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The first molecular identification of Egyptian Miocene petrified dicot woods (Egyptians’ dream becomes a reality)

Shaimaa S. Sobieh, Botany Department, Faculty of Women for Arts, Science and Education, Ain Shams University, Cairo, Egypt.

Accepted: 2020-01-24 | Published Online: 2020-04-27 | DOI: 10.13128/caryologia-750


This is the first work on Egyptian ancient DNA (aDNA) from plant fossil remains. Two aDNA extracts from Miocene petrified dicot woods were successfully obtained, amplified, sequenced and recorded for the first time in the world using a DNA barcoding technique. Internal transcribed spacers (ITS) barcoding is a technique for delimiting and identifying specimens using standardized DNA regions. The two Miocene dicot woods: Bombacoxylon owenii (Malvaceae/Bombacoideae) and Dalbergioxylon dicorynioides (Leguminosae/Papilionoideae) were collected from the Wadi Natrun area in Egypt and were identified by palaeobotanists on the basis of wood anatomy. The molecular identification by ITS region of Bombacoxylon owenii did not match the wood taxonomic assignation. The molecular identification of Bombacoxylon owenii suggested that it is more related to the extant genus Ceiba rather than to the extant genus Bombax. In contrast, the molecular identification by ITS of Dalbergioxylon dicorynioides matched the identification of the palaeobotanist (related to extant genus Dalbergia). Therefore, we suggest that this region should be used as a starting point to identify several plant fossil remains and this work will be helpful in solving problems related to the identification of plant fossils.

Gene flow patterns reinforce the ecological plasticity of Tropidurus hispidus (Squamata: Tropiduridae).

R. A. Torres, LAGEA – Laboratório de Genômica Evolutiva & Ambiental, Departamento de Zoologia, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Brazil

Accepted: 2020-03-27 | Published Online: 2020-04-28 | DOI: 10.13128/caryologia-672


The analysis of gene flow patterns can provide important insights into population dynamics in the context of landscape ecology. In lizards, this approach has been used to evaluate patterns related to climate change, habitat fragmentation, and taxonomic uncertainties. Tropidurus hispidus is an ecologically plastic species, which presents some evidence of population structuring. In the present study, we investigated the potential structuring of T. hispidus populations across a gradient of tropical biomes, including the Amazon and Atlantic rainforests, the Caatinga dry forest, the Caatinga-Atlantic Forest transition zone (Agreste), coastal Restinga, and urban environments. Nuclear ISSR markers were obtained by PCR/electrophoresis, and a number of population parameters were estimated and analyzed. Despite the extreme environmental discontinuities found across the vast study area, the results revealed a high degree of genetic connectivity among the different demes. This pattern indicates that the species can be considered to be a single evolutionary taxon with gene flow among all populations, despite the marked environmental discontinuities. Tropidurus hispidus clearly has a marked capacity for dispersal, which may be favored by its intrinsic genetic diversity.

The technique of Plant DNA Barcoding: potential application in floriculture

Antonio Giovino,Council for Agricultural Research and Economics (CREA), Research Centre for Plant Protection and Certification (CREA-DC), Bagheria, Italy

Federico Martinelli, Department of Biology, University of Florence, Sesto Fiorentino, Florence, 50019, Italy


Accepted: 2020-04-01 | Published Online: 2020-04-28 | DOI: 10.13128/caryologia-730


The objective of this work was to assess the ability of the DNA barcoding approach to identify different taxonomic groups from two flowering plant collections: 1) the most relevant commercial taxa (nursery production) and 2) Mediterranean plants with ornamental attitude (new emerging species). “Core markers”, rbcL and matK, were adoptedthe identification step of 100 taxa belonging to 20 families. A third marker, the intergenic spacer trnH-psbA, was also tested, on 74 taxa, when the core markers were not able to discriminate well the analysed germplasm.DNA barcode fragments were recovered for all the total taxa investigated (100%). The rbcL showed the best performances: the greatest amplification success, the best sequencing performance both in terms of the number of sequences obtained and in terms of quality of the sequences obtained. Despite having recorded greater amplification difficulties, according to numerous other studies, matK has shown a good success in sequencing and quality of the obtained sequences (de Vere et al. 2012), unlike what is indicated in some protocols that suggests for this region the need for further primers to be adopted for the sequencing phase (Hollingsworth et. al 2011). Results showed that sixty-one taxa overall (61%) were totally resolved at specific or subspecific level, by at least one of the three markers. The matK and rbcL locus respectively resolved 44% and 35% of the taxa. The core markers in multilocus approach led to the discrimination of a total of 49% taxa. The trnH-psbA was able to discriminate 52% of taxa analysed and resulting determinant in the discrimination of 14 taxa. Four families, including the major number of taxa (Arecaeae, Fabaceae, Euphorbiaceae, Asteraceae), were evaluated in terms of genetic distance (K2P% value). This work highlighted the potential of the barcoding approach for a rapid identification of plant species in order to solve taxonomic disputes and support commercial traceability of floreal products.

Cytogenetic of Brachyura (Decapoda): testing technical aspects for obtaining metaphase chromosomes in six mangrove crab species

Stefano Cannicci, School of Biological Sciences, Faculty of Science, HKU

Accepted: 2020-04-01 | Published Online: 2020-04-29 | DOI: 10.13128/caryologia-791


Brachyura is one of the most specious infra-order belonging to Decapoda and it plays a central role from an ecological and economic point of view. Despite its importance, cytogenetic studies on Brachyura (Decapoda) are extremely limited due to the difficulties in obtaining chromosome preparations of good quality. Molecular cytogenetic have proven to provide basic knowledge on the genome organization of species and in methods for manipulating genomes. It is also very useful to support aquaculture.
In this study, we focused on six semi-terrestrial mangrove crab species to test several variations of technical steps to produce chromosome preparations in Brachyura. The best results were obtained using cells from early stage embryos incubated with 0.0005% nocodazole or 0.05% colchicine and hypotonized with 0.075 M KCl solution. The best method to analyze the chromosomes was the use of confocal microscope after DAPI staining. We recorded a high chromosome number for the six study species. Similar chromosome morphology was recorded for allied species likely due to phylogenetic relationship. Variable results with cytogenetic treatments in different species suggest that there may be a species-specific response to the techniques we tested. Chromosome number ranges reported in this study will be useful in future genome sequencing studies, i.e. to assess the quality of sequencing assemblies.

Comparison of the Evolution of Orchids with that of Bats

Antonio Lima-de-Faria, Lund University, Lund, Sweden

Accepted: 2020-04-02 | Published Online: 2020-04-29 | DOI: 10.13128/caryologia-891


The evolution of orchids and bats is an example of DNA’s own evolution which has resulted in structures and functions which are not necessarily related to any obvious advantage to the organism. The flowers of orchids resemble: humans, apes, lizards, frogs and even shoes. The faces of bats resemble plant leaves but also horseshoes. These similarities are not accidental because they emerge repeatedly in different genera and different families. This evolutionary situation bewildered botanists and zoologists for many years, but is now elucidated by the molecular unification of plants and animals derived from the following evidence: (1) Contrary to expectation, plant and animal cells (including those of humans) could be fused and the human chromosomes were seen dividing in the plant cytoplasm. (2) Orchids, bats and humans have about the same number of genes: orchids, 21,841; bats, 21,237 and humans circa 20,000. (3) These three groups contain the same homeotic genes which decide: flower formation (orchids), body segmentation (bats) and body segmentation (humans). The leaf pattern, is formed in plants by the LEAFY master gene, but this pattern even appears in minerals, which have no genes, an indication that pure atomic processes are responsible for its emergence at the organism level.

Identification of the differentially expressed genes of wheat genotypes in response to powdery mildew infection

Mehdi Zahravi, Seed and Plant Improvement Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.

Accepted: 2020-04-01 | Published Online: 2020-04-29 | DOI: 10.13128/caryologia-752


Bread wheat (Triticum aestivum L.) is the most widely grown crop worldwide. Powdery mildew caused by fungal pathogen Blumeria graminis is one of the most devastating diseases of wheat. The present study aimed to identify differentially expressed genes and investigate their expression in response to B. graminis in susceptible (Bolani) and resistant (KC2306) wheat genotypes, using publicly available microarray data set and qRT-PCR analysis. A total of 5760 and 5315 probe sets were detected which 5427 and 4630 by adjusted P-value < 0.05 and 168 and 144 genes based on e-value < 1 × 10–5 cut-off were differentially expressed in susceptible and resistant wheat genotypes, respectively. Among exclusively up regulated genes in the resistant genotype 12 hpi compared to its control, fifteen potential genes that may be responsible for B. graminis inoculation resistance were detected. The results of real time PCR for the candidate genes showed that the genes were upregulated in the resistant genotype 12 hpi compared to its control, which validated the results of microarray analysis. The bZIP, ERF, and ARF1 genes may play an important role in B. graminis resistance. The powdery mildew responsive genes identified in the present study will give us a better understanding on molecular mechanisms involved in B. graminis resistance in different wheat genotypes.

Populations genetic study of the medicinal species Plantago afra L. (Plantaginaceae)

Saeed Mohsenzadeh, Faculty of Life Sciences & Biotechnology, Shahid Beheshti University, Tehran, Iran.

Accepted: 2020-02-23 | Published Online: 2020-04-29 | DOI: 10.13128/caryologia-135


Plantago afra (Plantaginaceae) is the most medicinally important species in genus Plantago and it is native to the western Mediterranean region, West Asia and North Africa, and cultivated extensively in Asia and Europe for seed husk known as black Psyllium. We have no data on the population genetic structure of this species in the world. Therefore a population genetic and morphological investigation was performed through light on genetic and morphological variability in this taxa. We used ISSR molecular markers for population genetic investigation. Genetic diversity analyses revealed a moderate genetic variability within Plantago afra, while PCoA showed some degree of genetic admixture among populations. AMOVA produced significant genetic difference among populations. The Mantel test showed a positive significant correlation between the genetic and geographic distance of the studied populations. STRUCTURE analysis showed that there are different genetic groups in the studied populations. Morphometric analysis showed that one population differed in seed color and mean stem diameter. The same population contained specific allele combinations and differed genetically from the rest of the studied populations. Therefore, we considered it as a new variety within Plantago afra.

A comparative karyo-morphometric analysis of Indian landraces of Sesamum indicum using EMA-giemsa and fluorochrome banding

Timir Baran Jha,Department of Botany, Maulana Azad College, Rafi Ahmed Kidwi Road, Kolkata- 700113,
5 West Bengal, India

Accepted: 2020-04-03 | Published Online: 2020-04-29 | DOI: 10.13128/caryologia-580


Sesamum indicum commonly known as ‘Sesame’, ‘Til’ or ‘Gingli’ is an age-old high valued oil crop. With distinct seed and floral diversity and no detailed chromosomal analysis is available on Indian landraces of S. indicum (2n= 26). The present study demonstrates standardization of enzymatic maceration and air drying (EMA) method of chromosome preparation and comparative karyometric analysis in four Indian of S indicum. All the landraces were characterized by very small chromosomes, length ranging from 1.24 ± 0.02 to 2.87 ± 0.09 µm. The EMA- Giemsa based karyotype analysis revealed nine pairs of chromosomes with nearly median primary constriction, three pairs were submedian and a single satellite pair in each of the studied landrace. The CMA staining of Sesamum chromosomes revealed the presence of distinct CMA positive (CMA+ve) signals in all the studied landraces. The Black seeded til (BT) and White seeded til (WT) were characterized by six chromosomes with distal CMA+ve signal on short arm, while the Dark brown seeded til (DBT) showed ten chromosomes with distal CMA+ve signal on short arm. The Light brown seeded til (LBT) was characterized by eight chromosomes with distal CMA+ve signal on short arm. The results obtained from the scatter plot of A1 versus A2 and PCA analysis provide a strong relationship with that of the fluorochrome banding analysis. The present research offers an explicit karyo-morphometric characterization of four Indian landraces of S. indicum for the first time.

Chromosome Count, Male Meiotic Behaviour and Pollen Fertility Analysis in Agropyron thomsonii Hook.f. And Elymus nutans Griseb. (Triticeae: Poaceae) from Western Himalaya, India

Jaswant Singh, Department of Botany, Punjabi University Patiala, India, 147002

Accepted: 2020-01-15 | Published Online: 2020-04-29 | DOI: 10.13128/caryologia-618


Present cytological study records existing chromosome number diversity, their male meiotic course and pollen fertility analysis in the two wheatgrass species of tribe Triticeae Dumort. (Poaceae) From Western Himalaya, India. Agropyron thomsonii Hook. f. is an endemic grass of alpine zones of Western Himalaya and Elymus nutans Griseb., a widely distributed grass in sub-alpine to glacial regions of Himalaya. The gametophytic chromosome number count of n=21 (Jadh Ganga Valley, Uttarkashi) is a pioneer count for A. thomsonii. During the male meiotic course of A. thomsonii, 14.03-16.29% and 2.97-4.17% pollen mother cells, respectively at prophase-I and metaphase-I, observed to be involved in phenomenon of cytomixis. Seven accessions of E. nutans collected from Bhagirathi Valley and Jadh Ganga Valley of Uttarkashi district, and Pangi Valley of Chamba district, recorded with gametophytic chromosome number count of n=21 and record of 1B-chromosome in PUN61958 is a new record for the species. And 5.56-9.41% and 2.5% pollen mother cells at prophase-I and metaphase-I, respectively were also noted with phenomenon of cytomixis. In addition to phenomenon of cytomixis, during meiotic course of both species pollen mother cells also depicted associated meiotic course irregularities viz. non-synchronous disjunction of bivalents, chromatin bridges, laggards, micronuclei in sporads and shriveled microspores. These species are growing in cold climatic condition habitats. So, cold stress seems to be a preferential inductor for cytomixis and associated meiotic abnormalities in the gametic cells of stamens of A. thomsonii and E. nutans that ultimately leads to reduction in pollen fertility.

Population genetic and phylogeographic analyses of Ziziphora clinopodioides Lam., (Lamiaceae), “kakuti-e kuhi”: An attempt to delimit its subspecies

Raheleh Tabaripour, Faculty of Biological Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran

Accepted: 2020-04-13| Published Online: 2020-04-30 | DOI: 10.13128/caryologia-573


Ziziphora clinopodioides Lam., (Lamiaceae), is a perennial herb which is used as traditional medicine in Iran. Different authors disagree on the number of subspecies. In general, taxonomic and biosystematic studies of Ziziphora clinopodioides have been limited and no molecular phylogenetic or biogeographic study of the species has been carried out. Therefore, the aims of this study were (1) to determine the number of subspecies, (2) to produce information on the species’ genetic structure and intra-specific genetic variability, and (3) to produce data on the probable date of appearance of Ziziphora clinopodioides in Iran. We used a combination of morphological and molecular data to study plants randomly collected from 5 geographical regions. Both analyses revealed a high level of within population variability and grouping of the studied provinces produced an admixture that indicated the absence of any subspecies within the species. STRUCTURE analysis and K-Means clustering identified two gene pools within the country. The probable date of divergence obtained was 5-10 Mya for the appearance of this species in the mountainous regions of Qazvin and Mazandaran.

Karyotype studies on the genus Origanum (Lamiaceae) species and some hybrids defining homoploidy

TUNCAY DİRMENCİ,Department of Biology Education, Necatibey Education Faculty, Balıkesir University, Balıkesir, Turkey

Accepted: 2020-04-03| Published Online: 2020-05-01 | DOI: 10.13128/caryologia-579


In this study, chromosome numbers and structures of some Origanum taxa growing in Turkey were identified. Using the Image Analysis System, the complements of plant accessions belonging to eight sections, namely Amaracus (Gleditsch) Vogel, Anatolicon Benth., Brevifilamentum Ietsw., Longitubus Ietsw., Chilocalyx (Briq.) Ietsw., Majorana (Miller) Ietsw., Origanum Ietsw., and Prolaticorolla Ietsw. were determined, by classification with the cytogenetic method. The chromosome number of all taxa except O. sipyleum (2n = 28) and O. rotundifolium (2n = 28) is 2n = 30. In addition, the hybrids and their parental species have 2n = 30 chromosome numbers. Also, the smallest chromosome length is 0.32 μm in O. sipyleum. The largest chromosome length is 2.02 μm in O. munitiflorum. The smallest total haploid length is 10.08 μm in O. vulgare subsp. hirtum and the largest value is 22.00 μm in O. haussknechtii. The smallest mean length is 0.33 μm in O. vulgare subsp. hirtum and O. saccatum. The largest mean length is 0.74 μm in O. sipyleum. The chromosome numbers obtained in this study support the speciation of Origanum members via homoploid hybridization. Finally, the somatic chromosome numbers of 10 taxa (including 2 hybrids), chromosome measurements of 22 taxa (including 2 hybrids), and ideograms of 19 taxa (including 2 hybrids) were for the first time performed in this study.

Induced cytomictic crosstalk behaviour among micro-meiocytes of cluster bean [Cyamopsis tetragonoloba (L.) Taub.]: Reasons and repercussions

Shefali Singh, Plant Genetics Laboratory, Department of Botany, University of Allahabad, India.

Accepted: 2020-04-13| Published Online: 2020-04-30 | DOI: 10.13128/caryologia-544


Cytomictic behaviour of chromosomes among pollen mother cells was observed in mutagenic studies in cluster bean (Cyamopsis tetragonoloba (L.) Taub.). The study of pollen mother cells (PMC) revealed various chromosomal aberrations among which cytomixis was notified due to its obtrusive peculiarity and is therefore given description in this article. Cytoplasmic and chromatin transmigration were discernible among contiguous or slightly distant PMCs through recreation of passage via direct cell-to-cell fusion or channel formation. This cytomictic phenomenon was invariably more pronounced at meiosis I as compared to meiosis II. Plasmodesmatal connections play a paramount role in aiding this behaviour by establishing intercellular crosstalks. The cellular intermingling resulted in syncyte cells which were identified due to their doubled size. Syncyte or unreduced PMC formation leading to unreduced fertile gametes is speculated to act as a possible way out for infraspecific polyploidization of species. Pollen fertility was computed, alongwith this heterosized of varying diameter were segregated. Large sized pollens were 2n pollens; where size difference is a consequence of cytomixis. Cytoplasmic connections among pollens were also observed sporadically. It is opined that syncyte formation and 2n pollen production have evolutionary significance.

Karyotype diversity of stingless bees of the genus Frieseomelitta (Hymenoptera, Apidae, Meliponini)

Marco Antonio Costa, Departamento de Ciências Biológicas, Universidade Estadual de Santa Cruz, Ilhéus, BA, Brazil.

Accepted: 2020-04-01| Published Online: 2020-05-01 | DOI: 10.13128/caryologia-610


Frieseomelitta (Ihering, 1912) is a genus of stingless bees (Hymenoptera, Apidae, Meliponini), distributed in the Nearctic and Neotropical regions. Specimens can be found in forests, cerrado, caatinga and mountainous regions. This genus has 16 species, of which 13 are recorded in Brazil. Cytogenetics has contributed to evolutionary studies of some Hymenoptera groups and although many Frieseomelitta species have been described, few species have been studied cytogenetically. The present study aims to contribute to the knowledge of the karyotype diversity of this genus, seeking to understand the possible evolutionary mechanisms that occurred in the diversification of the karyotype of this genus. Frieseomelitta portoi and Frieseomelitta trichocerata and Frieseomelitta doederleini showed diploid karyotypes with 2n = 30 chromosomes, similarly to all the species previously analyzed in the genus. Unprecedentedly, Frieseomelitta longipes showed 2n = 34. These results confirm that the frequent diploid number of 30 chromosomes is typical of this genus. The finding of 2n = 34 chromosomes in F. longipes comprises the first record of a diploid chromosome number different from 2n=30 in this group, which suggests that it can be the result of a recent chromosome change event. An interspecific comparative analysis was developed involving present and previous studies, as well as a discussion on the mechanisms involved in the karyotypic evolution in the genus.

Effects of high temperature on mitotic index, microtubule and chromatin organization in rye (Secale cereale L.) root-tip cells

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

Accepted: 2020-03-27| Published Online: 2020-05-02 | DOI: 10.13128/caryologia-788


Stressful high temperatures on plants can limit whole-plant function and decrease crop productivity. However, little is known regarding heat stress effects on microtubule cytoskeleton and chromatin in roots from intact plants. Here we studied high-temperature effects on cell division, microtubule, and chromatin organization patterns in rye root tips from intact plants subjected to 40ºC for 4 h and after different recovery periods (0RT, 7RT, 24 RT). We showed that heat stress-induced changes in nuclear morphology as detected by the unusual presence of interphase cells with irregularly shaped nuclei, probably associated with changes in chromosome segregation at anaphase, leading to micronuclei formation as well as changes in the mitotic index.  These alterations were associated with differential effects in microtubules organization in both heat-stressed interphase and mitotic cells at 0RT and 7RT. Although no changes in the distribution of H3 phosphorylation of Ser 10 residues on chromatin were found in cells from heat-stressed plants, marked alterations in chromatin DNA methylation patterns were detected. These effects included higher agglutination of 5-methylcytosine domains in both interphase and metaphase cells compared to controls. Taken together these results seem to suggest that alterations in microtubule conformation upon heat stress influences nuclear chromatin organization and cell cycle progression. However, when seedlings recovered from stress (24RT), root tip cells presented microtubule configurations and chromatin organization patterns similar to controls. We conclude that in spite of heat stress markedly altered cell cycle progression and distribution of epigenetic marks, these responses are transient to cope with such stress conditions in the roots.