Dynamic organization of the β-heterochromatin in the Drosophila melanogaster polytene X chromosome

Region 20 of the polytene X chromosome of Drosophila melanogaster was studied in salivary glands (SG) and pseudonurse cells (PNC) of otu mutants. In SG chromosomes the morphology of the region strongly depends on two modifiers of position effect variegation: temperature and amount of heterochromatin. It is banded in XYY males at 25° C and β-heterochromatic in X0 males at 14° C, i.e. it shows dynamic transitions. In PNC chromosomes region 20 is not heterochromatic, but demonstrates a clear banding pattern. Some molecular markers of mitotic heterochromatin were localized by means of in situ hybridization on PNC chromosomes: DNA of the gene su(f) in section 20C, the nucleolar organizer and 359-bp satellite in 20F. The 359-bp satellite, which has been considered to be specific for heterochromatin of the mitotic X chromosome, was found at two additional sites on chromosome 3L, proximally to 80C. The right arm of the X chromosome in SG chromosomes was localized in the inversion In(1LR)pn2b: the telomeric HeT-A DNA and AAGAG satellite from the right arm are polytenized, having been relocated from heterochromatin to euchromatin. Received: 1 July 1998 / Accepted: 7 September 1998     

Genetic and morphological variation in the venus clam Cyclina?sinensis along the coast of China

Six geographic samples of Cyclina sinensis were collected from the coast along China and analyzed to reveal morphological and genetic variation by using nine allozyme loci and 11 morphological variables. The discriminant function analysis (DFA) of morphology suggested a clear separation between the southern and northern populations. Polymorphism was detected at nine loci across all six populations. The mean allele number ranged from 2.44 to 2.78, and the mean observed heterozygosity ranged from 0.218 to 0.296. High level of genetic differentiation was found between three northern populations and three southern populations. The marked genetic differentiation can be explained by the upwelling of the Zhejiang province and the freshwater outflow of Yangtze River. The results obtained in this study indicate that the northern and southern populations of C. sinensis should be treated as separate units for conservation management.     

The Role of dinB in the Appearing of Antibiotics Resistance in E.coli

The role of dinB gene in the appearing of antibiotics resistance was studied. Plasmid containing multi-copy dinB gene was transfected into E. coli to create an overexpression. The strains carrying multi-copies of dinB gene demonstrate a significant survival advantage over the wild strain. In vitro experiment, the dinB-overexpressed strain evolved resistance within 8 hours, while wild strain could not.In vivo experiment with mice model infected with dinB-overexpressed strain, resistant clones emerged significantly earlier and demonstrated significant higher level of resistance than those infected with the wild control strain. The results showed that dinB gene made a contribution in the appearing of the antibiotics resistance and has a potential as a target for prevention from the appearing of antibiotic resistance.     

Structural variability in the genome of phage ?H of Halobacterium halobium

Summary The partially circularly permuted, terminally redundant structure of the DNA of phage H has been confirmed by a cleavage map for the restriction enzymes PstI, ClaI, BglII, HindIII, and, partially, BamHI.Six variants of phage H have been isolated from 71 single plaques. Their genomes differ by several insertions, a deletion, and an inversion of a DNA segment with a minimal length of 11 kb. The inversion occurs with high frequency in variants carrying at the flanks of the invertible DNA in verted repeats of a 1.8 kb DNA element which shares sequence homology with the DNA of H. halobium and may be involved in the extreme variability of its genome.     

The initiation site for recombination cog is at the 3′ end of the his-3 gene in Neurospora crassa

Summary The response of Nicotiana tabacum to tentoxin (chlorosis) is inherited with chloroplasts. N. tabacum var. Xanthi, a tentoxin-resistant line, was used to pollinate tentoxin-sensitive N. tabacum line 92, an alloplasmic male-sterile line containing N. undulata plastids. The seeds were mutagenized with nitrosomethylurea and germinated in the presence of tentoxin. Two percent of the seedlings had green sectors in their first true leaves. These plants were grown to maturity under non-selective conditions. Homogeneous tentoxin-resistant lines were obtained in the third generation. DNA analysis indicated, however, that selection for paternal plastids, rather than mutagenesis of maternal ones, had occurred in the tentoxin-resistant progeny. Mitochondria, which were not under selection pressure, were inherited maternally as expected. Inheritance of tentoxin-resistant paternal plastids did not require seed mutagenesis. Normally germinated seedlings that were kept under tentoxin selection consistently produced a low level of resistant green sectors in their first true leaves. Thus, normal, low-frequency transmission of paternal plastids in N. tabacum can be directly revealed by using tentoxin.     

Population dynamics of the mushroom coral Heliofungia?actiniformis in the Spermonde Archipelago, South Sulawesi, Indonesia

The fungiid Heliofungia actiniformis is one of the most popular coral species in the Indonesian aquarium trade, yet information on the biology of this species is limited. H. actiniformis growth rates, population size–frequency distributions and the seasonality of recruitment rates were measured at three replicate sites in the Spermonde Archipelago, South Sulawesi. Growth and population models were applied to estimate coral ages, mortality rates and the size of maximum yield. Growth decreased linearly with polyp size. High numbers of attached polyps budded from clusters of stalks attached to the reef, with each cluster originating from the settlement of a sexually produced larva. Neither the settlement of sexual recruits, nor their asexual budding, showed seasonality. The overall population structure reflected the high mortality rates of young, attached polyps (Z = 0.5–0.6 yr⁻¹), and the much lower mortalities of free-living individuals (Z = 0.05–0.08 yr⁻¹). There were no statistically significant differences in overall mortality rates and the age–frequency distributions of polyps aged 0–15 years between the sites. Differences in the abundance of large H. actiniformis polyps at the three replicate sites were correlated with percent cover of coral rubble. The application of the Beverton and Holt model revealed the highest biomass per H. actiniformis recruit was 12 cm, corresponding to a polyp age of 20 years.     

Suppressors of thermosensitive mutations in the DNA polymerase δ gene of Saccharomyces cerevisiae

DNA polymerases (Pol) , and are necessary for replication of nuclear DNA. Po1 interacts permanently or transiently with numerous accessory proteins whose identification may shed light on the function(s) of Po18. In vitro mutagenesis was used to induce thermosensitive (ts) mutations in the DNA polymerase gene (POL3). We have attempted to clone two recessive extragenic suppressors of such is mutants (sdp1 for mutation pol3-14 and sdp5-1 for mutation pol3-11) by transforming thermoresistant haploid strains pol3-14 sdpl and pol3-11 sdp5-1 with wild-type genomic libraries in singlecopy or multicopy vectors. None of the thermosensitive transformants so obtained was identified as being sdp1 or sdp5-1. Instead, three genes were cloned whose products interfere with the activity of suppressors. One of them is the type 1 protein phosphatase gene, D1S2. Another is a novel gene, ASM4, whose gene product is rich in asparagine and glutamine residues.     

Apyrase and α-glucosidase in the salivary glands of Aedes albopictus

An apyrase and an α-glucosidase were detected in the salivary glands extracts of adult Aedes albopictus. The apyrase is a 61,000 Da secreted protein that hydrolyses ATP and ADP. This protein is synthesized in adults and is preferentially accumulated in the distal lateral lobes of the female salivary glands. The α-glucosidase is a secreted 67,000 Da protein. This enzyme is synthesized during adult life and accumulated in the proximal-lateral lobes of both males and females. The results are discussed and compared with data previously obtained with Aedes aegypti salivary glands.     

Suppressors of the temperature sensitivity of DNA polymerase α mutations in Saccharomyces cerevisiae

We have isolated two high copy, allele-specific suppressors of the temperature sensitivity of mutations in POL1, the gene that encodes the catalytic subunit of DNA polymerase α in the yeast Saccharomyces cerevisiae. Both genes, PSP1 and PSP2, also partially suppressed a mutation in POL3 which encodes DNA polymerase δ, and both also affected a mutation in CDC6, which acts in initiation of DNA replication. Suppression was not general, since ts mutations in several genes unrelated to replication were not affected. PSP1 was partially effective on low-copy-number vectors, while PSP2 required high copy numbers. The presence of suppressing plasmids did not alter the steady-state level of Pol1 protein, so suppression does not appear to be due to an increase in production or stability of Pol1p. Deletion of either PSP gene or both in combination resulted in apparently normal viable cells. While neither gene is homologous to genes with known functions, PSP1 and PSP2 both have unusual amino acid compositions: PSP1 is rich in asparagine and glutamine, while PSP2 is rich in asparagine and contains “RGG” motifs that have been associated with RNA-binding proteins. We also describe a transposon-mediated strategy that should be generally effective for rapid characterization of multicopy suppressors. Received: 20 July 1997 / Accepted: 1 October 1997     

Does uptake of Alexandrium fundyense cysts contribute to the levels of PSP toxin found in the sea scallop, Placopecten magellanicus?

Atlantic sea scallops, Placopecten magellanicus, in most areas of the Bay of Fundy, New Brunswick, Canada, have year-round concentrations of paralytic shellfish posioning (PSP) toxins greater than the regulatory concentration of 80 μg STX eq. 100 g⁻¹ wet weight. Scallops (mean shell height of 10.7 cm, age 3–5 years) were collected by SCUBA and individually tagged near Parker Island, Bay of Fundy. Half were hung 2 m below the low tide water level and the remainder were placed on the bottom (11 m depth at low tide) under the scallops held at 2 m. Scallop, water and sediment samples were collected monthly for determination of concentrations of PSP toxins and Alexandrium fundyense.In October, 1993, mean concentrations of PSP toxins in digestive gland, and mantle were 3205 and 1018 μg STX eq. 100 g⁻¹ wet weight, respectively. Eight months later (June 1994), PSP concentrations in digestive glands from the surface and bottom had declined to 504 and 682 μg STX eq. 100 g⁻¹ wet weight, respectively, whereas those in the mantle had declined to 802 and 681 μg STX eq. 100 g⁻¹ wet weight. During July 1994, A. fundyense concentrations observed at Parker Island and offshore were 320 cells l⁻¹ and 14,200 cells l⁻¹, respectively. Subsequently, toxin concentrations in surface and bottom scallop digestive glands increased to 12,720 and 11,408 μg STX eq. 100 g⁻¹ wet weight, whereas concentrations in mantles increased to 2126 and 1748 μg STX eq. 100 g⁻¹ wet weight, respectively. Concentrations of PSP toxins in these tissues in October 1994 were similar to those measured in October 1993. Concentrations of PSP toxin were less than the regulatory concentration in the gonads and non-detectable in adductor muscles of all scallops sampled.There were no statistically significant differences in profiles for uptake and depuration of PSP toxins in scallops held at the surface compared to those from bottom, suggesting that A. fundyense cysts at the concentrations found in the sediment (45 cysts cm⁻³) did not contribute significantly to the year-round presence of PSP toxins within scallop tissues. The year-round occurrence of PSP toxin is probably due to accumulation during summer blooms followed by a very slow rate of depuration.     

Cloning and comparison of Aα mating-type alleles of the basidiomycete Schizophyllum commune

Summary An A mating-type allele (A4) was isolated by walking the chromosome from the closely linked PAB1 gene. A cosmid clone containing the A1 allele isolated from the walk was used as a probe to recover the A1 allele from another cosmid library. Cosmids encoding mating-type activity were identified by transforming Schizophyllum cells and screening for activation of A-regulated development. Putative mating-type transformants were confirmed in mating tests and genetic analyses of progeny. The identity of the specific alleles isolated was demonstrated by showing that their effectiveness in transforming for mating type is limited to recipient strains possessing an A allele different from the one encoded by the cloned sequences. Transforming DNA is active in trans, suggesting that A encodes a diffusible product. Restriction mapping shows that A1 and A4 are coded in the same physical region of the genome, but within a subregion that contains extensive sequence divergence. In addition, Southern analyses show that there is only one copy of A1 or A4 per haploid genome, and that they do not cross-hybridize to one another or to any of the other A alleles. A1 and A4 were subcloned as 2.8 and 1.2 kb fragments, respectively, retaining in transformation all the mating-type activity demonstrated of the original cosmids.     

In?vitro evaluation of the effect of the nematophagous fungi Duddingtonia flagrans , Monacrosporium sinense and Pochonia chlamydosporia on Schistosoma mansoni eggs

The in vitro effect of four isolates of the nematophagous fungi Duddingtonia flagrans (AC 001), Monacrosporium sinense (SF 53) and Pochonia chlamydosporia (VC 1 and VC 4) on eggs of Schistosoma mansoni was examined. One thousand S. mansoni eggs were plated on 2% water–agar with the grown isolates and control without fungus. After 7, 14 and 21 days, the eggs were removed and classified according to the following parameters: type 1, lytic effect without morphological damage to eggshell; type 2, lytic effect with morphological alteration of embryo and eggshell; and type 3, lytic effect with morphological alteration of embryo and eggshell, besides hyphal penetration and internal egg colonization. Significant differences (P < 0.01) were found among the studied fungal isolates for ovicidal activity, confirming type 3 effect for the isolates VC 1 and VC 4, which characterizes the ovicidal activity of a fungus. Type 3 effect was only found for P. chlamydosporia (VC 1 and VC 4) with 26.6 and 17.2%, 25.6 and 22.6%, 27.4 and 23.9% in the 7, 14 and 21 days respectively (P < 0.01). P. chlamydosporia can thus be a potential biological control agent for S. mansoni eggs.     

Circular structure of the genome of phage ?H in a lysogenic Halobacterium halobium

Summary Phage H is a temperate phage, i.e., it can establish lysogeny in the archaebacterium Halobacterium halobium. H-lysogens are immune to phage infection and phage production is spontaneously induced at a rate of about 10^-7. In the prophage state. H DNA exists as a covalently closed circle of 57 kb.The frequent occurrence of clones carrying the phage genome but unable to produce phage is another proof of the high variability of DNA in H. halobium. In one such strain, R₁-3, the phage genome has undergone a structural change which may have abolished an essential phage gene.     

Dinophysis blooms in the deep euphotic zone of the Baltic Sea: do they grow in the dark?

In situ growth rates of the toxin-producing dinoflagellate Dinophysis norvegica collected in the central Baltic Sea were estimated during the summers of 1998 and 1999. Flow cytometric measurements of the DNA cell cycle of D. norvegica yielded specific growth rates (μ) ranging between 0.1 and 0.4 per day, with the highest growth rates in stratified populations situated at 15–20 m depth. Carbon uptake rates, measured using ¹⁴C incubations followed by single-cell isolation, at irradiances corresponding to depths of maximum cell abundance were sufficient to sustain growth rates of 0.1–0.2 per day. The reason for D. norvegica accumulation in the thermocline, commonly at 15–20 m depth, is thus enigmatic. Comparison of depth distributions of cells with nutrient profiles suggests that one reason could be to sequester nutrients. Measurements of single-cell nutrient status of D. norvegica, using nuclear microanalysis, revealed severe deficiency of both nitrogen and phosphorus as compared to the Redfield ratio.It is also possible that suitable prey or substrate for mixotrophic feeding is accumulating in the thermocline. The fraction of cells containing digestive vacuoles ranged from 2 to 22% in the studied populations. Infection by the parasitic dinoflagellate Amoebophrya sp. was observed in D. norvegica in all samples analysed. The frequency of infected cells ranged from 1 to 3% of the population as diel averages, ranging from 0.2 to 6% between individual samples. No temporal trends in infection frequency were detected. Estimated loss rates based on observed infection frequencies were 0.5–2% of the D. norvegica population daily, suggesting that these parasites were not a major loss factor for D. norvegica during the periods of study.