scholarly journals Satellitome Analysis in the Ladybird Beetle Hippodamia variegata (Coleoptera, Coccinellidae)

Genes ◽  
2020 ◽  
Vol 11 (7) ◽  
pp. 783 ◽  
Author(s):  
Pablo Mora ◽  
Jesús Vela ◽  
Francisco J. Ruiz-Ruano ◽  
Areli Ruiz-Mena ◽  
Eugenia E. Montiel ◽  
...  
Keyword(s):  
Biological Control ◽  
In Situ Hybridization ◽  
Repeat Unit ◽  
Pest Species ◽  
Dapi Staining ◽  
Ladybird Beetle ◽  
Sequencing Technologies ◽  
Chromosomal Markers ◽  
Hippodamia Variegata

Hippodamia variegata is one of the most commercialized ladybirds used for the biological control of aphid pest species in many economically important crops. This species is the first Coccinellidae whose satellitome has been studied by applying new sequencing technologies and bioinformatics tools. We found that 47% of the H. variegata genome is composed of repeated sequences. We identified 30 satellite DNA (satDNA) families with a median intragenomic divergence of 5.75% and A+T content between 45.6% and 74.7%. This species shows satDNA families with highly variable sizes although the most common size is 100–200 bp. However, we highlight the existence of a satDNA family with a repeat unit of 2 kb, the largest repeat unit described in Coleoptera. PCR amplifications for fluorescence in situ hybridization (FISH) probe generation were performed for the four most abundant satDNA families. FISH with the most abundant satDNA family as a probe shows its pericentromeric location on all chromosomes. This location is coincident with the heterochromatin revealed by C-banding and DAPI staining, also analyzed in this work. Hybridization signals for other satDNA families were located only on certain bivalents and the X chromosome. These satDNAs could be very useful as chromosomal markers due to their reduced location.

Tandem repeat DNA localizing on the proximal DAPI bands of chromosomes in Larix, Pinaceae

Genome ◽  
2002 ◽  
Vol 45 (4) ◽  
pp. 777-783 ◽  
Author(s):  
Masahiro Hizume ◽  
Fukashi Shibata ◽  
Ayako Matsumoto ◽  
Yukie Maruyama ◽  
Eiji Hayashi ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Dna ◽  
Genomic Dna ◽  
Repeat Unit ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Repeat Sequence ◽  
Proximal Region

Repetitive DNA was cloned from HindIII-digested genomic DNA of Larix leptolepis. The repetitive DNA was about 170 bp long, had an AT content of 67%, and was organized tandemly in the genome. Using fluorescence in situ hybridization and subsequent DAPI banding, the repetitive DNA was localized in DAPI bands at the proximal region of one arm of chromosomes in L. leptolepis and Larix chinensis. Southern blot hybridization to genomic DNA of seven species and five varieties probed with cloned repetitive DNA showed that the repetitive DNA family was present in a tandem organization in genomes of all Larix taxa examined. In addition to the 170-bp sequence, a 220-bp sequence belonging to the same DNA family was also present in 10 taxa. The 220-bp repeat unit was a partial duplication of the 170-bp repeat unit. The 220-bp repeat unit was more abundant in L. chinensis and Larix potaninii var. macrocarpa than in other taxa. The repetitive DNA composed 2.0–3.4% of the genome in most taxa and 0.3 and 0.5% of the genome in L. chinensis and L. potaninii var. macrocarpa, respectively. The unique distribution of the 220-bp repeat unit in Larix indicates the close relationship of these two species. In the family Pinaceae, the LPD (Larix proximal DAPI band specific repeat sequence family) family sequence is widely distributed, but their amount is very small except in the genus Larix. The abundant LPD family in Larix will occur after its speciation.Key words: AT-rich tandem repetitive DNA, fluorescence in situ hybridization, Larix, proximal DAPI band.

Molecular and cytological characterization of a highly repeated DNA sequence in Raphanus sativus

Genome ◽  
1995 ◽  
Vol 38 (6) ◽  
pp. 1237-1243 ◽  
Author(s):  
K. Hirai ◽  
K. Irifune ◽  
R. Tanaka ◽  
H. Morikawa
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Raphanus Sativus ◽  
Repeat Unit ◽  
Repeated Dna ◽  
Dna Base Composition ◽  
C Banding ◽  
Highly Repeated Dna ◽  
Repeated Dna Sequence

A highly repeated DNA sequence with a repeat unit of ca. 180 bp was found in genomic DNA HindIII-digests of Raphanus sativus. The repeating units of six isolated, independent clones were sequenced. These units have 177 or 178 bp, are 36% G+C in their DNA base composition, and show 90% sequence homology. The copy number of this 180-bp repeat unit is about 0.5 × 106 per diploid genome. In situ hybridization analysis with the repeating unit as the probe and C-banding analysis indicated that the repeated DNA sequence of R. sativus is closely associated with the major C-heterochromatins in the proximal regions of all 18 chromosomes at mitotic metaphase.Key words: Raphanus sativus, repeated DNA sequence, nucleotide sequence, in situ hybridization, C-banding.

Chromosomal characterization in native populations of Elymus scabrifolius from Argentina through classical and molecular cytogenetics (FISH–GISH)

Genome ◽  
2012 ◽  
Vol 55 (8) ◽  
pp. 591-598 ◽  
Author(s):  
P.A. Tomas ◽  
G.E. González ◽  
G.E. Schrauf ◽  
L. Poggio
Keyword(s):  
In Situ Hybridization ◽  
Chromosome Pair ◽  
Molecular Cytogenetics ◽  
Chromosome Morphology ◽  
Genomic In Situ Hybridization ◽  
Dapi Staining ◽  
Accurate Identification ◽  
Native Populations ◽  
Strong Hybridization

The karyotype of Elymus scabrifolius (Döll) J.H. Hunz. (2n = 4x = 28) was investigated by DAPI staining and in situ hybridization. All the accessions studied presented a symmetric and uniform karyotype constituted by 9m+2m–sm+3sm. DAPI stain showed 1–7 conspicuous bands in all the chromosomes and polymorphisms between accessions. FISH experiments carried out with 45S rDNA as probe (pTa71) showed strong hybridization signals on the metacentric SAT-chromosome pair 8; the submetacentric SAT-chromosome pair 13 presented weaker hybridization. FISH using pSc119.2 clone as probe identified five chromosome pairs. Then, the combination of chromosome morphology, DAPI-staining, and FISH enabled the accurate identification of each chromosome pair in E. scabrifolius. Genomic in situ hybridization (GISH) experiments using Hordeum DNA as probe on mitotic metaphases confirmed unequivocally the presence of the H genome in E. scabrifolius, allowing us to observe six uniformly labeled chromosome pairs and two chromosome pairs with only one arm labeled. The remaining six chromosome pairs were weakly labeled. The rehybridization of FISH slides with Hordeum DNA as probe allow us to assign the genomic provenance of most of the chromosomes in the studied accessions. Moreover, intergenomic rearrangement was detected between genome H and the still unknown progenitor genome.

Molecular analyses of a repetitive DNA sequence in wheat (Triticum aestivum L.)

Genome ◽  
2000 ◽  
Vol 43 (3) ◽  
pp. 556-563 ◽  
Author(s):  
P P Ueng ◽  
A Hang ◽  
H Tsang ◽  
J M Vega ◽  
L Wang ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
In Situ Hybridization ◽  
Long Terminal Repeat ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Sequence ◽  
Genomic Dna ◽  
Repeat Unit ◽  
Terminal Repeat ◽  
Genomic Clones

A repetitive sequence designated WE35 was isolated from wheat genomic DNA. This sequence consists of a 320-bp repeat unit and represents approximately 0.002% of the total wheat DNA. It is unidirectionally distributed either continuously or discretely in the genome. Ladder-like banding patterns were observed in Southern blots when the wheat genomic DNA was restricted with endonuclease enzymes EcoRI, HincII, NciI, and NdeI, which is characteristic for tandemly organized sequences. Two DNA fragments in p451 were frequently associated with the WE35 repetitive unit in a majority of λ wheat genomic clones. A 475-bp fragment homologous to the 5'-end long terminal repeat (LTR) of cereal retroelements was also found in some λ wheat genomic clones containing the repetitive unit. Physical mapping by fluorescence in situ hybridization (FISH) indicated that one pair of wheat chromosomes could be specifically detected with the WE35 positive probe p551. WE35 can be considered a chromosome-specific repetitive sequence. This repetitive unit could be used as a molecular marker for genetic, phylogenetic, and evolutionary studies in the tribe Triticeae.Key words: repetitive sequence, genomic DNA, Triticum aestivum, fluorescence in situ hybridization, long terminal repeat.

scholarly journals Molecular cytogenetic studies in Chenopodium quinoa and Amaranthus caudatus

2014 ◽  
Vol 70 (2) ◽  
pp. 85-90 ◽  
Author(s):  
Jolanta Małuszyńska ◽  
Luz Gomez Pando ◽  
Bożena Kolano
Keyword(s):  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Chenopodium Quinoa ◽  
45S Rdna ◽  
Fluorescence Staining ◽  
Dapi Staining ◽  
Amaranthus Caudatus ◽  
Homologous Chromosomes ◽  
Molecular Cytogenetic

<em>Chenopodium quinoa</em> Wild. and <em>Amaranthus caudatus</em> L., two plant species from South America, have small and numerous chromosomes. Looking for chromosome markers to distinguish pairs of homologous chromosomes double fluorescence staining, in situ hybridization with 45S rDNA and silver staining were applied. Fluorescent in situ hybridization with 45S rDNA has shown two sites of hybridization occurring on one pair of chromosomes in qunion genre (lines PQ-1, PQ-8). The number of RDA loci in Amaranth's caudate L. genre depends on the accession. Kiwicha 3 line has one pair of chromosomes with signals and Kiwicha Molinera cultivar two pairs. All observed rDNA loci were active. After chromomycin/DAPI staining in all cases, except Kiwicha Molinera cultivar, the CMA<sub>3</sub> positive bands co-localized with signals of in situ hybridization with rDNA. In Kiwicha Molinera the number of CMA<sup>+</sup> bands was higher than the number of 45S rDNA signals after FISH.

scholarly journals Karyological and cytogenetic research of the conifers of boreal zone by classic and new methods

2019 ◽  
Vol 25 ◽  
pp. 74-79
Author(s):  
E. N. Muratova ◽  
T. S. Sedel’nikova ◽  
A. V. Pimenov ◽  
O. V. Goryachkina
Keyword(s):  
In Situ Hybridization ◽  
Biological Diversity ◽  
Natural Populations ◽  
Chromosomal Anomalies ◽  
Extreme Conditions ◽  
Dapi Staining ◽  
Wide Range ◽  
Karyotypic Diversity ◽  
Chromosome Mutations

Aim. Establishing of karyological features and conducting of cytogenetic analysis on conifer plants for biological diversity studies, solving of problems of taxonomics, evolutionary and population genetics. Methods. Classic methods with acetohematoxylin staining of slides and fluorescent in situ hybridization (FISH). Results. More than 150 populations and provenances of representatives of different conifer genera from the Pinaceae and Cupressaceae families were studied. The studies were carried out in natural populations and during the introduction, in optimal and extreme conditions, in disturbed ecosystems, botanical gardens and parks; in addition, various intraspecific forms have been studied. The variability of chromosome numbers and a wide range of chromosomal mutations have been revealed. Fluorescence in situ hybridization (FISH) with the 45S and 5S ribosomal RNA gene probes and DAPI staining allows to identify of homologous chromosome pairs in the karyotypes of conifers and to facilitate the comparative karyotype analysis of these species. Conclusions. The studies of chromosomes in species of the Pinaceae and Cupressaceae families showed a karyotypic diversity and chromosomal anomalies in extreme conditions and under introduction. The use of molecular cytogenetic markers made it possible to obtain new information on the structure of conifer chromosomes. Keywords: chromosomes, nucleolar loci, chromosome mutations, Pinaceae, Cupressaceae.

Allopolyploid origin and genome differentiation of the parasitic species Cuscuta veatchii (Convolvulaceae) revealed by genomic in situ hybridization

Genome ◽  
2019 ◽  
Vol 62 (7) ◽  
pp. 467-475 ◽  
Author(s):  
Amália Ibiapino ◽  
Miguel A. García ◽  
Maria Eduarda Ferraz ◽  
Mihai Costea ◽  
Saša Stefanović ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Molecular Data ◽  
High Rate ◽  
Hybrid Origin ◽  
Dapi Staining ◽  
Rdna Sites ◽  
35S Rdna ◽  
Genomic Probes ◽  
Haploid Karyotype

Interspecific hybridization and genome duplication to form allopolyploids are major evolutionary events in angiosperms. In the parasitic genus Cuscuta (Convolvulaceae), molecular data suggested the existence of species of hybrid origin. One of them, C. veatchii, has been proposed as a hybrid between C. denticulata and C. nevadensis, both included in sect. Denticulatae. To test this hypothesis, a cytogenetic analysis was performed with CMA/DAPI staining and fluorescent in situ hybridization using 5S and 35S rDNA and genomic probes. Chromosomes of C. denticulata were small with a well-defined centromeric region, whereas C. nevadensis had larger, densely stained chromosomes, and less CMA+ heterochromatic bands. Cuscuta veatchii had 2n = 60 chromosomes, about 30 of them similar to those of C. denticulata and the remaining to C. nevadensis. GISH analysis confirmed the presence of both subgenomes in the allotetraploid C. veatchii. However, the number of rDNA sites and the haploid karyotype length in C. veatchii were not additive. The diploid parentals had already diverged in their chromosomes structure, whereas the reduction in the number of rDNA sites more probably occurred after hybridization. As phylogenetic data suggested a recent divergence of the progenitors, these species should have a high rate of karyotype evolution.

Development and Application of Transposable Element-Based Chromosomal Markers for the St Genome in Triticeae

2019 ◽  
Vol 159 (4) ◽  
pp. 215-224
Author(s):  
Ruijuan Liu ◽  
Feng Yu ◽  
Linna Wei ◽  
Bo Liu ◽  
Demei Liu ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Genomic Dna ◽  
Dna Analysis ◽  
Repetitive Sequences ◽  
Distribution Patterns ◽  
Signal Distribution ◽  
Sequence Composition ◽  
Plasmid Library ◽  
Chromosomal Markers

The St genome, originating from Pseudoroegneria (Nevski) Á. Löve, plays an important role in Triticeae. In this study, the Pseudoroegneria stipifolia genome (2n = 2x = 14, StSt) was screened to identify sequences that could be used for FISH. A total of 163 effective clones were obtained from the genomic plasmid library which was constructed by DNase I digestion of P. stipifolia nuclear genomic DNA. Analysis of these clones identified 112 with characteristics of transposable elements (TEs), 13 with characteristics of tandem repetitive sequences, 8 with characteristics of mRNA sequences, and 30 unknown sequences. Fluorescent signals were detected for 11 of 41 TE sequences on P. stipifolia chromosomes after in situ hybridization and were divided into 4 types according to signal distribution patterns: over the whole St genome chromosomes, telomere to pericentromeric regions, centromere to pericentromeric regions, and terminal regions. The affinity between St and Y genomes was studied using the 11 TE probes in 3 StStYY species. Five TE probes showed no obvious difference between subgenomes, 2 probes displayed divergence only in 2 StStYY species, and 4 probes exhibited significant differences among 3 StStYY species. These results provide a preliminary understanding of the sequence composition of the St genome and enabled 11 novel TE probes to be developed and applied.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

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