Karyological Studies Research Articles

Karyological studies of four species of Lady’s Slipper Orchids (Paphiopedilum) collected in the Bogor Botanical Garden, Indonesia

Paphiopedilum is one of the most widely grown and hybridized of all orchid genera due to its distinctive flower morphology. This genus consists of 139 accepted species and is native to southern China to tropical Asia. Karyological studies on the genus Paphiopedilum have been reported by many cytologists in different countries. However, many Indonesian members of Paphiopedilum have remained comparatively limited in investigated cytologically. This study aimed to analyze karyological characters of four species of Paphiopedilum collected in the Bogor Botanical Garden, Indonesia, namely P. armeniacum, P. hirsutissimum, P. primulinum, and P. superbiens. Karyological studies were conducted by root tips squash method. The results showed that four species of Paphiopedilum have a basic chromosome number of x = 15. They are diploid with 2n=30, instead of the common diploid chromosome number of Paphiopedilum (2n = 26). Paphiopedilum hirsutissimum and P. primulinum possess 16 m + 14 sm chromosome formulae. Whereas Paphiopedilum armeniacum and P. superbiens revealed 16 m + 14 sm and 18 m + 12 sm chromosome formulae, respectively.A Robertsonian change in chromosome number generated by the fission of chromosomes would best explain the origin of the new diploid chromosome number and karyotypes of these species.

The role of karyosystematics in the taxonomy of the genera Sorex and Crocidura (Mammalia: Soricidae), and the state of chromosomal studies of shrews in the Russian Far East

Literature data on the chromosomal characteristics of Soricidae, genera Crocidura and Sorex are analyzed. The importance of karyological studies for shrew taxonomy at the species level in the Russian Far East is shown. For each species, information on taxonomy, distribution, collection site of studied samples and the structure of karyotypes is provided. The intraspecifi c variability of shrew karyotypes in the Russian Far East is discussed based on a comparison of chromosomal characteristics from diff erent populations located in Russia and other geographical regions.

Karyological Study of Acanthocephalus lucii (Echinorhynchida): The Occurrence of B Chromosomes in Populations from PCB-Polluted Waters

In this study, we performed a cytogenetic analysis of Acanthocephalus lucii specimens from three sites with different levels of environmental pollution. Standard and fluorochrome staining (CMA3/DAPI), fluorescence in situ hybridization (FISH) with 18S rDNA and histone H3 probes, and silver impregnation were performed. Chromosome complements of 2n = 7/8 (male/female), n = 1m + 2sm + 1a (X), and CMA3-positive bands in all chromosomes were found in all three populations. FISH revealed one 18S rDNA locus on the X chromosome and one locus of H3 histone genes on the first chromosome pair. At the intraspecific level, the populations differed in the presence of supernumerary B chromosomes, which were found in all specimens from Zemplínska Šírava and in 89.4% of specimens from the Laborec River, but not at the reference site. The first two sites are considered to be water bodies with high toxin contamination. Based on this fact, we assume an increased frequency of chromosome breaks leading to the formation of DNA fragments that have the potential to form B chromosomes. The present results add to the very limited data on the organization of multigene families in the genome of Acanthocephala and suggest a possible causal link between water pollution and the occurrence of B chromosomes in fish parasites.

Karyological Study of the Isolated Scots Pine (<i>Pinus sylvestris</i>) Population of the Shirinskaya Steppe of the Republic Of Khakasia

The results of a karyological study of a peripheral isolated population of Scots pine (Pinus sylvestris L.) growing in the Shirinskaya steppe of Khakassia are presented. In the seed progeny of trees, disturbances of the number of chromosomes (myxoploidy) and a wide range of chromosomal rearrangements, including unique multiple aberrations, were revealed. For the first time at the population level, the highest occurrence of genomic and chromosomal rearrangements for this species was diagnosed. In fact, all seed progeny of the studied Scots pine population is mutant, which indicates probable adaptive changes in genotypes under isolation.

Chromosomal analysis of Sander lucioperca (L., 1758) (Perciformes: Percidae) from Turkey

Sander lucioperca (pikeperch) is a percid fish species of high commercial value and potential for being aquaculture in Turkey. However, karyological studies are deficient for population of Turkey. So, the aim of this study is to carry out diploid chromosome number, karyotype formula, fundamental arm number and chromosomal banding properties (with C-banding and Ag-NOR staining) of S. lucioperca. Specimens of S. lucioperca were captured from Konya, Turkey and alive specimens carried to the laboratory. Chromosome obtaining was provided by using air-drying technique from the head kidney. Chromosome slides were prepared and banding procedures were applied. Result of the analysis, diploid chromosome number was found as 48 and karyotype of the pikeperch consist of 32 biarmed and 16 uniarmed chromosomes. Constitutive heterochromatin regions were observed on the pericentromeres of some of the chromosomes. Ag-NORs were determined on one pair of submetacentric chromosome. This report is the first that determines chromosomal properties of S. lucioperca from Turkey. This study may contribute the cytogenetic information of this species.

Intraspecific karyomorphological and genome size variations of in vitro embryo derived Iranian endemic Asafoetida (Ferula assa-foetida L., Apiaceae)

Asafoetida (Ferula assa-foetida L.) is one of the endemic medicinal plants in Iran. Analysis of karyomorphology and 2Cx DNA measurements (monoploid genome size) of 18 Iranian endemic Ferula assa-foetida populations were performed. The in vitro embryo-derived root tips were examined for karyological studies, via technique of squash and stain with 2% (w/v) aceto-orcein. Seeds of the Ferula samples and leaves of Solanum lycopersicum as standard reference (2C DNA = 1.96 pg) were stained with propidium iodide (PI), using flow cytometric (FCM) technique. All the studied populations were diploids (2n = 2x = 22) with mean chromosome length (CL) of 3.95 μm, varied from 3.05 μm (P7) to 4.94 μm (P18). The mean total chromosome volume (TCV) was 0.98 μm3, ranged from 0.47 μm3 (P7) to 1.57 μm3 (P3). Two-typed chromosomes (“m”, “sm”) formed three classes of karyotype formula. Karyotypes were mostly symmetrical and fell in 1A and 2A Stebbins category. The monoploid genome size of Iranian endemic Ferula assa-foetida populations is being stated for the first time; its mean value was 4.51 pg, ranged from 4.09 (P4) to 4.69 pg (P16). Intraspecific karyomorphological and genome size variations were clearly confirmed in studied Ferula assa-foetida.

Karyological studies of Iranian Allium (Amaryllidaceae) species with a focus on sect. Acanthoprason. 2. Meiotic chromosomes

Meiosis in pollen mother cells (PMCs) was studied of 23 Iranian Allium species (33 accessions, 105 individuals) that belong to two subgenera and six sections. Materials of 13 species were sampled from (near) type locations. Gametic chromosome numbers, chromosome configurations at metaphase I, chiasma frequency, as well as type and percentages of abnormalities were recognized. The basic chromosome number for all taxa investigated was x = 8. Most taxa were diploid and showed eight bivalents or in rare cases two or four pairs of univalents, but in A. subakaka, A. ubipetrense and A. zagricum tetravalents also occurred. Meiosis in less than 10% of PMCs of diploid accessions was disturbed displaying lagging chromosomes, chromatid bridges, micronuclei, or unbalanced chromosome segregations, but very rarely more than one kind of irregularities were found within one dividing cell. One to three B chromosomes were found in 11 accessions, and were recognized for the first time in A. alamutense, A. elburzense and A. iranshahrii. Our data showed no correlation between the occurrence of B chromosomes and the chiasma frequency, and also no noticeable effect of habitat factors on meiotic chromosome behavior. The studied accessions of A. atroviolaceum and A. sabalense were tetraploid (n = 16) showing irregular meiosis in 20–69% of the PMCs which is regarded as sign of autopolyploidy. Since only two out of 31 investigated accessions belonging to subg. Melanocrommyum were tetraploid, we may suggest a trivial role of polyploidy in the evolution of this subgenus.

Genetic Differentiation of the Fourhorn Sculpin <i>Myoxocephalus quadricornis</i> (Linnaeus, 1758) and Its Position in the Tribe Myoxocephalini Taranetz (Cottidae: Myoxocephalinae)

A molecular genetic and karyological study of the fourhorn sculpin Myoxocephalus quadricornis (Linnaeus, 1758) was carried out. The karyotype was studied for the first time and the results of Ag-staining of the chromosomes of specimens of the species from the East Siberian Sea are presented. The karyotype of M. quadricornis is stable, contains 32 (10 metacentrics, 2 submetacentrics, 20 subtelocentrics) chromosomes, the number of chromosome arms is 44. Nucleolar organizers were found in the telomeric regions of the short arms of two homologous medium-sized subtelocentrics. Variability in the number of nucleolarforming chromosomes and stained blocks of nucleolar organizers was found. Common features in the chromosome sets of M. quadricornis and Megalocottus platycephalus indicate their greater similarity compared to species of the genus Myoxocephalus. The results of the karyological analysis are consistent with the molecular genetic data about the sister relationships between M. quadricornis and Megalocottus platycephalus. A complex of karyological and molecular-genetic features has been identified that allow one to reliably distinguish M. quadricornis from species of the tribe Myoxocephalini and indicate the need for its removal from the genus Myoxocephalus.

The Balkan endemic Centaurea grbavacensis (Asteraceae) ─ New evidence on variation, typification, and distribution, with karyological studies

The Balkan endemic Centaurea grbavacensis (Asteraceae) ─ New evidence on variation, typification, and distribution, with karyological studies

Mixed-Ploidy and Dysploidy in Hypericum perforatum: A Karyomorphological and Genome Size Study.

Karyomorphology and genome size of 15 St John's wort (Hypericum perforatum L.) populations are reported for the first time. Root tips and fresh young leaves were used for karyological studies and flow cytometric (FCM) measurements, respectively. The chromosome length varied from 0.81 µm to 1.16 µm, and chromosome types were determined as "m". Eight different somatic chromosome numbers were found (2n = 16, 22, 24, 26, 28, 30, 32, 38). Based on the observed basic (x) chromosome numbers of x = 8, 11, 13, 14, 15, 19, this may correspond to diploid (2x), triploid (3x), tetraploid (4x), respectively. Interestingly, we found mixoploidy (3x - 4x) in the root tips of one of the populations. Hybridization, polyploidy and dysploid variation may be the main factors associated with the chromosome number evolution of this species. FCM showed that 2C DNA contents vary from 0.87 to 2.02 pg, showing more than a 2-fold variation. The mean amount of 2C DNA/chromosome and the mean of monoploid genome size were not proportional to ploidy.

Morphological, Anatomical, Palynological and Karyological Studies on Endemic Alyssum kaynakiae (Brassicaceae) from Southwest Region of Turkey

Alyssum kaynakiae is an endemic biennial species growing in Southwest Anatolia, Turkey. In this study, the morphology of this taxon was revised, and its anatomical, palynological, and karyological characteristics were analysed for the first time. Alyssum kaynakiae has been considered part of the traditional Alyssum sect. Gamosepalum. Alyssum kaynakiae has diploid chromosome number 2n=16. It has a primary structure in the root and stem, collenchyma located under the epidermis in the root and under the cortex layer in the stem, and vascular bundles varying between 9 to 11. Leaves are isolateral, stomata are at the same level as the epidermis, and both sides of the leaves have lepidote hairs. Its palynological features are compatible with the Brassicaceae family, with its pollen type being tricolpate, its pollen shape being prolate, and its ornamentation being reticulate.

Karyology and Genome Size Analyses of Iranian Endemic Pimpinella (Apiaceae) Species.

Pimpinella species are annual, biennial, and perennial semibushy aromatic plants cultivated for folk medicine, pharmaceuticals, food, and spices. The karyology and genome size of 17 populations of 16 different Pimpinella species collected from different locations in Iran were analyzed for inter-specific karyotypic and genome size variations. For karyological studies, root tips were squashed and painted with a DAPI solution (1 mg/ml). For flow cytometric measurements, fresh leaves of the standard reference (Solanum lycopersicum cv. Stupick, 2C DNA = 1.96 pg) and the Pimpinella samples were stained with propidium iodide. We identified two ploidy levels: diploid (2x) and tetraploid (4x), as well as five metaphase chromosomal counts of 18, 20, 22, 24, and 40. 2n = 24 is reported for the first time in the Pimpinella genus, and the presence of a B-chromosome is reported for one species. The nuclear DNA content ranged from 2C = 2.48 to 2C = 5.50 pg, along with a wide range of genome sizes between 1212.72 and 2689.50 Mbp. The average monoploid genome size and the average value of 2C DNA/chromosome were not proportional to ploidy. There were considerable positive correlations between 2C DNA and total chromatin length and total chromosomal volume. The present study results enable us to classify the genus Pimpinella with a high degree of morphological variation in Iran. In addition, cytological studies demonstrate karyotypic differences between P. anthriscoides and other species of Pimpinella, which may be utilized as a novel identification key to affiliate into a distinct, new genus – Pseudopimpinella.

Somaclonal variation in pomegranate (Punica granatum L.): ISSR and cytological evidences

New advancements in tissue culture techniques have resulted in vitro propagation of several regenerations of horticultural crops. The somaclonal variation which occurs during plant tissue culture is a potential source for inducing new genetic variability in such crops. Several approaches have been utilized to ascertain the occurrence of somaclonal variation. The pomegranate plant can adapt to various environmental conditions and grow well in different kinds of soil and climate. Pomegranate Zagheh cultivar is mainly cultivated in the state of Esfehan, Kashan city in Iran. The present study was performed to investigate the potential occurrence of somaclonal variation in tissue culture regenerated plants of the cultivar Zagheh and provide morphological, cytological and molecular evidences for its occurrence. Nodal segments were chosen for the in vitro propagation of pomegranate. Explants were cultured on MS basal medium. We carried out both molecular (ISSR) and karyological studies in propagation collected. In the present study, we observed variation in both morphological (length of shoot, branch, internode, leaf and root and number of branch and root) and genetic features of the tissue culture regenerated plants. The regenerated plants of the third sub-culture differed from mother plants in the ISSR marker. We observed frequent occurrence of cells with different chromosome numbers, ranging from normal diploid 2n = 2x = 16 to tetraploid 2n = 32 chromosome number.

The morphological, karyological and phylogenetic analyses of three Artemisia L. (Asteraceae) species that around the Van Lake in Turkey

Artemisia is one of the biggest genera in the family Asteraceae, with around 500-600 taxa at specific and sub-specific levels and organised in five subgenera. Due to the high number of taxa, a lot taxonomists are trying to solve the problem of its classification and phylogeny but its natural classification still has not been achieved. The aim of this study is to try to solve the problematic systematic relationship between three different Artemisia species growing in close proximity to each other in the light of morphological, karyological and molecular data. The roots, stems, leaves, flowers structures of the plant samples collected from different populations belong to these species were investigated within the framework of morphological studies. Additionally, the chromosome counts and karyotype analysises of these species were made and idiograms were drawn in the karyological studies. In the context of phylogenetic studies, ITS (ITS1-5.8S-ITS2) and trnT - trnL3’ regions of 22 individuals belonging to 3 taxa were studied. According to results of phylogenetic anlysis, it has been found that there is completed speciation genetic isolation mechanism between the species Artemisia spicigera, Artemisia taurica and Artemisia fragrans that inhibit gene flow. Also Artemisia fragrans and Artemisia spicigera species are very similar to each other in terms of morphological characteristics. However, since populations of the species Artemisia fragrans are autopolyploid, the dimensional values of their morphological squares are larger than those of the species Artemisia spicigera. This study is important as it is the first molecular based study relating with some species of Artemisia growing naturally in Turkey.

A Karyological Study of Tenualosa Ilisha (Hamilton, 1822) From the Confluence of Padma and Meghna River of Bangladesh

Tenualosa ilisha (Hamilton, 1822), commonly known as Hilsha shad is a valuable and highly acceptable species in terms of their high flavored properties. Hilsha shad has striking morpho-genetical adaptation to heterogeneous habitats across their migratory routes. Cytogenetic analysis demonstrates the changes in chromosomes. But none was focused on the cytogenetic analysis of T. ilisha in Bangladesh. T. ilisha was found to possess 2n = 42 number of chromosomes along with a karyotype formula: 1M + 31m + 8sm + 2st using giemsa staining technique. The results demonstrated ‘diffuse type of interphase nuclei, co-existence of continuous type and interstitial type of prophase chromosomes respectively. No heteromorphic sex chromosomes were determined cytologically. The presence of diverse types of chromosomes based on centromeric position, gradual decrease in total haploid chromosome complement, mean centromeric asymmetry, coefficient of variation of chromosome length and Stebbins’s classification highlighted its asymmetry in karyotype with advance nature. Therefore, the elemental karyological data will offer information for the proper identification, cytotaxonomical classification, expanding productivity and preservation of genetic resources ofT. ilisha. Bangladesh J. Zool. 49 (2): 243-255, 2021

A new species of Calomyscus Thomas, 1905 (Calomyscidae: Rodentia) from western Iran

Eight extant species have been so far recognized for Calomyscus Thomas, 1905 which is distributed in the west of Asia. The only species which was reported from the Zagros Mountains in western and southern Iran was C. bailwardi. However, based on the integrative methods such as molecular, morphometric and karyological studies a new species is worth to be described other than C. bailwardi from the Zagros Mountains. In the present survey a new species which has been distributed in western regions of Iran was described as Calomyscus behzadi sp. nov. The new species constituted a monophyletic clade in phylogenetic tree which its relationships are unresolved. A new karyotype was reported for the new species as 2N=44 and FNa=48. The results of multivariate statistical analyses also separated the new species from C. bailwardi and other recognized species of the genus Calomyscus. Based on the size + shape data, C. behzadi sp. nov. has shorter nasal than C. bailwardi, group D and C. grandis and narrower than C. baluchi but wider than C. hotsoni and C. elburzensis. Moreover, according to the shape data, the new species has shorter m2 than C. elburzensis, C. bailwardi, C. hotsoni, C. grandis and group D and narrower than C. bailwardi, C. grandis and C. baluchi. The new species was described from regions which had been known already as distribution area of C. bailwardi.

Karyological studies of Siberian species of the genus Viola L.

The article presents an analysis of chromosome numbers in Siberian species of the genus Viola (40 speciesfrom 12 sections) based on numerous literary sources and original data. In addition, hybrid forms from the collection site“Rare and endangered Plant Species” of the Central Siberian Botanical Garden (Novosibirsk) were analyzed. Just as theroots of germinated seeds, so the roots of adult plants from an open ground area were studied. Samples of different originswere investigated: Siberia, the Far East, Mongolia, Belarus, Korea, Japan, the Caucasus, in some taxa from remote points ofthe area (for example, V. selkirkii). This allowed us to reveal the variability of the chromosome numbers for some species.Chromosomes in species of this genus are small, 0.8–1.2 mkm long. This involves difficulties to count them. The presenceof mixoploids in some sections and the polybasic state of chromosome numbers for a number of species were established.Polymorphic species in the Violidum section are characterized by polyploid races. The variability of chromosome numbersin taxa reflects their ability to hybridize, as well as the processes of speciation. However, the predominance of autogamy formost species of the genus prevents intensive hybridization. Endemic and tertiary relicts are characterized by a stable number of chromosomes and they are tetraploids.

SPECIES DELIMITATION IN NICOTIANA SECT. SUAVEOLENTES (SOLANACEAE): RECIPROCAL ILLUMINATION LEADS TO RECOGNITION OF MANY NEW SPECIES

SummaryIn this volume of Curtis's Botanical Magazine, we increase the number of species recognised in Nicotiana sect. Suaveolentes to 38, up from the 21 recorded in the Flora of Australia published 39 years ago, but we estimate the final number is likely to exceed 60. We examine the reasons why so many unrecognised species exist. Several Australian and American botanists have previously specialised in Nicotiana but did not detect such diversity. We have carried out many fieldtrips, seen most known species in the wild and have grown these accessions from diverse areas side‐by‐side in cultivation to avoid basing decisions about species characteristics on features heavily influenced by environmental plasticity. In the end, we conclude that it is a mixture of focused and extensive fieldwork and more recent genomic, genetic and karyological studies combined in the process of reciprocal illumination, that has enabled us to describe so many new species. We also briefly focus on the topic of why Nicotiana is so species‐rich in Australia.

Karyotype Analysis of Populations in Five Satureja Species from Iran

A karyological study of five taxa (15 populations) of the genus Satureja L. from different geographic origins is presented. The haploid levels were different among species (n= 12 and 15). The work deals with chromosome number and morphometry. We found two of the usual basic chromosome numbers in this study for satureja species, x=12 and x=15. All populations were diploid and were located in 1B or 2B Stebbins classes. The karyotypic results showed the diversity among the species as mostly displayed median region (m) and submedian region (sm) and the chromosome lengths were determined between 0.60 µm in S. sahendica (1041) to 2.27 µm in S. macrantha (3). Detailed karyotype analysis allows us to group the different populations and determine the relationships among them. It also pointed out the possibility of inter and intra-species cross to improve plants.

Morpho-taxonomical and karyological study of Gopi banana: an indigenous palatable cultivated variety of Musa × paradisiaca

Morpho-taxonomical and karyological study of Gopi banana: an indigenous palatable cultivated variety of Musa × paradisiaca