.02 mg/ml) induced increase in the frequency of necrosis as compared to the positive control. Our results demonstrated that methanol extracts of P. major, collected from Tetovo
and Smetovi, showed cyto/genotoxic effects on A. cepa meristem cells.

Genetic diversity of Rhododendron simsii Planch. natural populations at different altitudes in Wujiashan Mountain (central China)

Yuanping FangHubei Key Laboratory of Economic Forest Germplasm Improvement and Resources Comprehensive Utilization; Hubei Collaborative Innovation Center for the Characteristic Resources Exploitation of Dabie Mountains; College of Life Science, Huanggang Normal University, Huanggang, 438000, Hubei Province, P.R. China

Accepted: 2019-11-18 | Published Online: 2019-12-12 | DOI: 10.13128/caryologia-760


Altitude could greatly influence species distribution and even their genetic diversity. However, it is unclear how altitude has affected the genetic diversity and population structure of Rhododendron simsii Planch., an dominant forestry species in north temperate forest. In this research, 22 polymorphic EST-SSR markers were utilized to assess the genetic diversity of R. simsii population distributed at different altitudes of Wujiashan Mountain, a major peak of Dabie Mountains (central China). Totally, 203 alleles were obtained, and each locus gave out 5 to 19 alleles. High genetic diversity existed, as Nei’s gene diversity (h) and Shannon’s Information index (I) ranged from 0.728 to 0.920 and 1.430 to 2.690, with the mean value of 0.821 and 1.916, respectively. In particular, 11.1% of genetic differentiation was maintained between populations, while 88.9% occurred within populations. Moreover, moderate gene flow (2.001) among populations was observed, which could effectively resist genetic drift. The genetic diversity of all these five R. simsii populations varied significantly with elevation, basically showing high-low-high pattern with elevation increase. Without human intervention, genetic diversity of R. simsii populations might increase with the altitude. At the significance level (p < 0.05), negative correlation was found between genetic diversity and attenuation rate of light intensity (r=-0.873). Soil of Wujiashan Mountain was acid (the pH value ranged from 4.33 to 4.70), which was rich in organic matter, available phosphorus, available potassium, and alkali hydrolysable nitrogen, as these soil factors interacted with each other to affect the growth of R. simsii population. This research would contribute to the knowledge of evolutionary history of R. simsii species and be benefit for corresponding management and conservation actions.

Nonreduction via meiotic restitution and pollen heterogeneity may explain residual male fertility
in triploid marine halophyte Limonium algarvense (Plumbaginaceae)

Ana D. CapertaLinking Landscape, Environment, Agriculture and Food (LEAF), Instituto Superior de Agronomia (ISA), Universidade de Lisboa, Tapada da Ajuda, 1349-017 Lisboa, Portugal

Accepted: 2019-11-18 | Published Online: 2019-12-12 | DOI: 10.13128/caryologia-761


The cosmopolitan halophylic genus Limonium (Plumbaginaceae) presents high cytogenetic interest because of the natural occurrence of diploid and polyploid variants. Natural triploids are very rare in nature but common in this genus, including the widespread triploid Limonium algarvense found in the Iberian Peninsula and in Morocco. This study describes male sporogenesis and gametogenesis, pollen formation and germination, and seed production in triploid L. algarvense and diploid Limonium ovalifolium using various cytological approaches.
The diploid species presented regular meiosis. The triploid species was defective in male meiosis due to unpaired chromosomes, trivalent and tetravalent pairing, unbalanced chromosome segregation in meiosis I, and meiotic restitution in both meiosis I and I. These results may be explained by indeterminate and broad first meiotic restitution. Dyads and restitution nuclei at meiosis I were the most frequent meiotic products in the triploid species. Cytomixis was observed in both species, and callose deposition did not differ among them. In the diploid species, regular, tricolpate pollen grains, which germinated in vitro were found. Contrastingly, in the triploid species produced heterogeneous pollen in morphology and size, with moderate to no viability that poorly germinated in vitro. We conclude that even if most triploids male gametes are non-functional, they seem to generate small numbers of viable gametes via nonreduction of chromosomes. Flow cytometric seed screening demonstrated that the diploid presented a diploid progeny whereas triploids only showed triploid
progenies. Triploids low pollen fertility coupled with viable seed production may assure their persistence in natural populations.

Active chemical constituents of Cynanchum viminale and its cytotoxic effects via apoptotic signs on Allium cepa root meristematic cells

Neethu Kannan Bhagyanathan,  Cell and Molecular Biology Division, Department of Botany, University of Calicut, Kerala, India

Accepted: 2019-11-18 | Published Online: 2019-12-13 | DOI: 10.13128/caryologia-769


The present study evaluates the cytotoxic efficacy of methanolic extract of C. viminale on A. cepa root meristematic cells. DAPI staining was used to study the chromosomal aberrations indued by extract of C. viminale. Evans blue staining method was employed to estimate the cell death of root cells of A. cepa. The plant extract was found to impart severe cytological damges, specifically chromosomal aberrations at interphase and prophase stage of cell division. Various apoptotic signs such as apoptotic body formation, nuclear budding, micronucleus, nuclear disintegration, nuclear breakage etc. were observed in meristematic cells of A. cepa. The results suggest the cytotoxic, preferably genotoxic effect of methanolic extract of C. viminale as evidenced by various apoptotic symptoms on A. cepa root cells.