Changes in the levels of wheat- and barley-germ agglutinin during embryogenesis in vivo,in vitro and during germination

Radioimmunoassay has been used to measure levels of wheat-germ agglutinin and barley-germ agglutinin during embryogenesis and germination. The two lectins exhibited similar patterns of accumulation during grain maturation in vivo and both decreased to low levels after imbibition of harvest-ripe grains for 3 d. Precocious germination of immature wheat and barley embryos excised and cultured in vitro could be prevented either by inclusion of abscisic acid or mannitol in the culture medium. Changes in the level of wheat-germ agglutinin induced by in vitro culture depended on the maturation stage of the embryo. No direct correlation was found between application of exogenous abscisic acid and accumulation of the lectin.     

Construction of improved bacteriophage phi 105 vectors for cloning by transfection in Bacillus subtilis

A series of improved phage vectors have been constructed, based on Bacillus subtilis bacteriophage phi 105, which can be used to clone genes in B. subtilis by direct transfection of protoplasts. The new vectors, designated phi 105J23, phi 105J24, phi 105J27 and phi 105J28, show frequencies of plaque formation that are equal to those of wild-type phi 105. This represents at least a 10-fold improvement over phi 105J9, the vector used in previous cloning experiments. Two of the new vectors phi 105J27 and phi 105J28 incorporate a mutation, cts-52, that renders the prophage temperature inducible. This has made it possible to devise a rapid small-scale procedure for screening progeny phage for the presence of inserted DNA. The usefulness of the new vectors is illustrated in the accompanying paper by cloning more than 20 B. subtilis sporulation genes.     

Protistan microbial observatory in the Cariaco Basin,Caribbean. I. Pyrosequencing vs Sanger insights into species richness

Microbial diversity and distribution are topics of intensive research. In two companion papers in this issue, we describe the results of the Cariaco Microbial Observatory (Caribbean Sea, Venezuela). The Basin contains the largest body of marine anoxic water, and presents an opportunity to study protistan communities across biogeochemical gradients. In the first paper, we survey 18S ribosomal RNA (rRNA) gene sequence diversity using both Sanger- and pyrosequencing-based approaches, employing multiple PCR primers, and state-of-the-art statistical analyses to estimate microbial richness missed by the survey. Sampling the Basin at three stations, in two seasons, and at four depths with distinct biogeochemical regimes, we obtained the largest, and arguably the least biased collection of over 6000 nearly full-length protistan rRNA gene sequences from a given oceanographic regime to date, and over 80 000 pyrosequencing tags. These represent all major and many minor protistan taxa, at frequencies globally similar between the two sequence collections. This large data set provided, via the recently developed parametric modeling, the first statistically sound prediction of the total size of protistan richness in a large and varied environment, such as the Cariaco Basin: over 36 000 species, defined as almost full-length 18S rRNA gene sequence clusters sharing over 99% sequence homology. This richness is a small fraction of the grand total of known protists (over 100 000–500 000 species), suggesting a degree of protistan endemism.     

The effect of exogenous hormone treatment on spermiation and vitellogenesis in the grey mullet, Mugil cephalus L.

The effects of mammalian gonadotropins, methyltestosterone and partially purified salmon gonadotropin on spermiation and oocyte maturation were studied in adult grey mullet. Methyltestosterone was a potent spermiating agent in both prespawning and spawning trials. The most effective dose was 5 mg/100 g body weight. Human chorionic gonadotropin (HCG) was moderately effective at a dose rate of 20 IU/100 g body weight. Both hormones blocked resorption of milt in captive animals. The HCG, FSH/LH combination, Synahorin+mullet pituitary homogenate, and partially purified salmon gonadotropin were equally effective, at the doses used, in inducing oocyte maturation and preventing onset of atrophy before completion of vitellogenesis. Atrophy occurred in all experimental fish at mean oocyte diameter of 750 μ. The significance of this finding is discussed. (Oceanic Institute Contribution No. 98).     

Staining Neuroglia with Silver Diammino-Hydroxide after Sensitizing with Sodium Sulfite and Embedding in Paraffin

Pieces of brain are fixed in formalin ammonium bromide for about 4 days at room temperature, and given the following treatment: After washing, dehydrating and clearing, embed in paraffin, section and mount. Deparaffinize sections and pass through graded alcohols to water. Sensitize in 5% sodium sulfite for 2 hours, wash in distilled water and impregnate with silver diamminohydroxide solution 2-5 minutes at room temperature. Reduce in 2% formalin, wash in distilled water and tone in gold chloride. Fix in 5% hypo, counterstain with 1% picric acid, dehydrate and cover in balsam. Equally good results are obtained by impregnating with Hortega's strong silver carbonate. The microglia, oligodendroglia, fibrous and protoplasmic astrocytes in the cat, rabbit, newborn and adult human are successfully stained by this method.