Mitotic Recombination in the Heterochromatin of the Sex Chromosomes of DROSOPHILA MELANOGASTER

The frequency of spontaneous and X-ray-induced mitotic recombination involving the Y chromosome has been studied in individuals with a marked Y chromosome arm and different XY compound chromosomes. The genotypes used include X chromosomes with different amounts of X heterochromatin and either or both arms of the Y chromosome attached to either side of the centromere. Individuals with two Y chromosomes have also been studied. The results show that the bulk of mitotic recombination takes place between homologous regions.     

Dual-color FISH karyotype analyses using rDNAs in three Cucurbitaceae species

Dual-color fluorescence in situ hybridization (FISH) analysis of three Cucurbitaceae species from different genera was conducted using 5S and 45S rDNA probes. In Benincasa hispida (Thunb.) Cogn. (2n=24), the 45S rDNA probe hybridized on two chromosomes, one in the short arm of a medium-sized metacentric chromosome and another at the satellite of a chromosome. The 5S rDNA hybridized at a site proximal to the centromere of the same short arm of the 45S rRNA gene locus that occupied almost the entire short arm. For Citrullus lanatus (Thunb.) Matsum & Nakai (2n=22), the 45S rDNA probe hybridized at sites in the short arms of two chromosomes and the 5S rDNA probe was co-localized with the 45S rRNA locus at the region proximal to the centromere in one chromosome. The 45S rRNA loci occupied almost all of the short arms in both chromosomes. In Cucurbita moschata Duch. (2n=40), the 45S rDNA probe hybridized in five chromosomes in which the 45S rRNA genes occupied almost two-thirds of the chromosomes in two large chromosomes and the entire short arm of a medium-sized chromosome. Two other loci were present in two medium-sized chromosomes, one in the proximal region in the short arm of a chromosome and another at the tip of the long arm of a chromosome. Chromosomes of B. hispida were relatively larger than those of the other two species. The karyotype of B. hispida is composed of two metacentrics and 10 submetacentrics, while that of C. lanatus is composed of seven metacentrics and four submetacentrics and that of C. moschata is composed of 18 metacentrics and two submetacentrics. Comparative chromosome evolution among the three Cucurbitaceae species was attempted using the karyotypes and the chromosomal distribution patterns of the 5S and 45S rDNAs. The results presented herein will be useful in elucidating the phylogenetic relationships among Cucurbitaceae species, and will provide basic data for their breeding programs.     

Linkage relationships of 19 enzyme Loci in maize

Linkage relationships of 19 enzyme loci have been examined. The chromosomal locations of eight of these loci are formally reported for the first time in this paper. These localizations should assist in the construction of additional useful chromosome marker stocks, especially since several of these enzyme loci lie in regions that were previously poorly mapped. Six loci are on the long arm of chromosome 1. The arrangement is (centromere)—Mdh4-mmm-Pgm1-Adh1-Phi-Gdh1, with about 46% recombination between Mdh4 and Gdh1.—Linkage studies with a2 and pr have resulted in the localization of four enzyme genes to chromosome 5 with arrangement Pgm2-Mdh5-Got3-a2-(centromere)-pr-Got2. Pgm2 lies approximately 35 map units distal to a2 in a previously unmapped region of the short arm of 5, beyond ameiotic.—Approximately 23% recombination was observed between Mdh4 and Pgm1 on chromosome 1, while 17% recombination occurred between Mdh5 and Pgm2 on chromosome 5. Similarly, linkages between Idh1 and Mdh1, about 22 map units apart on chromosome 8, and between Mdh2 and Idh2, less than 5 map units apart on chromosome 6, were observed. Thus, segments of chromosomes 1 and 5 and segments of 6 and 8 may represent duplications on nonhomologous chromosomes.     

Investigation of Homologous Crossing over and Sister Chromatid Exchange in the Wheat Nor-B2 Locus Coding for Rrna and Gli-B2 Locus Coding for Gliadins

Recombination was investigated within the Nor-B2 locus of wheat chromosome 6B that contains several thousand of the 18S-5.8S-26S rRNA (rDNA) repeated units. Additionally, recombination was assessed for several chromosome regions, in arm 6Bq between the centromere and the B2 locus (awn suppressor) and in arm 6Bp between the centromere and Nor-B2, between Nor-B2 and a distal C-band and between Nor-B2 and Gli-B2 coding for gliadins. The experimental design permitted the distinction between crossing over between homologous chromosomes and exchange between sister chromatids. No homologous crossing over within the Nor-B2 locus was found in a sample of 446 chromosomes, but one exchange with the attributes of unequal sister chromatid exchange was identified. The molecular characteristics of this presumed sister chromatid exchange indicate that the spacer variants present in the Nor-B2 locus are clustered. No homologous recombination was detected within the distal Gli-B2 locus containing repeated genes coding for gliadin seed-storage proteins. Both arms of chromosome 6B showed low crossing-over frequency in the proximal regions. The distance from the centromere to Nor-B2 was only from 0.3 to 2.2 cM although it accounts for about two-thirds of the metaphase chromosome arm, which shows a great distortion of the metaphase map of the arm. The level of homologous recombination within the Nor-B2 locus is lower than in the chromosome region immediately distal to it. Whether it is comparable to that in the chromosome region proximal to it could not be determined. Recombination frequencies of different pairs of chromosome 6B in all but one interval paralleled the frequencies of their metaphase I pairing: Lower pairing at metaphase I was paralleled by lower crossing-over frequency. This relationship indicated that reduced metaphase I pairing between 6B chromosomes from different populations is due to impaired crossing-over and not due to precocious chiasma terminalization.     

Molecular Cytogenetic Characterization of the Dioecious Cannabis sativa with an XY Chromosome Sex Determination System

Hemp (Cannabis sativa L.) was karyotyped using by DAPI/C-banding staining to provide chromosome measurements, and by fluorescence in situ hybridization with probes for 45 rDNA (pTa71), 5S rDNA (pCT4.2), a subtelomeric repeat (CS-1) and the Arabidopsis telomere probes. The karyotype has 18 autosomes plus a sex chromosome pair (XX in female and XY in male plants). The autosomes are difficult to distinguish morphologically, but three pairs could be distinguished using the probes. The Y chromosome is larger than the autosomes, and carries a fully heterochromatic DAPI positive arm and CS-1 repeats only on the less intensely DAPI-stained, euchromatic arm. The X is the largest chromosome of all, and carries CS-1 subtelomeric repeats on both arms. The meiotic configuration of the sex bivalent locates a pseudoautosomal region of the Y chromosome at the end of the euchromatic CS-1-carrying arm. Our molecular cytogenetic study of the C. sativa sex chromosomes is a starting point for helping to make C. sativa a promising model to study sex chromosome evolution.     

Nature and distribution of chromosomal intertwinings in Saccharomyces cerevisiae.

To elucidate yeast chromosome structure and behavior, we examined the breakage of entangled chromosomes in DNA topoisomerase II mutants by hybridization to chromosomal DNA resolved by pulsed-field gel electrophoresis. Our study reveals that large and small chromosomes differ in the nature and distribution of their intertwinings. Probes to large chromosomes (450 kb or larger) detect chromosome breakage, but probes to small chromosomes (380 kb or smaller) reveal no breakage products. Examination of chromosomes with one small arm and one large arm suggests that the two arms behave independently. The acrocentric chromosome XIV breaks only on the long arm, and its preferred region of breakage is approximately 200 kb from the centromere. When the centromere of chromosome XIV is relocated, the preferred region of breakage shifts accordingly. These results suggest that large chromosomes break because they have long arms and small chromosomes do not break because they have small arms. Indeed, a small metacentric chromosome can be made to break if it is rearranged to form a telocentric chromosome with one long arm or a ring with an "infinitely" long arm. These results suggest a model of chromosomal intertwining in which the length of the chromosome arm prevents intertwinings from passively resolving off the end of the arm during chromosome segregation.     

Chromosomes and DNA of Mus

Using C-banded preparations of Mus dunni it is possible to study the behavior of constitutive heterochromatin in early stages of meiotic prophase. The X and the Y chromosomes, both of which contain a large amount of heterochromatin, lie apart in leptotene but move toward each other during zygotene. They then form the sex vesicle at late zygotene. In autosomes zygotene pairing appears to start from the telomeric ends. The centromere of the Y chromosome associates end-to-end with the terminal end of the long arm of the X chromosome. The autosomal heterochromatic short arms show forked morphology in certain bivalents at pachytene, suggesting probable incomplete synapsis.     

Recombination map of the common shrew, Sorex araneus (Eulipotyphla, Mammalia)

The Eurasian common shrew (Sorex araneus L.) is characterized by spectacular chromosomal variation, both autosomal variation of the Robertsonian type and an XX/XY(1)Y(2) system of sex determination. It is an important mammalian model of chromosomal and genome evolution as it is one of the few species with a complete genome sequence. Here we generate a high-precision cytological recombination map for the species, the third such map produced in mammals, following those for humans and house mice. We prepared synaptonemal complex (SC) spreads of meiotic chromosomes from 638 spermatocytes of 22 males of nine different Robertsonian karyotypes, identifying each autosome arm by differential DAPI staining. Altogether we mapped 13,983 recombination sites along 7095 individual autosomes, using immunolocalization of MLH1, a mismatch repair protein marking recombination sites. We estimated the total recombination length of the shrew genome as 1145 cM. The majority of bivalents showed a high recombination frequency near the telomeres and a low frequency near the centromeres. The distances between MLH1 foci were consistent with crossover interference both within chromosome arms and across the centromere in metacentric bivalents. The pattern of recombination along a chromosome arm was a function of its length, interference, and centromere and telomere effects. The specific DNA sequence must also be important because chromosome arms of the same length differed substantially in their recombination pattern. These features of recombination show great similarity with humans and mice and suggest generality among mammals. However, contrary to a widespread perception, the metacentric bivalent tu usually lacked an MLH1 focus on one of its chromosome arms, arguing against a minimum requirement of one chiasma per chromosome arm for correct segregation. With regard to autosomal chromosomal variation, the chromosomes showing Robertsonian polymorphism display MLH1 foci that become increasingly distal when comparing acrocentric homozygotes, heterozygotes, and metacentric homozygotes. Within the sex trivalent XY(1)Y(2), the autosomal part of the complex behaves similarly to other autosomes.     

B Chromosome Nondisjunction in Corn: Control by Factors near the Centromere

B chromosomes of corn are stable at all mitotic and meiotic divisions of the plant except the second pollen mitosis. In the latter division, B chromosomes undego mitotic nondisjunction at rates as high as 98%. Studies by several workers on B-A translocation chromosomes have provided evidence for the existence of four factors on the B chromosome that control nondisjunction and are separable from the centromere. Two of these factors, referred to here as factors 3 and 4, flank the B chromosome centromere. Factor 3 is the centromere-adjacent heterochromatin in the long arm of the B chromosome; factor 4 is located in the minute short arm. Evidence is presented here supporting the existence of factors 3 and 4. Deficiencies that include each factor were identified following centromeric misdivision events, with breaks at or near the centromere of a B-translocation chromosome. B chromosomes lacking factors 3 or 4 show much less nondisjunction than do chromosomes containing them. The possible function of factor 4 in nondisjuntion is also discussed.     

Patterns of replication in the neo-sex chromosomes ofDrosophila nasuta albomicans

Drosophila nasuta albomicans (with 2n = 6), contains a pair of metacentric neo-sex chromosomes. Phylogenetically these are products of centric fusion between ancestral sex (X, Y) chromosomes and an autosome (chromosome 3). The polytene chromosome complement of males with a neo-X- and neo-Y-chromosomes has revealed asynchrony in replication between the two arms of the neo-sex chromosomes. The arm which represents the ancestral X-chromosome is faster replicating than the arm which represents ancestral autosome. The latter arm of the neo-sex chromosome is synchronous with other autosomes of the complement. We conclude that one arm of the neo-X/Y is still mimicking the features of an autosome while the other arm has the features of a classical X/Y-chromosome. This X-autosome translocation differs from the other evolutionary X-autosome translocations known in certain species ofDrosophila.     

The origin of tertiary monosomics in the tomato

Six aneuploid tomato plants with 2n–1=23 chromosomes were observed in populations grown from the seedlings treated with thermal neutrons and from seeds treated with X-rays. Four of the aneuploids were tertiary monosomics in which, as a result of centromeric interchanges between two different chromosomes, two whole arms were missing from the complement and two arms connected at the centromere. In one aneuploid, as a result of centromeric breakage, the two short arms of a homologous pair were missing from the complement and the two long arms connected to the long arm and the short arm respectively of another chromosome in which breakage had occurred also at the centromere. In one aneuploid, the interchange has occurred in the arms, and not in the centromere. Here the aneuploid condition is due to the loss of an arm with a centromere and a short piece of the other arm.In most of the tertiary monosomics the missing arms were either the short arms of sub-metacentric chromosomes or any of the arms of metacentric chromosomes. However, in one case the long arms of two submetacentric chromosomes were lost from the complement. That in spite of such large chromosomal deletions the sporophyte can survive, may be due to the fact that the aberrant plants are mostly chimeras.This study was part of a project resulting from a contract between the Association Euratom-I.T.A.L., and the Agricultural University of Wageningen.     

The karyotype of the golden monkey (Rhinopithecus r. roxellanae)

In this paper, the karyotype and G-banding pattern of the chromosomes of cultured peripheral blood lymphocytes in R. r. roxellanae were investigated. The chromosome number of this species is 44 in both sexes. In R. r. roxellanae, as in other monkeys, sex is determined by specific sex chromosomes, i.e. the male is XY and the female is XX. The 21 pairs of autosomes consist of 7 pairs of metacentric chromoomes, 13 pairs of submetacentric chromosomes and one acrocentric pair. Chromosome measurements were made from highly enlarged photographic prints. They included the relative length, arm ratio and centromere index of each chromosome. Both chromosomal and chromatid aberrations were observed. They were 0·67 and 2%, respectively. Finally, G-banding pattern analysis of chromosomes of R. r. roxellanae were carried out. The results show that each homologous pair has its own special banding pattern, so that each of them is easily recognizable. Idiograms of chromosome complements with the Giemsa banding pattern are constructed.     

Embryonic Development in House Fly Eggs Fertilized with Genetically Deficient Sperm

In house flies, Musca domestica L., eggs fertilized with sperm that have chromosome deficiencies and duplications do not hatch, but develop to a stage where a fully differentiated, prehatch larva is formed. Fifteen different chromosome translocations involving most of the 10 arms of the 5 autosomes were studied by crossing male translocation heterozygotes to normal females. Egg hatch was reduced to 36–66% depending on the translocation used. Eggs that did not hatch after 24 hours were fixed, stained, and examined for stage of development. Several translocations involving the right arm of chromosome 4 indicate that the region closest to the centromere contains genes that affect the process of syngamy or early cleavage divisions, but do not reduce the ability of the sperm to compete for egg fertilization. Approximately 70% of the autosomal genes can be absent from sperm (not simultaneously but in different crosses) without inhibiting embryonic development.     

水貂的染色体研究

采用复制带、C带和硝酸银染色等分带技术研究了水貂的核型和带型。结果表明,2n=30,枝型为10（M）＋16（SM）＋2（A）,XX（M）。C-带显示该水貂的一些染色体的结构异染色质比较丰富,从着丝粒区域延伸到两臂上,No.5染色体着丝粒结构异染色质有些弱化;X染色体的结构异染色质较常染色体的丰富。Ag-NORs有3个,分布在No.8染色体的次缢痕区域和一条No.2染色体长臂接近着丝粒的区域。     

Localisation of several chromosome I genes of Aspergillus nidulans: implications for mitotic recombination

Summary Another laboratory previously reported that the vast majority of mitotic recombinants in chromosome I disomics of Aspergillus nidulans arise from double exchange events involving the centromeric region and a far distal, possibly telomeric, region. This conclusion was based on the assumption that the camC gene is located in a position far distal to the centromere on the left arm of chromosome. I. As a left arm location for camC distal to the centromere was possibly in conflict with mapping data obtained in the context of an unrelated project, camC was partially mapped along with three other previously unlocated chromosome I genes, davA, ornD and uapA. The data presented here indicate that camC is located in a position far distal to the centromere but on the right arm of chromosome I, a conclusion also supported by the previous data. The positioning of uapA and camC in far distal locations on the right arm of chromosome I indicates the existence of a vast, otherwise nearly unmapped region on this chromosome arm.     

The chromosome complement of the Rhesus monkey (Macaca mulatta) determined in kidney cells cultivated in vitro

Summary The chromosome complement of male and female Rhesus monkey has been investigated in kidney cells cultivatedin vitro for 3 to 6 days. The chromosome number is 42. The Y chromosome of the heterogametic male is the smallest element in the complement, and it is acrocentric. The X chromosome ranks eigth in decreasing order of size and typically has an arm ratio of 1.4. The autosomes form a graded size series of metacentric chromosomes, 3–15μ long in early metaphase, and with arm ratios from 1.1 to 3.3. Chromosome IX carries a large secondary constriction near the centromere; it is presumed to be the main nucleolar chromosome. A smaller secondary constriction is found consistently in the long arm of chromosome I. The X chromosome and chromosome XXI appear to be dimorphic in the limited population studied, the alternative forms differing in arm ratios but not in total length. An idiogram of the haploid chromosome complement is presented incorporating measurements of 10 completely analyzed nuclei, five from male monkeys and five from females. On the basis of relative length, arm ratio, and occurrence of secondary constrictions, most chromosomes of the complement can be individually identified. Supported in part by grants from the National Cancer Institute of Canada; the National Institutes of Health of the United States, Public Health Service; and the National Foundation for Infantile Paralysis.     

MULTIPLE GENE SITES FOR 5S AND 18 + 28S RNA ON CHROMOSOMES OF GLYPTOTENDIPES BARBIPES (STAEGER)

Ribosomal RNAs (28 + 18S and 5S) and 4S RNA extracted from the chironomid Glyptotendipes barbipes were iodinated in vitro with ¹²⁵I and hybridized to the salivary gland chromosomes of G. barbipes and Drosophila melanogaster. Iodinated 18 + 28 S RNA labeled three puffed sites with associated nucleoli on chromosomes IR, IIL, and IIIL of G. barbipes and the nucleolar organizer of Drosophila. Labeled 5S RNA hybridized to three sites on chromosome IIIR, two sites on chromosome IIR and one site in a Balbiani ring on chromosome IV of Glyptotendipes. Most of the label produced by this RNA was localized seven bands away from the centromere on the right arm of chromosome III, and we consider this to be the main site complementary to 5S RNA in the chironomid. This same RNA preparation specifically labeled the 56 EF region of chromosome IIR of Drosophila which has been shown previously to be the only site labeled when hybridized with homologous 5S RNA. Hybridization of G. barbipes chromosomes with iodinated 4S RNA produced no clearly localized labeled sites over the exposure periods studied.     

X heterochromatin subdivision and cytogenetic analysis in Sciara coprophila (Diptera,Sciaridae)

Five new translocations recovered from irradiated sperm, each having a break-point in the proximal X heterochromatin, have been designated T23, T29, T32, T40, and T70. These translocations, together with Oak Ridge T1, make possible the precise localization of several genetic components including the X centromere, the controlling element, and the ribosomal cistrons. Only the data pertaining to centromere location are presented here. The ribosomal cistrons and controlling element will be dealt with separately. Full cytological details are given for each of the five translocations. The break-points on the X define three blocks of heterochromatin designated H₁, H₂, and H₃. Together they comprise the right arm of the X. H₃ is the smallest and forms the very end of the chromosome. H₁ lies immediately to the right of the centromere. In T23 and T70 the breakpoints are located between H₂ and H₃; in T29 and T32 between H₂ and H₁. In Oak Ridge T1 the break-point lies between H₁ and the centromere and in T40 between the centromere and the whole left arm of the chromosome. For the first time it has been possible to determine the exact breakpoints of the long paracentric inversion that is found on the X homologue.This series of papers is dedicated to Professor Sally Hughes-Schrader —cytologist, naturalist, scholar — who in her eighty-second year is still exulting in the wonders of chromosome behavior and still endowed with that understanding and grace which have heightened immeasurably the lives of those who have known her     

Temperature-Sensitive Lethal Mutations on Yeast Chromosome I Appear to Define Only a Small Number of Genes

A method was developed for isolating large numbers of mutations on chromosome I of the yeast Saccharomyces cerevisiae. A strain monosomic for chromosome I (i.e., haploid for chromosome I and diploid for all other chromosomes) was mutagenized with either ethyl methanesulfonate or N-methyl-N'-nitro-N -nitrosoguanidine and screened for temperature-sensitive (Ts^- ) mutants capable of growth on rich, glucose-containing medium at 25° but not at 37°. Recessive mutations induced on chromosome I are expressed, whereas those on the diploid chromosomes are usually not expressed because of the presence of wild-type alleles on the homologous chromosomes. Dominant ts mutations on all chromosomes should also be expressed, but these appeared rarely. — Of the 41 ts mutations analyzed, 32 mapped on chromosome I. These 32 mutations fell into only three complementation groups, which proved to be the previously described genes CDC15, CDC24 and PYK1 (or CDC19). We recovered 16 or 17 independent mutations in CDC15, 12 independent mutations in CDC24 and three independent mutations in PYK1. A fourth gene on chromosome I, MAK16, is known to be capable of giving rise to a ts-lethal allele, but we recovered no mutations in this gene. The remaining nine mutations isolated using the monosomic strain appeared not to map on chromosome I and were apparently expressed in the original mutants because they had become homozygous or hemizygous by mitotic recombination or chromosome loss. — The available information about the size of chromosome I suggests that it should contain approximately 60–100 genes. However, our isolation in the monosomic strain of multiple, independent alleles of just three genes suggests that only a small proportion of the genes on chromosome I is easily mutable to give a Ts^--lethal phenotype. — During these studies, we located CDC24 on chromosome I and determined that it is centromere distal to PYK1 on the left arm of the chromosome.     

Chromosomal location of genes controlling flavonoid production in hexaploid wheat

Two-dimensional paper chromatography was performed on methanol extracts of leaves of hexaploid bread wheat, Triticum aestivum L. em. Thell. cultivar Chinese Spring, and of the available nullisomic-tetrasomic compensating lines, the tetrasomic lines and the ditelocentric lines. The chromatograms had 27 spots identified as flavonoids and six representing phenolic acids. Some of the areas were complex and contained more than one compound. Four flavonoids were identified as under the control of gene(s) on chromosome arms 1DS, 4DL, 5AS and 6BS. A phenolic glycoside was concluded to be controlled by a gene(s) on chromosome arm 7BL. Gene(s) on chromosome arm 4DL affected the amount of compounds in two other spots, and gene(s) on chromosome arm 4BS reduced the level of all flavonoid compounds. The individual compounds in some of the complex spots may be under the control of gene(s) on homoeologous chromosomes.