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Species delimitation in the genus Cousinia Cass. (Family Asteraceae), sections Cynaroideae Bunge and Platyacanthae Rech. f.: morphometry and molecular analysis

Neda Atazadeh, Faculty of Life Sciences & Biotechnology, Shahid Beheshti University, Evin, Tehran, Iran

Accepted: 2020-04-05| Published Online: 2020-07-26| DOI: 10.13128/caryologia-275


The genus Cousinia of the tribe Cardueae with about 700 species is one of the most diverse genera in Central and southwest Asia. The section Cynaroides with 89 species is the largest section of the genus. Due to the controversy in the number of Cousinia species and their delineation, the first step in studying the genus is to identify and delimit presumed species. Species delimitation is usually difficult in the species with overlaps in their morphological features. Therefore, we used a combination of morphological and molecular markers (ISSRs) to carry out delimitation in 204 taxa of 68 Cousinia species within the Cynaroideae and Platyacanthae sections. The species delineation based on morphometry and ISSR data were done by UPGMA clustering. The samples of each species were placed close to each other and formed a single sub-cluster, separated from the other studied Cousinia species. In the present study, the studied Cousinia species within Cynaroideae and Platyacanthae sections could be delimited from each other based on ISSR and morphological data. Therefore, using ISSR and morphological data can be useful in identifying and delineating crucial species. The Mantel test performed between morphological distance and Nei genetic distance produced a non-significant correlation. This result also supports distance analyses of the trees and reveals that the two dendrograms are not correlated. Some possible reasons for this incongruence are proposed: the high number of taxa in the genus Cousinia, morphological traits homoplasious, convergent evolution, and incomplete lineage sorting.

Investigation of Benzyl Benzoate Toxicity with Anatomical, Physiological, Cytogenetic and Biochemical Parameters in In Vivo

Ali Akar, Vocational School of Health Services, Department of Medical Services and Techniques, Giresun University, Giresun, Turkey.

Accepted: 2020-04-13| Published Online: 2020-08-03| DOI: 10.13128/caryologia-167


In this study, the toxic effects of benzyl benzoate, which is widely used in the food, cosmetics, agriculture, and pharmaceutical sectors, have been investigated using Allium cepa L. test material. In the determination of toxicity, physiological parameters with the determination of root lengths, weight gains, and germination percentages; cytogenetic changes with the determination of chromosomal abnormalities formation, micronucleus (MN) frequency and mitotic index ratio (MI);  anatomical changes with the determination of anatomical differentiation in root tip cells; biochemical changes with lipid peroxidation and antioxidant enzyme analysis were determined and the obtained data were evaluated statistically. The bulbs were divided into four groups consisting of one control and three application groups, bulbs of the control group were treated with tap water and the bulbs of the application groups were treated with Benzyl benzoate at doses of 10,000, 25,000 and 50,000 mg/L for 72 hours. At the end of the study, it was determined that germination percentage, weight gain and root length and MI ratio decreased, chromosomal abnormalities, MN formation, MDA, SOD and CAT levels increased dose-dependent in the application groups when compared with the control group. Depending on the application, it has been determined that root cells have chromosomal abnormalities such as fragments, sticky chromosomes, chromosome bridges, unequal distribution of chromatins and c-mitosis. Furthermore, when compared with the control group, it was determined that benzyl benzoate administration caused anatomical changes in root tip cells. It was determined that these changes were in the form of necrosis, cell deformation, flattened cell nuclei, cortex cell deformation, accumulation of certain substances in cortex cells, wall thickening in cortex cells and unclear vascular tissue. In conclusion, it was determined by physiological, anatomical, cytogenetic and biochemical parameters that benzyl benzoate showed a dose-dependent toxic effect in Allium cepa L. root cells. Also, the parameters used in the study were determined to be useful biomarkers for the determination of toxicity.

Karyological variability and chromosomal asymmetry in highland cultivars of Chenopodium quinoa Willd. (Amaranthaceae)

Sreetama Bhadra and Zhi–Quan Cai, CAS Key Laboratory of Tropical Plant Resources and Sustainable Use, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Mengla 666303, China;

Department of Horticulture, Foshan University, Foshan 528000, China.

Accepted: 2020-04-27| Published Online: 2020-08-10| DOI: 10.13128/caryologia-625


Chenopodium quinoa Willd. is rapidly gaining importance worldwide as a superfood. However, structural diversity and asymmetry analyses of chromosomes of different cultivars of the species are largely understudied primarily owing to their small chromosomes. In this paper, karyomorphological investigations were performed on 21 cultivars of C. quinoa with varying seed morphology cultivated widely in the highland regions of the Andes, which is the center of domestication of this species. Somatic chromosome number was found to be 2n= 36 in all cultivars with no occurrence of mixoploidy. Lengths of individual chromosomes varied between 0.63–6.53 μm, with their short arms ranging from 0.25–2.95 μm and long arms between 0.38–3.58 μm. Types of primary constriction ranged from median to sub–terminal. One pair of chromosome in each complement possessed a secondary constriction. Chromosome complements of all cultivars belonged to the asymmetry class 2B with an average asymmetry index value of 3.21±0.61. Values of intra– and inter–chromosomal asymmetry indices were 0.30±0.02 and 0.20±0.02 respectively across all cultivars. The average coefficients of variation of chromosome lengths was 19.77±2.11 and average centromeric index was 16.28±2.12. Arm ratio of the chromosomes varied from 0.34 to 5.76. The mean values of karyotypic asymmetry, symmetry index and karyotype asymmetry index percentage were 17.69±1.53, 147.69±5.54 and 58.71±0.86 respectively. Pearson correlation revealed strong correlation within inter– and intra–chromosomal asymmetry indices. Our analyses uncovered higher chromosomal variation in quinoa than previously found with high inter–varietal similarities among the studied cultivars, revealed from scattered diagrams between asymmetry indices.

Cytogenetic and molecular studies of the Egyptian Capsella bursa-pastoris (Brassicaceae)  

Rania A. Hassan, Botany and Microbiology Department, Science Faculty, Cairo University, Egypt.

Accepted: 2020-04-19| Published Online: 2020-08-11| DOI: 10.13128/caryologia-867


Capsella bursa-pastoris (Brassicaceae) is one of the most successful tetraploid species in the world. It showed high morphological diversity within Egyptian populations. Morphological investigations of herbarium specimens and fresh collected populations grouped them under three distinguished morphotypes (Lobed "L"; Simple "S" and Lobed-Simple "LS") depending mainly on the basal leaves structure. This high degree of phenotypic variation has received our critical attention. Until recently, the previous studies on C. bursa-pastoris attributed its phenotypic variation to environmental factors. But in Egypt, these three morphotypes were traced in mixed populations along with the species geographical range, so the environmental factors have no influence on their distribution or phenotypic variation. Accordingly, our primary concern in this study was to determine the factors controlling this variation. The cytogenetic studies revealed that the three identified morphotypes are three distinct genotypes with three different chromosome numbers: 2n=2x=16 (diploid) for "L"; 2n=3x=24 (triploid) for "S"; 2n=4x=32 (tetraploid) for "LS". The triploid genotype "S" showed rare occurrence among the studied populations and is postulated to be a new record of a hybrid in Egypt. Karyotyping of the three genotypes showed significant differences in the genome and chromosomes relative lengths. A molecular study using cpSSR technique supported the cytogenetic results and differentiated the three studied genotypes. The retrieved results revealed that the phenotypic diversity within the Egyptian C. bursa-pastoris populations is genetically controlled.

Genome size and chromosome number of Psidium friedrichsthalianum Ndz (“Costa Rican Guava”) in six populations of Costa Rica  

Mónica Rojas-Gómez, Centro Nacional de Innovaciones Biotecnológicas (CENIBiot), CeNAT-CONARE,1174-1200 San José, Costa Rica.

Accepted: 2020-05-31| Published Online: 2020-08-11| DOI: 10.13128/caryologia-646


Psidium friedrichsthalianum (O. Berg) Nied is a species found from southern Mexico, Central America; and there are reports that it is also found in Venezuela and Ecuador. It is a common fruit component of the Costa Rican diet, and it is valued industrially for its high content of polyphenols, mainly proanthocyanidins (PACs). This crop is not completely domesticated and there are no improved varieties produced through plant breeding. Genome size or ploidy levels have not been investigated in Costa Rican populations of Psidium friedrichsthalianum. Information about chromosome number and genome size is paramount for plant breeding strategies. Therefore, the main objective of our study was to determine chromosome number using pollen meiocytes and genome size by flow cytometry in six populations of P. friedrichsthalianum in Costa Rica. We found x = 11 bivalent chromosomes in all meiocytes analysed, classifying these populations as diploid. All populations had an average nuclear DNA content of 2C = 1.960 ± 0.005 pg. No statistically significant differences in nuclear DNA content were found among populations. We conclude that the consistency in chromosome number and genome size among populations suggests a common origin among them. Our estimates of the number of chromosomes and genome size of P. friedrichsthalianum determined in this study will be essential for future breeding programs, hybridization practices and development of QTL (Quantitative Trait Loci).

Karyotype analysis of Trichogramma embryophagum Htg. (Hymenoptera: Trichogrammatidae) using a new method and estimate its karyotype symmetry

Jamasb Nozari, Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran.

Accepted: 2020-06-02| Published Online: 2020-08-11| DOI: 10.13128/caryologia-203


Different methods of chromosome preparation for insects are now being used across the world. Well- spread chromosomes with explicit morphology, in addition to no cell wall debris are required for karyotype investigations. Cytogenetic knowledge in Trichogramma is extremely limited. In the article, chromosome characters, karyotype and monoploid ideogram of Trichogramma embryophagum Htg. (Hymenoptera: Trichogrammatidae) were evaluated using a new method. Also, karyotype symmetry/asymmetry of this species was calculated for the first time as one of the Trichogramma species. Our results showed that the diploid chromosome number of the wasp was 2n = 10. The karyotype formula was 6m + 4a. The symmetry/asymmetry index value was 1.8. The new method resulted in higher quality metaphase plates spread and provided an ideal karyomorphology for this parasitoid which has small chromosomes.

Assessment of anti-cytotoxic, anti-genotoxic and antioxidant potentials of Bulgarian Rosa alba L. essential Oil

Svetla Gateva, Institute of Biodiversity and Ecosystem Research, Bulgarian Academy of Sciences, 2 Gagarin Str., Bulgarian Academy of Sciences, Sofia 1113, Bulgaria

Accepted: 2020-06-02| Published Online: 2020-08-11| DOI: 10.13128/caryologia-260


Bulgarian Rosa alba L. essential oil is widely used in perfumery, cosmetics and pharmacy. The scarce data about its cytotoxic/genotoxic effect and anti-cytotoxic/anti-genotoxic potential gave us a reason to set our aim: i) to study its cytotoxic/genotoxic activities, iii) to explore its cytoprotective/genoprotective potential against the experimental mutagen N-methyl-N’-nitro-N-nitrosoguanidine (MNNG) in two experimental test-systems - barley and human lymphocytes using appropriate endpoints and iii) to assess its antioxidant properties. Findings about chemical composition of rose essential oil would help us to explain its activities. Chromatogaphic profile of rose essential oil was obtained by Gas Chromatography-Mass Spectrometry and quantificaton of particular constituents was done with a Gas Chromatography-FID system. Superoxide anion radical scavenging, DPPH inhibition and iron ion chelating activity were used to study a possible antioxidant potential of the rose oil. Its defense potential was investigated by induction of chromosome aberrations and micronuclei in both test-systems. Cytogenetic analysis showed a low cytotoxic effect in both test-systems and no high genotoxic effect in human lymphocytes in vitro in a dose-dependent manner. Rose oil possessed a well-expressed anti-cytotoxic/anti-genotoxic potential against MNNG manifested by decreasing both of chromosome aberrations and micronuclei regardless of the experimental schemes used. A well-expressed concentration-depended free radical scavenging activity of the essential oil was obtained. Current data suggest a promising ethnopharmacological potential of Bulgarian white rose essential oil.

Display of Sukkula Distributions on Barley Roots via in Situ Hybridization

Elif Karlik, University of Istinye, Faculty of Arts and Sciences, Department of Molecular Biology and Genetics, Istanbul, Turkey.

Accepted: 2020-06-19| Published Online: 2020-08-12| DOI: 10.13128/caryologia-191


Retrotransposons are an abundant and ancient part of the plant genomes that especially LTR retrotransposons influence the genome size and evolution. Sukkula is a non-autonomous and active, relatively high copy-number retroelement. In this study, we performed fluorescence in situ hybridization (FISH) to observe the distributions of Sukkula elements (LTRs and internal-domain) by using labelled-PCR products. The localization of Sukkula elements (LTRs and internal-domain) was observed under a confocal microscope on Hordeum vulgare L. cv. Hasat root preparations. Our results revealed that Sukkula elements are still active and spread through the whole barley chromosomes. Additionally, the re-sequencing analysis of Sukkula LTRs demonstrated that LTRs sequences had ~65 bp gain. These analyses represent a valuable resource to reveal the genome organization of barley and large-sized plants.

Karyotype Analysis of Lilium lancifolium and Four Related Cultivars

Lei-Feng Xu and Jun Ming, The Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China.

Accepted: 2020-06-19| Published Online: 2020-08-12| DOI: 10.13128/caryologia-218


Lilium lancifolium is one of the most important species of genus Lilium. Besides ornamental value, it also has highly edible and medicinal properties. We investigated the karyotypes of L. lancifolium and four related cultivars. The results indicated that the ploidies of four cultivars varied from diploid to tetraploid. Both ‘Flore Pleno’ and ‘Red Velvet’ were triploid (2n=3x=36), consistent with the wild species L. lancifolium. ‘Sweet Surrender’ was diploid (2n=2x=24), and ‘Red Life’ was tetraploid. All karyotypes of candidates belonged to 3B type except ‘Flore Pleno’, which belonged to 3A type. Karyotype symmetry analysis revealed that the wild species L. lancifolium had a middle value of A1, A2, and TF%, which meant that the cultivars related to L. lancifolium had different tendencies to symmetry compared to L. lancifolium, but whether they were higher or lower was unclear.

An adverse effect of meiotic abnormalities on spore fitness in medicinal fern Glaphyropteridopsis erubescens (Wall. ex Hook.) Ching

Mandeep Kaur Aulakh, Department of Botany, Punjabi University, Patiala-147002, Punjab, India.

Accepted: 2020-06-23| Published Online: 2020-08-13| DOI: 10.13128/caryologia-252


Glaphyropteridopsis erubescens plants were collected from Parvati valley, Himachal Pradesh, India. Meiotic investigations in the accessions collected from different areas of Himachal Pradesh revealed normal meiosis and good spore fertility. However, the accessions collected from the Parvati valley have depicted meiotic anomalies and high spore abortion index. All populations shared the same cytological status of 2n = 72, which is in line with previous records. Individuals depicted normal behavior at diakinesis and metaphase-I but later stages were abnormal. It includes chromatin stickiness, interbivalent connections, early and late disjunction. The formation of laggards at the anaphase-I lead to a series of abnormalities such as chromatin bridges, random sub-grouping of chromosomes at anaphase-II, chromatin fragmentation, and irregular sporogenesis. Large numbers of micronuclei were present in triads and tetrads along with pycnotic nuclei. Empty and some abnormal sporangia with heterogenous spores were also observed.

Toxicity of Aristolochia decoction: a relevant herbal in folk medicine

Tatiana da Silva Souza,Departamento de Biologia, Centro de Ciências Exatas, Naturais e da Saúde, Universidade Federal do Espírito Santo - UFES, Alto Universitário, s/ número Guararema, 29500-000, Alegre, ES Brasil.

Accepted: 2020-07-14| Published Online: 2020-08-13| DOI: 10.13128/caryologia-880


Ethnopharmacology studies report the use of Aristolochia (Aristolochiaceae) species as medicinal plants in various parts of the world. However, the acids aristolochic (AAs), secondary metabolites present in all species of Aristolochia, have cytogenotoxic activity and they are a potent carcinogen to rodents and humans. The aim of the current research was to perform initial screening for the toxicity of Aristolochia labiata Willd and Aristolochia triangularis Cham decoctions through germination and growth rate, flow cytometry, mitotic index, and cytogenetics analysis in Allium cepa. The decoctions were prepared from 2, 4, 8, 16, and 32 g L-1. Decoctions at concentrations 4 g to 16 g L-1 significantly reduced the germination rate of Allium cepa. Seeds exposed to 32 g L-1 decoctions did not germinate. All as decoctions reduced the growth rate of onion seedlings. Decoctions at 4 g L-1 to 16 g L-1 inhibited the mitotic index. The highest concentrations of decoctions (8 g L-1 and 16 g L-1 for Aristolochia labiata; 16 g L-1 for Aristolochia triangularis) showed a statistically significant increase in the frequency of Allium cepa nuclei in the G0/G1 phase. Both decoctions induced the formation of heteropycnotic nuclei. Qualitative phytochemical prospecting of decocts was performed and alkaloids secondary compounds were the largest presence in both species, indicating that the AAs may be related to the observed toxicity. Caution is recommended in the consumption of decoctions from Aristolochia labiata and Aristolochia triangularis stems.

Physical mapping of 45S and 5S rDNA in two Sprekelia formosissima cytotypes (Amaryllidaceae) through Fluorescent In Situ Hybridization (FISH)

Rodrigo Barba-Gonzalez, Unidad de Biotecnología Vegetal, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, Guadalajara, Jalisco, México.

Accepted: 2020-07-14| Published Online: 2020-08-13| DOI: 10.13128/caryologia-578


Chromosome number and position of rDNA were studied in plants of Sprekelia formosissima (Amaryllidaceae) collected in two populations with different ploidy levels (2n=2x=60 and 2n=5x=150). The 5S and 45S rRNA loci were localized and physically mapped using two-color fluorescence in situ hybridization probes. The diploid (2n=2x=60) cytotype showed four loci for the 45S rDNA in two chromosome pairs (11 and 25) in telomeric position. The 5S rDNA was present in six loci of three homologous chromosome pairs (3, 13, and 19) in subtelomeric and telomeric positions. The chromosomes of the pentaploid cytotype (2n=5x=150) showed five loci for the 45S rDNA in telomeric position and five loci for the 5S rDNA in subtelomeric position. The karyotypic formula is 13m + 16sm + 1 st and the karyotype symmetry/asymmetry index is TF % = 34.67, AsK % = 65.32 and Syi % = 54.81, concluding that it is an asymmetric karyotype, bimodal with one distinctively large pair of chromosomes (10.42 µm) and a gradual decrease in the size of the other chromosome pairs, from the longest of 6.84 µm to the shortest of 2.61 µm.

The Role of Oleuropein Against Nanocomposite Toxicity in Fruit Fly: Evidence For Lifespan Extension

D. Altun Çolak, Department of Biology, Faculty of Art and Science, Erzincan Binali Yıldırım University, Erzincan, 24100 Turkey

Accepted: 2020-07-22| Published Online: 2020-08-13| DOI: 10.13128/caryologia-226


The effect of zinc oxide/titanium dioxide nanocomposite the lifespan of the fruit fly Drosophila melanogaster and the protective role of oleuropein, a strong antioxidant, against the zinc oxide/titanium dioxide were investigated. Chemicals prepared in different concentrations (0.005, 0.1, 0.5, and 1 g/L for zinc oxide/titanium dioxide; 0.1 mmol/L for oleuropein) have been separately applied to female and male populations of D. melanogaster for the control and application groups. In both female and male populations, it has been observed that zinc oxide/titanium dioxide has decreased the lifespan and oleuropein has increased the lifespan according to the control group, depending on the concentration. These findings demonstrate the beneficial effect of oleuropein, suggested as a protective role in the prevention of zinc oxide/titanium dioxide induced developmental toxicity.