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PubMed1996

Self-sterility in Arabidopsis due to defective pollen tube guidance.

In flowering plants, a series of cell-cell interactions govern the delivery of sperm to the ovules through precise guidance of pollen tubes. Two Arabidopsis genes, POP2 and POP3, were found that mediate pollen tube guidance and are critical for self-fertility in diploid reproductive cells. The pop2 and pop3 mutations exhibited genetic redundancy: Self-sterility occurred only when male and female tissues were defective in both genes. This phenotype resembles that found in many self-incompatible species.

Arabidopsis↗
PubMed1988

DNAs of the two mating-type alleles of Neurospora crassa are highly dissimilar.

The mating-type alleles A and a of Neurospora crassa control mating in the sexual cycle and function in establishing heterokaryon incompatibility in the vegetative cycle. The A and a alleles were cloned, and they were shown to encode both the sexual functions and vegetative incompatibility. The mating-type clones contain nonhomologous DNA segments that are flanked by common DNA sequences. Neurospora crassa and all heterothallic and pseudohomothallic Neurospora species contain a single copy of one mating-type sequence or the other within each haploid genome. The six known self-fertile homothallic isolates contain an A homolog, but only one species also contains a homologous sequences. Homothallism in these species is not due to mating-type switching, as it is in Saccharomyces cerevisiae.

Chromosome Mapping↗
PubMed1986

Delay of disease development in transgenic plants that express the tobacco mosaic virus coat protein gene.

A chimeric gene containing a cloned cDNA of the coat protein (CP) gene of tobacco mosaic virus (TMV) was introduced into tobacco cells on a Ti plasmid of Agrobacterium tumefaciens from which tumor inducing genes had been removed. Plants regenerated from transformed cells expressed TMV mRNA and CP as a nuclear trait. Seedlings from self-fertilized transgenic plants were inoculated with TMV and observed for development of disease symptoms. The seedlings that expressed the CP gene were delayed in symptom development and 10 to 60 percent of the transgenic plants failed to develop symptoms for the duration of the experiments. Increasing the concentration of TMV in the inoculum shortened the delay in appearance of symptoms. The results of these experiments indicate that plants can be genetically transformed for resistance to virus disease development.

DNA↗
PubMed1979

Osmotic shock prevents nuclear exchange and produces whole-genome homozygotes in conjugating Tetrahymena.

Exposure of conjugating Tetrahymena to a hyperosmotic shock blocks the exchange of gametic nuclei and produces self-fertilized exconjugants that are homozygous for their whole genome. Cells are sensitive to this induction during a brief period after meiosis. The high efficiency of the treatment and the fertility of the progeny make this a useful method for the isolation of induced recessive mutations and enhances the value of Tetrahymena as an animal-cell model system in which genetic dissection is practical. The sharp peak of sensitivity is useful in the study of those cellular mechanisms responsible for the independent handling of several functionally distinct nuclei during conjugation.

Animals↗
PubMed1981

Mating types and macrocyst formation in Dictyostelium.

Macrocysts have been found among strains of Dictyostelium rosarium Raper and Cavender (K. B. Raper and J. C. Cavender, J. Elisha Mitchell Sci. Soc. 84:31-47, 1968). Strains that exhibit mating capabilities could be classified into one of three mating types: A1, A2, or A3. Each mating type was self-incompatible but cross-compatible with the other two types in most cases. No self-fertile strain was observed. Wet conditions, darkness, and moderate nutrient particularly favored macrocyst production; opposite conditions generally favored asexual sorocarp formation. Macrocyst development followed the same morphogenetic pattern reported previously for other species of Dictyostelium. Cultures exposed to light in stages prior to precyst appearance could be switched from macrocyst to sorocarp development. In contrast, precysts always developed into macrocysts despite the presence of light. There appeared to be an "all-or-none" phenomenon in macrocyst production, suggesting that transformation to macrocysts occurred once certain light-sensitive critical factors had reached a threshold level. Intimate association of cells of mating pairs appeared to be essential for macrocyst formation, and no mating hormones were detected.

Crosses, Genetic↗
PubMed1993

Unidirectional dominance of cytoplasmic inheritance in two genetic crosses of Plasmodium falciparum.

Malarial parasites have two highly conserved cytoplasmic DNA molecules: a 6-kb tandemly arrayed DNA that has characteristics of a mitochondrial genome, and a 35-kb circular DNA that encodes functions commonly found in chloroplasts. We examined the inheritance pattern of these elements in two genetic crosses of Plasmodium falciparum clones. Parent-specific oligonucleotide probes and single-strand conformation polymorphism analysis identified single nucleotide changes that distinguished the parental 6- and 35-kb DNA molecules in the progeny. In all 16 independent recombinant progeny of a cross between a Central American clone, HB3, and a Southeast Asian clone, Dd2, the 6- and 35-kb DNAs were inherited from the Dd2 parent. In all nine independent recombinant progeny of a cross between clone HB3 and a likely African clone, 3D7, the 6-kb DNA was inherited from the 3D7 parent. Inheritance of cytoplasmic genomes of the Dd2 and 3D7 parents was, therefore, dominant over that of the HB3 parent. Cytoplasmic DNA molecules were found almost exclusively in the female gametes of malarial parasites; hence, clone HB3 did not appear to have served as a maternal parent for the progeny of two crosses. Defective differentiation into male gametes by clone Dd2 is likely to be a reason for the cytoplasmic inheritance pattern seen in the HB3 x Dd2 cross. However, incompetence of male or female gametes is unlikely to explain the uniparental dominance in recombinant progeny of the HB3 x 3D7 cross, since both parents readily self-fertilized and completed the malaria life cycle on their own. Instead, the data suggest unidirectional parental incompatibility in cross-fertilization of these malarial parasites, where a usually cosexual parental clone can participate only as a male or as a female. Such an incompatibility may be speculated as indicating an early phase of reproductive isolation of P. falciparum clones from different geographical regions.

Animals↗
PubMed1997

Roles of Wee1 and Nim1 protein kinases in regulating the switch from mitotic division to sexual development in Schizosaccharomyces pombe.

In self-fertile strains of the fission yeast Schizosaccharomyces pombe, nitrogen starvation initiates a program of sexual development in which cells express mating pheromones and receptors, arrest cell cycle progression in G1, and conjugate. This process is dependent on Rum1, an inhibitor of the Cdc2-Cdc13 and Cdc2-Cig2 cyclin B kinases. The M-phase induction activity of Cdc2-Cdc13 is inhibited by Wee1 tyrosine kinase, which phosphorylates Cdc2 on tyrosine-15. We report here that Wee1 activity is also important for mating. This discovery arose from studies of Nim1, a kinase which promotes mitosis by inhibiting Wee1. Nim1 was previously thought to have an important role in promoting mitosis during nitrogen starvation, but our studies revealed that Nim1 protein drops to an undetectable level within 15 min of nitrogen depletion. In contrast, Wee1 remains abundant, and tyrosine-phosphorylated Cdc2 is detected for at least 4 h after resuspension of cells in nitrogen-free medium. This suggested that maintenance of Wee1 activity may be important during the early stages of nitrogen starvation, a proposal confirmed by the observation that mating efficiency is reduced ca. fivefold in wee1- cells. Transcriptional induction of genes encoding mating factors and receptors is also delayed in wee1- cells. The wee1- mating defect is suppressed by deletion of cig2+, which encodes a B-type cyclin that promotes the onset of S and inhibits conjugation. These findings indicate that Wee1 and Rum1 act jointly to inhibit Cdc2 and promote sexual development in nitrogen-starved cells.

Ammonium Chloride↗
PubMed1985

Direct and indirect gene replacements in Aspergillus nidulans.

We performed three sets of experiments to determine whether cloned DNA fragments can be substituted for homologous regions of the Aspergillus nidulans genome by DNA-mediated transformation. A linear DNA fragment containing a heteromorphic trpC+ allele was used to transform a trpC- strain to trpC+. Blot analysis of DNA from the transformants showed that the heteromorphic allele had replaced the trpC- allele in a minority of the strains. An A. nidulans trpC+ gene was inserted into the argB+ gene, and a linear DNA fragment containing the resultant null argB allele was used to transform a trpC- argB+ strain to trpC+. Approximately 30% of the transformants were simultaneously argB-. The null argB allele had replaced the wild-type allele in a majority of these strains. The A. nidulans SpoC1 C1-C gene was modified by removal of an internal restriction fragment and introduced into a trpC- strain by transformation with a circular plasmid. A transformant containing a tandem duplication of the C1-C region separated by plasmid DNA was self-fertilized, and trpC- progeny were selected. All of these had lost the introduced plasmid DNA sequences, whereas about half had retained the modified C1-C gene and lost the wild-type copy. Thus, it is possible with A. nidulans to replace chromosomal DNA sequences with DNA fragments that have been cloned and modified in vitro by using either one- or two-step procedures similar to those developed for Saccharomyces cerevisiae.

Aspergillus nidulans↗
PubMed1999

Microsatellite retrieval in lettuce (Lactuca sativa L.).

By using enriched genomic libraries, microsatellite-containing sequences were isolated from lettuce (Lactuca sativa) with high efficiency. With this approach, a sizeable fraction (up to 55%) of the clones contained a microsatellite. In about half of these clones, primers could be designed for PCR amplification of the microsatellite. This yielded 28 primer sets amplifying unambiguously scorable products, of which 26 showed polymorphisms in a test set of six lettuce varieties. Practically all microsatellite-amplifying primer sets yielded products in lettuce's nearest relative, L. serriola, but only half of the primer sets yielded products in the more distant species L. saligna and L. virosa. An average polymorphism information content (PIC) value of 0.55 and an average number of 3.5 alleles per locus were in the normal range for a self-fertilizing species like lettuce. In addition, the incidental cloning of a microsatellite-containing repeat family, apparently specific for Lactuca, is reported and the implications for the efficient retrieval of single-locus microsatellite sequences are discussed. The microsatellite loci isolated will be useful for distinguishing lettuce cultivars and for screening diversity of genetic resources.

Alleles↗
PubMed1993

Analysis of ploidy and sexual characteristics of natural isolates of Cryptococcus neoformans.

We report here an extensive examination of the ploidy of a basidiomycetous yeast. The ploidy of 34 strains of Cryptococcus neoformans var. neoformans, isolated mostly from Japanese patients, was determined by means of quantitative fluorescence microscopy after DNA staining with propidium iodide. The sexual characteristics of the strains including the two tester strains were as follows. Twenty-four strains were of the heterothallic alpha type, 1 was of the heterothallic a type, and 2 were self fertile; the remaining 7 showed no sexual response. The following strains were found to be haploid: the heterothallic a strain, 20 of the 24 heterothallic alpha strains, and 3 of the 7 asexual strains. Thus, the C. neoformans strains were mostly haploid, as the sexual response of this organism suggests. Both of the self-fertile strains and four of the seven asexual strains were diploid. Unexpectedly, 4 of the 24 heterothallic alpha strains were diploid.

Cryptococcus neoformans↗
PubMed1995

A predicted membrane protein, TRA-2A, directs hermaphrodite development in Caenorhabditis elegans.

The nematode C. elegans naturally develops as either an XO male or XX hermaphrodite. The sex-determining gene, tra-2, promotes hermaphrodite development in XX animals. This gene encodes a predicted membrane protein, named TRA-2A, which has been proposed to provide the primary feminising activity of the tra-2 locus. Here, we show that transgenic TRA-2A driven from a heat shock promoter can fully feminise the somatic tissues of XX tra-2 loss-of-function mutants, which would otherwise develop as male. TRA-2A is thus likely to provide a component of the tra-2 locus that is both necessary and sufficient to promote female somatic development. Transgenic TRA-2A driven by the heat shock promoter can also transform XO animals from male to self-fertile hermaphrodite. This result establishes the role of tra-2 as a developmental switch that controls somatic sexual cell fate. We show that a carboxy-terminal region of TRA-2A, predicted to be intra-cellular, can partially feminise XX tra-2 loss-of-function mutants and XO tra-2(+) males. We suggest that this intra-cellular domain of TRA-2A promotes hermaphrodite development by negatively regulating the FEM proteins.

Animals↗
PubMed1983

Bioassay of genotoxic effects of environmental particles in a feeding ciliate.

The ciliated protozoan, Paramecium, can be used to quantitate cytotoxic and genotoxic effects of ingested complex environmental particles. Cytotoxicity is quantitated by the increased proportion of treated versus control cells which do not retain their capacity for normal cell replication. Genotoxic effects are assessed by the increased fraction of nonviable offspring from treated versus control parent cells after the self-fertilization process of autogamy. Since these cells ingest nonnutrient respirable-sized particles, biological activity of intracellular extraction of dusts and fly ash can be compared before and after extraction with polar and nonpolar solvents. Previous studies indicated that coal fly ash was mutagenic in these eukaryotic cells. Mutagenicity of coal fly ash was not detectable after extraction with a concentration of HCl known to remove nonmatrix trace elements. These results suggested that this ciliate bioassay might be a detector of mineral mutagens. Fine particles of the carcinogenic nickel compounds, alpha-nickel subsulfide, and beta-nickel sulfide were compared for their biological activity in this bioassay. Both nickel compounds were ingested by the ciliates and induced heritable damage in the progeny of the treated parent cells.

Animals↗
PubMed1999

Ploidy of serotype AD strains of Cryptococcus neoformans.

Twenty-four serotype AD strains of Cryptococcus neoformans were tested for ploidy and mating type. These included 13 natural isolates (1 from a patient, 1 from peach juice, 11 from pigeon droppings), 2 single clones from one of these natural isolates, 7 F1 progeny from a self-fertile strain and 2 F1 progeny of a cross between a strain from a patient (serotype A, alfa-mating type: MATalfa) and a tester strain (serotype D, a-mating type: MATa). Six strains (2 of natural isolates, 1 of single clone and 3 of F1 progeny) were MATalfa, 1 of F1 progeny was MATa, 4 (1 of natural isolate, 1 of single clone and 2 of F1 progeny) were a-alfa-mating type (MATa-alfa) and 13 (10 of natural isolates and 3 of F1 progeny) were untypable. Most strains, regardless of mating type, were diploid.

Cryptococcus neoformans↗
PubMed1992

Mating behavior in the pulmonate small melampus: can regeneration restore function?

Previous anatomical observations have established that the penial complex of adult Melampus bidentatus is specifically reinnervated and that the penial complex itself can regenerate. This review describes experiments to determine whether a reinnervated penial complex and a regenerated penial complex can function in mating. The snails are not self-fertile, so the ability to transfer sperm, evaluated by production of fertile eggs, was the test of successful regeneration. Snails with nerve transections produced fertile eggs within the time-span expected for axonal regeneration to the target organ. Penial complex ablation interfered with sperm transfer for two egg-laying cycles but fertile eggs were laid within a time-span consistent with regeneration of a penial complex.

Animals↗
PubMed1990

Sexuality and hermaphroditism in fishes. I. Synchronous functional hermaphroditism in the serranid fish Serranus scriba L.

Anatomical and histological examination of Serranus scriba L. showed the existence of primary females (67%), hermaphrodites (31%) and primary males (2%). Synchronous functional hermaphroditism is described on the basis of an anatomical and histological study of the gonads. Although they function simultaneously, the testicular and ovarian parts of hermaphrodite gonads have completely separate ducts. Females and hermaphrodites have the same annual reproduction cycle. In hermaphrodites, the testicular part matures one month sooner than the ovarian part. Cross fertilization between primary females and hermaphrodite individuals and between two different hermaphrodites probably occurs, while self-fertilization is less likely. The testicular tissues of primary males are of the acinar type and those of hermaphrodites are of the radial type. It is possible that primary males do not take part in reproduction. Serranus scriba in Egyptian Mediterranean waters is a longperiod spawner, which spawns from June to the end of October, i.e. it is a summer-autumn spawner.

Animals↗
PubMed1986

Diethylnitrosamine-induced pancreatic neoplasms in the fish Rivulus ocellatus marmoratus.

Diethylnitrosamine [(DEN) CAS: 55-18-5] caused neoplasms of the exocrine pancreas in rivulus, Rivulus ocellatus marmoratus, a self-fertilizing hermaphroditic fish, after a single exposure to 810, 270, or 130 mg DEN/liter or after a continuous exposure to 50, 17, or 9 mg DEN/liter. Rivulus developed pancreatic adenomas after a single exposure. After continuous exposure, pancreatic adenomas, cystadenomas, and adenocarcinomas developed. Adenocarcinoma developed in fish first exposed as larvae but not in those first exposed as juveniles. Adenocarcinoma of pancreatic or biliary origin invaded the intestinal muscularis layer and mucosa. Dedifferentiation of acinar cells preceded the formation of adenomas, and atypical acinar cells formed parts of some of the ductlike structures in the pancreatic neoplasms.

Adenocarcinoma↗
PubMedDate not supplied

Population genetics of Biomphalaria straminea in Hong Kong. A neotropical schistosome-transmitting snail recently introduced into China.

Since its introduction in the early 1970s Biomphalaria straminea (Gastropoda: Planorbidae) has spread rapidly and is locally the most abundant fresh water snail in Hong Kong. Studies of 19 electrophoretically detected loci in four populations show that the colonists retain high levels of variability (P = 0.26, H = 0.056 - 0.097), comparable with those found in autochthonous samples of related species. Genotype frequencies at the five polymorphic loci, and a comparison of maternal and progeny genotypes of individual field-collected snails, revealed no evidence for self-fertilization in these functional hermaphrodites. F statistics indicated minimal genetic structuring, presumably because of outcrossing and recency of origin of the populations. Geographic distribution of various alleles and their frequencies suggest that two southern populations were derived from the original colonists by dispersal but that a northern population represents a second introduction in about 1982. This interpretation (based on genetics) is consistent with the known history of the various populations. The Asian populations of this South American snail are interpreted as being in the "flush" phase of the colonization process. Finally, the probability of the secondary spread of this snail from Hong Kong, and the probability of its parasite, the human blood fluke Schistosoma mansoni, being introduced to Asia are discussed.

Alleles↗
PubMed1984

Covariances of relatives stemming from a population undergoing mixed self and random mating.

We consider covariances of all parent and first-generation relatives from outcrossing or self-fertilization in a parent population that is in equilibrium with respect to these processes. The results, which are for any number of alleles and loci with additive and dominance effects, are phrased in terms of six quadratic genetic components whose coefficients are given by descent measures for equilibrium populations. Because of the variation in the inbreeding coefficients for this system of mating, the expressions include joint contributions of loci to the variances and covariances of relatives. By inclusion of the full complement of relatives, all quadratic components can be estimated. The findings of Ghai (1982, Biometrics 38, 87-92) for compound functions of the covariances with two alleles at a single locus are analyzed in terms of the more general model.

Alleles↗