Synthesis of an attached autosome,C(3)RM,inDrosophila pseudoobscura

Drosophila pseudoobscura has three acrocentric autosomes. In the experiments reported, homologous arms of the third chromosome were attached to the same centromere. This is a reversed metacentric compound third chromosome, denoted by C(3)RM. This compound chromosome is relatively fertile in within-strain crosses (ca. 50% egg hatch) but sterile when outcrossed to a normal karyotype. When constructing translocations for this experiment, the behavior of the Y-autosome translocations suggested that this species can tolerate more Y chromosome deficiency while retaining fertility than canDrosophila melanogaster. Finally, there were no Robertsonian exchanges observed among the 96 autosome-autosome translocations analyzed cytologically.     

Karyotype of the Snow Sculpin <Emphasis Type="Italic">Myoxocephalus brandti</Emphasis> Steindachner (Cottidae) from Peter the Great Bay,Sea of Japan

The karyotype of the snow sculpin Myoxocephalus brandti, 2n = 44, NF = 46, from Peter the Great Bay was studied. Two-armed chromosomes were presented by one pair of metacentric chromosomes of medium size; one-armed chromosomes included two pairs of large subtelocentric chromosomes and a pair of large acrocentric chromosomes. Ag-NOR-staining in the telomere vicinity revealed nucleolus-organizing regions in one metacentric chromosome and in one medium size acrocentric chromosome in one of the fishes, in two homological small acrocentric chromosomes in three fishes, and in one acrocentric chromosome of average size in six fishes. No difference between the male and female karyotypes and any type of variability was revealed. The karyotypes of the snow sculpin M. brandti and the frog sculpin M. stelleri were compared. Their distinctions and similarities were displayed.Original Russian Text Copyright ¢ 2005 by Biologiya Morya, Ryazanova.     

Heteromorphic sex chromosomes of lizardfish (Synodontidae): focus on the ZZ-ZW1W2 system in Trachinocephalus myops

The karyotype and DNA content of four lizardfish species (family Synodontidae), that is, Saurida elongata, Synodus ulae, Synodus hoshinonis and Trachinocephalus myops, were analyzed. The karyotype of T. myops significantly differed from that of the other three species having diploid chromosome number of 48 with mainly acrocentric chromosomes and the ZZ-ZW sex chromosome system. The chromosome number of male T. myops was 2n=26, while that of female T. myops was 2n=27. The karyotype consisted of 11 pairs of metacentrics, one pair of acrocentrics and, in addition, two large metacentrics in the male and a single large metacentric, a distinctly small subtelocentric and a microchromosome in the female. C-banding demonstrated that in the female the subtelocentric chromosome and the microchromosome were heterochromatic. The karyotype of T. myops was thought to be derived from a 48 chromosome type synodontid fish through the involvement of Robertsonian rearrangement; the rearrangement of the sex chromosomes proceeded during karyotype evolution. Among the chromosomes, the large metacentrics were determined to be neo-Z (a fusion of the original Z and an autosome), the microchromosomes the W₁ (originally W), and the subtelocentric chromosomes the W₂ (derived from an autosome pair). The miniaturization of W₁ and W₂ chromosomes and their heterochromatinization suggested that sex chromosomes in this species have been already highly differentiated. The findings on DNA content implied that the karyotype of T. myops evolved by centric fusion events without loss in DNA amount.     

Chromosomal homologies betweenDrosophila lebanonensis andD. melanogaster determined by in situ hybridization

Twelve biotin-labelled recombinant DNA probes were hybridized to polytene chromosomes ofDrosophila melanogaster andD. lebanonesis. Probes were chosen in order to cover the whole chromosomal complement. Six probes correspond to known genes fromD. melanogaster (RpII215, H3–H4, MHC, hsp28/23, hsp83, hsp70), four probes are clones isolated from aD. subobscura library (Xdh, DsubS3, DsubG3, DsubG4) and the remaining two probes correspond to the Adh gene ofD. lebanonensis and to one sequence (262), not yet characterized, from the same species. The chromosomal homologies obtained from the in situ hybridization results allow us to determine that Muller's C and D chromosomal elements are fused in the karyotype ofD. lebanonensis and constitute the large metacentric chromosome. Single pericentric inversions in theE andB elements have generated the medium and small metacentric chromosomes, respectively. No great changes are detected in Muller'sA element, which remains acrocentric. The changes detected in the karyotypic evolution ofD. lebanonensis are frequently observed inDrosophila evolution, as deduced from chromosomal homologies of severalDrosophila species. The results are also consistent with Muller's proposal that chromosomal elements have been conserved during the evolution ofDrosophila.     

Physical mapping of the 5S ribosomal RNA genes in Arabidopsis thaliana by multi-color fluorescence in situ hybridization with cosmid clones

The 5S ribosomal RNA genes were mapped to mitotic chromosomes of Arabidopsis thaliana by fluorescence in situ hybridization (FISH). In the ecotype Landsberg erecta, hybridization signals appeared on three pairs of chromosomes, two of which were metacentric and the other acrocentric. Hybridization signals on one pair of metacentric chromosomes were much stronger than those on the acrocentric and the other pair of metacentric chromosomes, probably reflecting the number of copies of the genes on the chromosomes. Other ecotypes, Columbia and Wassilewskija, had similar chromosomal distribution of the genes, but the hybridization signals on one pair of metacentric chromosomes were very weak, and detectable only in chromosomes prepared from young flower buds. The chromosomes and arms carrying the 5S rDNA were identified by multi-color FISH with cosmid clones and a centromeric 180 bp repeat as co-probes. The metacentric chromosome 5 and its L arm carries the largest cluster of the genes, and the short arm of acrocentric chromosome 4 carries a small cluster in all three ecotypes. Chromosome 3 had another small cluster of 5S rRNA genes on its L arm. Chromosomes 1 and 2 had no 5S rDNA cluster, but they are morphologically distinguishable; chromosome 1 is metacentric and 2 acrocentric. Using the 5S rDNA as a probe, therefore, all chromosomes of A. thaliana could be identified by FISH. Chromosome 1 is large and metacentric; chromosome 2 is acrocentric carrying 18S-5.8S-25S rDNA clusters on its short arm; chromosome 3 is metacentric carrying a small cluster of 5S rDNA genes on its L arm; chromosome 4 is acrocentric carrying both 18S-5.8S-25S and 5S rDNAs on its short (L) arm; and chromosome 5 is metacentric carrying a large cluster of 5S rDNA on its L arm.     

Cytogenetic characterization and description of an XX/XY1Y2 sex chromosome system in catfish Harttia carvalhoi (Siluriformes, Loricariidae)

Karyotypic and cytogenetic characteristics of catfish Harttia carvalhoi (Paraíba do Sul River basin, S?o Paulo State, Brazil) were investigated using differential staining techniques (C-banding, Ag-staining) and fluorescent in situ hybridization (FISH) with 18S and 5S rDNA probes. The diploid chromosome number of females was 2n = 52 and their karyotype was composed of nine pairs of metacentric, nine pairs of submetacentric, four pairs of subtelocentric and four pairs of acrocentric chromosomes. The diploid chromosome number of males was invariably 2n = 53 and their karyotype consisted of one large unpaired metacentric, eight pairs of metacentric, nine pairs of submetacentric, four pairs of subtelocentric, four pairs of acrocentric plus two middle-sized acrocentric chromosomes. The differences between female and male karyotypes indicated the presence of a sex chromosome system of XX/XY1Y2 type, where the X is the largest metacentric and Y1 and Y2 are the two additional middle-sized acrocentric chromosomes of the male karyotype. The major rDNA sites as revealed by FISH with an 18S rDNA probe were located in the pericentromeric region of the largest pair of acrocentric chromosomes. FISH with a 5S rDNA probe revealed two sites: an interstitial site located in the largest pair of acrocentric chromosomes, and a pericentromeric site in a smaller metacentric pair of chromosomes. Translocations or centric fusions in the ancestral 2n = 54 karyotype is hypothesized for the origin of such multiple sex chromosome systems where females are fixed translocation homozygotes whereas males are fixed translocation heterozygotes. The available cytogenetic data for representatives of the genus Harttia examined so far indicate large kayotype diversity.     

Synaptonemal complex analysis of heteromorphic trivalents in Lemur hybrids

Synaptonemal complexes (SCs) in surface spread pachytene spermatocytes of Lemur resemble those in other mammals and are of two types: metacentric (or submetacentric) and acrocentric, with a very short second arm. In autosomal SC and mitotic karyotypes of Lemur fulvus (2n=60) a 11 proportionality in relative length is observed as in other mammals. In an intraspecific lemur hybrid (2n=55) obtained by mating L. fulvus rufus (2n=60) x L. fulvus collaris (2n=51), G-band patterns show that 10 single acrocentric mitotic chromosomes correspond to the arms of 5 single metacentrics, implying homology. It is inferred that the metacentrics have evolved by centric (Robertsonian) fusion of the acrocentrics. In the SC karyotype of the hybrid all SCs are normal except for five which have the configurations expected of metacentric-acrocentric trivalents. Similarly, in L. f. collaris (2n= 51), with one unpaired metacentric and two unpaired acrocentrics, one such SC trivalent is present in the complement. In an SC trivalent, each of the acrocentric long axes is synapsed with an arm of the metacentric axis, confirming the homology predicted from banding similarities. At late zygotene, the acrocentric short arms, which are non-homologous, are the last to pair, demonstrating that synapsis of the homologous arms occurs first. At later pachytene the acrocentric short arms are fully synapsed, producing a short SC side arm. This subsequent non-homologous synapsis is taken to be an instance of the synaptic adjustment phenomenon which has been shown to lead to non-homologous synapsis in a duplication and several inversions in the mouse. The kinetochore of the metacentric is the same size as those of the acrocentrics, and thus is unlikely to have arisen by true centromeric fusion, but rather by a translocation. The kinetochores of the acrocentrics always lie together on the same side of the metacentric kinetochore (cis configuration), implying a single pairing face on the metacentric axis. The observed trivalent configuration may well constitute a prerequisite for proper meiotic disjunction in metacentric-acrocentric heterozygotes. Such a mechanism is consistent with fertility regularly observed in such hybrid lemurs.     

Flow karyotyping and sorting of Vicia faba chromosomes

Summary Chromosome suspensions were prepared from formaldehyde-fixed, synchronized Vicia faba root tips. After staining with the DNA intercalating fluorochrome propidium iodide, the suspensions were analysed with a flow cytometer. The resulting histograms of integral fluorescence intensity contained peaks similar to those of theoretical V.faba flow karyotypes. From V. Faba cv Inovec (2n = 12) only one peak, corresponding to a single chromosome type (metacentric chromosome), could be discriminated. However, it was found that the peak also contained doublets of acrocentric chromosomes. Bivariate analysis of fluorescence pulse area (chromosome DNA content) and fluorescence pulse width (chromosome length) was necessary to distinguish the metacentric chromosome. To achieve a high degree of purity, a two-step sorting protocol was adopted. During a working day, more than 25 000 metacentric chromosomes (corresponding to 0.2 g DNA) were sorted with a purity of more than 90%. Such chromosomes are suitable for physical gene mapping by in situ hybridization or via the polymerase chain reaction (PCR) and allow the construction of chromosome-specific DNA libraries. While it was only possible to distinguish and sort one chromosome type from V. Faba cv Inovec with the standard karyotype, it was possible to sort with a high degree of purity five out of six chromosome types of the line EFK of V. Faba, which has six pairs of morphologically distinct chromosomes. This result confirmed the possibility of using reconstructed karyotypes to overcome existing problems with the discrimination and flow sorting of individual chromosome types in plants.     

Heterochromatin banding in the genus Paris

The heterochromatin content of new accessions of the genus Paris have been studied using C banding and Q banding techniques. Supernumerary chromosomes in two accessions are shown to differ in their heterochromatin content. The results are related to previous cytogenetic work in the genus Paris.     

Molecular characterization and cytogenetic analysis of highly repeated DNAs of lake trout,Salvelinus namaycush

The chromosomes of lake trout (Salvelinus namaycush) contain a considerable amount of heterochromatin located at the centromeres and/or telomeres of several chromosomes, including a sex-specific block located distally on the X chromosome. In order to investigate further the repetitive DNAs of lake trout, genomic DNA from a female was size fractionated (<600 bp) with the restriction endonuclease AluI and fragments were cloned into the bacteriophage M13. A total of 42 clones were isolated. Relative copy number of individual inserts within the lake trout genome was estimated by Southern analysis. Twelve clones were determined to be highly repetitive and were chosen for further investigation. Inserts of these clones contained sequences similar to the AluI/RsaI, EcoRI/DraI, DraI/BstEII, and MboI/BglII families reported from Arctic char (Salvelinus alpinus). The chromosomal location of several of these fragments was determined in lake trout by fluorescence in situ hybridization (FISH). Two related AluI/RsaI sequences (Type A, 140 bp, and Type B, 120 bp) showed differential hybridization. Type A hybridized to the centromeres of all metacentric as well as several acrocentric chromosomes. Type B hybridized to the centromeres of most acrocentric chromosomes. A sequence with homology to the EcoRI/DraI family hybridized to the centromeres of several acrocentric chromosomes. Sequences with partial similarity to the DraI/BstEII family hybridized to the major rDNA sites (nucleolar organizer regions, NORs) and several minor telomeric sites. The interstitial and telomeric heterochromatin of lake trout, including that of the X chromosome, appears to comprise sequences belonging to the MboI/BglII family.     

Notes on chromosomes of ten species of the genus chrysolina mots. (Coleoptera:Chrysomelidae)

SixChrysolina species from Catalonia and the Canary Islands (Spain), viz.americana, gemina, femoralis, cerealis, menthastri andpolita, have similar diploid karyotypes of 24 (sub)metacentric chromosomes, and show Xy sex-determining system.C. banksi andobsoleta have 2n ()=23; their karyotype is presumably derived from that of the former group by loss of the y chromosomes. InC. haemoptera andC. carnifex 40 elements appear in the diploid set. It seems that 2n=24 is the most frequent number in theChrysolina. Higher chromosome numbers have possibly originated through centric fissions, as the acrocentric shape ofC. carnifex chromosomes seems to suggest. The 2n=23 and 24 species feed onLabiatae, while the two higher chromosome number species are associated with plants belonging to other families.     

Complex sex-determining mechanisms in three species of South American grasshoppers (Orthoptera,Acridoidea)

The karyotype of three species of South American grasshoppers are studied in this paper. Leiotettix sanguineus has two chromosome races, one of them with 2n=23 and an XO sex mechanism and the other, as far as we know limited to the Cerro Chato population, with 2n=22 and an XY sex mechanism. Leiotettix politus has two kinds of individuals, one with 2n=14 and XY sex chromosomes and the other 2n=13 and an X₁X₂Y mechanism. Dichroplus dubius presents 2n=21 and an X₁X₂Y sex chromosomes. One of the three specimens studied shows aberrant behaviour in the meiotic process.     

Unequal X Chromosomes in Bradysia Hygida (Diptera:Sciaridae) Females: Karyotype Assembly and Morphometric Analysis

The mitotic chromosomes of Bradysia hygida(Diptera:Sciaridae) neuroblast cells are described together with their morphometric data. Giemsa-stained neuroblast chromosomes from female and male larvae confirm the chromosome number of this species, 2n=8 (XX) and 2n=7 (XO), respectively. The karyotype assembly reveals two metacentric autosomic pairs, the A and B chromosome; a subtelocentric, the C chromosome, the smallest one; and a sexual unequal metacentric pair, X chromosome, in female karyotype and a one sexual metacentric X chromosome in male. The implications of the unequal X chromosome pair are discussed.     

Proximity of the CTLA-1 serine esterase and Tcr α loci in mouse and man

The serine esterase CTLA-1 gene was shown by in situ hybridization to map to the D segment of mouse chromosome 14, the same localization as a member of the immunoglobulin super family, Tcr . To further demonstrate the proximity of CTLA-1 and Tcr , genetic linkage was tested in mouse using restriction fragment length polymorphisms and a backcross progeny, and no recombination was observed in the 100 backcross products studied. Recombination events between Tcr /CTLA-1 and the markers Gdh-X and NP-1 show that the most probable order of these loci in the mouse 14D region is NP-1-Tcr /Ctla-1-Gdh-X. In man, the human homologue of CTLA-1 was shown by in situ hybridization to map on chromosome 14, at 14q11-q12, where Tcr also maps. Using the human cell line SUP-Tl, bearing the inversion inv(14)(q11;q32), we further demonstrated the loci order in man to be centromere-NP-1-Tcr -CTLA-1. To complement the cytogenetic and genetic mapping data, we tried to determine the physical distance between the two genes by pulsed field gel electrophoresis (PFGE). DNA prepared from various cell types, both mouse and human, were digested with a panel of rare cutter enzymes and hybridized first with CTLA-1, then with Tcr probes. None of the bands identified hybridized with both Tcr and CTLA-1 probes for either mouse or human cells. Although the physical mapping by PFGE is inconclusive, the cytogenetic and genetic data support close linkage of the Tcr and CTLA-1 genes in both mouse and man, suggesting homology between the D region of mouse chromosome 14 and the q11–q12 region of human chromosome 14, encompassing the Tcr and CTLA-1 loci. These findings also provide another example of proximity of genes coding for a member of the Ig super-family and a serine esterase.     

Karyological study of four Japanese Myotis bats (Chiroptera,Mammalia)

Karyological investigations of four Japanese Myotis species were made based on Gand C-banding pattern analysis. It was revealed that the four species, M. nattereri, M. hosonoi, M. frater kaguyae and M. macrodactylus have all 2n=44 and their karyotypes are, excepting one chromosome pair, identical each other. The only difference in their karyotypes was found on the morphology of the chromosome no. 5. A minute acrocentric (A) was observed in M. nattereri, and a polymorphic (A) and an (M^h) which is a minute metacentric with totally heterochromatic arm was found in M. hosonoi. In M. f. kaguyae, pair no. 5 was a small submetacentric with a totally heterochromatic long arm (SM^h). Polymorphic (SM^h) and (M) which is a small metacentric derived from (SM^h) by a pericentric inversion was seen in M. macrodactylus. Such morphological differentiations of no. 5 were interpreted by assuming an increase of constitutive heterochromatin and also an inversion. The evolutionary pathway in the genus Myotis is assumed to be as follows: (A)(M^h)(SM^h)(M). This assumption was supported by the geographical evidence that the species with the (A) type no. 5 pair is widely distributed in the whole world but the others are restricted to Asia (M^h type) or only to Japan (SM^h and M types).     

A high-resolution karyotype of <Emphasis Type="Italic">Brassica rapa</Emphasis> ssp. <Emphasis Type="Italic">pekinensis</Emphasis> revealed by pachytene analysis and multicolor fluorescence in situ hybridization

A molecular cytogenetic map of Chinese cabbage (Brassica rapa ssp. pekinensis, 2n=20) was constructed based on the 4-6-diamino-2-phenylindole dihydrochloride-stained mitotic metaphase and pachytene chromosomes and multicolor fluorescence in situ hybridization (McFISH), using three repetitive DNA sequences, 5S rDNA, 45S rDNA, and C11-350H. The lengths of mitotic metaphase chromosomes ranged from 1.46 m to 3.30 m. Five 45S and three 5S rDNA loci identified were assigned to different chromosomes. The C11-350H loci were located on all the mitotic metaphase chromosomes, except chromosomes 2 and 4. The pachytene karyotype consisted of two metacentric (chromosomes 1 and 6), five submetacentric (chromosomes 3, 4, 5, 9 and 10), two subtelocentric (chromosomes 7 and 8), and one acrocentric (chromosome 2) chromosome(s). The mean lengths of ten pachytene chromosomes ranged from 23.7 m to 51.3 m, with a total of 385.3 m, which is 17.5-fold longer than that of the mitotic metaphase chromosomes. In the proposed pachytene karyotype, all the chromosomes of B. rapa ssp. pekinensis can be identified on the basis of chromosome length, centromere position, heterochromatin pattern, and the location of the three repetitive sequences. Moreover, the precise locations of the earlier reported loci of 5S rDNA, 45S rDNA, and Chinese cabbage tandem DNA repeat C11-350H were established using McFISH analysis. We also identified a 5S rDNA locus on the long arm of pachytene bivalent 7, which could not be detected in the mitotic metaphase chromosomes in the present and earlier studies. The deduced karyotype will be useful for structural and functional genomic studies in B. rapa.     

Father and son with karyotype 47,XY,?Yq-

Summary A 22-year-old man and his father, both with 47 chromosomes, an extra small metacentric chromosome and the presumptive karyotype of 47,XY, ?Yq-, are presented.Literature concerning cases with 47 chromosomes and a small metacentric chromosome is reviewed and discussed. It is concluded that it is unlikely that persons with 47 chromosomes and an extra small metacentric chromosome constitute a new cytogenetic disease the syndrome of the metacentric microchromosome, as suggested by Abbo and Zellweger (1970).Persons with 47 chromosomes and an extra small metacentric chromosome have most probably a great variety of chromosomal aberrations, some of autosomal and some of sex chromosomal origin.

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Zusammenfassung Es wird über einen 22jährigen Mann und seinen Vater berichtet. Beide hatten 47 Chromosomen: Ein zusätzliches metazentrisches kleines Chromosom und den angenommenen Karyotyp 47,XY, ?Yq-.In diesem Zusammenhang wird die Literatur über Fälle mit 47 Chromosomen und einem kleinen metazentrischen Extrachromosom diskutiert. Die Autoren gelangen zu der Schlußfolgerung, es sei unwahrscheinlich, daß eine neue cytogenetisch charakterisierte Krankheit, das Syndrom des metazentrischen Mikrochromosoms, wie Abbo u. Zellweger (1970) es annehmen, in der Tat existiert. Personen mit 47 Chromosomen und einem kleinen metazentrischen Extrachromosom haben sehr wahrscheinlich eine große Anzahl verschiedener Chromosomenaberrationen, einige autosomalen und andere geschlechtschromosomalen Ursprungs.

     

A comparative study of karyotypes of muntjacs by chromosome painting

We have used a combination of chromosome sorting, degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), chromosome painting and digital image capturing and processing techniques for comparative chromosome analysis of members of the genus Muntiacus. Chromosome-specific paints from a female Indian muntjac were hybridised to the metaphase chromosomes of the Gongshan, Black, and Chinese muntjac by both single and three colour chromosome painting. Karyotypes and idiograms for the Indian, Gongshan, Black and Chinese muntjac were constructed, based on enhanced 4, 6-diamidino-2-phenylindole (DAPI) banding patterns. The hybridisation signal for each paint was assigned to specific bands or chromosomes for all of the above muntjac species. The interspecific chromosomal homology was demonstrated by the use of both enhanced DAPI banding and comparative chromosome painting. These results provide direct molecular cytogenetic evidence for the tandem fusion theory of the chromosome evolution of muntjac species.Deceased     

A new mechanism for altering chromosome number during karyotype evolution

Summary A new mechanism for changing chromosome numbers (preserving the fundamental number of long chromosome arms) during karyotype evolution is suggested. It includes: 1) Occurrence of individuals heterozygous for two interchanges between different arms of three chromosomes (a metacentric and two acrocentric ones). 2) Formation in heterokaryotypes of multivalents during meiosis between the chromosomes involved in the interchanges and their unchanged homologues. 3) Mis-segregation of chromosomes from these multivalents resulting in hypoploid (n-1) and hyperploid (n+1) simultaneously instead of euhaploid gametes. 4) Fusion of n-1 or n+1 gametes which gives rise to (zygotes and) individuals representing homokaryotypes with changed number of chromosomes (2n+2 or 2n-2), but preserves (as compared to the parental karyotypes) the number of long chromosome arms. Under definite conditions, chromosome numbers of the progeny may be changed by this process in both directions (upwards and downwards). The mechanism is free of the difficulties associated with the explanation for such changes by direct Robertsonian interchanges (see Discussion), which are usually considered to be responsible for such alterations in chromosome number. The above-mentioned process has been experimentally documented in Vicia faba and it probably also occurred naturally within the Vicia sativa group.     

Sex identification by male-specific growth hormone pseudogene (GH-Ψ) in Oncorhynchus masou complex and a related hybrid

It is often difficult to identify sexes of many fish species by conventional cytological method because of the lack of heteromorphic sex chromosomes. Isolation of sex-specific molecular markers is thus important for sexing and for understanding sex chromosome evolution in these species. We have identified genetic sexes by PCR-based male-specificity of a growth hormone pseudogene (GH-) in masu and Biwa salmon, two subspecies of the Oncorhynchus masou complex, and their hybrid Honmasu. PCRs with primers designed from sequences of chinook salmon GH genes amplified GH-I and GH-II fragments in both sexes, but a third GH- fragment was detected in predominant proportion of males and very few phenotypic females. The consistency of phenotypic sex with genetic sex identified by GH- for masu salmon, Biwa salmon and Honmasu was 93.1, 96.7 and 94%, respectively. The remaining individuals showed inconsistency or deviation from sex-specificity: a few phenotypic males lacked the GH-, whereas a few phenotypic females possessed the GH-. Sequence of the putative GH- fragment from such females was identical to that from genetic males, and shared about 95% homology with the corresponding GH- fragment from chinook salmon. This result confirmed that these females were really GH--bearing individuals. PCR analyses with primers designed from masu salmon GH- gave identical results, indicating that the absence of GH- in a few males was not resulted from primer mismatching. These GH--bearing females and GH--absent males were more likely to originate from spontaneous sex reversion than from crossing-over between GH- and the sex determination gene/region.