Aneuploid Progeny Research Articles

Completion of meiosis in male zebrafish (Danio rerio) despite lack of DNA mismatch repair gene mlh1

Mlh1 is a member of DNA mismatch repair (MMR) machinery and is also essential for the stabilization of crossovers during the first meiotic division. Recently, we have shown that zebrafish mlh1 mutant males are completely infertile because of a block in metaphase I, whereas females are fertile but have aneuploid progeny. When studying fertility in males in a two-fold more inbred background, we have however observed low numbers of fertilized eggs (approximately 0.4%). Histological examination of the testis has revealed that all spermatogenic stages prior to spermatids (spermatogonia, primary spermatocytes, and secondary spermatocytes) are significantly increased in the mutant, whereas the total weight of spermatids and spermatozoa is highly decreased (1.8 mg in wild-type vs. 0.1 mg in mutants), a result clearly different from our previous study in which outbred males lack secondary spermatocytes or postmeiotic cells. Thus, a delay of both meiotic divisions occurs rather than complete arrest during meiosis I in these males. Eggs fertilized with mutant sperm develop as malformed embryos and are aneuploid making this male phenotype much more similar to that previously described in the mutant females. Therefore, crossovers are still essential for proper meiosis, but meiotic cell divisions can progress without it, suggesting that this mutant is a suitable model for studying the cellular mechanisms of completing meiosis without crossover stabilization.

Genetic basis for dosage sensitivity in Arabidopsis thaliana.

Aneuploidy, the relative excess or deficiency of specific chromosome types, results in gene dosage imbalance. Plants can produce viable and fertile aneuploid individuals, while most animal aneuploids are inviable or developmentally abnormal. The swarms of aneuploid progeny produced by Arabidopsis triploids constitute an excellent model to investigate the mechanisms governing dosage sensitivity and aneuploid syndromes. Indeed, genotype alters the frequency of aneuploid types within these swarms. Recombinant inbred lines that were derived from a triploid hybrid segregated into diploid and tetraploid individuals. In these recombinant inbred lines, a single locus, which we call SENSITIVE TO DOSAGE IMBALANCE (SDI), exhibited segregation distortion in the tetraploid subpopulation only. Recent progress in quantitative genotyping now allows molecular karyotyping and genetic analysis of aneuploid populations. In this study, we investigated the causes of the ploidy-specific distortion at SDI. Allele frequency was distorted in the aneuploid swarms produced by the triploid hybrid. We developed a simple quantitative measure for aneuploidy lethality and using this measure demonstrated that distortion was greatest in the aneuploids facing the strongest viability selection. When triploids were crossed to euploids, the progeny, which lack severe aneuploids, exhibited no distortion at SDI. Genetic characterization of SDI in the aneuploid swarm identified a mechanism governing aneuploid survival, perhaps by buffering the effects of dosage imbalance. As such, SDI could increase the likelihood of retaining genomic rearrangements such as segmental duplications. Additionally, in species where triploids are fertile, aneuploid survival would facilitate gene flow between diploid and tetraploid populations via a triploid bridge and prevent polyploid speciation. Our results demonstrate that positional cloning of loci affecting traits in populations containing ploidy and chromosome number variants is now feasible using quantitative genotyping approaches.

Progenies of allotriploids of Oriental × Asiatic lilies (Lilium) examined by GISH analysis

With the aim of utilizing allotriploid (2n = 3x = 36) lily hybrids (Lilium) in introgression breeding, different types of crosses were made. First, using diploid Asiatic lilies (2n = 2x = 24), reciprocal crosses (3x − 2x and 2x − 3x) were made with allotriploid hybrids (AOA) obtained by backcrosses of F1 Oriental × Asiatic hybrids (OA) to Asiatic cultivars (A). Secondly, the AOA allotriploids were crossed with allotetraploid (OAOA, 2n = 4x = 48), in 3x − 4x combination. Finally, the AOA allotriploids where crossed to 2n gamete producer F1 OA hybrids (3x − 2x (2n)). Two types of triploids were used as parents in the different types of crosses, derived from: (a) mitotic polyploidization and (b) sexual polyploidization. Ploidy level of the progeny was determined by estimating the DNA values through flowcytometry as well as chromosome counting. The aneuploid progeny plants from 3x − 2x and reciprocal crosses had approximate diploid levels and in 3x − 4x crosses and 3x − 2x (2n) the progeny had approximate tetraploid levels. Balanced euploid gametes (x, 2x and 3x) were formed in the AOA genotypes. Recombinant chromosomes were found in the progenies of all crosses, except in the case of 2x − 3x crosses through genomic in situ hybridization (GISH) analyses. Recombinant chromosomes occurred in the F1 OA hybrid when the triploid AOA hybrid was derived through sexual polyploidization, but not through mitotic polyploidization with two exceptions. Those recombinant chromosomes were transmitted to the progenies in variable frequencies.

Asexual reproduction in a close relative of Arabidopsis: a genetic investigation of apomixis in Boechera (Brassicaceae)

Understanding apomixis (asexual reproduction through seeds) is of great interest to both plant breeders and evolutionary biologists. The genus Boechera is an excellent system for studying apomixis because of its close relationship to Arabidopsis, the occurrence of apomixis at the diploid level, and its potentially simple inheritance by transmission of a heterochromatic (Het) chromosome. Diploid sexual Boechera stricta and diploid apomictic Boechera divaricarpa (carrying a Het chromosome) were crossed. Flow cytometry, karyotype analysis, genomic in situ hybridization, pollen staining and seed-production measurements were used to analyse the parents and resulting F1, F2 and selected F3 and test-cross (TC) generations. The F1 plant was a low-fertility triploid that produced a swarm of aneuploid and polyploid F2 progeny. Two of the F2 plants were fertile near-tetraploids, and analysis of their F3 and TC progeny revealed that they were sexual and genomically stabilized. The apomictic phenotype was not transmitted by genetic crossing as a single dominant locus on the Het chromosome, suggesting a complex genetic control of apomixis that has implications for future genetic and evolutionary analyses in this group.

Mechanisms and Risk of Chemically Induced Aneuploidy in Mammalian Germ Cells

Aneuploidy is a pathological condition that affects 35% of human spontaneous abortions and 0.3% of livebirths. In spite of the increasing knowledge about molecular mechanisms of meiosis and chromosome segregation, maternal age remains the only ascertained aetiological factor. Genetically modified mouse models have been produced that show increased incidence of aneuploid gametes or abnormalities in meiotic recombination and synapsis. They suggest that genetic polymorphisms might also be involved in the aetiology of human germ cell aneuploidy. Experimental studies in the mouse have identified chemicals that can induce aneuploidy in male and female germ cells. Compounds affecting spindle assembly/dynamics are potent aneugens for oocytes and less so also for spermatocytes. They are active at acute doses during a short time interval preceding the metaphase-to-anaphase transition. Topoisomerase inhibitors are also meiotic aneugens which act on the recombination process; for the first time, the production of viable aneuploid mouse progeny was shown after paternal treatment with etoposide. A comparison between in vitro and in vivo effects of suspect aneugens demonstrates that there are biological mechanisms protecting mammalian oocytes from acute exposures to exogenous chemicals. Endocrine disruptors are a novel group of compounds that might affect chromosome segregation at meiosis. Data on bisphenol-A suggest that such chemicals could be active at low chronic exposure levels, but this hypothesis needs to be confirmed by further experiments. Experiments on cultured mouse oocytes treated with inhibitors of biochemical reactions involved in the regulation of chromosome segregation point to possible new mechanisms of action of environmental aneugens.

Decoding the links between mitosis, cancer, and chemotherapy: The mitotic checkpoint, adaptation, and cell death

Disrupted passage through mitosis often leads to chromosome missegregation and the production of aneuploid progeny. Aneuploidy has long been recognized as a frequent characteristic of cancer cells and a possible cause of tumorigenesis. Drugs that target mitotic spindle assembly are frequently used to treat various types of human tumors. These lead to chronic mitotic arrest from sustained activation of the mitotic checkpoint. Here, we review the linkage between the mitotic checkpoint, aneuploidy, adaptation from mitotic arrest, and antimitotic drug-induced cell death.

Genetic Basis for Dosage Sensitivity in A. thaliana

Aneuploidy, the relative excess or deficiency of specific chromosome types, results in gene dosage imbalance. Plants can produce viable and fertile aneuploid individuals, while most animal aneuploids are inviable or developmentally abnormal. The swarms of aneuploid progeny produced by Arabidopsis triploids constitute an excellent model to investigate the mechanisms governing dosage sensitivity and aneuploid syndromes. Indeed, genotype alters the frequency of aneuploid types within these swarms. Recombinant inbred lines that were derived from a triploid hybrid segregated into diploid and tetraploid individuals. In these recombinant inbred lines, a single locus, which we call SENSITIVE TO DOSAGE IMBALANCE (SDI), exhibited segregation distortion in the tetraploid subpopulation only. Recent progress in quantitative genotyping now allows molecular karyotyping and genetic analysis of aneuploid populations. In this study, we investigated the causes of the ploidy-specific distortion at SDI. Allele frequency was distorted in the aneuploid swarms produced by the triploid hybrid. We developed a simple quantitative measure for aneuploidy lethality and using this measure demonstrated that distortion was greatest in the aneuploids facing the strongest viability selection. When triploids were crossed to euploids, the progeny, which lack severe aneuploids, exhibited no distortion at SDI. Genetic characterization of SDI in the aneuploid swarm identified a mechanism governing aneuploid survival, perhaps by buffering the effects of dosage imbalance. As such, SDI could increase the likelihood of retaining genomic rearrangements such as segmental duplications. Additionally, in species where triploids are fertile, aneuploid survival would facilitate gene flow between diploid and tetraploid populations via a triploid bridge and prevent polyploid speciation. Our results demonstrate that positional cloning of loci affecting traits in populations containing ploidy and chromosome number variants is now feasible using quantitative genotyping approaches.

Transmission and recombination of homeologous Solanum sitiens chromosomes in tomato.

The goal of the present experiments was to transfer the chromosomes of Solanum sitiens (syn. Solanum rickii) into cultivated tomato ( Lycopersicon esculentum). By crossing an allotetraploid L. esculentum x Solanum sitiens hybrid to sesquidiploid L. esculentum x S. lycopersicoides, a trigenomic hybrid (2n+14=38) was obtained. Analysis of the latter by GISH (genomic in situ hybridization) indicated it contained a full set of 12 S. sitiens chromosomes, plus two extras from S. lycopersicoides. This and other complex hybrids were pollinated with Lycopersicon pennellii-derived bridging lines to overcome unilateral incompatibility. A total of 40 progeny were recovered by embryo rescue, including diploids and aneuploids (up to 2n+8). In order to determine the origin of chromosomes and the location of introgressed segments, progeny were genotyped with RFLP markers. S. sitiens-specific markers on all chromosomes, except 6 and 11, were detected in the progeny. Several S. sitiens chromosomes were transmitted intact, either through chromosome addition (i.e., trisomics) or substitution (i.e., disomics). Recombination between S. sitiens and L. esculentum was detected on most chromosomes, in both diploid and aneuploid progeny. A monosomic alien addition line for S. sitiens chromosome 8 was identified, and the extra chromosome was stably transmitted to approximately 13% of the backcross progeny. This study demonstrates the feasibility of gene transfer from S. sitiens to L. esculentum through chromosome addition, substitution, and recombination in the progeny of complex aneuploid hybrids.

Non-Mendelian Inheritance Revealed in a Genetic Analysis of Sexual Progeny of Phytophthora cinnamomi with Microsatellite Markers

We report the development of four microsatellite loci into genetic markers for the diploid oomycete plant pathogen Phytophthora cinnamomi and that (AC)n and (AG)n microsatellites are significantly less frequent than in plant and mammal genomes. A minisatellite motif 14 bp long was also discovered. The four microsatellite loci were used to analyze sexual progeny from four separate crosses of P. cinnamomi. A large proportion of non-Mendelian inheritance was observed across all loci in all four crosses, including inheritance of more than two alleles at a locus and noninheritance of alleles from either parent at a locus. The aberrant inheritance is best explained by nondisjunction at meiosis in both the A1 parent and the A2 trisomic parents, resulting in aneuploid progeny. Two loci on the putative trisomic chromosome showed linkage and no loci were linked to mating type. One aneuploid offspring was shown to have lost alleles at two loci following subculture over 4 years, indicating that aneuploid progeny may not be mitotically stable.

Aberrant meioses and viable aneuploid progeny of induced triploid loach ( Misgurnus anguillicaudatus) when crossed to natural tetraploids

In induced triploid loach Misgurnus anguillicaudatus, both bivalents and univalents were observed in the early pachytene stage of oocytes and they then degenerated in the late pachytene stage. Only a few yolk-laden oocytes were occasionally observed in the ovaries of triploids. Spermatocytes of triploid males formed both bivalents and univalents, and they developed to aneuploid spermatozoa. Flow cytometry revealed that the modal DNA content of these spermatozoa was 1.3C. These spermatozoa were able to fertilize the eggs of natural tetraploids (4 n×3 n) and the resultant aneuploid progeny showed chromosome numbers and DNA contents between triploidy and tetraploidy. These aneuploid progeny exhibited poorer survival than control triploids and tetraploids and most fry died before feeding due to severe malformation. However, those fry which began to feed survived. The average body length of one-month-old survivors was not different between aneuploids and tetraploids, but aneuploids showed significantly larger variance than the control.

Mapping a telomere using the translocation eT1(III;V) in Caenorhabditis elegans.

In Caenorhabditis elegans, individuals heterozygous for a reciprocal translocation produce reduced numbers of viable progeny. The proposed explanation is that the segregational pattern generates aneuploid progeny. In this article, we have examined the genotype of arrested embryonic classes. Using appropriate primers in PCR amplifications, we identified one class of arrested embryo, which could be readily recognized by its distinctive spot phenotype. The corresponding aneuploid genotype was expected to be lacking the left portion of chromosome V, from the eT1 breakpoint to the left (unc-60) end. The phenotype of the homozygotes lacking this DNA was a stage 2 embryonic arrest with a dark spot coinciding with the location in wild-type embryos of birefringent gut granules. Unlike induced events, this deletion results from meiotic segregation patterns, eliminating complexity associated with unknown material that may have been added to the end of a broken chromosome. We have used the arrested embryos, lacking chromosome V left sequences, to map a telomere probe. Unique sequences adjacent to the telomeric repeats in the clone cTel3 were missing in the arrested spot embryo. The result was confirmed by examining aneuploid segregants from a second translocation, hT1(I;V). Thus, we concluded that the telomere represented by clone cTel3 maps to the left end of chromosome V. In this analysis, we have shown that reciprocal translocations can be used to generate segregational aneuploids. These aneuploids are deleted for terminal sequences at the noncrossover ends of the C. elegans autosomes.

Identification, transmission, and cytological behavior ofSolanum lycopersicoidesDun. monosomic alien addition lines in tomato (Lycopersicon esculentumMill.)

The wild nightshade Solanum lycopersicoides possesses a number of disease- and arthropod-resistance and environmental stress tolerance traits of potential value for the cultivated tomato, Lycopersicon esculentum. F1hybrids are readily obtained by sexual crosses, but backcrosses to tomato are hindered by sterility and incompatibility barriers. In the present studies, intergeneric sesquidiploid hybrids (2 genomes of L. esculentum and 1 of S. lycopersicoides) were pollinated with Lycopersicon pennellii derived bridging lines, yielding diploid, aneuploid (up to 2n + 6), and polyploid (3x, 4x) progeny. After diploids (58% of total progeny), the group most frequently obtained was 2n + 1 (31%), which included all 12 possible monosomic alien addition lines (MAALs) containing a single extra chromosome from S. lycopersicoides in a tomato background. MAAL groups were identified with a combination of allozyme, RFLP, and monogenic visual markers. Transmission rates of the extra chromosomes in MAALs varied from 0%, for chromosome 6, to 24%, for chromosome 10. At diakinesis and metaphase I of meiosis, the alien chromosome was usually unpaired (in an average of 90% of PMCs), but association of homoeologous chromosomes was observed in each of the 12 MAAL groups (in an average of 5.7% of PMCs). The MAALs showed a striking morphological resemblance to the corresponding tomato primary trisomic. In addition, each MAAL expressed S. lycopersicoides specific leaf, flower, and (or) fruit characters. This set of MAALs, the first of its kind in Lycopersicon, is expected to be useful for a variety of gene-mapping and introgression studies.

Occurrence of aneuploid progenies from an asynaptic amphidiploid ofScilla scilloides (lindley) druce II. Mechanism of production of the various aneuploid progenies

The mechanism of production of the various aneuploid progenies was clarified in the asynaptic amphidiploid plants (2n=34+4f+2F, AABB) ofScilla scilloides. Its asynaptic nature and chromosomal stickiness lead to the unequal segregation at anaphase I (AI) in PMC's. The observed values in 18 segregation patterns, 17:17 to 0: 34, were different from the expected values estimated from random segregation of chromosomes. Nevertheless, the preferential transmission of special chromosomes among genomes A (x=8=a1−a8) and B (x=9=b1−b9) had not occurred. As the result of unequal segregation, the pollen grains with various chromosome numbers were observed. Almost all of the 200 pollen grains contained chromosome numbers more than 17 (range 8 to 34). The observed values of each chromosome number were roughly similar to the expected values of containing the complete set of genome A or B in the random distribution without preferential segregation of chromosomes at AI. The difference between the index of polien mitosis and the pollen fertility was significant in the Wilcoxon matched-pairs signed rank test and suggested the selection for some genomically unbalanced pollen grains during maturation. Consequently, viable pollen grains with various chromosome constitutions are a few (mean pollen fertility of 5.8%) but might produce many aneuploids by self- and cross-pollination.

Detection of nondisjunction and recombination in meiotic and postmeiotic cells from XYSxr [XY,Tp(Y)1Ct] mice using multicolor fluorescence in situ hybridization.

Current meiotic dogma holds that synapsis is required for recombination and that recombination is required for proper disjunction. The mouse chromosome aberration XYSxr [sex reversal; redesignated XY,Tp(Y)1Ct] appears to challenge this assumption, for although chromosomes X and Y often fail to synapse and recombine, there is no dramatic increase in aneuploid progeny. An explanation of this conundrum might be that X-Y univalent spermatocytes do not survive. The phenotype of sex reversal is generated by the "obligatory" crossover between the X and Y chromosomes, which always occurs proximal to a duplicated copy of the testis-determining gene Sry and transfers one copy from one chromatid of the Y chromosome to one chromatid of the X. Animals that inherit an X chromosome with the Sry gene are chromosomally female but phenotypically male. We have used fluorescence in situ hybridization (FISH) to visualize probes for the X and Y chromosomes and for the Sry sequence and chromosome 8 to track the fate of both recombinant and nonrecombinant chromosomes through metaphases I and II into spermatids and sperm. In the 219 gametes examined by multicolor FISH, the frequency of aneuploid products (XY or "O") was low (3.7%) despite a high frequency (66%) of X-Y separation at metaphase I. In balanced gametes, X and Y recombinant chromosomes slightly exceeded nonrecombinants. Both of these observations support the earlier proposal that asynapsis and nondisjunction in primary spermatocytes lead to their developmental arrest and degeneration.

Fertility of Aneuploids between the 5x and 6x Levels in Blueberry: The Potential for Gene Transfer from 4x to 6x Levels

Vaccinium ashei (6x) /V. corymbosum (4x) pentaploid hybrids backcrossed to V. ashei yield aneuploid progeny ranging in chromosome number from 5x to 6x levels. Six backcross aneuploids having chromosome numbers of 2n = 61, 62, 64, 66, 68, and 70 were selected from this backcross and crossed in a complete diallel mating design and backcrossed (as female parents) to two V. ashei cultivars and an interspecific hexaploid hybrid. Fertility variables measured were percent fruit set, total seed per berry, developed seed per berry, percent developed seed per berry, percent seed germination, developed seed per pollination, and seedlings per pollination. A significant linear and positive relationship was found between chromosome number and all seven fertility variables. However, regression accounted for 30% or less of the variation among crosses. Diallel analysis revealed that general combining ability was the major contributing effect for all seven variables, followed by reciprocal effects. Specific combining ability was not significant. The second backcross to the hexaploid level suggested significant effects due to both the BC1 aneuploid and hexaploid genotypes and to a significant genotype × genotype interaction for three of the variables. All six aneuploids were either fully or partially self-sterile. The findings of this study substantiate earlier suggestions that pentaploids in blueberry can be used to facilitate bilateral transfer of characteristics between the tetraploid and hexaploid levels in blueberry.

Haploid Oat Plants Produced by Application of Maize Pollen to Emasculated Oat Florets

Only six haploids in oat (Avena sativa L.) have been previously reported, five of spontaneous origin and one from anther culture. Our objective was to develop more efficient methods for producing oat haploids to use in selecting mutants, recovering aneuploids, and producing doubled‐haploid lines for genetic and breedings tudies. In a series of experiments, pollen from maize (Zea mays L.) was applied to previously emasculated oat florets. Twelve to 15 d later excised ovaries/caryopses, or embryos taken from them, were placed onto an amino acid‐supplemented Murashige and Skoog medium containing 7% sucrose for embryo rescue. Recovered plantlets were potted in soil and grown to maturity. Root tips and meiotic tissues were sampled for cytological analyses. Overall, 14 haploid oat plants were recovered by embryo rescue following application of maize pollen to approximately 3300 emasculated oat florets. Root tip cells in each of the recovered plants had the oat haploid chromosome number of 21. Presumably these oat haploids originated from interspecies hybrid zygote formation followed by elimination of maize chromosomes during initial cell divisions, as has been described in haploid wheat (Triticum aestivum L.) formation in wheat ✕ maize hybridizations. In the initial experiment, which involved combinations of various oat and maize genotypes, each of the four oat haploids recovered was from a different oat cultivar and each involved a different source of maize pollen; thus, indicating that the process is not genotype unique. Meiotic cells of the recovered haploid plants were characterized by aberrant chromosome behavior and numerous micronuclei, as expected in a haploid. Occasional seed were set on haploid plants and both euploid (2n) and aneuploid (2n‐1 and 2n‐2) progeny were obtained. The use of maize pollinations provides a new approach for obtaining haploid oat plants for genetic and breeding studies.

The influence of the Robertsonian translocation Rb(X.2)2Ad on anaphase I non-disjunction in male laboratory mice.

A Robertsonian translocation in the mouse between the X chromosome and chromosome 2 is described. The male and female carriers of the Rb(X.2)2Ad were fertile. A homozygous/hemizygous line was maintained. The influence of the X-autosomal Robertsonian translocation on anaphase I non-disjunction in male mice was studied by chromosome counts in cells at metaphase II of meiosis and by assessment of aneuploid progeny. The results conclusively show that the inclusion of Rb2Ad in the male genome induces non-disjunction at the first meoitic division. In second metaphase cells the frequency of sex-chromosomal aneuploidy was 10.8%, and secondary spermatocytes containing two or no sex chromosome were equally frequent. The Rb2Ad males sired 3.9% sex-chromosome aneuploid progeny. The difference in aneuploidy frequencies in the germ cells and among the progeny suggests that the viability of XO and XXY individuals is reduced. The pairing configurations of chromosomes 2, Rb2Ad and Y were studied during meiotic prophase by light and electron microscopy. Trivalent pairing was seen in all well spread nuclei. Complete pairing of the acrocentric autosome 2 with the corresponding segment of the Rb2Ad chromosome was only seen in 3.2% of the cells analysed in the electron microscope. The pairing between the X and Y chromosome in the Rb2Ad males corresponded to that in males with normal karyotype. Reasons for sex-chromosomal non-disjunction despite the normal pairing pattern between the sex chromosomes may be seen in the terminal chiasma location coupled with the asynchronous separation of the sex chromosomes and the autosomes.(ABSTRACT TRUNCATED AT 250 WORDS)

Aneuploids induced by deficiencies of chromosome 9 and analysis of the time of nondisjunction in cotton

Plants of allotetraploid cotton, Gossypium hirsutum L. (2n = 4x = 52), which are monosomic for chromosome 9 or haplodeficient for the long arm of the chromosome, an Ah genome chromosome, produce a high frequency of aneuploid progeny. The aneuploids include monosomes, trisomes, and multiple monosomic and trisomic combinations that are the result of chromosome nondisjunction, which is induced by the deficiency of the long arm of chromosome 9. Loss of chromosomes occurs far more frequently than additions, and chromosomes of the Ah genome undergo nondisjunction at a significantly greater rate than do chromosomes of the Dh genome. Stained paraffin sections of ovules at different stages of development of monotelodisomic 9 and normal plants (TM1) were analyzed to determine the time and probable causes of nondisjunction during megasporogenesis. The most significant difference between mono 9 and TM1 was observed in ovules in stages of development just before and at the time of flowering. The mono 9 female gametophytes had supernumerary nuclei, indicating that chromosome nondisjunction occurs at the mitotic divisions of the embryo sac.Key words: chromosome nondisjunction, aneuploids, megasporogenesis, cotton, Gossypium hirsutum cytogenetics.

The induction of rho− mutants by UV or γ-rays is independent of the nuclear recombinational repair pathway in Saccharomyces cerevisiae

In order to discover whether the nuclear recombinational repair pathway also acts on lesions induced in mitochondrial DNA (mtDNA), the possible role of the RAD50, −51, −52, −55 and −56 genes on the induction of rho − mutants by radiations was studied. Such induction appeared to be independent of this pathway. Nevertheless, an efficient induction of respiration-deficient mutants was observed in γ-irradiated rad52 diploids. We demonstrate that these mutants do not result from a lack of mtDNA repair, but from chromosome losses induced by γ-rays. Such an impairment of the respiratory ability of diploids by chromosome losses was effectively observed in the aneuploid progeny of unirradiated RAD + cdc6 diploids incubated at the restrictive temperature.

Nuclear fusion-defective phenocopies in Chlamydomonas reinhardtii: mating-type functions for meiosis can act through the cytoplasm.

Nuclear fusion in newly formed Chlamydomonas reinhardtii zygotes can be inhibited by drugs that affect microtubule stability, which include colchicine, amiprophosmethyl, oryzalin, and taxol. This inhibition can be monitored genetically by the production of haploid meiotic products from conjugations between haploid and diploid parents. Such zygotes would normally produce aneuploid progeny. Inhibition of nuclear fusion by colchicine requires treatment of gametic cells both before conjugation and after formation of the zygotes. These results suggest that nuclear fusion requires dynamic microtubules. Treated zygotes formed from a haploid-diploid mating can produce six spores, but only four spores germinate to form viable haploid colonies. No contribution from the nuclear genome of the haploid parent is recovered, whereas all loci from the diploid parent are recovered. The four viable products from the diploid parent of inhibited zygotes show normal segregation of loci located on linkage groups segregating according to Mendelian laws. Levels of meiotic recombination were tested for pairs of loci on linkage groups XVIII and XIX and found to be unchanged by inhibition of nuclear fusion. Thus, similar to Saccharomyces cerevisiae, C. reinhardtii mating-type functions necessary for nuclear fusion are not nuclear limited and can act through the cytoplasm. Inhibition of nuclear fusion can be used to analyze diploid Chlamydomonas that cannot enter meiosis. This technique permits direct analysis of dominant mutations, dominant suppressors and enhancers, and new alleles of identified loci that have been isolated in diploid strains.