Obstruction of Blastodisc Formation by Cytochalasin B in the Zebrafish, Brachydanio rerio

The blastodisc formation in the zebrafish, Brachydanio rerio , was obstructed by treatment with 1.0 μg/ml of cytochalasin B (CB), but not by 1.0 μg/ml of colchicine. The cortex in normal eggs contained a meshwork of microfilaments associated with the plasma membrane. The cortex was thicker at the vegetal pole and thinner at the animal pole of the egg. In CB treated eggs the cortex contained masses of microfilaments detached in places from the plasma membrane. Microtubules were never observed in the cortex of eggs with or without CB treatment. These results suggest that ooplasmic segregation, which results in blastodisc formation, is carried out by activity of the cortex, which contains CB sensitive microfilaments.     

Primers Designed for Amplification of Echinococcus multilocularis DNA Amplify the DNA of Encephalitozoon-Like Spores in the Polymerase Chain Reaction

ABSTRACT. Microsporidian spores were developed from cells which were grown in vitro from a human liver lesion which was due to larval Echinococcus multilocularis . The microsporidian spores developed in the same fashion as an Encephalitozoon cuniculi . The Encephalitozoon -like spores were completely separated on Percoll gradients. The separated spores contained DNA capable of amplification by two different primer sets designed for the polymerase chain reaction (PCR) of E. multilocularis DNA. However, the cell DNA from which microsporidium developed was thoroughly insensitive to the PCR using the E. multilocularis primer sets. The results strongly suggested that Encephalitozoon should be taken into consideration, when DNA isolated from larval E. multilocularis is analyzed.     

Avoidance of egg parasitism through submerged oviposition by tandem pairs in the water strider, Aquarius paludum insularis (Heteroptera: Gerridae)

Abstract.  1. Females of the water strider Aquarius paludum insularis (Motschulsky) (Heteroptera: Gerridae) carry males on their backs and oviposit under water after copulation. This study focuses on the benefit  A. paludum insularis receives by ovipositing in tandem.
2. Males guarded females in tandem through to the end of oviposition in 85% of copulations. Females in tandem dived deeper than single females, and the density of A. paludum insularis eggs increased with water depth. The proportion of eggs parasitized by a scelionid wasp, Tiphodytes gerriphagus Marchal (Hymenoptera: Scelionidae) decreased with increasing water depth.
3. These results suggest that during oviposition guarding by males is beneficial for females, because it enables pairs to dive and lay eggs deeper and in oviposition sites where the risk of egg parasitism is lower.     

Demographic genetics of the American beech, Fagus grandifolia. II. Genet substructure of populations for the Blue Ridge, Piedmont and the Great Smoky Mountains

Populations of American beech in Virginia and the Great Smoky Mountain National Park in Tennessee and North Carolina were investigated for demographic genetic substructurings. Two Virginia populations, one on the Blue Ridge (WG1) and the other on the Piedmont (WG2) occur over an elevational gradient of several hundreds meters. One of the Great Smoky Mountain populations (GS1) was in a 'beech gap' and the other (GS2) in a 'cove forest' along a creek. The populations in the Great Smoky Mountain National Park were only separated by a few hundred meters in elevation, but both on the same physiographic province. The populations had two growth forms. Trees produced extensive root suckers at WG1, GS1 and GS2, but WG2 had no root suckers and all individuals had obviously been established from seeds. A total of 1335 shoots were mapped at the four sites, their size measured [diameter at breast height (DBH) or diameter at ground height (DGH)], and genotypes were determined for each locus using allozyme analysis. F_IS among five different size-classes revealed an excess of homozygotes in WG1, GS1 and GS2, and an excess of heterozygotes in WG2. The offshoot formation from root suckers obviously contributed to the abundance of intermediate size-classes in WG1, GS1 and GS2. Exceedingly localized patchiness of different multilocus genotypes reveals genetic clustering of shoots that have obviously originated from root suckers in WG1, GS1 and GS2. The Piedmont population (WG2), on the other hand, showed loose localization of genetically related trees at a scale of 35–40 m in area, suggesting broader ranges of pollen and seed dispersal. The data are discussed in the light of the differences in growth form and mode of reproduction, and also in relation to the post-glacial migration and the current geographic distribution of the species.     

RNA SYNTHESIS IN CULTURED CHICK EMBRYO CELLS IN GROWING AND CONFLUENT PHASES

RNA synthesis at the growing phase in monolayer cultures of chick embryo fibroblasts was compared with that at confluent phases by zonal sedimentation, base composition and hybridization experiments. The nuclei were isolated by treatment with Nonidet p-40. The ratio of RNA/DNA in isolated nuclei was higher at the growing phase than that of confluent. The rate of RNA synthesis was reduced in the cells at confluent phase to 15.1% of that at the growing phase. The sucrose density gradient sedimentation pattern of nuclear RNA was on the whole the same in both phases. According to the distribution of ¹⁴C-uridine incorporated into nuclear RNA, 45S ribosomal precursor RNA was more distinct for the growing cell, while the radioactivities were found to be polydispersed, including the RNA which sedimented faster than 28S RNA in the cells at confluent phase. The base compositions and hybridization analyses indicated that ribosomal RNA was synthesized more actively in the growing cells. About 50% of newly synthesized RNA was ribosomal in the growing cells but 35% in the confluent.
It was found that newly synthesized 18S and 28S ribosomal RNAs appeared in cytoplasm after 21 and 33 min lag periods respectively. These times were exactly same in both growing and confluent phases.     

RNA SYNTHESIS IN THE DIFFERENT PHASES OF CELL CYCLE OF CHICK EMBRYO CELLS

RNA synthesis was studied at different phases of the cell cycle of chick embryo fibroblasts, which were synchronized by medium replacement in the confluent phase. The synthesis of DNA started at 4 hr and continued for 8 hr. RNA synthesis increased with time after medium change. The ratio of total amount of radioactivity in nuclear RNA prepared at 0, 2 and 8 hr was 1.0:1.03:5.05. The distribution of radioactive RNA in the sedimentation pattern was similar, showing remarkable incorporation in 45S region of ribosomal precursor RNA. The base composition of newly synthesized RNA, however, varied at different time intervals after medium replacement. Even within the G₁ phase, the molar percentage of G and C was quite different. Treatment with actinomycin D at a concentration of 0.02 μg/ml for 1 hr specifically inhibited ribosomal RNA synthesis. At 2 hr after medium change, ribosomal and AU-rich RNA including larger than 28S were synthesized in about equal amounts.     

Life-history monographs of Japanese plants. 12: Asarum caulescens Maxim. (Aristolochiaceae)

The life-history characteristics of Asarum caulescens Maxim. (Aristolochiaceae) are described here. This typical summer-green perennial of the Aristolochiaceae is a unique woodland element distributed in central Honshu, that is, south of Fukushima Prefecture to Shikoku and Kyushu, Japan. It grows in the somewhat shady understory of temperate montane deciduous broad-leaved or mixed forests associated with conifers, such as Cryptomeria japonica and Abies firma . One of the unique features of A. caulescens is its rhizome structures in combination with aerial shoots, which play an important role in vegetative propagation. The perennation strategy of A. caulescens is exceedingly complex, producing various combinations of linear or branched rhizomes bearing different numbers of leaves. Rhizomes consist of two or three segments, each 5 mm to 5 cm long as a unit, connected linearly, but also occasionally branched. At the tip of the newly formed rhizome segment, a single leaf or a pair of cordate leaves are formed. Exceedingly complex branching patterns of the rhizome segments were also recognized, forming ramets in late June to early July. Vegetative propagation by ramet formation obviously plays a very important role in the maintenance of local populations.