Relationship between secondary constrictions and nucleolus organizing regions inAllium sativum chromosomes

Summary Allium sativum L. (2 n=16) had three types of clones with regard to the number of chromosomes carrying well-defined secondary constrictions: the first type had two secondary constricted chromosomes (type I), the second had three (type II) and the third had four (type III). Silver staining was applied to these three types of cells to determine the number of nucleolus organizing regions (NORs) per cell and to study the relationship between the morphological appearance of the secondary constrictions and the ability of the chromosomes to form nucleoli. Ag-positive regions appeared on two chromosomes in type I, on three in type II and on four in type III. The comparison of Giemsa and Feulgen stained chromosomes with the silver stained ones clearly indicated that the positive reaction with silver occurred exclusively on the secondary constricted regions that responded negatively to both Giemsa and Feulgen staining, indicating that the size of the achromatic secondary constrictions directly reflects the volume of the Ag-positive materials. However, all three types of clones had a maximum of four nucleoli at interphase. Of the four nucleoli, either two or one was extremely small (less than 1 m in diameter) in types I and II respectively. The size variations of the other nucleoli seemed to be positively correlated with those of the Ag-positive regions. This and the observation that the maximum number of nucleoli per cell did not coincide with the number of Ag-positive regions on the metaphase chromosome complement suggest strongly that the NORs responsible for the minute nucleoli cannot be detected on the metaphase chromosomes. The present observations indicate that not all NORs are indicated by the morphological appearance of secondary constrictions.     

Chromosome complement and nucleolar organization in Podophyllum hexandrum Royle

The karyomorphology of Podophyllum hexandrum Royle (2n=2x=12) is described for the first time. The haploid set comprises one metacentric, three sub-metacentric and two acrocentric chromosomes. Metacentrics are the longest and acrocentrics the smallest in the complement. Secondary constrictions are located in the short arm of two meta-/submetacentrics and in the long arm of the two acrocentric chromosomes. The number of NORs is in agreement with the number of nucleoli organized per root tip nucleus.     

Chromosomal organization of ribosomal genes and NOR-associated heterochromatin, and NOR activity in some populations of Allium commutatum Guss. (Alliaceae)

The position and the number of 18S-5.8S-26S and 5S rDNA loci, characterization of nucleolar organizing region (NOR)-associated heterochromatin and NOR activity assessment are given for six south-eastern Adriatic populations of Allium commutatum Guss. The karyotype characteristics were identical for all the populations studied, even those of distant islands. Diploid karyotypes (2 n = 16) always possessed two NOR-bearing chromosome pairs with pericentric and median secondary constrictions (SCs) on the short arm of the chromosomes VII and VIII. Fluorescent in situ hybridization (FISH) confirmed that these were the only sites of 18S-5.8S-26S rRNA genes. NOR-associated heterochromatin was of the constitutive character as shown after C-banding. Differential fluorochrome banding with Chromomycin A3 (CMA) and 4,6-diamidino-2-phenylindole (DAPI) revealed that this heterochromatin comprises both GC- and AT-rich DNA segments. Heteromorphism of C- and CMA-bands was noticed between homologous NOR-bearing chromosomes. The maximum number of four active NORs was correlated with the maximum number of four nucleoli in interphase. Variability of NOR-activity, expressed as number and size of silver stained NORs, existed between cells and between individuals of the same population. The different size of homologous and nonhomologous silver stained NORs was correlated with the extension of SCs. The only 5S rDNA locus was in an intercalary position on short arm of the chromosome VI, at the region of AT-rich constitutive heterochromatin. Dimorphism of C-bands and DAPI/Hoechst(H)-fluorescent bands was noticed between homologous chromosomes VI. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 139 , 99–108.     

Dimorphic nucleolar organizer regions in the frog Rana blairi.

The nucleolar organizer-specific staining procedure, ammoniacal silver (Ag-AS), has been used to study the distribution and size of the nucleolar organizer regions (NORs) in chromosomes of the frog Rana blairi (Mecham, Littlejohn, Oldham, Brown and Brown). The somatic metaphase karyotype of this frog is similar to that of other frogs of the Rana pipiens species complex, numerically (2n=26) and morphologically. Secondary constrictions are detectable in untreated Giemsa-stained metaphase preparations as achromatic gaps in the long arms of a pair of submetacentric chromosomes (no. 10). These constrictions are the only regions which are deeply stained with the Ag-AS method and are thus identified as the nucleolar organizer regions (Ag-NORs). In each of the three individuals, the Ag-NORs as visualized on the homologues are of unequal length.     

Chromosomal diversity and evolution in the ground beetle genus Bembidion and related taxa (Coleoptera: Carabidae: Trechitae)

Chromosome numbers and sex chromosome systems of 154 previously unstudied Bembidion species are described. The genus is nearly uniform: males of 176 of 205 species are 2n=22+XY. Karyotypes are presented for 30 species. There is some variation among species in size of Y and size of autosomes. Within most species autosomes are subequal in size, and metacentric or submetacentric. Subterminal secondary constrictions and B chromosomes are reported from several species.The supertribe Trechitae (Zolini + Trechini + Pogonini + Bembidiini) is hypothesized to be primitively male 2n=22+X or 24+X, and the ancestral Bembidion stock 2n=22+XY. Conclusions are based on the most parsimonious hypothesis of ancestral state given an inferred phylogeny of the group, rather than the widespread-is-primitive arguments used previously. Evolution within Bembidion away from the presumably-primitive 2n=22+XY is discussed. Six lineages have lost Y chromosomes; seven have undergone changes in autosome number. It is not known why such changes are so scarce, nor what particular rearrangements led to the observed diversity. Nonetheless, the cytogenetic data can be used to infer a monophyletic origin of groups possessing derived chromosome numbers or sex chromosomes, and to help resolve species limits.     

Light and electron microscopy of Ag-NORs in domestic horse chromosomes identified after R-banding

Silver staining shows the presence in the domestic horse of six NORs located on chromosomes 1, 26 and 31 as identified after R-banding. Following electron microscopy, the argyrophilic material was observed outside the terminal secondary constrictions (satellite stalks) on the terminal portion of the short arm of chromosome 1, outside the secondary constrictions on the proximal region of the long arms of chromosome 31, and beside the proximal region of the long arms of chromosome 26. Satellite staining applied to these chromosomes appears to reveal only the active NORs.     

Cytogenetics of Batrachyla species (Anura: Neobatrachia: Ceratophryidae) of southern South America,with phylogenetics comments

Abstract

A comparative cytogenetic analysis showed that B. antartandica, B. leptopus, B. nibaldoi and B. taeniata share the same diploid number 2n = 26, but not the same fundamental number (FN), which was 52 in B. leptopus and 50 in the other three species. The karyotype of B. nibaldoi, including C‐band patterns and locations of the NORs, are described for the first time. The C‐band patterns were species‐specific and were helpful in distinguishing taxa. Batrachyla leptopus can be distinguished from its congeners by the absence of telocentric chromosomes and by secondary constrictions in the long arms of pair 6 and short arms of pairs 7 and 11. The chromosomal data suggest that Batrachyla might be a paraphyletic genus.     

The karyotype of Lacerta horváthi (Reptilia,Sauria, Lacertidae)

The chromosomes of Lacerta horváthi have been studied by means of conventional, C-banding, and silver-NOR techniques. The karyotype of this species, characterized by 36 acrocentric macrochromosomes, lacks the typical pair of microchromosomes shared by all other lacertid lizards. It is hypothesized that the microchromosomes could have been translocated to the large elements of the karyotype. The occurrence of such a rearrangement in the chromosome complement of L. horváthi underlines its isolation from the other species of the subgenus Archaeolacerta. The C-banding analysis evidences the existence of a female sex heteromorphism in which the W-chromosome has the same shape and size of the Z, but differs from it in being completely heterochromatic. The nucleolar organizer regions (NORs) are located on a pair of medium size chromosomes in subtelomeric position, where the standard Giemsa-staining reveals secondary constrictions.     

Karyotype analysis and heterochromatin differentiation with Giemsa C-banding and fluorescent counterstaining inCephalanthera (Orchidaceae)

Detailed studies of the chromosomes of the three Austrian species of the genusCephalanthera showed them all to have basically similar karyotypes. BothC. damasonium (2n = 36) andC. longifolia (2n = 32) have three large and several classes of smaller chromosome pairs. The karyotype ofC. rubra (2n = 44) is composed of four large and several groups of smaller pairs. The heterochromatin in these species amounts to about 10% of total karyotype length. All the chromosomes have Giemsa-positive centromeres, but only a few have intercalary or terminal bands. Using differential fluorescent staining with DAPI/actinomycin D, quinacrine/actinomycin D (both A-T specific), and chromomycin A₃/distamycin A (G-C specific) three different types of major heterochromatic bands can be characterized in respect of their satellite DNA composition: highly A-T rich, slightly A-T rich, and very G-C rich. The chromosomes ofC. longifolia contain more A-T rich C-bands than those ofC. damasonium, while the latter's have more G-C rich heterochromatin. In both species several C-bands appear as secondary constrictions or gaps in the Feulgen-stained chromosomes, but most likely, in each species there is only one pair of chromosomes where the secondary constrictions function as nucleolus organizing regions. No major intraspecific variation could be observed except on one small chromosome pair ofC. longifolia which had a heteromorphic C-band in most individuals. Possible pathways of karyotype evolution involving polyploidy and Robertsonian events are discussed.     

Evidence for chromosome number reduction and chromosomal homosequentiality in the 24-chromosome Korean frog Rana dybowskii and related species

The karyotype of the Korean frog Rana dybowskii with its pattern of C-band heterochromatin distribution was numerically analyzed. There are 2n = 24 chromosomes in the karyotype representing a reduction in number from the typical 2n = 26 chromosome karyotype of Rana. The karyotype shows other evidence of reorganization relative to 26-chromosome species. The chromosomes grade smoothly in size from largest to smallest without the two size classes that are characteristic for 26-chromosome species. In contrast to many 26-chromosome species, there are few centromeric C-bands but many interstitial ones. C-bands for each homologous chromosome pair are distinctive. A prominent secondary constriction is located on one of the smallest chromosomes, chromosome 11, in a position similar to that seen in most 26-chromosome species. The karyotype of R. dybowskii is compared to those of other species of Rana known to have 2n = 24 chromosomes; it is most similar to that of R. chensinensis, less so that of R. ornativentris and less still to that of R. arvalis in terms of the positions of centromeres and secondary constrictions. C-bands as well as secondary constrictions in the karyotypes of these frogs show evidence of chromosomal homosequentiality. The process and possible consequences of chromosome number reduction from an ancestral 26-chromosome karyotype is also evident in the karyotypes of these closely allied palearctic frogs. Pericentric inversions followed by fusion of two small elements apparently produced a new chromosome, chromosome 6, occurring originally among northeast Asian populations.     

南山茶Camellia semiserrata Chi染色体核型的分析

<正> 引言 南山茶(Camellia semiserrata Chi)又名广宁红花油茶,属山茶属(Camellia L.)山茶亚属(Subg.Camellia)红山茶组(Sect.Camellia),分布于我国广东和广西。南山茶的种子油可供食用,为我国南方主要油料经济树种之一,花红色,形大而艳丽,可供观赏。 山茶属植物约共二百种,但已做过染色体计数者仅47种,做过核型分析者则不超过10种。本文提供的南山茶染色体核型的资料将有助于山茶属植物的遗传育种工作和属内系     

浙江红山茶色染体核型的分析

<正> 引言 山茶属植物将近二百种,分布于东南亚热带和亚热带地区,其中近90％以上的种集中在我国的南部。浙江红山茶(Camellia chekiangoleoso Hu)又名浙江红花油茶,属于山茶属(Camellia)山茶亚属(Subgen.Camellia)的山茶组(Sect.Camellia),是我国特有的树种,分布于浙江、安徽、湖南、江西和福建北部海拔600—1400米的山地。这种植物具有硕大而美丽的红花,为庭园观赏佳品;其种子含油量较高,可供食用。目前已有不少     

不同地理区域鲫鱼染色体银染核仁组织者的比较研究

本文对不同地理区域的鲫鱼(Carassius auratus)—滇池高背鲫、低背鲫、方正银鲫(C.auratusgibelio)的核型及核仁组织者NORs进行了比较研究,并对高背鲫来源作些初步探讨,结果如下: 1.低背鲫Carassius auratus (back low type):2n=100,22m+30sm+48t.st,NORs=4,出现于第5—6对亚中着丝粒染色体短臂。 2.滇池高背鲫Carassius auratus(back high type):2n=156,30m+46sm+80t.st,NORs=6,出现于第5—7对亚中着丝粒染色体短臂。 3.方正银鲫C.auratus gibelio:2n=162,32m+52sm+78t.st NORs=4,出现于第5—6对亚中着粒染色体短臂。     

Karyotype analysis of Eucinostomus argenteus, E. gula, E. harengulus, and Eugerres plumieri (Teleostei, Gerreidae) from Florida and Puerto Rico

The karyotypes of four gerreids of the western Atlantic Ocean are documented. A diploid chromosome complement of 48 telocentric chromosomes was found in the four species (2N=48t, fundamental number FN=48). No differences were detected either in the number of chromosomes of the standard karyotype, in their karyotype size, or between the karyotypes derived from male or female specimens of any of the species. Chromosome length decreased progressively and slightly from pair 1 to pair 24. The Ag–NOR karyotypes of E. argenteus and E. harengulus were characterized by the position of the nucleolar organizer regions next to the centromere in chromosome pair 1, whereas in E. gula and E. plumieri Ag–NORs were detected in pair 4. The other 46 chromosomes showed a light staining of the centromere with no terminal or intermediate heterochromatic blocks. All Eucinostomus species showed Ag–NORs of similar size, while Eugerres plumieri showed Ag–NORs 10–20% larger than Eucinostomus species. A combination of size and position of the Ag–NORs identified E. gula, while size alone identified E. plumieri. However, the ancestral state for size and position of Ag–NORs could not be established. There was no differential staining of the chromosomes by G-banding. The karyotype of the gerreids appears similar to the hypothetical ancestral karyotype of fish. The phylogenetic relationships among these species could not be established because of the lack of chromosome G-bands. Most likely this indicates a homogeneous distribution of GC nucleotides in the chromosomes.     

Localisation of NORs and counterstain-enhanced fluorescence studies inPerca fluviatilis (Pisces,Percidae)

The karyotype of the perch (Percafluviatilis L.) was analysed by means of silver-staining, the chromomyc in A/3 Distamycin A/DAPI and Actinomycin D fluorescence technique in order to locate active NORs and to selectively highlight certain heterochromatic regions. The ribosomal RNA genes are localized at the secondary constrictions of the short arms of chromosomes no. 16. Additionally, bright chromomycin fluorescence was observed in the same regions. No DAPI/ Distamycin A bright heterochromatic block was detected in the perch karyotype.     

Polyploidy in the Australian leptodactylid frog genus Neobatrachus

Karyotypic analysis of six species of the Australian leptodactylid frog genus Neobatrachus showed that N. pictus, N. centralis, N. pelobatoides and N. wilsmorei are diploid (2n=24) while N. sudelli and N. sutor are tetraploid (4n=48). Polyploidy has not been reported previously among Australian anurans. Idiograms of the six species indicate that they are similar to the other Australian leptodactylids so far discribed. DNA values of the tetraploids are approximately double the values for diploids. Tetraploid nuclear and cell sizes are greater compared with diploids but total body size shows no increase. At diakinesis in primary spermatocytes of tetraploids, mainly tetravalents together with a few bivalents are present. Silver staining of metaphase spreads clearly demonstrates the location of NORs at the secondary constrictions and their frequent association in the tetraploid N. sutor. Nucleolar number in interphase nuclei provides a reliable guide for distinguishing tetraploid from diploid frogs in the absence of chromosome analysis and can be determined for both living and preserved specimens. The possible origins and relationships of the tetraploid species are discussed.     

Location and variation of the constitutive heterochromatin in Petunia hybrida

The location of heterochromatin in the chromosomes of Petunia hybrida (2n=14) is presented. C-banded mitotic metaphase chromosomes and carmine-stained pachytene bivalents have been studied. It is shown that the heterochromatin is predominantly located near the centromeres and at the secondary constrictions of the satellite chromosomes. The distribution of chromomeres in pachytene bivalents also reveals that heterochromatin is not restricted to distinct blocks, as is the case in tomato, but occurs in smaller chromomeres which gradually decrease in size towards the ends. Conspicuous telomeres have not been observed. Both C-banding technique and pachytene analysis demonstrate large variation of heterochromatin between different lines of Petunia. The study of pachytene morphology has been hampered by a high degree of non-specific stickiness of the bivalents. Both techniques prove to be unsuitable tools for large-scale chromosome identification of Petunia lines.     

An attempt to define 1qh+, 9qh+, and 16qh+

Summary The lengths of the secondary constrictions of chromosomes 1, 9, and 16 vary with the degree of contraction of the chromosomes but these constrictions contract to a lesser degree than the euchromatic portions of the chromosomes. The regression coefficient for the regression of the length of the secondary constriction on the length of the euchromatic part of the chromosomes is shown to be larger for large constrictions. It is furthermore shown that there is a linear correlation between the regression coefficient and the size of the secondary constriction in question. This linear correlation makes it possible to correct the lengths of the secondary constrictions to the lengths expected when contraction is average. The correction method is used in a sample of 30 couples, and on the basis of this sample, the normal limits for the lengths of the secondary constrictions in chromosomes 1, 9, and 16 are defined.     

Chromosome banding and essential oils composition of Brazilian accessions of <Emphasis Type="Italic">Lippia alba</Emphasis> (Verbenaceae)

The essential oil components and a karyotypic analysis of five Lippia alba (Verbenaceae) accessions from Brazil were performed with the objective of investigating the variation among different populations. The chemistry analysis allowed the grouping of the accessions in two main chemotypes: neral chemotype (LaCat, LaJF and LaRJ) and linalool chemotype (LaGua and LaVC). However, large karyotypic differences, verified by different chromosome banding techniques, were not detected. The results presented the same chromosome number for all accessions (2n = 30) with 10 metacentric chromosomes and 5 submetacentric. The chromosome banding showed great blocks of constitutive heterochromatin (C-bands) around the centromeric region, which was rich in AT bases (DAPI₊), while the CMA bands were observed only in terminal regions of six chromosomes. Through Ag-NOR techniques, only two active pairs of NORs were detected on the three pairs of secondary constrictions (the NOR activity is discussed). This work relates the pattern of heterochromatin for Lippia alba for the first time.