Cytogenetic variability in genus odontocheila (Coleoptera,Cicindelidae): karyotypes,C-banding,NORs and localisation of ribosomal genes of O. confusa and O. nodicornis

Two species of Odontocheila, O. confusa and O. nodicornis, from the Neotropical Region were studied regarding their karyotypes, localisation and activity of ribosomal genes and C-banding. The species, although belonging to the same genus, have quite distinct karyotypes. O. confusa has 10 pairs of autosomes and a single sex chromosome mechanism of the XY/XX type, thus a diploid value of 2n = 22 in males and females. One aneuploid male with a diploid number of 2n = 20 and one male with three B chromosomes were found in a total of eight males studied. O. nodicornis has 17 autosomal pairs and also a single chromosome system but of the X0/XX type, thus a diploid value of 2n = 35 in males and 2n = 36 in females. Fluorescence in situ hybridisation (FISH) revealed the presence of rDNA clusters in two autosomes in both species in mitotic and meiotic figures. Silver staining of male interphase nuclei confirmed the FISH results and showed that all rDNA genes were active. C-banding analysis revealed the presence of constitutive heterochromatin in the centromeres of all chromosomes in the two species plus two pairs in O. nodicornis with terminal positive C-bands. These results are discussed from the cytogenetic and evolutionary point of view.     

Gene-centromere mapping of medaka sex chromosomes using triploid hybrids between Oryzias latipes and O. luzonensis

Four sex-linked genetic markers (SL1, SL2, B2.38 and stsOPQ05-1) on the sex chromosomes of the medaka, O. latipes, were mapped in relation to the centromere by means of triploid hybrids between O. latipes and O. luzonensis. Female F₁ hybrid O. latipes of two inbred strains, Hd-rR and HNI, were crossed with male O. luzonensis. Triploidization was induced by heat-shock treatment. Hatching rate of heat-shock treated eggs was 59%, and that of untreated hybrid eggs was 2%, indicating that most of the hatched fry were triploid. Using these triploid hatched fry, the map distances between the four loci and the centromere were examined. The order was SL2 – centromere – SL1 – B2.38 – stsOPQ05-1 and the map distances were: SL2 – centromere, 1%; centromere – SL1, 18%; SL1 – B2.38, 19%; B2.38 – stsOPQ05-1, 9%. Previous studies using FISH showed that SL2 is located on the short arm of large submetacentric chromosomes, and SL1 was closely linked to SDF (sex-determining factor). The results of gene-centromere mapping of this study show that SL1, B2.38 and stsOPQ05-1 are located on the long arm, and that, SDF is thus also on the long arm of the sex chromosomes.     

Numerical uniformity and structural variation of moss chromosomes

Abstract

In a population of Atrichum undulatum with n = 14, gametophytic interphase nuclei included two unequal, but substantial, blocks of heterochromatin sensu lato. Positive C-banding within each was limited to the terminal portion. Only one of these two chromosomes, together with its homologue, entered meiosis with a similarly extensive distribution of heterochromatin. The bivalent involved was metacentric and usually achiasmate in the heterochromatic region, which amounted to almost the whole of one arm. A single proximal chiasma was rare. In addition to this behavioural difference between the two basically haploid sets of seven chromosomes included in what has been regarded as auto diploid A. undulatum, evidence of a morphological distinction is presented. Six pairs were recognizable on morphological grounds but two chromosomes were unique. Meiosis in triploid A. undulatum with n = 21 also included only a single heterochromatic bivalent.

Variation between basically haploid complements is also presented for Philonotis fontana, in a population of which a dimorphic bivalent was consistently present during meiosis. The dimorphism was related to the presence or absence of a short heterochromatic arm and, hence, of synapsis between telocentric and acrocentric homologues. Sampling to date suggests an association with dioecism.     

Comparative karyotype analysis ofCeratozamia mexicana andMicrocycas calocoma (Zamiaceae) using fluorochrome banding (CMA/DAPI) and fluorescence in situ hybridization of ribosomal DNA

Orcein staining, differential staining with CMA and DAPI, and FISH with an rDNA probe were used to compare somatic chromosomes ofCeratozamia mexicana andMicrocycas calocoma. CMA-positive dots and hybridization signals appeared on chromosomes at early interphase and mitotic prophase, but in significantly different number in the two species. InCeratozamia mexicana, the CMA-positive and DAPI-negative bands and the hybridization signal were located at the terminal region of the long arm of three median-centromeric chromosomes, the terminal region of the short arm of two median-centromeric chromosomes and the terminal region of the long arm of two subterminal-centromeric chromosomes. InMicrocycas calocoma, they were located at the pericentric region of two median-centromeric chromosomes. These chromosome data suggested thatMicrocycas has no simple Robertsonian relationship toCeratozamia.     

Induced apogamous sporophytes inPteris dispar andP. semipinnata,and the meiotic behavior in their sporocytes

Spores ofPteris dispar andP. semipinnata were aseptically cultured in flasks for apogamous sporophyte induction. Calli or cell colonies similar to calli were induced in cultures supplemented with hormones. Sporophytic leaves subsequently developed from them in hormone-free medium and the young sporophytes were raised into plants with sporangia. Since the wild-type plants having 116 chromosomes are tetraploid, the sporophytic plants originating from spores would appear to be diploid (dihaploid). In induced sporophytes ofP. semipinnata, non-homologous chromosomes (58 univalents) were found during the meiotic process in sporocytes. InP. dispar, however, the meiotic cells showed many bivalent chromosomes (maximum 29ll). These results suggest thatP. semipinnata is allotetraploid, whereasP. dispar is autotetraploid.     

The Effect of W Chromosome Origin on Sex-Chromosome Pairing in ZZWW Tetraploid Females of the Domesticated Silkworm, Bombyx Mori, and the Congenic Wild Silkworm, Bombyx Mandarina

To analyze the degree of pairing of the Z and W chromosomes in ZZWW tetraploid female silkworms that have the W chromosomes of the domesticated silkworm, Bombyx mori, and those of the wild silkworm, Bombyx mandarina, we induced two types of ZZWW tetraploid female silkworms (Cr4n, Wr4n) through cold treatment of the eggs. The Wr4n female is congenic to the Cr4n female for W chromosomes; namely, the W chromosomes of the Wr4n female are derived from those of B. mandarina. Each of the sex ratios (/) in filial triploids from the Cr4n females was shown to be in the range of 3.9–5.3 (4.6 as an average of six cases). On the other hand, each of the sex ratios (/) in filial triploids from the Wr4n females was shown to be in the range of 6.2–9.0 (6.9 as an average of nine cases). The results of a t-test indicated that the difference in sex ratios in the two groups is highly significant (at the 0.1% level). These results suggest that, in the meiosis of the ZZWW tetraploid female, the frequency of pairing of the W chromosome of B. mandarina and the Z chromosome of B. mori is lower than that of the pairing of the W and Z chromosomes of B. mori. Furthermore, the t-test results are evidence that the W chromosomes have undergone significant evolutional change.     

Chromosome painting supports lack of homology among sex chromosomes in Oncorhynchus, Salmo, and Salvelinus (Salmonidae)

The sex chromosome pair has been identified previously as the largest submetacentric pair in the genome in several species of the genus Salvelinus (eastern trouts and chars) including S. namaycush (lake trout) and as a large subtelocentric/acrocentric pair in several species of the genus Oncorhynchus (Pacific trouts and salmon). Sex chromosomes have not been identified in Salmo (Atlantic salmon and brown trout). Two paint probes, one specific for the short arm (Yp) and the other for the long arm (Yq) of the sex chromosome pair in Salvelinus namaycush were hybridized to chromosomes of Oncorhynchus mykiss (rainbow trout) and O. tshawytscha (chinook salmon) and Salmo salar (Atlantic salmon) and S. trutta (brown trout). The two probes hybridized to two different autosomal pairs in each of the Oncorhynchus species, supporting lack of homology between the sex chromosomes in the two genera. The Yp probe hybridized to interstitial regions on two different chromosome pairs in S. salar and one pair in S. trutta. The Yq probe hybridized to a different pair in both species.     

Cytogeographical study of four aneuploids ofCarex oxyandra Kudo in Japan

Chromosome numbers were determined for 342 clones ofCarex oxyandra collected from 35 localities in Hokkaido, Honshu, Shikoku and Kyushu, Japan. Four intraspecific aneuploids, 2n=18, 20, 24 and 26, were found. In meiotic division, only bivalent chromosomes were observed in all clones at metaphases I and II, suggesting that the aneuploids are established gamodemes. In the mitotic metaphase chromosomes, trimodal variation in chromosome length was observed. The 2n=26 clones found on Mt. Hiko had two particularly small chromosomes. The cytodemes with higher number of chromosomes are distributed in more southern areas of Japan.Carex oxyandra, therefore, accompanied with chromosome fragmentations, might spread the geographical distribution to the southern parts. The morphological characters of leaves, spikes, scales, perigynia and nuts were similar among the four cytodemes, except for the small leaves on plants from Yaku Island.     

Reticulate evolution of the hybrid produced artificially by crosses between Osmunda banksiifolia and Osmunda lancea

Although ferns have been developed by hybridization and chromosome doubling, no natural polyploidy has yet been recorded in Osmundaceae. So, we produced hybrids artificially by crosses between Osmunda banksiifolia (2n = 2x = 44) and Osmunda lancea (2n = 2x = 44), and investigated their sporogenesis. From the O. banksiifolia × O. lancea hybrid with 44 univalent chromosomes, allotetraploids with 44 bivalent chromosomes were produced by chromosome doubling, and allotriploids with 22 univalent chromosomes and 22 bivalent chromosomes were then produced by back crosses. The results show when and how chromosome doubling occurs in hybrids. The success of artificial hybridization between O. banksiifolia and O. lancea, did not, however, reflect any product of natural hybridization between the two species.     

Ultrastructure,meiotic behavior,and evolution of sex chromosomes of the genus Ellobius

The whole-mount SC preparations from males of three species of the genus Ellobius (Ellobius fuscocapillus, Ellobius lutescens), and Ellobius tancrei were studied by electron microscopy. In the males of Ellobius fuscocapillus, behavioral peculiarities of the sex bivalent (viz. the normal male heterozygosity) are characterized by early complete desynapsis of sex chromosomes (X, Y), occurring at late pachytene-early diplotene. The karyotype of species Ellobius lutescens is unique for mammals. In both sexes it is characterized by an odd number of chromosomes (2n=17). At prophase I the unpaired chromosome 9 is not involved in synapsis with other chromosomes and forms a sex body at the end of pachytene.The complete Robertsonian fan has been described for superspecies Ellobius tancrei. As shown on the basis of G-band patterns the male and female sex chromosomes are cytologically indistinguishable.Analysis of whole-mount SC preparations revealed the formation of a closed sex SC bivalent and showed some morphological differences in the axes of sex chromosomes at meiotic prophase I. A number of assumptions are made about the relationship between the behavior of sex chromosomes, their evolution and the sex determination system in the studied species of genus Ellobius.

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Genetic mapping of Y-chromosomal DNA markers in Pacific salmon

Sex chromosomes in fish provide an intriguing view of how sex-determination mechanisms evolve in vertebrates. Many fish species with single-factor sex-determination systems do not have cytogenetically-distinguishable sex chromosomes, suggesting that few sex-specific sequences or chromosomal rearrangements are present and that sex-chromosome evolution is thus at an early stage. We describe experiments examining the linkage arrangement of a Y-chromosomal GH pseudogene (GH-Y) sequence in four species of salmon (chum, Oncorhynchus keta; pink, O. gorbuscha; coho, O. kisutch; chinook, O. tshawytscha). Phylogenetic analysis indicates that GH-Y arose early in Oncorhynchus evolution, after this genus had diverged from Salmo and Salvelinus. However, GH-Y has not been detected in some Oncorhynchus species (O. nerka, O. mykiss and O. clarki), consistent with this locus being deleted in some lineages. GH-Y is tightly linked genetically to the sex-determination locus on the Y chromosome and, in chinook salmon, to another Y-linked DNA marker OtY1. GH-Y is derived from an ancestral GH2 gene, but this latter functional GH locus is autosomal or pseudoautosomal. YY chinook salmon are viable and fertile, indicating the Y chromosome is not deficient of vital genetic functions present on the X chromosome, consistent with sex chromosomes that are in an early stage of divergence.     

Diversity of centromeric repeats in two closely related wild rice species, Oryza officinalis and Oryza rhizomatis

Oryza officinalis (CC, 2n=24) and Oryza rhizomatis (CC, 2n=24) belong to the Oryza genus, which contains more than 20 identified wild rice species. Although much has been known about the molecular composition and organization of centromeres in Oryza sativa, relatively little is known of its wild relatives. In the present study, we isolated and characterized a 126-bp centromeric satellite (CentO-C) from three bacterial artificial chromosomes of O. officinalis. In addition to CentO-C, low abundance of CentO satellites is also present in O. officinalis. In order to determine the chromosomal locations and distributions of CentO-C (126-bp), CentO (155 bp) and TrsC (366 bp) satellite within O. officinalis, fluorescence in situ hybridization examination was done on pachytene or metaphase I chromosomes. We found that only ten centromeres (excluding centromere 7 and 2) contain CentO-C arrays in O. officinalis, while centromere 7 comprises CentO satellites, and centromere 2 is devoid of any detectable satellites. For TrsC satellites, it was detected at multiple subtelomeric regions in O. officinalis, however, in O. rhizomatis, TrsC sequences were detected both in the four centromeric regions (CEN 3, 4, 10, 11) and the multiple subtelomeric regions. Therefore, these data reveal the evolutionary diversification pattern of centromere DNA within/or between close related species, and could provide an insight into the dynamic evolutionary processes of rice centromere.     

The chromosomes ofCunninghamia konishii,C. lanceolata,andTaiwania cryptomerioides (Taxodiaceae)

Karyotype analyses were conducted onCunninghamia konishii, Cunninghamia lanceolata, andTaiwania cryptomerioides, all members ofTaxodiaceae. The somatic chromosome number was found to be 2n = 2x = 22 in all species which concurrs with previous reports. The karyotypes are generally asymmetrical with the smaller chromosomes being more submedian than the larger ones. Chromosomes with unusual or specific structures, thought to be associated with the nucleolar organizing region, were found in each species.Cunninghamia species have a marker chromosome pair with an unusually long secondary constriction.Taiwania has an unusually long kinetochore region present in a submedian chromosome pair.     

银白色葡萄球菌的发现、分布与致病性

银白色葡萄球菌(Staphylococcus argenteus)是金黄色葡萄球菌(S. aureus)的近缘新种,于2015年正式被鉴定、命名。在此之前,银白色葡萄球菌一直被认为是金黄色葡萄球菌种内的一个远源支系。该物种绝大多数菌株分离自人体,基因组上含有大量与金黄色葡萄球菌同源的毒力基因,可导致与金黄色葡萄球菌类似的临床感染和食物中毒症状。因为与人类健康相关,银白色葡萄球菌受到了越来越多的关注,目前已在20多个国家被报道。但是由于出现较晚,与金黄色葡萄球菌难以区分,银白色葡萄球菌的全球分布状况、原始生境和传播过程尚不清楚。本文从发现过程、分布、致病性和鉴定方法等方面综述了银白色葡萄球菌的最新研究进展。     

Cytogenetic studies on Apareiodon affinis (Pisces, Characiformes) from Paraná river basin: sex chromosomes and polymorphism

Comparative cytogenetic studies were carried out on Apareiodon affinis from an important hydrographic system at South America, the Paraná river basin. Two distant regions were chosen, which were separated by Guaíra Falls (formerly Sete Quedas); the region in the upper part of the hydrographic basin is called Upper Paraná (Brazil), whereas and the other in the lower part is called Lower Paraná (Argentina). Individuals from Upper Paraná have diploid numbers of 2n = 54 (NF = 108) for males and 2n = 55 (NF = 110) for females, showing female heterogamety with a ZZ/ZW₁W₂ multiple sex chromosomes system that is endemic for the region. In different localities at Lower Paraná, the specimens presented diploid number of 2n = 54 for both sexes, without any sex chromosomes heteromorphism. However, they have an accentuated polymorphism characterized by variation in number of acrocentric chromosomes, constituting something new for family Parodontidae. The most likely hypothesis to explain the origin of such polymorphism is based on successive pericentric inversions giving rise to acrocentric chromosomes. Thus, it was possible to detect 10 cytotypes along the Lower Paraná basin. Such chromosomal variations possibly are the consequence of an adaptative process. Our data probably indicate the occurrence of distinct species in each region that share the same denomination. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparative studies of isozymes in Oryza sativa,O. minuta,and their interspecific derivatives: evidence for homoeology and recombination

Enzyme electrophoresis was used to compare the isozyme phenotypes of Oryza sativa, IR31917 (AA genome), and two O. minuta accessions (Om 101089 and Om101141; BBCC genome) for ten enzyme systems. Between the two species, two systems were monomorphic (isocitrate dehydrogenase and alcohol dehydrogenase) and eight were polymorphic (shikimate dehydrogenase, phosphogluconate dehydrogenase, phosphoglucose isomerase, malate dehydrogenase, glutamate oxaloacetate transaminase, esterase, aminopeptidase, and endopeptidase). Polymorphism between O. minuta accessions was detected for shikimate dehydrogenase and glutamate oxaloacetate. As expected, the quaternary structure of the O. minuta isozymes was comparable to that of O. sativa. Possible allelic relationships with known O. sativa alleles and their genomic designation are discussed. Combined with chromosome data, the interspecific variation was exploited to monitor the relative genetic contribution of the two parents in the IR31917/Om101141 F₁ hybrids and recurrent (IR31917) backcross progenies. The isozyme content of F₁ hybrid reflected its triploid nature (ABC genome composition), while that of the backcross progenies paralleled the duplication of the A genome and the gradual loss of O. minuta chromosomes during the backcrossing process. Evidence is provided for a degree of homoeology between the A, B, and C genomes, and for introgression from O. minuta into O. sativa.     

Inactivation and injury of Escherichia coli O157:H7 and Staphylococcus aureus by pulsed electric fields

Two pathogenic microorganisms Escherichia coli O157:H7 and Staphylococcus aureus, suspended in peptone solution (0.1% w/v) were treated with 12, 14, 16 and 20 kV/cm electric field strengths with different pulse numbers up to 60 pulses. Pulsed electric field (PEF) treatment at 20 kV/cm with 60 pulses provided nearly 2 log reduction in viable cell counts of E. coli O157:H7 and S. aureus. S. aureus cells were slightly more resistant than E.coli O157:H7 cells. The results related to the effect of initial cell concentration of E. coli O157:H7 on the PEF inactivation showed that more inactivation was obtained by decreasing initial cell concentration. Any possible injury by PEF was also investigated after applying 20 kV/cm electric field to the microorganisms. As a result, it was determined that there was 35.92 to 43.36% injury in E. coli O157:H7 cells, and 17.26 to 30.86% injury in S. aureus cells depending on pulse number. The inactivation results were also described by a kinetic model.     

Characterization of sex chromosomes in rainbow trout and coho salmon using fluorescence in situ hybridization (FISH)

With the aim of characterizing the sex chromosomes of rainbow trout (Oncorhynchus mykiss) and to identify the sex chromosomes of coho salmon (O. kisutch), we used molecular markers OmyP9, 5S rDNA, and a growth hormone gene fragment (GH2), as FISH probes. Metaphase chromosomes were obtained from lymphocyte cultures from farm specimens of rainbow trout and coho salmon. Rainbow trout sex marker OmyP9 hybridizes on the sex chromosomes of rainbow trout, while in coho salmon, fluorescent signals were localized in the medial region of the long arm of one subtelocentric chromosome pair. This hybridization pattern together with the hybridization of a GH2 intron probe on a chromosome pair having the same morphology, suggests that a subtelocentric pair could be the sex chromosomes in this species. We confirm that in rainbow trout, one of the two loci for 5S rDNA genes is on the X chromosome. In males of this species that lack a heteromorphic sex pair (XX males), the 5S rDNA probe hybridized to both subtelocentrics This finding is discussed in relation to the hypothesis of intraspecific polymorphism of sex chromosomes in rainbow trout.     

Karyotype analysis and physical mapping of 45S rDNA in eight species of Sophora, Robinia , and Amorpha

The karyotype analysis and physical locations of 45S rDNA were carried out by means of fluorescence in situ hybridization in three species, and two forms of Sophora, two species of Robina, and one species of Amorpha. S. japonica L., S. japonica L. f. oligophylla Franch., S. japonica L. f. pendula Loud., and S. xanthantha C. Y. Ma. are all tetraploids with 2n = 28. There were four 45S rDNA sites in pericentromeric regions of two pairs of chromosomes in each of them. S. rubriflora Tsoong. is a triploid with 2n = 21, and three sites were located in each satellite of group 5 chromosomes. In R. pseudoacacia L. (2n = 2x = 22), we examined four intensive signals in telomeric regions of two pairs of satellite chromosomes. In R. hispida L. (2n = 2x = 30), there were four other signals in centromeric regions besides those like in R. pseudoacacia. Amorpha fruticosa L. has most chromosomes (2n = 40) among the eight materials, however, there were only six 45S rDNA loci and they laid in centromeric regions, and satellites of three pairs of chromosomes. 45S rDNA is a valuable chromosomal landmark in karyotype analysis. The distribution and genomic organization of rDNA in the three genera were also discussed. __________ Translated from Acta Botanica Yunnanica, 2005, 27(3): 261–268 [译自: 云南植物研究, 2005, 27(3): 261–268]     

A single dominant gene for Fusarium wilt resistance in protoplast-derived tomato plants

Summary Autotetraploids were established from 8 diploid wild species of section Arachis. In all the autotetraploids the chromosomes paired largely as bivalents even though they possess the ability to pair as multivalents. Pollen and pod fertility in the C₁ generation were not directly associated with chromosome pairing. The C₂ generation autotetraploids showed a gradual increase in bivalent associations and pollen and pod fertility. The identification of two genomes, A and B, in the diploid species and in the tetraploid, A. hypogaea, of the section Arachis, a fairly good crossability, and the type of chromosome associations observed in hybrids between A. hypogaea and the autotetraploids of wild Arachis species indicated good prospects of utilizing autotetraploids as genetic bridges in transferring desired traits from these taxa into groundnut.Submitted as Journal Article No. 516 by International Crops Research Institute for the Semi-Arid Tropics (ICRISAT)