大麦G—显带核型的研究

本文报道了 ASG 法处理的三个栽培大麦(Hordeum Vulgare)品种 G-带的核型研究。结果表明无论是早中期或中期染色体都显示出了密切邻近的、多重的 G-带带纹。在有丝分裂过程中染色体愈浓缩带纹数目愈少。同源染色体之间带纹分布的位置、染色深浅以及带纹数目都基本一致,可以较为准确地进行配对。同一分裂时期不同染色体的 G-带带纹各具一定的特点,可以作为鉴别的标记。讨论了显带技术和中期染色体的 G-带等问题。     

Studies on G-Banding Karyotype in Barley (Hordeum vulgare)

G-banding karyotypes of three cultivars in barley were analyzed. Multiple closely adjacent G-bands were able to be observed in each early metaphase or metaphase chromosome treatted by an ASG method. The more concentrated the chromosome, the less was the number of G-bands during mitosis. The position of band distribution, staining degree and band numbers between homologous chromosomes were basically identical. Chromosome pairing for karyotype analysis could be carried out more accurately. G-banding patterns of different chromosome pairs were not the same, they could be used as the markers to distinguish one from another chromosome pair. During the same mitotic stage the banding patterns including number, relative position and staining degree of the bands between different cultivars were basically the same, but they had differences in the size and staining degree of some bands near centromeres. G-banding technique and G-banding of metaphase chromosomes were discussed.     

The characterization of high-resolution G-banded chromosomes of man

The detailed characterization of G-banding patterns of high resolution human chromosomes has been possible with the utilization of a refined cell synchronization technique which routinely yields a large number of excellent quality cells in late prophase, prometaphase, early metaphase, and mid-metaphase. These mitotic cells exhibit up to a 400% increase in the number of bands previously visualized by standard methods. From studies of the banding patterns, it has become evident that the G-positive and, to some extent, the G-negative bands of mid-metaphase results from a coalescence of finer subbands of earlier stages and that each band and its corresponding subbands maintain a constant location throughout the process of chromosome condensation. A precise schematic representation of the number, position, height and staining intensity of bands is presented for the five largest chromosomes of the complement at the four mitotic stages.     

水稻染色体G—带的研究

用改良的ASG法首次在籼稻(O.sativa subsp.indica)品种珍汕97和粳稻(O.subsp.iaponica)品种秀岭的有丝分裂染色体上显示了G-带,并作了相应的G-带核型分析。就同一材料来说,随着有丝分裂时期的推进,染色体上带纹数目逐渐减少。籼、粳亚种间相对应的同源染色体上G-带带纹特征彼此相似。讨论了水稻G-带带型与染色体不同区域分化的关系;G-带带型与籼、粳稻分歧的关系;以及G-显带的方法。     

玉米染色体G—带带型的研究

本文对3个玉米自交系,及其中两个自交系的杂交F_1有丝分裂早中期染色体的G-带带型进行了比较研究。所有的供试材料G-显带的染色体上都具有两种类型的带纹,我们称A型带和B型带。A型带为沿染色体长轴分布,较细的,密切邻近的多重带纹。不同自交系的A型带带型基本相同,杂交F_1的A型带无明显的异型性。非同源染色体间带型各不相同,某些染色体具有易于识别,特征性较强的A型带标记。B型带一般为深染色的大带,位于染色体的近端区。同一自交系每两个同源染色体的B型带可以配对,不同自交系B型带带型互有不同。杂交F_1某些染色体上的B型带带型异型性明显。具异型性的染色体对中一成员的带型与一个亲本相似,另一成员与另一亲本相似。比较对同一细胞先后作G-和C-显带处理的结果表明,B型带和C-带是相同的。     

Trypsin G- and C-banding for interchange analysis and sex identification in the chicken

The trypsin banding methods were applied to feather pulp and embryonic material of the chicken. Two contrasting types of chromosomal banding patterns were obtained by varying the duration of trypsin treatment. A short time treatment shows a G-banding pattern which has characteristic and distinctive bands along the chromosome arms. Prolonging the trypsin treatment causes the G-banding pattern to disappear, and only the centromeres and the W chromosome remained heterochromatic which is characteristic of the C-banding pattern. The application of the G-banding pattern analysis was used to identify regions of chromosomes involved in rearrangements. The simplified trypsin technique which produces the C-banding pattern makes it relatively easy to identify the W sex-chromosome and determine sex in avian species.     

The effect of chromosome banding techniques on the proteins of isolated chromosomes

Experiments were undertaken to determine the effect of various chromosome banding treatments on the histone and nonhistone proteins of isolated, fixed, air-dried metaphase chromosomes. Chromosome preparations were exposed to G-banding (SSC, urea, NaCl-urea, or trypsin), R-banding (Earle's balanced salt solution), and C-banding (NaOH or Ba(OH)₂) treatments, and the extracted and residual proteins were examined by SDS polyacrylamide gel electrophoresis. The results indicate that each of the banding treatments induce characteristic alterations in the chromosomal proteins. The residual proteins left in chromosomes after the diverse G-banding treatments were generally similar to one another, indicating that treatments inducing the same type of banding have similar effects on the chromosomal proteins. This was also true for the two different C-banding treatments. On the other hand, the residual protein patterns seen after the G-banding treatments were strikingly different from those seen after R-banding, which in turn differed from those seen after C-banding. The treatments inducing different types of banding therefore produce markedly different effects on the chromosomal proteins. These protein alterations may have an important influence on the induction of chromosome bands.     

Chromosome banding and compaction

Summary We have described a characteristic substructure of mitotic chromosomes, the chromosomal unit fibre, with lengths about five times the length of the corresponding metaphase chromosomes and a uniform diameter of 0.4 m. In order to study the relationship of chromosome banding to chromosome compaction, methods have been devised to obtain banding patterns on chromosomal unit fibres, similar to G-band patterns of intact mitotic chromosomes. The total number of bands plus interbands per haploid human karyotype is estimated at about 3000. The banding pattern of chromosomal unit fibres indicates a certain resemblance to the normal G-banding pattern of human chromosomes even if the details indicate a short-range random distribution.     

Intra- and interspecific chromosome polymorphisms in cultivated Cichorium L. species (Asteraceae)

Endive (Cichorium endivia L.) and chicory (C. intybus L.) both have 2n = 18, but until now, there has been no detailed karyomorphological characterization. The present work evaluated five accessions of each species using FISH with rDNA probes and fluorochrome staining with CMA and DAPI. Both species presented distinct banding patterns after fluorochrome staining: while endive had proximal CMA⁺⁺/DAPI⁻ bands in the short arms of pairs 1, 2 and 3, chicory had proximal CMA-positive bands in chromosomes 1 and 3 and interstitial in the short arm of chromosome 8. Among endive accessions, FISH procedures revealed conserved position and number of 5S and 45S rDNA sites (two and three pairs, respectively), associated with the CMA-positive bands. Notwithstanding, polymorphisms were detected within chicory accessions regarding the number and the distribution of rDNA sites in relation to the most frequent karyotype (two pairs with 45S and one with 5S rDNA). The karyological markers developed allowed karyotypic differentiation between both species, uncovering peculiarities in the number and position of rDNA sites, which suggest chromosome rearrangements, such as translocations in chicory cultivars. The interspecific and intraspecific polymorphisms observed emphasize the potential of karyomorphological evaluations, helping our understanding of the relationships and evolution of the group.     

Chromosome organisation in the Australian plague locust,Chortoicetes terminifera

In Chortoicetes terminifera, G-banding, produced by the trypsin treatment of air-dried slides followed by Giemsa staining, leads to light staining gaps at the secondary constrictions on autosomal pair 6 and regions proximal to the centromere on the long arms of pair 4. The variable short arms of two of the three smallest pairs were usually flared and lightly stained after treatment. In contrast to the relatively minor response of the normal chromosome set to G-banding, the large supernumerary chromosomes of C. terminifera show a spectacular series of dark bands alternating with lightly stained gaps. Two G-band variants of the B-chromosome were found in a laboratory stock. These patterns of G-banding are discernable both at mitosis in adults and embryos of both sexes and at all stages of male meiosis. Some regions which are gaps after G-banding appear as dark bands after C-banding. Consequently the supernumerary chromosome is mainly darkly stained with C-banding. In addition the centromeres and some telomeres are C-banded along with narrow interstitial bands and polymorphic heterochromatic blocks. — C-banding was not always successful, the technique often yields a mixture of G- and C-banding. The disparity of banding between the normal complement and the B-chromosome implies that whatever the source of origin of the B it has undergone spectacular changes in organisation since its origin.     

Karyotypic characterization of Iheringichthys labrosus (Pisces, Pimelodidae): C-, G- and restriction endonuclease banding

Various chromosomal banding techniques were utilized on the catfish, Iheringichthys labrosus, taken from the Capivara Reservoir. C-banding regions were evidenced in telomeric regions of most of the chromosomes. The B microchromosome appeared totally heterochromatic. The restriction endonuclease AluI produced a banding pattern similar to C-banding in some chromosomes; the B microchromosome, when present, was not digested by this enzyme and remained stained. G-banding was conspicuous in almost all the chromosomes, with the centromeres showing negative G-banding. When the restriction endonuclease BamHI was used, most of the telomeres remained intact, while some centromeres were weakly digested. The B chromosome was also not digested by this enzyme. The first pair of chromosomes showed a pattern of longitudinal bands, both with G-banding and BamHI; this was more evident with G-banding. This banding pattern can be considered a chromosomal marker for this population of I. labrosus.     

Distribution of radiation induced lesions in human chromosomes and dose-effect relation analysed with G-banding

Summary Human female lymphocytes were exposed to X-rays in vitro at 7 different doses between 40–280 R. In 830 metaphases chromosome analyses were carried out with either conventional staining or G-banding, respectively. 486 breakpoints are non-randomly distributed between chromosomes and chromosome arms. An excess of lesions was present in chromosomes 1 and 5 or in lp. 85% of the lesions were located in G-negative bands (pale G-bands). 29% of all lesions appeared in either the last terminal pale band (21%) or in the centromere region (8%).With regard to an application of G-banding for a biological dose-estimation, the dose-response relations of dic and ace were analysed. Although G-banding enables detailed analysis of the whole karyotype it cannot be recommended for cytogenetic routine analyses in medical radioprotection monitoring, without suitable automated scoring techniques. Dose estimations based on the frequency of dic and carried out with conventional staining cannot be essentially improved at present with banding. Nevertheless, by banding criteria for a correct evaluation of other aberration types, e.g. ace, can be provided. This is a prerequisite for the calculation of representative dose-effect curves.     

Chromosome homology and heterochromatin in goat,sheep and ox studied by banding techniques

Peripheral blood lymphocyte metaphase chromosomes of three Bovoidean species have been studied using Quinacrine fluorescence and Giemsa banding techniques to give Q-, G-, and C-banding patterns. Q- and G-banding characteristics, coupled with chromosome length, enabled all of the chromosomes in each of the chromosome complements to be clearly distinguished, although some difficulties were encountered with the very smallest chromosomes. A comparison of G-banding patterns between the species revealed a remarkable degree of homology of banding patterns. Each of the 23 different acrocentric autosomes of the domestic sheep (2n=54) was represented by an identical chromosome in the goat (2n=60) and the arms of the 3 pairs of sheep metacentric autosomes were identical matches with the remaining 6 goat acrocentrics. A similar interspecies homology was evident for all but two of the autosomes in the ox (2n=60). This homology between sheep metacentric and goat acrocentric elements confirms a previously suggested Robertsonian variation. The close homology in G-banding patterns between these related species indicates that the banding patterns are evolutionarily conservative and may be a useful guide in assessing interspecific relationships. —The centromeric heterochromatin in the autosomes of the three species was found to show little or no Q-or G-staining, in contrast to the sex chromosomes. This lack of centromeric staining with the G-technique (ASG) contrasts markedly with results obtained with other mammalian species. However, with the C-banding technique these regions show a normal intense Giemsa stain and the C-bands in the sex chromosomes are inconspicuous. The amount of centromeric heterochromatin in the sheep metacentric chromosomes is considerable less than in the acrocentric autosomes or in a newly derived metacentric element discovered in a goat. It is suggested that the pale G-staining of the centromeric heterochromatin in these species might be related to the presence of G-Crich satellite DNA.     

Two phases of DNA replication in human cells

The course of DNA synthesis in the chromosomes was studied in synchronized human lymphocyte cultures, by means of the BrdU-Hoechst-Giemsa method. In comparing replication patterns and G-banding it was found that with regard to banding the process of DNA replication can be divided into two separate phases, an early replication period which is characterized by DNA synthesis in R bands of the autosomes and active X chromosome, and a late replication period which concerns the G-positive regions of the autosomes and all the bands of the heterochromatic X and Y chromosomes. No overlapping was found between the two phases mentioned. The possible role of regulatory mechanisms was discussed.     

A relation between G-, C-, and N-band patterns as revealed by progressive oxidation of chromosomes and a note on the nature of N-bands

Near-ultraviolet irradiation of chromosome preparations mounted in a hydrogen peroxide solution resulted in an oxidative disintegration of the structure of fixed metaphase chromosomes with concomitant production of various band patterns appearing after staining with Giemsa. Neither irradiation nor hydrogen peroxide alone could produce banding. After irradiation in the presence of hydrogen peroxide the gradually increasing effect of oxidation on the chromosomes along the gradient of light intensities from the periphery of the slide towards the radiation focus in the centre of the slide became visible as G-, C-, and N-banding, respectively. Close to the centre only contours of chromosomes were left after this treatment. Although G-banding and differential DNA-extraction often went together, extraction of DNA was not an absolute requirement to obtain a G-band pattern. N-bands appeared to be the chromosomal regions that were most resistant to destruction. Staining methods specific for DNA failed to demonstrate these bands, although with Giemsa an intense staining reaction occurred. On the analogy of the staining behaviour of model protein preparations with Giemsa a phosphoprotein nature is suggested for the N-band material in the chromosomes.     

Functional implications of differential chromosome banding.

Information on DNA content and banding patterns has been utilized in the construction of a novel ideogram of the human complement. Correspondence between certain features of this ideogram and the clinical record of human autosomal imbalance supports the idea that banding patterns reflect structural as well as functional heterogeneity of chromosomes.     

High-resolution analysis of DNA replication domain organization across an R/G-band boundary.

Establishing how mammalian chromosome replication is regulated and how groups of replication origins are organized into replication bands will significantly increase our understanding of chromosome organization. Replication time bands in mammalian chromosomes show overall congruency with structural R- and G-banding patterns as revealed by different chromosome banding techniques. Thus, chromosome bands reflect variations in the longitudinal structure and function of the chromosome, but little is known about the structural basis of the metaphase chromosome banding pattern. At the microscopic level, both structural R and G bands and replication bands occupy discrete domains along chromosomes, suggesting separation by distinct boundaries. The purpose of this study was to determine replication timing differences encompassing a boundary between differentially replicating chromosomal bands. Using competitive PCR on replicated DNA from flow-sorted cell cycle fractions, we have analyzed the replication timing of markers spanning roughly 5 Mb of human chromosome 13q14.3/q21.1. This is only the second report of high-resolution analysis of replication timing differences across an R/G-band boundary. In contrast to previous work, however, we find that band boundaries are defined by a gradient in replication timing rather than by a sharp boundary separating R and G bands into functionally distinct chromatin compartments. These findings indicate that topographical band boundaries are not defined by specific sequences or structures.     

Location of the mouse complement factor H gene (cfh) by FISH analysis and replication R-banding.

A technique for replication R- and G-banding of mouse lymphocyte chromosomes was developed, and the replication R-banding pattern was analyzed. Optimal banding patterns were obtained with thymidine- and BrdU-treatment of lymphocytes in the same cell cycle. This produced replication R-band patterns that were the complete reverse of the G-band patterns on all chromosomes. Replication R-banding methods can be used in conjunction with nonisotopic, fluorescence in situ hybridization (FISH) to localize cloned probes to specific chromosomal bands on mouse chromosomes. with these methods the mouse complement factor H gene (cfh) was localized to the terminal portion of the F region of Chromosome 1. Q-banding patterns were also obtained by the replication R-banding method and may be useful for rapid identification of each chromosome.     

小麦属核型分析和BG染色体组及4A染色体的起源

应用植物有丝分裂染色体标本制备新方法和N—带技术对小麦属(Triticum)9个六倍体种(AABBDD),8个四倍体种(AABB,AAGG),3个二倍体种(AA,A~uA~u)及B组的可能供体沙融山羊草(Ae. shronensis)体细胞核型和N—带进行了分析。结果表明,小麦属全部为具中部或次中部着丝点染色体,核型属于“2A”类型,不对称性随倍性提高而有所增加。种问核型有一定差异。所有小麦B染色体组、G染色体组和4A染色体均显N—带,其它染色体则不显带或只显很浅的着丝点带。六倍体种B染色体组带型基本相同,四倍体小麦B组N—带种间有一定差异。提莫菲维小麦(T.Timopheevi)G组带纹数目和分布与B梁色体组有显著差别,作者认为两者非同源。沙融山羊草核型和带型都与小麦B组相近,是B组的可能供体。一粒系小麦A染色体组基本不显N—带,其中无与4A带型相同的染色体,4A起源尚待研究。