Heterochromatin heterogeneity in the holocentric X chromatin of Megoura viciae (Homoptera, Aphididae)

Genome ◽  
1996 ◽  
Vol 39 (2) ◽  
pp. 465-470 ◽  
Author(s):  
G. C. Manicardi ◽  
D. Bizzaro ◽  
E. Galli ◽  
U. Bianchi

Holocentric chromosomes, prepared by spreading embryo cells obtained from Megoura viciae parthenogenetic females, have been C-banded, enzymatically digested in situ using the specific endonucleases DdeI (C↓TNAG), DraI (TTT↓AAA), Tru9I (TT↓AA), and CfoI (GCG↓C), and subsequently stained with Giemsa, DAPI, CMA3, and AgNO3. We observed that the X chromosome had the best defined banding patterns. In the M. viciae X chromosome there is a certain amount of heterogeneity in heterochromatic DNA composition. In fact, the GC-rich NOR-associated heterochromatin differs from other heterochromatic bands that are characterized by AT-rich DNAs. Our data also indicate that, in M. viciae holocentric chromosomes, all heterochromatic blocks are accessible to in situ enzyme attack, the only limit to the digestion being the presence or absence of recognition targets. This is an interesting point, since, in monocentric chromosomes, it is well known that in situ endonuclease digestion is heavily affected not only by DNA base composition but also by chromatin compactness that may limit enzyme accessibility to their specific targets. Key words : heterochromatin, holocentric chromosomes, aphids.


Genome ◽  
1991 ◽  
Vol 34 (4) ◽  
pp. 661-665 ◽  
Author(s):  
G. C. Manicardi ◽  
D. C. Gautam ◽  
D. Bizzaro ◽  
E. Guicciardi ◽  
A. M. Bonvicini Pagliai ◽  
...  

The holocentric mitotic chromosomes of Megoura viciae, a species that has been little studied cytogenetically to date, have been characterized by applying G, C, AluI, and HaeIII banding techniques. C bands have shown the best defined patterns, particularly on the X chromosome. This chromosome, on the other hand, behaved as the most reactive to the various treatments. Uncondensed, prometaphase X chromosomes showed a number of heterochromatic bands, interspersed among the euchromatin, which fused together during metaphase condensation. AluI and HaeIII treatments also produced reproducible banding patterns. These data permit an accurate identification of the X chromosome as well as of the autosomal pairs 1 and 2, and facilitate the construction of nonambiguous karyotypes. They will also stimulate studies on the organization of chromatin in holocentric, holokinetic chromosomes. Finally they could also promote research on chromosomal rearrangements that have occurred during the course of speciation and evolution of aphids, since these kinds of events may be significantly affected by the condition of chromosomal holocentrism.Key words: aphids, holocentric chromosomes, chromosome banding, heterochromatin.



Genome ◽  
1998 ◽  
Vol 41 (2) ◽  
pp. 169-172 ◽  
Author(s):  
Gian Carlo Manicardi ◽  
Mauro Mandrioli ◽  
Davide Bizzaro ◽  
Umberto Bianchi

Using the in situ nick translation technique, we looked for the presence of DNase I sensitive sites in Megoura viciae chromosomes, to study the distribution of active or potentially active genes in aphids, a group of insects possessing holocentric chromosomes. Cytological preparations obtained by the spreading of embryo cells were treated in situ with increasing concentrations (ranging from 5 to 200 ng/mL) of DNase I. At DNase I concentrations below 50 ng/mL, only one hypersensitive site was observed, and this was located on a telomeric region of the X chromosome that contains transcriptionally active nucleolar organizing regions, as assayed by silver staining. Interestingly, at intermediate concentrations of DNase, the incorporation of biotinylated nucleotide occurred uniformly throughout all chromosomes, whereas at concentrations above 100 ng/mL, a C-like banding pattern was produced. Our data differ from results obtained with mammalian, frog, and grasshopper chromosomes, where it was found that DNase I nicking is concentrated at the distal regions of all chromosomes.Key words: aphids, holocentric chromosomes, DNase I sensitivity, nick translation.



Genome ◽  
1994 ◽  
Vol 37 (4) ◽  
pp. 625-630 ◽  
Author(s):  
G. C. Manicardi ◽  
D. Bizzaro ◽  
P. Azzoni ◽  
U. Bianchi

Chromosomal and purified DNA methylation patterns were determined in the holocentric chromosomes of Megoura viciae by treatment with MspI and HpaII. Both enzymes produced a clear C-like banding pattern but widely digested one telomere of the X chromosome, which appeared as heterochromatic after C-banding treatment and brightly fluorescent after chromomycin A3 staining. Quantitative microfluorometric evaluations of DNA extraction performed on cytological preparations showed that both isoschizomers resulted in the same DNA extraction (about 30%). Contrary to what was found by in situ endonuclease treatment, the electrophoretic patterns of purified and digested DNA showed that digestion with MspI was slightly more extensive than that with HpaII in a zone of fragments ranging from 23 to 9 kb. This result indicates that aphid chromatin is not wholly unmethylated. The discrepancy between electrophoretic and cytological data has been explained by taking into consideration that DNA fragments with high molecular weights could be cleaved in situ by the enzymes but not extracted from the chromatin.Key words: aphids, DNA methylation, holocentric chromosomes, heterochromatin, restriction enzyme bandings.



Genome ◽  
1989 ◽  
Vol 32 (2) ◽  
pp. 208-215 ◽  
Author(s):  
C. Sentís ◽  
J. Santos ◽  
J. Fernández-Piqueras

Fixed chromosomes of Baetica ustulata were treated with restriction endonucleases and subsequently stained with either Giemsa or the DNA-specific dye propidium iodide. Each enzyme produces a specific banding pattern with both dyes, which demonstrates the value of restriction endonucleases for chromosome banding in this species. The results obtained agree with the hypothesis that the action of restriction enzymes is based on cutting and extraction of DNA and is essentially determined by DNA base composition rather than by chromatin structure. However, strictly centric bands could be an exception. At least nine chromatin types can be distinguished in B. ustulata according to their different response to enzyme digestion and to the fluorescence banding patterns. Differential digestion of specific heterochromatic areas in band 4.4 and in the supernumerary segment of M5 by the HpaII – MspI enzyme pair suggests a high level of DNA methylation at these regions. The distribution of the different classes of DNA repeated sequences in the chromosome complement of B. ustulata indicates that some bands seem to follow the general principles of heterochromatin equilocality, while conceited evolution of heterochromatic DNA in other regions might have different rates of convergence in this species.Key words: restriction endonucleases, chromatin heterogeneity, methylated bands, equilocality, concerted evolution.



1971 ◽  
Vol 11 (2) ◽  
pp. 91-95 ◽  
Author(s):  
G. F. Gause ◽  
A. V. Laiko ◽  
M. V. Bibikova ◽  
L. I. Kusovkova ◽  
T. I. Selesneva ◽  
...  


Genetics ◽  
1996 ◽  
Vol 144 (2) ◽  
pp. 647-656
Author(s):  
William B Eggleston ◽  
Nac R Rim ◽  
Johng K Lim

Abstract The structure of chromosomal inversions mediated by hobo transposable elements in the Uc-1 X chromosome was investigated using cytogenetic and molecular methods. Uc-1 contains a phenotypically silent hobo element inserted in an intron of the Notch locus. Cytological screening identified six independent Notch mutations resulting from chromosomal inversions with one breakpoint at cytological position 3C7, the location of Notch. In situ hybridization to salivary gland polytene chromosomes determined that both ends of each inversion contained hobo and Notch sequences. Southern blot analyses showed that both breakpoints in each inversion had hobo-Notch junction fragments indistinguishable in structure from those present in the Uc-1 X chromosome prior to the rearrangements. Polymerase chain reaction amplification of the 12 hobo-Notch junction fragments in the six inversions, followed by DNA sequence analysis, determined that each was identical to one of the two hobo-Notch junctions present in Uc-1. These results are consistent with a model in which hobo-mediated inversions result from homologous pairing and recombination between a pair of hobo elements in reverse orientation.





1996 ◽  
Vol 8 (3) ◽  
pp. 229-237 ◽  
Author(s):  
Donald F. Kapraun ◽  
Juan Lopez-Bautista ◽  
Kimon T. Bird


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