Self-cloning baker's yeasts that accumulate proline enhance freeze tolerance in doughs

We constructed self-cloning diploid baker's yeast strains by disrupting PUT1, encoding proline oxidase, and replacing the wild-type PRO1, encoding gamma-glutamyl kinase, with a pro1(D154N) or pro1(I150T) allele. The resultant strains accumulated intracellular proline and retained higher-level fermentation abilities in the frozen doughs than the wild-type strain. These results suggest that proline-accumulating baker's yeast is suitable for frozen-dough baking.     

Leaf angle in Chenopodium album is determined by two processes: induction and cessation of petiole curvature

Petiole curvature is important in regulating light interception by the leaf. To dissect the determination processes of leaf angle, we irradiated the lamina or petiole of Chenopodium album L. with either one or two spots of actinic light, after dark adaptation. When the abaxial side of the petiole was irradiated with blue light, the petiole curvature increased, and under continuous irradiation, the curvature continued for up to 6 h. The rate of curvature increased with increasing blue light intensity. The curvature induced by irradiation of the abaxial side with blue light ceased when the adaxial side of the petiole was simultaneously irradiated with either blue or red light. When an inhibitor for photosynthesis, 3-(3,4-dichlorophenyl)-1,1-dimethylurea, was applied to the adaxial side of the petiole, the cessation of curvature caused by blue light was only weakly inhibited, while the cessation caused by red light was markedly inhibited. When the adaxial side of the petiole was irradiated alternately with red and far-red light, the far-red light antagonized the cessation of curvature caused by the red light. These results clearly show that the petiole curvature is controlled by two processes, the induction and the cessation of curvature. At least three photoreceptor systems, blue-light receptor, photosynthesis and phytochrome, are involved in the reactions.     

Abortion of reproductive organs as an adaptation to fluctuating daily carbohydrate production

Excess flower production is a common phenomenon in hermaphrodite plants. The tropical pioneer shrub Melastoma malabathricum (Melastomataceae) frequently aborts not only young ovaries just after flowering, but also flower buds and developed ovaries. We tested a hypothesis that the excess production of reproductive organs and their abortion in this species is an adaptation to environmental fluctuations over shorter time scales than had previously been reported in other plants. To calculate the daily demand for carbohydrate and water by reproductive organs at the level of individual plants, we measured the respiration and transpiration of the reproductive organs at various stages and monitored their growth and abortion. To determine the daily supply of carbohydrate and water, we measured the photosynthetic productivity of leaf area, solar radiation and rainfall. The daily carbohydrate demands of the reproductive organs were significantly correlated with total photosynthetic productivity per leaf area during the previous 1, 3 and 5 days, but no correlations were found between the demands for water and accumulated rainfall or radiation. The daily abortion rates of the population were also correlated with demand for carbohydrates on the previous day per total photosynthetic productivity per leaf area. In brief, it was considered that this species produced and grew more reproductive organs when more resources were supplied and that the abortion occurred when demands for carbohydrate were large. Therefore our hypothesis was supported. We concluded that this reproductive strategy was an adaptation for pioneers characterized by continuous reproduction in aseasonal tropics. In our study, the adaptive consequence of excess production was determined by measuring natural environmental fluctuation.     

Identification of kaonashi mutants showing abnormal pollen exine structure in Arabidopsis thaliana

Exine, the outermost architecture of pollen walls, protects male gametes from the environment by virtue of its chemical and physical stability. Although much effort has been devoted to revealing the mechanism of exine construction, still little is known about it. To identify the genes involved in exine formation, we screened for Arabidopsis mutants with pollen grains exhibiting abnormal exine structure using scanning electron microscopy. We isolated 12 mutants, kaonashi1 (kns1) to kns12, and classified them into four types. The type 1 mutants showed a collapsed exine structure resembling a mutant of the callose synthase gene, suggesting that the type 1 genes are involved in callose wall synthesis. The type 2 mutant showed remarkably thin exine structure, presumably due to defective primexine thickening. The type 3 mutants showed defective tectum formation, and thus type 3 genes are required for primordial tectum formation or biosynthesis and deposition of sporopollenin. The type 4 mutants showed densely distributed baculae, suggesting type 4 genes determine the position of probacula formation. All identified kns mutants were recessive, suggesting that these KNS genes are expressed in sporophytic cells. Unlike previously known exine-defective mutants, most of the kns mutants showed normal fertility. Map-based cloning revealed that KNS2, one of the type 4 genes, encodes sucrose phosphate synthase. This enzyme might be required for synthesis of primexine or callose wall, which are both important for probacula positioning. Analysis of kns mutants will provide new knowledge to help understand the mechanism of biosynthesis of exine components and the construction of exine architecture.     

Identification of a novel matrix protein contained in a protein aggregate associated with collagen in fish otoliths

In the biomineralization processes, proteins are thought to control the polymorphism and morphology of the crystals by forming complexes of structural and mineral-associated proteins. To identify such proteins, we have searched for proteins that may form high-molecular-weight (HMW) aggregates in the matrix of fish otoliths that have aragonite and vaterite as their crystal polymorphs. By screening a cDNA library of the trout inner ear using an antiserum raised against whole otolith matrix, a novel protein, named otolith matrix macromolecule-64 (OMM-64), was identified. The protein was found to have a molecular mass of 64 kDa, and to contain two tandem repeats and a Glu-rich region. The structure of the protein and that of its DNA are similar to those of starmaker, a protein involved in the polymorphism control in the zebrafish otoliths [S?llner C, Burghammer M, Busch-Nentwich E, Berger J, Schwarz H, Riekel C & Nicolson T (2003) Science302, 282-286]. (45)Ca overlay analysis revealed that the Glu-rich region has calcium-binding activity. Combined analysis by western blotting and deglycosylation suggested that OMM-64 is present in an HMW aggregate with heparan sulfate chains. Histological observations revealed that OMM-64 is expressed specifically in otolith matrix-producing cells and deposited onto the otolith. Moreover, the HMW aggregate binds to the inner ear-specific short-chain collagen otolin-1, and the resulting complex forms ring-like structures in the otolith matrix. Overall, OMM-64, by forming a calcium-binding aggregate that binds to otolin-1 and forming matrix protein architectures, may be involved in the control of crystal morphology during otolith biomineralization.     

Transmembrane and ubiquitin-like domain-containing protein 1 (Tmub1/HOPS) facilitates surface expression of GluR2-containing AMPA receptors

Some ubiquitin-like (UBL) domain-containing proteins are known to play roles in receptor trafficking. Alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors (AMPARs) undergo constitutive cycling between the intracellular compartment and the cell surface in the central nervous system. However, the function of UBL domain-containing proteins in the recycling of the AMPARs to the synaptic surface has not yet been reported.Here, we report that the Transmembrane and ubiquitin-like domain-containing 1 (Tmub1) protein, formerly known as the Hepatocyte Odd Protein Shuttling (HOPS) protein, which is abundantly expressed in the brain and which exists in a synaptosomal membrane fraction, facilitates the recycling of the AMPAR subunit GluR2 to the cell surface. Neurons transfected with Tmub1/HOPS-RNAi plasmids showed a significant reduction in the AMPAR current as compared to their control neurons. Consistently, the synaptic surface expression of GluR2, but not of GluR1, was significantly decreased in the neurons transfected with the Tmub1/HOPS-RNAi and increased in the neurons overexpressing EGFP-Tmub1/HOPS. The altered surface expression of GluR2 was speculated to be due to the altered surface-recycling of the internalized GluR2 in our recycling assay. Eventually, we found that GluR2 and glutamate receptor interacting protein (GRIP) were coimmunoprecipitated by the anti-Tmub1/HOPS antibody from the mouse brain. Taken together, these observations show that the Tmub1/HOPS plays a role in regulating basal synaptic transmission; it contributes to maintain the synaptic surface number of the GluR2-containing AMPARs by facilitating the recycling of GluR2 to the plasma membrane.     

Functional analysis of spontaneous cell movement under different physiological conditions

Cells can show not only spontaneous movement but also tactic responses to environmental signals. Since the former can be regarded as the basis to realize the latter, playing essential roles in various cellular functions, it is important to investigate spontaneous movement quantitatively at different physiological conditions in relation to a cell's physiological functions. For that purpose, we observed a series of spontaneous movements by Dictyostelium cells at different developmental periods by using a single cell tracking system. Using statistical analysis of these traced data, we found that cells showed complex dynamics with anomalous diffusion and that their velocity distribution had power-law tails in all conditions. Furthermore, as development proceeded, average velocity and persistency of the movement increased and as too did the exponential behavior in the velocity distribution. Based on these results, we succeeded in applying a generalized Langevin model to the experimental data. With this model, we discuss the relation of spontaneous cell movement to cellular physiological function and its relevance to behavioral strategies for cell survival.     

A new species of the genus Malakichthys (Perciformes: Acropomatidae) from Japan

A new acropomatid, Malakichthys barbatus, is described on the basis of 27 specimens (76.3–180.1 mm SL) collected from Suruga Bay to Tosa Bay, along the southern Pacific coast of Japan. The new species is unique in having numerous pairs of spines on the chin (a pair of spines in other congeners). Although the new species resembles M. elegans in having a slender body (the depth less than 35% SL), the former is further distinguishable from the latter in having a slender first proximal radial of the anal fin with no hollow (broad with a cone-shaped hollow in M. elegans), 43–48 lateral line scales (48–51), modally 21 gill rakers on the lower arm (modally 23), and a dark blotch on the opercle (no remarkable blotch). Received: July 2, 2000 / Revised: September 27, 2000 / Accepted: January 30, 2001     

High‐Performance Thermomagnetic Generators Based on Heusler Alloy Films

Recent developments on Heusler alloys including Ni–Mn–X and Ni–Co–Mn–X (X = Ga, In, Sn,…) demonstrate multiferroic phase transformations with large abrupt changes in lattice parameters of several percent and corresponding abrupt changes in ferromagnetic ordering near the transition temperatures. These materials enable a new generation of thermomagnetic generators that convert heat to electricity within a small temperature difference below 5 K. While thermodynamic calculations on this energy conversion method predict a power density normalized to material volume of up to 300 mW cm^?3, experimental results have been in the range of µW cm^?3. Challenges are related to the development of materials with bulk‐like single‐crystal properties as well as geometries with large surface‐to‐volume ratio for rapid heat exchange. This study demonstrates efficient thermomagnetic generation via resonant actuation of freely movable thin‐film devices of the Heusler alloy Ni–Mn–Ga with unprecedented power density of 118 mW cm^?3 that compares favorably with the best thermoelectric generators. Due to the large temperature‐dependent change of magnetization of the films, a periodic temperature change of only 3 K is required for operation. The duration of thermomagnetic duty cycle is only about 12 ms, which matches with the period of oscillatory motion.