scholarly journals Comparative molecular cytogenetic characterization of five wild Vigna species (Fabaceae)

2020 ◽  
Vol 14 (2) ◽  
pp. 243-264
Author(s):  
Chao-Wen She ◽  
Ying Mao ◽  
Xiang-Hui Jiang ◽  
Chun-Ping He
Keyword(s):  
Genomic Dna ◽  
Interspecific Variation ◽  
Rrna Genes ◽  
Fluorochrome Banding ◽  
45S Rdna ◽  
Molecular Cytogenetic ◽  
Rdna Sites ◽  
Cytogenetic Characterization ◽  
Rdna Fish

To extend our knowledge on karyotype variation of the genus Vigna Savi, 1824, the chromosomal organization of rRNA genes and fluorochrome banding patterns of five wild Vigna species were studied. Sequential combined PI (propidium iodide) and DAPI (4',6-diamidino-2-phenylindole) (CPD) staining and fluorescence in situ hybridization (FISH) with 5S and 45S rDNA probes were used to analyze the karyotypes of V. luteola (Jacquin, 1771) Bentham, 1959, V. vexillata (Linnaeus, 1753) A. Richard, 1845, V. minima (Roxburgh, 1832) Ohwi & H. Ohashi, 1969, V. trilobata (Linnaeus, 1753) Verdcourt, 1968, and V. caracalla (Linnaeus, 1753) Verdcourt,1970. For further phylogenetic analysis, genomic in situ hybridization (GISH) with the genomic DNA of V. umbellata (Thunberg, 1794) Ohwi & H.Ohashi, 1969 onto the chromosomes of five wild Vigna species was also performed. Detailed karyotypes were established for the first time using chromosome measurements, fluorochrome bands, and rDNA-FISH signals. All species had chromosome number 2n = 2x = 22, and symmetrical karyotypes that composed of only metacentric or metacentric and submetacentric chromosomes. CPD staining revealed all 45S rDNA sites in the five species analyzed, (peri)centromeric GC-rich heterochromatin in V. luteola, V. trilobata and V. caracalla, interstitial GC-rich and pericentromeric AT-rich heterochromatin in V. caracalla. rDNA-FISH revealed two 5S loci in V. caracalla and one 5S locus in the other four species; one 45S locus in V. luteola and V. caracalla, two 45S loci in V. vexillata and V. trilobata, and five 45S loci in V. minima. The karyotypes of the studied species could be clearly distinguished by the karyotypic parameters, and the patterns of the fluorochrome bands and the rDNA sites, which revealed high interspecific variation among the five species. The V. umbellata genomic DNA probe produced weak signals in all proximal regions of V. luteola and all (peri)centromeric regions of V. trilobata. The combined data demonstrate that distinct genome differentiation has occurred among the five species during evolution. The phylogenetic relationships between the five wild species and related cultivated species of Vigna are discussed based on our present and previous molecular cytogenetic data.

scholarly journals Molecular cytogenetic characterization and phylogenetic analysis of four Miscanthus species (Poaceae)

2019 ◽  
Vol 13 (3) ◽  
pp. 211-230 ◽  
Author(s):  
Yan-Mei Tang ◽  
Liang Xiao ◽  
Yasir Iqbal ◽  
Jian-Feng Liao ◽  
Long-Qian Xiao ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Phylogenetic Trees ◽  
Repetitive Sequences ◽  
Comparative Genomic ◽  
45S Rdna ◽  
Molecular Cytogenetic ◽  
Rdna Sites ◽  
Molecular Phylogenetic ◽  
Cytogenetic Characterization

Chromosomes of four Miscanthus (Andersson, 1855) species including M. sinensis (Andersson, 1855), M. floridulus (Schumann & Lauterb, 1901), M. sacchariflorus (Hackel, 1882) and M. lutarioriparius (Chen & Renvoize, 2005) were analyzed using sequentially combined PI and DAPI (CPD) staining and fluorescence in situ hybridization (FISH) with 45S rDNA probe. To elucidate the phylogenetic relationship among the four Miscanthus species, the homology of repetitive sequences among the four species was analyzed by comparative genomic in situ hybridization (cGISH). Subsequently four Miscanthus species were clustered based on the internal transcribed spacer (ITS) of 45S rDNA. Molecular cytogenetic karyotypes of the four Miscanthus species were established for the first time using chromosome measurements, fluorochrome bands and 45S rDNA FISH signals, which will provide a cytogenetic tool for the identification of these four species. All the four have the karyotype formula of Miscanthus species, which is 2n = 2x = 38 = 34m(2SAT) + 4sm, and one pair of 45S rDNA sites. The latter were shown as strong red bands by CPD staining. A non-rDNA CPD band emerged in M. floridulus and some blue DAPI bands appeared in M. sinensis and M. floridulus. The hybridization signals of M. floridulus genomic DNA to the chromosomes of M. sinensis and M. lutarioriparius genomic DNA to the chromosomes of M. sacchariflorus were stronger and more evenly distributed than other combinations. Molecular phylogenetic trees showed that M. sinensis and M. floridulus were closest relatives, and M. sacchariflorus and M. lutarioriparius were also closely related. These findings were consistent with the phylogenetic relationships inferred from the cGISH patterns.

scholarly journals Establishment and Optimization of Molecular Cytogenetic Techniques (45S Rdna-FISH, GISH And Fiber-FISH) In Kiwifruit (Actinidia L.)

2021 ◽  
Author(s):  
Yang Zhao ◽  
Honghong Deng ◽  
Yao Chen ◽  
Jihan Li ◽  
Silei Chen ◽  
...  
Keyword(s):  
Fiber Quality ◽  
Molecular Cytogenetics ◽  
45S Rdna ◽  
Metaphase Chromosomes ◽  
Physical Size ◽  
Molecular Cytogenetic ◽  
Rdna Fish ◽  
Molecular Cytogenetic Techniques ◽  
First Time

Abstract Background: Kiwifruit has long been regarded as ‘the king of fruits’ for its nutritional importance. However, the molecular cytogenetics of kiwifruit has long been hampered because of the large number of basic chromosome (x=29), the inherent small size and highly similar morphology of metaphase chromosomes. Fluorescence in situ hybridization (FISH) is an indispensable molecular cytogenetic technique widely used in many plant species. Herein, the effects of post-hybridization washing temperature on FISH, blocking DNA concentration on genomic in situ hybridization (GISH), extraction method on nuclei isolation and the incubation time on the DNA fiber quality in kiwifruit were evaluated.Results: The post-hybridization washing in 2×SSC solution for 3×5 min at 37 ˚C ensured high stringency and distinct specific FISH signals in kiwifruit somatic chromosomes. The use of 50× blocking DNA provided an efficient and reliable means of discriminating between chromosomes derived from in the hybrids of A. chinensis var. chinensis (2n=2x=58) × A. eriantha Benth (2n=2x=58), and inferring the participation of parental genitors. The chopping method established in the present study were found to be very suitable for preparation of leaf nuclei in kiwifruit. A high-quality linear DNA fiber was achieved by an incubation of 20 min. The physical size of 45S rDNA signals was approximately 35-40 μmm revealed by the highly reproducible fiber-FISH procedures established and optimized in this study.Conclusions: The molecular cytogenetic techniques (45S rDNA-FISH, GISH, and high-resolution fiber-FISH) for kiwifruit was for the first time established and optimized in the present study, which is the foundation for the future genomic and evolutionary studies.

Comparative analysis of rDNA distribution in 29 species of Begonia sect. Coelocentrum Irmsch.

Phytotaxa ◽  
2018 ◽  
Vol 381 (1) ◽  
pp. 141 ◽  
Author(s):  
YAN-LI HAN ◽  
DAI-KE TIAN ◽  
NAI-FENG FU ◽  
YAN XIAO ◽  
ZONG-YUN LI ◽  
...  
Keyword(s):  
5S Rdna ◽  
Distribution Patterns ◽  
Hybrid Origin ◽  
45S Rdna ◽  
Species Relationships ◽  
Rdna Sites ◽  
Rdna Fish ◽  
5S And 45S Rdna ◽  
Chromosome Landmarks

The rDNA sites are useful chromosome landmarks and can provide valuable information for species identification and species relationships. In this study, we investigated the distribution of 5S and 45S rDNA sites in 29 species of Begonia sect. Coelocentrum Irmsch. using a two-colour fluorescence in situ hybridization (FISH) technique. This is the first report of chromosomal rDNA mapping in Begonia species. The analyzed species showed considerable diversity in rDNA distribution patterns. The 45S rDNA signals are always located in terminal regions on 1−4 chromosomes, while 5S rDNA signals are mainly located at proximal regions on 2−8 chromosomes, varying from specific major signals to highly dispersed minor signals. Based on rDNA FISH patterns, most of the investigated species could be distinguished from each other and species relationships were identified. In addition, the results provided clear proof that B. huangii is of hybrid origin and the triploid B. longgangensis was allotriploid rather than autotriploid as suggested before. The data will provide a useful reference for evaluation, conservation and utilization of the natural resources of the mega-diverse genus Begonia.

scholarly journals Insight into octoploid strawberry (Fragaria) subgenome composition revealed by GISH analysis of pentaploid hybrids

Genome ◽  
2016 ◽  
Vol 59 (2) ◽  
pp. 79-86 ◽  
Author(s):  
Bo Liu ◽  
Elizabeth G. Poulsen ◽  
Thomas M. Davis
Keyword(s):  
In Situ Hybridization ◽  
Genomic Dna ◽  
Genomic In Situ Hybridization ◽  
Molecular Cytogenetic ◽  
Phylogenetic Studies ◽  
Cytogenetic Studies ◽  
Rdna Fish ◽  
25S Rdna ◽  
Insight Into

As the product of interspecific hybridization between its two ancestral octoploid (2n = 8x = 56) species (Fragaria chiloensis and F. virginiana), the cultivated strawberry (F. ×ananassa) is among the most genomically complex of crop plants, harboring subgenomic components derived from as many as four different diploid ancestors. To physically visualize the octoploids’ subgenome composition(s), we launched molecular cytogenetic studies using genomic in situ hybridization (GISH), comparative GISH (cGISH), and rDNA-FISH techniques. First, GISH resolution in Fragaria was tested by using diploid and triploid hybrids with predetermined genome compositions. Then, observation of an octoploid genome was implemented by hybridizing chromosomes of pentaploid (2n = 5x = 35) hybrids from F. vesca × F. virginiana with genomic DNA probes derived from diploids (2n = 2x = 14) F. vesca and F. iinumae, which have been proposed by phylogenetic studies to be closely related to the octoploids yet highly divergent from each other. GISH and cGISH results indicated that octoploid-derived gametes (n = 4x = 28) carried seven chromosomes with hybridization affinities to F. vesca, while the remaining 21 chromosomes displayed varying affinities to F. iinumae, indicating differing degrees of subgenomic contribution to the octoploids by these two putatively ancestral diploids. Combined rDNA-FISH revealed severe 25S rDNA loss in both the F. vesca- and F. iinumae-like chromosome groups, while only the prior group retained its 5S loci.

Divergence between C. melo and African Cucumis Species Identified by Chromosome Painting and rDNA Distribution Pattern

2016 ◽  
Vol 150 (2) ◽  
pp. 150-155 ◽  
Author(s):  
Kunpeng Li ◽  
Huaisong Wang ◽  
Jiming Wang ◽  
Jianying Sun ◽  
Zongyun Li ◽  
...  
Keyword(s):  
Diploid Species ◽  
Distribution Patterns ◽  
45S Rdna ◽  
Rdna Sites ◽  
Cucumis Species ◽  
Cytogenetic Evidence ◽  
Rdna Fish ◽  
5S And 45S Rdna ◽  
Painting Probes

The 5S and 45S rDNA sites are useful chromosome landmarks and can provide valuable information about karyotype evolution and species interrelationships. In this study, we employed fluorescence in situ hybridization (FISH) to determine the number and chromosomal location of 5S and 45S rDNA loci in 8 diploid Cucumis species. Two oligonucleotide painting probes specific for the rDNA-bearing chromosomes in C. melo were hybridized to other Cucumis species in order to investigate the homeologies among the rDNA-carrying chromosomes in Cucumis species. The analyzed diploid species showed 3 types of rDNA distribution patterns, which provided clear cytogenetic evidence on the divergence between C. melo and wild diploid African Cucumis species. The present results not only show species interrelationships in the genus Cucumis, but the rDNA FISH patterns can also be used as cytological markers for the discrimination of closely related species. The data will be helpful for breeders to choose the most suitable species from various wild species for improvement of cultivated melon.

Comparative Karyotype Investigations in the European Crayfish Astacus astacus and A. leptodactylus (Decapoda, Astacidae)

Crustaceana ◽  
2011 ◽  
Vol 84 (12-13) ◽  
pp. 1497-1510 ◽  
Author(s):  
M. Pavlica ◽  
M. Mcžić ◽  
G. Klobučar ◽  
M. Šrut ◽  
I. Maguire ◽  
...  
Keyword(s):  
Chromosome Number ◽  
Chromosome Complement ◽  
Size Difference ◽  
45S Rdna ◽  
Astacus Astacus ◽  
Fluorochrome Staining ◽  
Rdna Sites ◽  
Freshwater Habitats ◽  
Acrocentric Chromosomes

AbstractThis study reports on the chromosome number and karyological characteristics of the endangered species of European crayfish, Astacus astacus and A. leptodactylus (Decapoda, Astacidae), both native to Croatian freshwater habitats. The karyotype of A. astacus and A. leptodactylus consists of 2n = 176 and 2n = 180 chromosomes, respectively. The haploid chromosome complement of A. astacus consists of 52 metacentric, 35 metacentric-submetacentric, and 1 acrocentric chromosomes. Fluorochrome staining with 4,6-diamino-2-phenylindole (DAPI) has revealed that the karyotypes of A. astacus and A. leptodactylus are characterized by large heterochromatic blocks located at centromeric and intercalary positions on the chromosomes. Interstitial heterochromatic blocks were more frequent in A. astacus than in A. leptodactylus. In both species pairing of chromosomes in meiosis was regular with the majority of bivalents in a ring- and a dumbbell-form. Fluorescence in situ hybridization (FISH) has revealed that two 45S rDNA loci were present in the investigated species. In A. astacus one of the two 45S rDNA-bearing chromosome pairs was highly heteromorphic, exhibiting a three-fold size difference between 45S rDNA sites on homologous chromosomes. Such a size difference was significantly less pronounced in A. leptodactylus. The karyotype differences between A. astacus and A. leptodactylus suggest changes in chromosome number as well as position of repetitive DNAs have played a role in the karyotype evolution of the species of Astacus.

Molecular Cytogenetic Characterization of the DiGeorge Syndrome Region Using Fluorescence in Situ Hybridization

Genomics ◽  
1993 ◽  
Vol 17 (2) ◽  
pp. 403-407 ◽  
Author(s):  
Elizabeth A. Lindsay ◽  
Stephanie Halford ◽  
Roy Wadey ◽  
Peter J. Scambler ◽  
Antonio Baldini
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Digeorge Syndrome ◽  
Molecular Cytogenetic ◽  
Cytogenetic Characterization

scholarly journals A populational survey of 45S rDNA polymorphism in the Jefferson salamander Ambystoma jeffersonianum revealed by fluorescence in situ hybridization (FISH)

2009 ◽  
Vol 55 (2) ◽  
pp. 145-149 ◽  
Author(s):  
Ke Bi ◽  
James P. Bogart ◽  
Jinzhong Fu
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Dispersal Ability ◽  
Habitat Isolation ◽  
45S Rdna ◽  
Isolated Populations ◽  
Ambystoma Jeffersonianum ◽  
Rdna Sites ◽  
Rdna Polymorphism

Abstract The chromosomal localization of 45S ribosomal RNA genes in Ambystoma jeffersonianum was determined by fluorescence in situ hybridization with 18S rDNA fragment as a probe (FISH-rDNA). Our results revealed the presence of rDNA polymorphism among A. jeffersonianum populations in terms of number, location and FISH signal intensity on the chromosomes. Nine rDNA cytotypes were found in ten geographically isolated populations and most of them contained derivative rDNA sites. Our preliminary study provides strong indication of karyotypic diversification of A. Jeffersonianum that is demonstrated by intraspecific variation of 45S rDNA cytotypes. rDNA cytotype polymorphism has been described in many other caudate amphibians. We predict that habitat isolation, low dispersal ability and decline of effective population size could facilitate the fixation and accumulation of variable rDNA cytotypes during their chromosome evolution.

Cytogenetic characterization by in situ hybridization techniques and molecular analysis of 5S rRNA genes of the European hazelnut (Corylus avellana)

Genome ◽  
2013 ◽  
Vol 56 (3) ◽  
pp. 155-159 ◽  
Author(s):  
E. Falistocco ◽  
G. Marconi
Keyword(s):  
Chromosome Structure ◽  
5S Rdna ◽  
Corylus Avellana ◽  
The Self ◽  
Rrna Genes ◽  
Additional Information ◽  
Cytogenetic Characterization ◽  
5S Rrna Genes ◽  
Corylus Avellana L

The European hazelnut (Corylus avellana L.) is widespread in Europe, where it has been cultivated for centuries. Despite progress in genetics, most of the cytogenetic aspects of this species have been overlooked. The aim of this study was to fill in this gap and obtain basic information on the chromosome structure of this species. Karyomorphological analysis confirmed the chromosome number 2n = 22 and showed that, despite their apparent uniformity, the chromosomes could be separated into three groups of different size: large (L), medium (M), and small (S). As a first step towards the physical mapping of the hazelnut chromosomes, we applied FISH to localize the position of rRNA genes (rDNA). The sites of 45S and 5S rDNA enabled us to identify two chromosome pairs belonging, respectively, to the L and S groups. The self-GISH procedure revealed that repetitive DNA is concentrated in the pericentromeric regions of the chromosomes, as with other species with rather small genomes. The analysis of 5S rDNA repeats offered additional information on the hazelnut genome by obtaining the whole sequence of the transcribed region so far unpublished. The overall results constitute a substantial advance in hazelnut cytogenetics. Further investigation of other species of Corylus could be an effective approach to understanding the phylogenesis of the genus and resolving taxonomic problems.

Comparative chromosomal localization of 45S and 5S rDNAs and implications for genome evolution in Cucumis

Genome ◽  
2016 ◽  
Vol 59 (7) ◽  
pp. 449-457 ◽  
Author(s):  
Zhen-Tao Zhang ◽  
Shu-Qiong Yang ◽  
Zi-Ang Li ◽  
Yun-Xia Zhang ◽  
Yun-Zhu Wang ◽  
...  
Keyword(s):  
Chromosomal Localization ◽  
5S Rdna ◽  
Chromosome Analysis ◽  
45S Rdna ◽  
Evolutionary Patterns ◽  
Site Number ◽  
Rdna Sites ◽  
Wild Accessions ◽  
Ribosomal Dnas

Ribosomal DNAs are useful cytogenetic markers for chromosome analysis. Studies investigating site numbers and distributions of rDNAs have provided important information for elucidating genome organization and chromosomal relationships of many species by fluorescence in situ hybridization. But relevant studies are scarce for species of the genus Cucumis, especially in wild species. In the present study, FISH was conducted to investigate the organization of 45S and 5S rDNA among 20 Cucumis accessions, including cultivars and wild accessions. Our results showed that the number of 45S rDNA sites varied from one to five pairs in different accessions, and most of these sites are located at the terminal regions of chromosomes. Interestingly, up to five pairs of 45S rDNA sites were observed in C. sativus var. sativus, the species which has the lowest chromosome number, i.e., 2n = 14. Only one pair of 5S rDNA sites was detected in all accessions, except for C. heptadactylus, C. sp, and C. spp that had two pairs of 5S rDNA sites. The distributions of 5S rDNA sites showed more variation than 45S rDNA sites. The phylogenetic analysis in this study showed that 45S and 5S rDNA have contrasting evolutionary patterns. We find that 5S rDNA has a polyploidization-related tendency towards the terminal location from an interstitial location but maintains a conserved site number, whereas the 45S rDNA showed a trend of increasing site number but a relatively conserved location.

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