scholarly journals Genome Mutation Revealed by Artificial Hybridization between Chrysanthemum yoshinaganthum and Chrysanthemum vestitum Assessed by FISH and GISH

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Magdy Hussein Abd El-Twab ◽  
Katsuhiko Kondo
Keyword(s):  
5S Rdna ◽  
45S Rdna ◽  
Multicolor Fish ◽  
Hybrid Plants ◽  
Artificial Hybrid ◽  
Yellow Color ◽  
Rdna Sites ◽  
Labeled Probe

Present study has been done to investigate artificial interspecific crossability between Japanese Chrysanthemum yoshinaganthum (2n=36) and Chinese C. vestitum (2n=54), which were cultured in vitro and in vivo and characterization of their artificial hybrid chromosomes and type of changes assessed by FISH and GISH. GISH was applied by using biotin-labeled total genomic DNA probe of C. vestitum, which were mixed with blocking DNA of C. yoshinaganthum. Approximately 18 yellow-green colored chromosomes of C. vestitum were detected by the probe, approximately 18 yellow-red- mixed colored chromosomes could be common chromosomes of the two species, and nine chromosomes were relatively red of Ch. yoshinaganthum that were not detected by the probe. The expected average of FISH six signals of 5S rDNA sites and ten of 45S rDNA were observed on the chromosomes of three and six hybrid plants, respectively. Multicolor FISH signals showed unexpected average of seven and 14 yellow signals of 5S rDNA sites on seven and thirteen chromosomes simultaneous with ten and 11 red signals of 45S rDNA sites on ten and 11 chromosomes which were detected by the probes respectively. FISH mapping of the 5S rDNA at terminal sites was detected in hybrid chromosomes, for the first time. Yellow-color signals of the telomeres were detected by the biotin-labeled probe of the PCR-amplified telomeric probe in interphase and terminal sites in metaphase. All chromosomes that showed terminal signals except four chromosomes showed subterminal sites of telomeres indicating the presence of translocations.

scholarly journals Simultaneous Visualization of 5S and 45S rDNAs in Persimmon (Diospyros kaki) and Several Wild Relatives (Diospyros spp.) by Fluorescent in situ Hybridization (FISH) and MultiColor FISH (MCFISH)

2003 ◽  
Vol 128 (5) ◽  
pp. 736-740 ◽  
Author(s):  
Young A Choi ◽  
Ryutaro Tao ◽  
Keizo Yonemori ◽  
Akira Sugiura
Keyword(s):  
Fluorescent In Situ Hybridization ◽  
Diploid Species ◽  
5S Rdna ◽  
Diospyros Kaki ◽  
45S Rdna ◽  
Multicolor Fish ◽  
Rdna Probe ◽  
Rdna Sites ◽  
5S And 45S Rdna

5S ribosomal DNA (rDNA) was visualized on the somatic metaphase chromosome of persimmon (Diospyros kaki) and ten wild Diospyros species by fluorescent in situ hybridization (FISH). The digoxigenin (DIG)-labeled 5S rDNA probe was hybridized onto the chromosomes and visualized by incubation with anti-DIG-fluorescein isothiocyanate (FITC). Strong signals of 5S rDNA probe were observed on several chromosomes of Diospyros species tested. Furthermore, multicolor FISH using 5S and 45S rDNA probes differently labeled with DIG and biotin, revealed separate localization of the two rDNA genes on different chromosomes of Diospyros species tested, suggesting that 5S and 45S rDNA sites can be used as chromosome markers in Diospyros. The number of 5S rDNA sites varied with the Diospyros species. More 5S rDNA sites were observed in four diploid species native to Southern Africa than in three Asian diploid species. The former had four or six 5S rDNA sites while the latter had two. Three Asian polyploidy species had four to eight 5S rDNA sites. Among the Asian species, the number of 5S rDNA sites seemed to increase according to ploidy level of species. These features of 5S rDNA sites were very similar to those of 45S rDNA sites in Diospyros. Phylogenetic relationship between D. kaki and wild species tested are discussed based on the number and chromosomal distribution of 5S and 45S rDNA.

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.

scholarly journals Analyses of the Updated “Animal rDNA Loci Database” with an Emphasis on Its New Features

2021 ◽  
Vol 22 (21) ◽  
pp. 11403
Author(s):  
Jana Sochorová ◽  
Francisco Gálvez ◽  
Roman Matyášek ◽  
Sònia Garcia ◽  
Aleš Kovařík
Keyword(s):  
Sex Chromosomes ◽  
5S Rdna ◽  
High Variability ◽  
Rrna Genes ◽  
45S Rdna ◽  
Data Analyses ◽  
Rdna Sites ◽  
Rdna Loci ◽  
Staining Methods ◽  
Active Nors

We report on a major update to the animal rDNA loci database, which now contains cytogenetic information for 45S and 5S rDNA loci in more than 2600 and 1000 species, respectively. The data analyses show the following: (i) A high variability in 5S and 45S loci numbers, with both showing 50-fold or higher variability. However, karyotypes with an extremely high number of loci were rare, and medians generally converged to two 5S sites and two 45S rDNA sites per diploid genome. No relationship was observed between the number of 5S and 45S loci. (ii) The position of 45S rDNA on sex chromosomes was relatively frequent in some groups, particularly in arthropods (14% of karyotypes). Furthermore, 45S rDNA was almost exclusively located in microchromosomes when these were present (in birds and reptiles). (iii) The proportion of active NORs (positively stained with silver staining methods) progressively decreased with an increasing number of 45S rDNA loci, and karyotypes with more than 12 loci showed, on average, less than 40% of active loci. In conclusion, the updated version of the database provides some new insights into the organization of rRNA genes in chromosomes. We expect that its updated content will be useful for taxonomists, comparative cytogeneticists, and evolutionary biologists. 

Chromosomal Mapping of Repeat DNA in Bergiaria westermanni (Pimelodidae, Siluriformes): Localization of 45S rDNA in B Chromosomes

2018 ◽  
Vol 154 (2) ◽  
pp. 99-106 ◽  
Author(s):  
Geovana C. Malimpensa ◽  
Josiane B. Traldi ◽  
Danyelle Toyama ◽  
Flávio Henrique-Silva ◽  
Marcelo R. Vicari ◽  
...  
Keyword(s):  
Chromosomal Rearrangements ◽  
Indirect Evidence ◽  
5S Rdna ◽  
B Chromosomes ◽  
Chromosomal Mapping ◽  
Interindividual Variation ◽  
Rrna Genes ◽  
45S Rdna ◽  
Rdna Sites ◽  
Repeat Dna

The occurrence of repetitive DNA in autosomes and B chromosomes of Bergiaria westermanni was examined using conventional and molecular cytogenetic techniques. This species exhibited 2n = 56 chromosomes, with intra- and interindividual variation in the number of heterochromatic B chromosomes (from 0 to 4). The 5S rDNA was localized in pairs 1 and 5, and histone probes (H1, H3, and H4) and U2 small nuclear RNA were syntenic with 5S rDNA in pair 5. Histone sequences were also located in chromosome pair 14. The (GATA)n sequence was dispersed throughout the autosomes and B chromosomes, with clusters (microsatellite accumulation) in some chromosome regions. The telomeric probe revealed no signs of chromosomal rearrangements in the genome of B. westermanni. The 45S rDNA sites were detected in the terminal region of pair 27; these sites corresponded to a GC-rich heterochromatin block. In addition, 3 of the 4 B chromosomes also contained 45S rDNA copies. Silver nitrate staining in interphase nuclei provided indirect evidence of the expression of these rRNA genes in B chromosomes, indicating the probable origin of these elements. This report shows plasticity in the chromosomal localization of repeat DNA in B. westermanni and features a discussion of genomic diversification.

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 Comparative chromosomal localization of 45S and 5S rDNAs in 76 purple-fleshed sweet potato cultivars

2020 ◽  
Author(s):  
Dan Su ◽  
Lei Chen ◽  
Jianying Sun ◽  
Luyue Zhang ◽  
Runfei Gao ◽  
...  
Keyword(s):  
Sweet Potato ◽  
Nutritional Value ◽  
Chromosome Structure ◽  
Chromosomal Localization ◽  
5S Rdna ◽  
Potato Cultivars ◽  
45S Rdna ◽  
Rdna Sites ◽  
Numerical Variation ◽  
Rdna Analysis

Abstract Background: In recent years, purple-fleshed sweet potato has been paid more and more attention because of its high nutritional value. However, the current studies on purple-fleshed sweet potato were still focused on the research and production of the related products. The research on its cytogenetics is relatively lagging behind, which cannot satisfy the study of genetic diversity of purple-fleshed sweet potato. Therefore, we carried out cytogenetic analysis on 76 purple-fleshed sweet potato cultivars, aim to analyze the chromosome structure and distribution of 45S rDNA and 5S rDNA in 76 purple-fleshed sweet potato cultivars.Results: We have found that only 62 purple-fleshed sweet potato cultivars with 90 chromosomes, and the others were aneuploid with 88, 89, 91, 92 chromosomes. The number of 45S rDNA in 76 purple-fleshed sweet potato cultivars varies from 16 to 21, with different signal sizes and intendities, and localized at the terminal or satellite of chromosomes. The number of 5S rDNA were relatively stable, 74 of the varieties investigated contained 6 sites, located at the terminal of chromosomes and near centromere. Only the Quanzishu 96 has 7 5S rDNA sites, and Yuzixiang 10 has 5 5S rDNA sites. In addition, rDNA analysis was also performed on two parents of Quanzishu 96. Both the two parents had 18 45S rDNA sites and 6 5S rDNA sites, which were different from the results of Quanzishu 96.Conclusions: For hexaploid sweet potato cultivars, there is genetic instability between purple-fleshed sweet potato cultivars. The 45S rDNA sites showed numerical variation, whereas conserved number of 5S rDNA sites were observed.

scholarly journals Molecular and cytogenetic description of somatic hybrids between Gentiana cruciata L. and G. tibetica King

2019 ◽  
Vol 61 (1) ◽  
pp. 13-24
Author(s):  
Karolina Tomiczak
Keyword(s):  
In Situ Hybridization ◽  
Somatic Hybrids ◽  
Nuclear Dna Content ◽  
5S Rdna ◽  
Genetic Constitution ◽  
Fusion Partner ◽  
Rdna Sites ◽  
Gentiana Cruciata

AbstractSomatic hybridization provides an opportunity to create cells with new genetic constitution. Here, the interspecific somatic hybrid plants regenerated in vitro following fusion of cell suspension–derived protoplasts of tetraploid Cross Gentian (Gentiana cruciata L., 2n = 52) with protoplasts released from mesophyll tissue of another tetraploid species, Tibetan Gentian (G. tibetica King, 2n = 52), were studied. According to the results of genome analyses with AFLP, ISSR, and CAPS markers, all somatic hybrids were genetically closer to “suspension” fusion partner G. cruciata than to “mesophyll” partner G. tibetica, but they got G. tibetica chloroplasts. Chromosome counting revealed little variation in the number of chromosomes in hybrid’s cells (2n = 88 or 2n = 90), although all plants possessed similar nuclear DNA content which remained stable even after 2 years of in vitro culture. Fluorescence in situ hybridization (FISH) showed that hybrids possessed 4 to 7 chromosomes bearing 5S rDNA sites and 6 or 7 chromosomes with 35S rDNA sites. A part of FISH signals was smaller than those observed in the parental species, which could indicate the loss of rDNA sequences. Genomic in situ hybridization (GISH) showed the predominance of the number of G. cruciata chromosomes over chromosomes of G. tibetica. However, a significant level of cross-hybridization was observed for about one-third of hybrid chromosomes, indicating a high degree of homeology between the genomes of G. cruciata and G. tibetica.

scholarly journals Chromosome behavior during meiosis in pollen mother cells from Saccharum officinarum × Erianthus arundinaceus F1 hybrids

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Xueting Li ◽  
Fei Huang ◽  
Jin Chai ◽  
Qiusong Wang ◽  
Fan Yu ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
5S Rdna ◽  
Saccharum Officinarum ◽  
F1 Hybrids ◽  
45S Rdna ◽  
Pollen Mother Cells ◽  
Erianthus Arundinaceus ◽  
Rdna Sites ◽  
Mother Cells

Abstract Background In recent years, sugarcane has attracted increasing attention as an energy crop. Wild resources are widely used to improve the narrow genetic base of sugarcane. However, the infertility of F1 hybrids between Saccharum officinarum (S. officinarum) and Erianthus arundinaceus (E. arundinaceus) has hindered sugarcane breeding efforts. To discover the cause of this infertility, we studied the hybridization process from a cytological perspective. Results We examined the meiotic process of pollen mother cells (PMCs) in three F1 hybrids between S. officinarum and E. arundinaceus. Cytological analysis showed that the male parents, Hainan 92–77 and Hainan 92–105, had normal meiosis. However, the meiosis process in F1 hybrids showed various abnormal phenomena, including lagging chromosomes, micronuclei, uneven segregation, chromosome bridges, and inability to form cell plates. Genomic in situ hybridization (GISH) showed unequal chromatin distribution during cell division. Interestingly, 96.70% of lagging chromosomes were from E. arundinaceus. Furthermore, fluorescence in situ hybridization (FISH) was performed using 45S rDNA and 5S rDNA as probes. Either 45S rDNA or 5S rDNA sites were lost during abnormal meiosis, and results of unequal chromosomal separation were also clearly observed in tetrads. Conclusions Using cytogenetic analysis, a large number of meiotic abnormalities were observed in F1. GISH further confirmed that 96.70% of the lagging chromosomes were from E. arundinaceus. Chromosome loss was found by further investigation of repeat sequences. Our findings provide insight into sugarcane chromosome inheritance to aid innovation and utilization in sugarcane germplasm resources.

Uniparental loss of ribosomal DNA in the allotetraploid grass Zingeria trichopoda (2n = 8)

Genome ◽  
2003 ◽  
Vol 46 (1) ◽  
pp. 156-163 ◽  
Author(s):  
Violetta Kotseruba ◽  
Dorota Gernand ◽  
Armin Meister ◽  
Andreas Houben
Keyword(s):  
Ribosomal Dna ◽  
5S Rdna ◽  
45S Rdna ◽  
Rdna Sequences ◽  
Genome Wide ◽  
Rdna Sites ◽  
Rdna Loci ◽  
Subsequent Elimination ◽  
Genome Level

Analysis of the grass Zingeria trichopoda (2n = 8, 2C = 5.3 pg) revealed a dynamic evolution with the following characteristics. (i) Genomic in situ hybridization (GISH) demonstrates that Z. trichopoda evolved from an interspecific hybrid involving a species like contemporary Zingeria biebersteiniana (2n = 4) and a second species with a similar low number of chromosomes. The nucleus of Z. trichopoda is spatially organized at the genome level and the two parental genomes occupy distinct and separate domains of lateral arrangements. (ii) The copy number of the Z. biebersteiniana specific pericentromeric tandem repeat family Zbcen1 is drastically reduced in Z. trichopoda. (iii) GISH in combination with labeled rDNA sequences simultaneously discriminated the two parental genomes and the corresponding 5S and 45S rDNA sites. Hence, following allopolyploidization of Z. trichopoda the Z. biebersteiniana like parental chromosomes probably underwent drastic loss of 45S rDNA. This could have arisen either through the loss ofZ. biebersteiniana derived 45S rDNA or through Z. trichopoda genome-wide homogenization of Z. biebersteiniana type 45S rDNA and subsequent elimination of 45S rDNA loci from Z. biebersteiniana derived chromosomes. Finally, 5S rDNA loci are present in both subgenomes of Z. trichopoda and the chromosomal position of these loci is similar for both Z. biebersteiniana and the Z. biebersteiniana like parental genome of Z. trichopoda.Key words: genome evolution, polyploidy, ribosomal DNA, Poaceae.

scholarly journals Substrate Specificity and Colorimetric Assay for Recombinant TrzN Derived from Arthrobacter aurescens TC1

2005 ◽  
Vol 71 (5) ◽  
pp. 2214-2220 ◽  
Author(s):  
Nir Shapir ◽  
Charlotte Rosendahl ◽  
Gilbert Johnson ◽  
Marco Andreina ◽  
Michael J. Sadowsky ◽  
...  
Keyword(s):  
Colorimetric Assay ◽  
Pyrimidine Ring ◽  
Triazine Herbicide ◽  
Ring Compounds ◽  
Arthrobacter Aurescens ◽  
Colorimetric Test ◽  
Yellow Color ◽  
Hydrolysis Of

ABSTRACT The TrzN protein, which is involved in s-triazine herbicide catabolism by Arthrobacter aurescens TC1, was cloned and expressed in Escherichia coli as a His-tagged protein. The recombinant protein was purified via nickel column chromatography. The purified TrzN protein was tested with 31 s-triazine and pyrimidine ring compounds; 22 of the tested compounds were substrates. TrzN showed high activity with sulfur-substituted s-triazines and the highest activity with ametryn sulfoxide. Hydrolysis of ametryn sulfoxide by TrzN, both in vitro and in vivo, yielded a product(s) that reacted with 7-chloro-4-nitrobenz-2-oxa-1,3-diazole (NBD-Cl) to generate a diagnostic blue product. Atrazine chlorohydrolase, AtzA, did not hydrolyze ametryn sulfoxide, and no color was formed by amending those enzyme incubations with NBD-Cl. TrzN and AtzA could also be distinguished by reaction with ametryn. TrzN, but not AtzA, hydrolyzed ametryn to methylmercaptan. Methylmercaptan reacted with NBD-Cl to produce a diagnostic yellow product having an absorption maximum at 420 nm. The yellow color with ametryn was shown to selectively demonstrate the presence of TrzN, but not AtzA or other enzymes, in whole microbial cells. The present study was the first to purify an active TrzN protein in recombinant form and develop a colorimetric test for determining TrzN activity, and it significantly extends the known substrate range for TrzN.

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