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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.