scholarly journals Molecular cytogenetic studies in rubber, Hevea brasiliensis Muell. Arg. (Euphorbiaceae)

Genome ◽  
1998 ◽  
Vol 41 (3) ◽  
pp. 464-467 ◽  
Author(s):  
Andrew R Leitch ◽  
K Yoong Lim ◽  
Ilia J Leitch ◽  
Michelle O'Neill ◽  
MeeLen Chye ◽  
...  
Keyword(s):  
Genetic Mapping ◽  
Ribosomal Dna ◽  
Hevea Brasiliensis ◽  
Consensus Sequence ◽  
Molecular Cytogenetics ◽  
5S Rdna ◽  
Single Pair ◽  
Interphase Nuclei ◽  
Molecular Cytogenetic

This paper reports the start of a molecular cytogenetics programme targeting the genome of the angiosperm tree species Hevea brasiliensis Muell. Arg. (rubber, 2n = 36), a major world crop about whose genetics very little is known. A metaphase karyotype of rubber is presented. In situ hybidization with the probe pTa71 for ribosomal DNA (rDNA) shows that there are four sites of probe hybidization occurring on two pairs of chromosomes called chromosomes 6 and 7 carrying sites NOR-1 and NOR-2, respectively. An examination of meristematic interphase nuclei shows that all four loci have the potential to be partially decondensed at interphase, although in many nuclei one or more loci appear fully condensed and apparently inactive. The probe pXVI revealed a single pair of chromosomes carrying 5S rDNA. The probes pTa71 and pXVI provide cytogenetic markers for three pairs of chromosomes that will be of use in genetic mapping programmes. The rubber chromosomes also have telomere sequences that hybridize with the Arabidopsis consensus sequence TTTAGGG. With the exception of the satellite region containing rDNA, which fluoresces brightly with chromomycin A3, fluorescence banding showed that there is no strong demarcation of the genome into GC- and AT-rich regions, as occurs in mammalian genomes.Key words: rubber, Hevea, genetic mapping, cytogenetics, ribosomal DNA, rDNA fluorescence banding.

scholarly journals Molecular cytogenetic study of Eranthis (Ranunculaceae)

2021 ◽  
Vol 20 (1) ◽  
pp. 305-308
Author(s):  
E. Yu. Mitrenina ◽  
A. S. Erst ◽  
E. D. Badaeva ◽  
S. S. Alekseeva ◽  
G. N. Artemov
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Ribosomal Dna ◽  
Cytogenetic Study ◽  
5S Rdna ◽  
45S Rdna ◽  
5.8S Rdna ◽  
Molecular Cytogenetic ◽  
Specific Hybridization

45S and 5S ribosomal DNA were originally localized on chromosomes of five species of winter aconits,namely, Eranthis cilicica, E. hyemalis (section Eranthis), E. pinnatifida, E. stellata и E. tanhoensis (section Shibateranthis).Fluorescence in situ hybridization was performed with oligonucleotide DNA probes Oligo-pTa71-2 and Oligo-5S rDNAof wheat that are complementary to 45S and 5S ribosomal DNA. In addition, oligonucleotide DNA probe (Oligo-5.8SrDNA-Ran, 50 b) for localization of 45S rDNA was designed and tested. This probe is based on the 5.8S rDNA sequencesof some species of fam. Ranunculaceae taken from GenBank. A specific hybridization of the Oligo-5S rDNA and Oligo5.8S rDNA-Ran probes with the chromosomes of Eranthis was shown. The use of the Oligo-pTa71-2 probe did not localizeclusters of 45S rDNA on chromosomes of studied species.

Direct FISH of 5S rDNA on tomato pachytene chromosomes places the gene at the heterochromatic knob immediately adjacent to the centromere of chromosome 1

Genome ◽  
1996 ◽  
Vol 39 (1) ◽  
pp. 216-221 ◽  
Author(s):  
Jie Xu ◽  
E. D. Earle
Keyword(s):  
Rare Event ◽  
5S Rdna ◽  
Heterochromatic Region ◽  
Conclusive Evidence ◽  
Chromosome 1 ◽  
Cytogenetic Map ◽  
Rdna Gene ◽  
Molecular Cytogenetic ◽  
Direct Fluorescence

We describe a molecular cytogenetic procedure for high resolution physical mapping of DNA markers, an essential step toward construction of an integrated molecular–classical–cytological map. Tomato was selected as material because its pachytene chromosomes are amenable for study and because detailed molecular, classical, and cytological maps are available. Karyotyping of acetocarmine-stained pachytene chromosomes showing detailed cytogenetic landmarks was combined with direct FISH of the 5S rDNA gene. This enabled us to pinpoint the 5S rDNA gene to the first heterochromatic knob immediately adjacent to the centromere in the short arm of chromosome 1. Thus the position of the 5S rDNA gene on the molecular map was related to the position of the 5S rDNA on the cytogenetic map. The results also provide conclusive evidence of the location of a functional gene in the pericentric heterochromatic region, a rare event to date in plants. We conclude that karyotyping of pachytene chromosomes can be combined with FISH to map a DNA sequence to a cytogenetically defined region and to determine the chromatin origin of an expressed gene. Key words : direct fluorescence in situ hybridization, 5S rDNA, pachytene chromosomes, heterochromatic gene.

Heterochromatin patterns and ribosomal DNA loci distribution in diploid and polyploid Crotalaria species (Leguminosae, Papilionoideae), and inferences on karyotype evolution

Genome ◽  
2011 ◽  
Vol 54 (9) ◽  
pp. 718-726 ◽  
Author(s):  
Mateus Mondin ◽  
Margarida L.R. Aguiar-Perecin
Keyword(s):  
Dna Sequence ◽  
Ribosomal Dna ◽  
Karyotype Evolution ◽  
5S Rdna ◽  
Rdna Locus ◽  
Variable Number ◽  
Chromosome 1 ◽  
Distamycin A ◽  
Rdna Sites

Most Crotalaria species display a symmetric karyotype with 2n = 16, but 2n = 14 is found in Chrysocalycinae subsection Incanae and 2n = 32 in American species of the section Calycinae. Seven species of the sections Chrysocalycinae, Calycinae, and Crotalaria were analyzed for the identification of heterochromatin types with GC- and AT-specific fluorochromes and chromosomal location of ribosomal DNA loci using fluorescent in situ hybridization (FISH). A major 45S rDNA locus was observed on chromosome 1 in all the species, and a variable number of minor ones were revealed. Only one 5S rDNA locus was observed in the species investigated. Chromomycin A3 (CMA) revealed CMA+ bands colocalized with most rDNA loci, small bands unrelated to ribosomal DNA on two chromosome pairs in Crotalaria incana, and CMA+ centromeric bands that were quenched by distamycin A were detected in species of Calycinae and Crotalaria sections. DAPI+ bands were detected in C. incana. The results support the species relationships based on flower specialization and were useful for providing insight into mechanisms of karyotype evolution. The heterochromatin types revealed by fluorochromes suggest the occurrence of rearrangements in repetitive DNA families in these heterochromatic blocks during species diversification. This DNA sequence turnover and the variability in number/position of rDNA sites could be interpreted as resulting from unequal crossing over and (or) transposition events. The occurrence of only one 5S rDNA locus and the smaller chromosome size in the polyploids suggest that DNA sequence losses took place following polyploidization events.

scholarly journals Comparative study of four Mystus species (Bagridae, Siluriformes) from Thailand: insights into their karyotypic diversity

2021 ◽  
Vol 15 (2) ◽  
pp. 119-136
Author(s):  
Pun Yeesin ◽  
Phichaya Buasriyot ◽  
Sukhonthip Ditcharoen ◽  
Patcharaporn Chaiyasan ◽  
Chatmongkon Suwannapoom ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Comparative Study ◽  
Fluorescence In Situ Hybridization ◽  
Chromosome Numbers ◽  
Single Pair ◽  
Molecular Cytogenetic ◽  
Karyotypic Diversity ◽  
Species Specific

Karyotypes of four catfishes of the genus Mystus Scopoli, 1777 (family Bagridae), M. atrifasciatus Fowler, 1937, M. mysticetus Roberts, 1992, M. singaringan (Bleeker, 1846) and M. wolffii (Bleeker, 1851), were analysed by conventional and Ag-NOR banding as well as fluorescence in situ hybridization (FISH) techniques. Microsatellite d(GC)15, d(CAA)10, d(CAT)10 and d(GAA)10 repeat probes were applied in FISH. The obtained data revealed that the four studied species have different chromosome complements. The diploid chromosome numbers (2n) and the fundamental numbers (NF) range between 52 and 102, 54 and 104, 56 and 98, or 58 and 108 in M. mysticetus, M. atrifasciatus, M. singaringan or M. wolffii, respectively. Karyotype formulae of M. mysticetus, M. atrifasciatus, M. singaringan and M. wolffii are 24m+26sm+4a, 26m+24sm+2a, 24m+18sm+14a and 30m+22sm+6a, respectively. A single pair of NORs was identified adjacent to the telomeres of the short arm of chromosome pairs 3 (metacentric) in M. atrifasciatus, 20 (submetacentric) in M. mysticetus, 15 (submetacentric) in M. singaringan, and 5 (metacentric) in M. wolffii. The d(GC)15, d(CAA)10, d(CAT)10 and d(GAA)10 repeats were abundantly distributed in species-specific patterns. Overall, we present a comparison of cytogenetic and molecular cytogenetic patterns of four species from genus Mystus providing insights into their karyotype diversity in the genus.

scholarly journals Application of Veterinary Cytogenetics in Domestic Animals: A Review

2019 ◽  
pp. 1-16
Author(s):  
Muhammad Sanusi Yahaya ◽  
Mohd Shahrom Salisi ◽  
Nur Mahiza Md. Isa ◽  
Abd Wahid Haron ◽  
Innocent Damudu Peter
Keyword(s):  
Chromosomal Abnormalities ◽  
Hybrid Sterility ◽  
Molecular Cytogenetics ◽  
Basic Research ◽  
Animal Production ◽  
Somatic Development ◽  
Molecular Cytogenetic ◽  
Molecular Cytogenetic Techniques

Cytogenetics is the study of chromosomes; their structure and properties, chromosome behavior during cell division, their influence on traits and factors which cause changes in chromosomes.  Veterinary cytogenetics is the application of cytogenetics to clinical problems that occur in animal production. It has been applied to understand problems such as infertility and its types, embryonic and fetal death, abnormality in sexual and somatic development and hybrid sterility and also prenatal sex determination and other forms of chromosomal abnormalities. These are achieved through conventional and banded karyotyping techniques and molecular cytogenetic techniques. Although conventional techniques are still useful and very widely applied, the nature of cytogenetics has gradually changed as a result of advances achieved in the molecular cytogenetic techniques for example fluorescent in situ hybridization and array-based techniques. These changes are evident in both molecular diagnostics and basic research. The combination of conventional and molecular cytogenetics has given rise to high resolution techniques which have enabled the study of fundamental questions regarding biological processes. It enables the study of inherited syndromes, the mechanisms of tumorigenesis at molecular level, genome organization and the determination of chromosome homologies between species. It allows the ease with which animals are selected in breeding programs and other important aspects of animal production. In this paper we discussed a number of techniques employed in cytogenetics and their methodologies, and recommend where future focus should be for the benefits of animal production.

Distribution of FISH oligo-5S rDNA and oligo-(AGGGTTT)3 in Hibiscus mutabilis L.

Genome ◽  
2021 ◽  
pp. 1-10
Author(s):  
Xiaomei Luo ◽  
Zhoujian He
Keyword(s):  
Chromosome Number ◽  
Physical Map ◽  
Molecular Cytogenetics ◽  
5S Rdna ◽  
Chromosome Length ◽  
Chromosome Identification ◽  
Karyotype Asymmetry ◽  
First Time ◽  
Distribution Of Fish

Hibiscus exhibits high variation in chromosome number both within and among species. The Hibiscus mutabilis L. karyotype was analyzed in detail using fluorescence in situ hybridization (FISH) with oligonucleotide probes for (AG3T3)3 and 5S rDNA, which were tested here for the first time. In total, 90 chromosomes were counted in prometaphase and metaphase, and all exhibited similarly intense (AG3T3)3 signals at both ends. (AG3T3)3 showed little variation and thus did not allow discrimination among H. mutabilis chromosomes, but its location at both ends confirmed the integrity of each chromosome, thus contributing to accurate counting of the numerous, small chromosomes. Oligo-5S rDNA marked the proximal/distal regions of six chromosomes: weak signals on chromosomes 7 and 8, slightly stronger signals on chromosomes 15 and 16, and very strong signals on chromosomes 17 and 18. Therefore, 5S rDNA could assist in chromosome identification in H. mutabilis. Metaphase chromosome lengths ranged from 3.00 to 1.18 μm, indicating small chromosomes. The ratios of longest to shortest chromosome length in prometaphase and metaphase were 2.58 and 2.54, respectively, indicating karyotype asymmetry in H. mutabilis. These results provide an exact chromosome number and a physical map, which will be useful for genome assembly and contribute to molecular cytogenetics in the genus Hibiscus.

Use of meiotic FISH for identification of a new monosome in Gossypium hirsutum L.

Genome ◽  
1997 ◽  
Vol 40 (1) ◽  
pp. 34-40 ◽  
Author(s):  
Yuanfu Ji ◽  
Dwaine A. Raska ◽  
M. Nurul Islam-Faridi ◽  
Charles F. Crane ◽  
Michael S. Zwick ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Gossypium Hirsutum ◽  
Fluorescence In Situ Hybridization ◽  
Molecular Cytogenetics ◽  
Clinical Diagnostics ◽  
Identification System ◽  
Gossypium Hirsutum L ◽  
Molecular Cytogenetic ◽  
Cytogenetic Methods

The extensive use of molecular cytogenetics in human genetics and clinical diagnostics indicates that analogous applications in plants are highly feasible. One sort of application would be the identification of new aneuploids, which traditionally involves either direct karyotypic identification, which is feasible in only a few plant species, or tests with markers (cytogenetic, genetic, or molecular), which require sexual hybridization and at least one subsequent seed or plant generation. We have used meiotic fluorescence in situ hybridization (FISH) to analyze a new monosome of cotton (Gossypium hirsutum L., 2n = 4x = 52, 2(AD)1) that had a phenotype which seemed to be distinct from monosomes in the Cotton Cytogenetic Collection. Painting with A2-genome DNA revealed the monosome's D-subgenome origin. DAPI–PI staining showed that the monosome carries a major NOR, delimiting it to the major NOR-bearing chromosomes of the D-subgenome, i.e., 16 or 23. Dual-color FISH with 5S and 18S–28S rDNAs indicated that the monosome contains separate major clusters of each of these two tandemly repeated rDNA elements, thus delimiting the monosome to chromosome 23, for which the Cotton Cytogenetic Collection has previously been devoid of any sort of deficiency. Of the 26 chromosomes in the cotton genome, the Collection now provides coverage for 16 (70%) in the form of monosomy, and 20 (77%) in the form of monosomy and (or) telosomy. Use of molecular cytogenetic methods to identify a new plant aneuploid in cotton exemplifies the fact that a physicochemical karyotypic chromosome identification system is not required a priori for application of new molecular cytogenetic methods, thus indicating their potential applicability to nearly all plant species.Key words: fluorescence in situ hybridization, monosome, aneuploid, Gossypium hirsutum.

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.

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