Gonadal morphogenesis and sex differentiation in cultured chub mackerel,Scomber japonicus

The timing of primordial germ‐cell (PGC) migration with regard to the gonadal anlagen, gonad formation and sex differentiation was examined histologically in the chub mackerel (Scomber japonicus) at 5–190 days post hatching (dph). At 5 dph, PGCs appeared on the peritoneal epithelium surface or in the mesentery, on the dorsal side of the abdominal cavity. By 10 dph, stromal cells around the PGCs proliferated. The gonadal primordium was formed by 15 dph. The gonadosomatic index was 0.01% at 30 dph and increased thereafter (0.32% in females and 0.04% in males at 160 dph). Ovarian differentiation occurred at 30–40 dph, indicated by ovarian cavity formation (elongation and fusion of the upper and lower ovarian edges). Meiosis was subsequently initiated. A few meiotic oocytes surrounded the cavity at 50 dph; most were in the perinucleolus stage at 60 dph and attained a diameter of 60–70 μm at 190 dph. Testicular differentiation occurred at 30 dph, indicated by the formation of the sperm duct primordium. Spermatogonia gradually proliferated, developing into spermatocytes at the chromatin–nucleolus stage (after 90 dph) and subsequently into spermatids and spermatozoa (160 dph). These data could aid the development of seeding and cell‐engineering technologies for scombrid fish.     

Molecular cloning and gene expression of the riboflavin-binding protein in the Nile tilapia, Oreochromis niloticus

Tilapia riboflavin-binding protein (RfBP) cDNA was isolated by library screening. Even though it shows only 31～35% similarity to the reported RfBPs, tilapia RfBP is conserved in amino acid residues that are known to be essential for its protein function. The exclusive expression of RfBP in the brain-pituitary-gonadal axis, as revealed by both RT-PCR and Northern blot, suggested a possible role for RfBP in fish reproduction.     

Sex change in coral reef fish

Gonadal differentiation can take many forms in fish, ranging from gonochorism, where individuals directly develop as male or female and finally possess only testis or ovaries at sexual maturation, to hermaphroditism where the same individuals can produce mature male and female gametes at some time in their lives. Hermaphrodite fish are, thus, an excellent model for studying the plasticity of sex determination and differentiation in vertebrates. We have shown that sex steroids play a principal role in sex differentiation and sex change in fish. Our laboratory implements several fish models that undergo sex change from female to male or male to female or in both directions. In this review, we will briefly discuss recent advances in our understanding of the mechanism of sex change in coral reef fish.     

Partial cloning of 17B-HSD1 from the Nile tilapia ovary and its expression pattern during spawning cycle

A 548 bp partial cDNA fragment of 17β-hydroxysteroid dehydrogenase (17β-HSD1) was obtained by RT-PCR from the ovary of Nile Tilapia. The expression of 17β-HSD1 was high from 0 to 11 days after spawning, but there was a sharp decline at the day of spawning (day 14) indicating its involvement in ovarian cycle.     

Comparison of flower color with anthocyanin composition patterns in evergreen azalea

In evergreen azaleas, major anthocyanins were detected from petals of wild species and cultivars by HPLC analysis. Depending on flower color, all samples were divided into three groups: red, purple or white, using the Japan color standard for horticultural plants. The chromatic components a* and b* values of red group samples showed a convergent distribution, whereas those of purple group samples showed a wider distribution. According to the HPLC analysis, red group samples had two to four major anthocyanins, and those of the purple group had two to six major ones. In contrast, no anthocyanins were detected in the white group petals, although anthocyanidins were detected. These results suggest that the anthocyanin constitution of the purple group flowers is more varied than that of the red group flowers, and this wider variety among purple flowers contributes to extending the diversity of flower color in evergreen azalea.     

A new primer for 16S rDNA analysis of microbial communities associated with <Emphasis Type="Italic">Porphyra yezoensis</Emphasis>

To construct high-quality 16S rDNA clone libraries for microbial communities associated with Porphyra yezoensis and to minimize the detection of rDNA from leafy gametophytes of P. yezoensis, we designed a new 16S rDNA universal primer (75F). Of the clones prepared using 75F, which was designed to distinguish between bacteria and P. yezoensis, 95% were classified into four groups, namely, β-proteobacteria, γ-proteobacteria, Lentisphaerae, and Flavobacteria. PCR-based analysis of the 16S rDNA primer constructed in this study can be used to implement 16S rDNA-based methodologies for the investigation of microbial community composition and diversity related to the Porphyra group.     

Carbon and nitrogen stocks and their mineralization potentials are higher under organic than conventional farming practices in Japanese Andosols

ABSTRACT

This study aimed at understanding whether and how long-term organic rice farming affects soil carbon (C) and nitrogen (N) stocks and their mineralization potentials in submerged rice paddies in Tochigi, Japan. An incubation experiment was carried out to assess the impacts of internal nutrient cycling after organic farming (OF) for 4–5 years (4OF), 8–9 years (8OF), and 12 years (12OF), compared with a conventional rice field (CF). Soil samples were collected at 0–15 cm and 15–20 cm in flooded rice fields after harvest in October 2013. pH and bulk density at 0–15 cm were significantly lower in 12OF fields than in CF fields (by 0.22 unit pH and 17.5%, respectively). Compared with CF, 12OF fields showed significant differences in soil organic carbon (SOC) and total nitrogen (TN), but 4OF and 8OF fields did not. In 8OF fields, the C decomposition (Co) and N mineralization (No) potentials were significantly higher (by 34.0% and 35.6%, respectively, at 0–15 cm, and by 67.1% and 24.5% at 15–20 cm) than in CF fields. Similarly, in 8OF fields at 0–15 cm, the Co:SOC and No:TN ratios were 19.8% and 23.2% higher, respectively, than in CF fields. Co, No, Co:SOC, and No:TN in 12OF fields were higher than those in CF fields, demonstrating the effects of prolonged organic rice farming. Additionally, in 12OF fields, C and N stocks were significantly higher (by 15.5% and 17.2%, respectively, at 0–15 cm, and by 4.8% and 12.1% at 15–20 cm) than in CF fields. Our findings suggest that long-term organic rice farming increases soil C and N stocks as well as C and N mineralization in Japanese Andosols.     

Attaining inter-subgeneric hybrids in fragrant azalea breeding and the inheritance of organelle DNA

Inter-subgeneric hybrids were successfully obtained in reciprocal cross combinations between evergreen azaleas (Rhododendron nakaharae and its hybrids) and fragrant deciduous azaleas (R. arborescens and R. viscosum) for the purpose of fragrant evergreen azalea breeding. Nuclear and organelle DNA of these hybrids was investigated using PCR-RFLP markers. Viable hybrid seedlings have nuclear ribosomal DNA (nrDNA) inherited biparentally, mitochondrial DNA (mtDNA) from the seed parent, and chloroplast DNA (cpDNA) from the deciduous azalea, regardless of cross combination. These results suggest that the chloroplast genome from deciduous azaleas and the nuclear genome from evergreen azaleas are compatible in viable hybrid progenies.     

Functional changes in beta-lactoglobulin by conjugation with cationic saccharides

Bovine beta-lactoglobulin (beta-LG) was conjugated to each of three cationic saccharides [glucosamine (GlcN), chitopentaose (CPO), and chitosan (CHS)] by means of a water-soluble carbodiimide or by the Maillard reaction in an effort to improve the functional properties of beta-LG. The molar ratios of beta-LG to the cationic saccharide in the beta-LG-GlcN, beta-LG-CPO, and beta-LG-CHS conjugates were 2:1, 2:5, and 2:1, respectively. Fluorescence studies indicated that the conformation around Trp had changed in each conjugate and that the surface of each of the conjugates was covered with a saccharide chain. Structural analysis using monoclonal antibodies indicated that the conformation around (15)Val-(29)Ile (beta-sheet region) in beta-LG-GlcN and beta-LG-CPO had changed but that in beta-LG-CHS was maintained, whereas the conformation around (125)Thr-(135)Lys (alpha-helix region) in the conjugates had changed. The emulsifying activity of beta-LG was improved by conjugation with CPO or CHS, and aggregation of beta-LG was suppressed by conjugation with CHS. Reduction of the antigenicity and immunogenicity of beta-LG was achieved by conjugation with CHS.     

The effect of tractor logging on forest soil compaction and growth ofShorea selanica seedlings in Indonesia

The hardness in soil was investigated in four forest areas in Indonesia after a crawler tractor passed over it several times. The hardness in soil, expressed by bulk density, was measured at various depth and various distances from the main track. Recovery of soil hardness was also examined. The bulk density increased markedly by the first and second pass of the tractor, but did not change after the fifth pass. The bulk density was highest just under the main and the log track, and decreased with an increase in distance from the track (0.5 to 1.5 m). The greatest increase in bulk density was observed at the depth of 0–5 cm. The soil hardness tended to show recovery within 9 years after logging, though not completely. Growth and root penetration of seedlingsShorea selanica decreased with the increase of soil hardness. We considered that the 2 tractor passes that caused bulk density of 1.3 g/cm³ is critical for the growth ofS. selanica’s seedling.