Identification of genes showing differential expression profile associated with growth rate in skeletal muscle tissue of Landrace weanling pig

Suppression subtractive hybridization was used to identify genes showing differential expression profile associated with growth rate in skeletal muscle tissue of Landrace weanling pig. Two subtracted cDNA populations were generated from musculus longissimus muscle tissues of selected pigs with extreme expected breeding values at the age of 100 kg. Three upregulated genes (EEF1A2, TSG101 and TTN) and six downregulated genes (ATP5B, ATP5C1, COQ3, HADHA, MYH1 and MYH7) in pig with genetic propensity for higher growth rate were identified by sequence analysis of 12 differentially expressed clones selected by differential screening following the generation of the subtracted cDNA population. Real-time PCR analysis confirmed difference in expression profiles of the identified genes in musculus longissimus muscle tissues between the two Landrace weanling pig groups with divergent genetic propensity for growth rate. Further, differential expression of the identified genes except for the TTN was validated by Western blot analysis. Additionally, the eight genes other than the ATP5C1 co-localized with the same chromosomal positions as QTLs that have been previously identified for growth rate traits. Finally, the changes of expression predicted from gene function suggested association of upregulation of expression of the EEF1A2, TSG101 and TTN genes and downregulation of the ATP5B, ATP5C1, COQ3, HADHA, MYH1 and MYH7 gene expression with increased growth rate. The identified genes will provide an important insight in understanding of the molecular mechanism underlying growth rate in Landrace pig breed.     

In Vitro Production of Stigma-like Structures from Stigma Explants of Crocus sativus L.

Stigma-like structures (TC stigmas) were produced in tissuecultures from stigma explants of Crocus sativus under definedconditions. MS medium supplemented with NAA (10 mg dm^–3)+BA (1.0 mg dm^–3) induced the optimum response. NAA wasfound to be an important addendum to achieve a good response.The TC stigma regeneration response as a function of explantage showed significant differences (except between stage 1 andstage 4). A culture temperature of 20 °C seems to be betterthan 25 °C with reference to all parameters. Crocin andpicrocrocin pigments, responsible for colour and bitter taste,respectively, were extracted, identified and quantified fromthe TC stigmas. Safranal was not detected in fresh samples. Key words: Crocus sativus, stigma-like structures, organ regeneration, crocin, picrocrocin, safranal     

DNA binding of citrus dehydrin promoted by zinc ion

Dehydrins are hydrophilic proteins that accumulate during embryogenesis and osmotic stress responses in plants. Here, we report an interaction between citrus dehydrin Citrus unshiu cold-regulated 15 kDa protein (CuCOR15) and DNA. Binding of CuCOR15 to DNA was detected by an electrophoretic mobility shift assay, a filter-binding assay and Southwestern blotting. The binding was stimulated by physiological concentrations of Zn²⁺, but little stimulation occurred when other divalent cations, such as Mg²⁺, Ca²⁺, Mn²⁺, Ni²⁺ and Cu²⁺, were substituted for Zn²⁺. Ethylenediaminetetraacetic acid cancelled the Zn²⁺-stimulated binding. A binding curve and competitor experiments suggested that the DNA binding of CuCOR15 exhibited low affinity and non-specificity. Moreover, tRNA competed with the DNA binding. Histidine-rich domains and a polylysine segment-containing domain participated in the DNA binding. These results suggest that CuCOR15 can interact with DNA, and also RNA, in the presence of Zn²⁺. Dehydrin may protect nucleic acids in plant cells during seed maturation and stress responses.     

Effect of the anthraquinones emodin and physcion on availability of selected soil inorganic ions

The effects of emodin and physcion, two anthraquinones involved in Polygonum sachalinense allelopathy, were studied in soils amended with 5, 10 and 15 mg 100 g^-1 soil of each compound, and with their mixtures. Recovery of each compound and its effects on availability of soil Mn⁺⁺, Na⁺, K⁺, Mg⁺⁺, Ca⁺⁺ and PO₄^3- were monitored. Total phenolic content of each soil was determined to study the role of degraded phenolics. Soil amended with 5 mg of emodin and physcion gave maximum recovery of the two anthraquinones. Soil amended with different amounts of two anthraquinones and their mixture had significant lower values for pH compared to control. In general, both anthraquinones affect availability of soil inorganic ions. Compared to unamended control soil, a significant decrease in Mn⁺⁺ availability, and an increase in Na⁺ and K⁺ availability when emodin and physcion were added to the soil was observed. A correlation among soil inorganic ions, relative recovery of two compounds, and total phenolic content of amended soils was observed. Results indicate that emodin and physcion influence the availability of soil inorganic ions. The significance of the effect of allelopathic compounds on soil inorganic ions is discussed.     

Over-wintering stage polymorphism of a bruchine beetle: geographical variation in optimal diapause strategy

Abstract.  1. Most temperate insects undergo diapause at a specific developmental stage to overcome severe winters. The bruchine beetle Bruchidius dorsalis in a warmer region in Japan, however, has diverse over-wintering stages – non-diapausing larvae, diapausing larvae, and diapausing adults, whereas in a cooler region, it over-winters only as the final instar larva or adult in diapause.
2. The geographical pattern of over-wintering stages in 12 populations was investigated over a wide range along the mainland of Japan. It revealed that a distinct geographical pattern of over-wintering stages exists in relation to temperature conditions. In regions with warmer climates, the proportion of non-diapausing larvae increased and B. dorsalis had a more complex over-wintering stage structure.
3. Life cycles were also compared between two areas of Japan by field experiments. In the cooler area, the first generation over-wintered in the diapausing larval or adult stage. Conversely, in the warmer area, diapause was induced later and some of the first-generation adults produced second-generation offspring before over-wintering.
4. Based on the geographical cline of climates and the differences in cold hardiness among stages, we can demonstrate that the over-wintering stage variation among and within populations results from an adaptive timing of diapause induction in each region, because the late larval or adult diapauses protect pupae or eggs – which unlike other stages are not cold hardy – from being produced late in the season.     

Detection of Mouse Tyrosinase With a Monoclonal Antibody MAT-1 Against Human Tyrosinase

In this study we explored the possible application of MAT-1, which has been established as a monoclonal antibody against human tyrosinase, for detection of mouse tyrosinase. The MAT-1 reacted with B16 mouse melanoma cells, but not with tyrosinase-negative NIH-3T3 mouse fibroblasts. In western blot analysis of the large granule fraction (LGF) of B16 cells, MAT-1 detected a single protein of 80 kDa, whose size was close to that of human tyrosinase detected with MAT-1 in extracts of human melanocytes. Furthermore, the 80 kDa band that was detected with MAT-1 in the LGF of B16 cells was also detected by DOPA reaction. In order to confirm that the protein detected with MAT-1 is tyrosinase, a transient expression assay was carried out. When mouse tyrosinase or mouse tyrosinase-related protein 1, which shares high homology with human tyrosinase, was transiently expressed in tyrosinase-negative K1735 mouse melanoma cells by cDNA transfection, MAT-1 reacted only with the cells expressing mouse tyrosinase. These results indicate that MAT-1 specifically reacts with mouse tyrosinase.     

Effects of the Mode of Competition on Stationary Size Distribution in Plant Populations

The effects of the mode of between-individual competition onthe stationary size distribution in uneven-aged and continuallyreproducing tree populations were investigated theoretically,based on the diffusion and continuity equation models. The B(t,x) function, which gives a measure proportional to total leafarea of individual trees larger than dbh (trunk diameter atbreast height) x at time t, expresses the effect of one-sidedcompetition on the subject individual of dbh x. The effect oftwo-sided competition on individuals of any size is equallygiven by B(t, x₀), i.e. total leaf area of the stand (or LAI),where x₀ is the minimum dbh. The following two points, whichwere shown by Kohyama (1991) from results of model simulation,were theoretically derived: (1) the effect of the diffusionterm [D(t, x) function] of the diffusion model on the stationarysize distribution is small for the range of parameter valuesobtained empirically for the warm-temperate and tropical rainforests; (2) the shape of the stationary size distribution underone-sided competition is less changed by a change in the seedlingsupply rate than under two-sided competition. The followingtwo points were further shown theoretically: (3) one-sided competitionis likely to lead to a monotonically decreasing size distributionof dbh with the peak at x = x₀, which is a characteristic ofthe rain forest; (4) the slope of size distribution of dbh steepensas one-sided competition intensifies. Relationships betweenthe mode of competition and the stability of population structure,and general functional forms of one-sided and two-sided competitionare discussed. Diffusion model, one-sided competition, stability of population structure, stationary size distribution, two-sided competition