Molecular cloning of two caspase-like genes from the hard tick Haemaphysalis longicornis

We identified two caspase-like genes from the midgut cDNA library of the hard tick, Haemaphysalis longicornis. Sequence analysis showed that these cDNAs encoded homologues of caspase-2 and caspase-8 that were categorized as apoptosis initiators. The H. longicornis caspase-2 (Hlcaspase-2) cDNA encodes 340 amino acid residues with a predicted molecular weight (Mw) of 38.5 kDa. Another cDNA identified as the H. longicornis caspase-8 (Hlcaspase-8) encodes 306 amino acid residues with an estimated Mw of 35.3 kDa. A catalytic active site was highly conserved in Hlcaspase-8 but not in Hlcaspase-2. RT-PCR analysis showed that both Hlcaspase-2 and Hlcaspase-8 were expressed in tick midgut and salivary glands. This is the first report of the molecular cloning of apoptosis-related genes in the tick.     

Inheritance of Seed Shattering in Lolium temulentum and L. persicum Hybrids

Lolium temulentum and L. persicum are non-crop species found in wheat and barley fields. L. temulentum has non-shattering seeds like the associated grain crops, whereas L. persicum seeds shatter after maturity. We analyzed the inheritance mode of shattering tendency by comparing the F₂ of L. temulentum and L. persicum hybrids. The selfed progeny of L. temulentum and L. persicum exhibited typical non-shattering (1.6% shattering) and shattering phenotypes (70.8%), respectively. F₁ hybrids of L. temulentum×L. persicum and its reciprocals were of the shattering phenotype (71.4% and 63.8%, respectively), indicating that shattering is dominant to non-shattering. When the phenotype ratio was assumed to be 15 shattering: 1 non-shattering, the χ² value for F₂ segregation was not significant at the 5% level, and the reciprocal effect was not detected. This indicates that the non-shattering tendency is controlled by two recessive genes. The two-gene inheritance model of shattering tendency suggests that harvest is the selector for seed shattering in cultivated fields, thus the alternative tendency for non-shattering seeds of L. temulentum or shattering of L. persicum would be better adapted to cultivated fields.     

Varietal difference in vitamin C content in the fruit of kiwifruit and other actinidia species

Vitamin C content in the fruit of various cultivars of kiwifruit and other Actinidia species was estimated by determination of L-ascorbic acid and L-dehydroascorbic acid using ion-pair reversed-phase high-performance liquid chromatography. Fruit of A. deliciosa cv. Hayward, the most common commercially available cultivar, contained 65.5 mg/100 g fresh weight (FW) vitamin C. Vitamin C content in A. deliciosa fruit varied from 29 mg/100 g FW to 80 mg/100 g FW. In most cultivars of A. chinensis, vitamin C content in fruit was higher than that of Hayward. In particular, vitamin C content in cv. Sanuki Gold fruit reached more than 3-fold that of Hayward on a weight for weight basis. In A. argutafruit, there was wide variation in vitamin C content, with concentrations ranging from 37 to 185 mg/100 g FW. In cv. Gassan, Issai, and Mitsuko, vitamin C content of the fruit was much higher than that of Hayward. In A. arguta fruit, the ratio of L-ascorbic acid to total ascorbic acid tended to be higher than that of other species.     

Aneuploidy drives genomic instability in yeast

Aneuploidy decreases cellular fitness, yet it is also associated with cancer, a disease of enhanced proliferative capacity. To investigate one mechanism by which aneuploidy could contribute to tumorigenesis, we examined the effects of aneuploidy on genomic stability. We analyzed 13 budding yeast strains that carry extra copies of single chromosomes and found that all aneuploid strains exhibited one or more forms of genomic instability. Most strains displayed increased chromosome loss and mitotic recombination, as well as defective DNA damage repair. Aneuploid fission yeast strains also exhibited defects in mitotic recombination. Aneuploidy-induced genomic instability could facilitate the development of genetic alterations that drive malignant growth in cancer.     

Vertical Changes in Bacterial Communities in Percolating Water of a Japanese Paddy Field as Revealed by PCR-DGGE

Percolating water was sampled from the plow layer and subsoil layer in a Japanese paddy field, and the bacterial communities were compared together with floodwater by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) targeting a partial 16S rRNA gene and subsequent sequencing. The number of DGGE bands ranged from 16 to 28 with no significant differences among the sampling sites and times. Only 2 bands were common for the three sources of water samples. DGGE bands specific for the floodwater samples and percolating water samples from the plow layer were identified, while percolating water samples from the subsoil layer did not show specific bands but displayed common bands to those of the floodwater samples (7 bands) and percolating water samples from the plow layer (1 band). Cluster analysis of the DGGE banding patterns showed a distinct clustering in the samples of percolating water from the plow layer and a closer relationship between the others. These results suggest that the bacterial communities in percolating water changed during downward movement through the plow layer and subsoil layer. Sequences of the DGGE bands specific for the samples of percolating water from the plow layer showed a close relationship with anaerobic bacteria such as iron-reducers or uncultured bacterial DNA isolated from environments that are considered to be less oxic. On the other hand, the sequences of the bands specific for the samples of floodwater and percolating water from the subsoil layer showed a close relationship with uncultured bacterial DNA isolated from freshwater environments.     

Developmental competence and glutathione content of maternally heat-stressed mouse oocytes and zygotes

The loss of developmental competence and the glutathione (GSH) content of maternally heat‐stressed mouse oocytes and zygotes were determined. In experiment 1, zygotes were collected from female mice that were heat‐stressed at 35°C for 10 h after hCG injection (oocyte maturation stage), or for 12 h on Day 1 of pregnancy (zygote stage), followed by in vitro culture. To minimize the effects of heat stress on the fertilization process, heat‐stressed oocytes that were fertilized in vitro were also included in this experiment. In experiment 2, heat‐stressed oocytes and zygotes were assayed for GSH content. The application of heat stress to the oocytes resulted in a significant decrease in the percentage of zygotes that developed to morulae or blastocysts, both for naturally fertilized oocytes (56.9% for heat‐stressed vs 85.4% for control) or in vitro‐fertilized oocytes (54.5%vs 73.6%). In the heat‐stressed zygotes, the disruption of embryonic development was more drastic (24.3%vs 90.3%), with the majority of zygotes being arrested at the two‐cell stage. In contrast, the GSH content decreased significantly in heat‐stressed zygotes, but not in heat‐stressed oocytes. These results demonstrate that the loss of developmental competence of early embryos is associated with a decrease in the GSH content of maternally heat‐stressed zygotes, but not of maternally heat‐stressed oocytes.