Cryopreservation of shoot apices of hawthorn in vitro cultures originating from East Asia

The objective of this study was to establish a cryopreservation protocol for hawthorn shoot apices (Crataegus pinnatifida Bge.). Cryopreservation was carried out via encapsulation–dehydration, vitrification, and encapsulation–vitrification on shoot apices excised from in vitro cultures. We began by showing that cold-acclimation enhanced the regrowth of cryopreserved apices from 10.0 to 65.5% in encapsulation–dehydration. We then decided that the encapsulation–dehydration method was an optimal cryopreservation method for hawthorn shoot apices in terms of its high recovery after cryopreservation as well as its ease of use compared with vitrification and encapsulation–vitrification. In encapsulation–dehydration, the protocol leading to optimal regrowth was as follows: after cold-acclimation at 5 °C in the dark for 2 weeks, excised shoot tips were pretreated for 24 h at 25 °C on hormone-free Murashige and Skoog [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15, 473–497] (MS) basal medium with 0.4 mol/L sucrose, then encapsulated and precultured in liquid MS medium with 0.8 mol/L sucrose for 16 h at 25 °C. Precultured beads were dehydrated for 6 h at 25 °C in the dessicator containing 50 g silica gel to a moisture content of 15.3% (fresh-weight basis) before cryostorage for 1 h. In addition, we examined the effect of adding glycerol to both the alginate beads and loading solution to enhance regrowth after cryopreservation in encapsulation–dehydration. In the present study, it was shown that adding 0.5 mol/L glycerol resulted in high regrowth percentages (82.5–90.0%) in four Crataegus species.     

Growth and yield of self-compatible and hybrid common buckwheat lines pollinated with and without flies

Common buckwheat is a self-incompatible, insect-pollinated allogamous plant. This study examined growth and yield of the common buckwheat self-compatible and hybrid lines pollinated with and without flies. Self-compatible ‘IH3’, hybrid ‘IP2/IH3’, and standard self-incompatible ‘Kitawasesoba’ were used in field and pot experiments. Self-compatibility of ‘IH3’ was shown to be of high purity. Approximately 10% segregation of pin plants from ‘IP2/IH3’ was observed. The harvest index of ‘Kitawasesoba’ pollinated without flies was considerably lower than that pollinated with flies in both field and pot experiments. The harvest index values of ‘IH3’ and ‘IP2/IH3’ were hardly affected by the presence or absence of flies. The morphological traits of ‘IH3’ were significantly lower than those of the other two genotypes. Large differences in these traits between ‘IP2/IH3’ and ‘Kitawasesoba’ were not observed in either the field or pot experiments. The seed yield of ‘Kitawasesoba’ pollinated without flies tended to be lower than that pollinated with flies. The seed yield of ‘IP2/IH3’ tended to be higher than that of ‘IH3’. The hybrid line ‘IP2/IH3’ showed a high fertilization rate, which was nearly as high as that of ‘IH3’. Rate of fertilization and percentage of ripe seeds were higher in ‘IH3’ and ‘IP2/IH3’, wherein they were hardly affected by the presence or absence of flies, than in ‘Kitawasesoba’. The ‘IP2/IH3’ hybrid line will be useful for understanding the stable high-yielding ability of self-compatibility common buckwheat.     

Evaluating the durability of wood-based panels using internal bond strength results from accelerated aging treatments

In this study, the durability of wood-based panels was evaluated by comparing the internal bond (IB) strength retention after five different laboratory-based accelerated aging tests with the IB retention after 5 years of outdoor exposure in Shizuoka City. In each accelerated aging test, the IB retention of MDI-bonded panels showed high retention compared to other panels. Outdoor exposure in Shizuoka City resulted in an IB retention value for particleboard (PF) and oriented strandboard (aspen) of less than 10% after the 5-year exposure period. Medium-density fiberboards maintained their initial IB strength over the same period. Calculation of the mean IB retention for all board types allowed comparison of the severity of aging between the accelerated test methods and outdoor exposure. The ASTM six-cycle test method was the most severe among the standard treatment cycles applied.     

Supplemental effect of cholyltaurine and soybean lecithin to a soybean meal-based fish meal-free diet on hepatic and intestinal morphology of rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

The supplemental effects of cholyltaurine and soybean lecithin to a defatted soybean meal (SBM)-based fish meal-free diet on the hepatic and intestinal morphology of rainbow trout Oncorhynchus mykiss were investigated. A fish meal-based diet (FM), three fish meal-free diets supplemented either with soybean lecithin (SL), bovine gall powder (BG) or cholyltaurine (C-tau) and an unsupplemented fish meal-free diet (NFM), were given to fish (−11 g) for 10 weeks. The growth was the lowest in fish fed diet NFM, intermediate in fish fed diet SL and the highest in fish fed diet FM, BG and C-tau. Hepatocytes of fish fed diet NFM were atrophied, and the distal intestine of these fish showed abnormal features: disintegrated microvilli and fatty degeneration in epithelial cells and increased amount of connective tissue in the submucosa. Hepatic and intestinal histological features in fish fed diets SL, BG and C-tau were similar to those in fish fed diet FM. These results indicate that soybean lecithin, bovine bile salts and cholyltaurine have similar effects on normalizing the hepatic and intestinal morphologies of rainbow trout fed the SBM-based diet, although growth promotion effect was limited in the soybean lecithin.     

Migration history of Sakhalin taimen <Emphasis Type="Italic">Hucho perryi</Emphasis> captured in the Sea of Okhotsk,northern Japan,using otolith Sr:Ca ratios

Sakhalin taimen Hucho perryi populations have decreased drastically to near extinction. It is urgent to establish an effective conservation strategy based on an understanding of the characteristics of migration and habitat use of this species. We examined the migration history of anadromous Sakhalin taimen captured off the Sarufutsu coast, northern Hokkaido, Japan, using otolith Sr:Ca ratios and also examined the relationship between their otolith Sr:Ca ratios during freshwater and seawater residence in a rearing experiment. Otolith Sr:Ca ratios of some fish from the Sarufutsu coast showed freshwater levels (0.5–4.0 × 10⁻³) near the core, which thereafter increased to brackish water levels (4.0–6.0 × 10⁻³), and then to seawater levels (6.0–10.0 × 10⁻³) in the outermost regions. Those findings indicate that specimens from the Sarufutsu coast migrated to the brackish water region or the sea and spent most of their lives there. The anadromous migration pattern including the timing of downstream migration seems to be flexible among individuals in the species. They migrate between freshwater and seawater or brackish water several times during their lives, showing extensive habitat use. It is essential to secure the continuity among the freshwater, brackish water, and seawater areas for their effective conservation.     

Performance of a closed recirculating system with foam separation, nitrification and denitrification units for intensive culture of eel: towards zero emission

The development of a closed recirculating aquaculture system that does not discharge effluents would reduce a large amount of pollutant load on aquatic bodies. In this study, eel were reared in a closed recirculating system, which consisted of a rearing tank, a foam separation unit, a nitrification unit and a denitrification unit. The foam separation unit has an inhalation-type aerator and supplies air bubbles to the rearing water. The growth of eel, which were fed a commercial diet, was satisfactory, with gross weight increases of up three times in 3 months. The survival rate under the congested experimental conditions was 91%. The foam separation unit maintained oxygen saturation in the rearing water at about 80%. Furthermore, fine colloidal substances were absorbed on the stable foam formed from eel mucus and were removed from the rearing water by foam separation. Ammonia oxidation and the removal of suspended solids were accomplished rapidly and simultaneously in the nitrification unit. The ammonia concentration and turbidity were kept at less than 1.2 mg of N per litre and 2.5 units, respectively. When the denitrification process was operated, nitrate that accumulated in the rearing water (151 mg of N per litre) was reduced to 40 mg of N per litre. The sludge was easily recovered from the nitrification and denitrification tanks, and the components were found suitable as compost. Based on these results, the intensive aquaculture of freshwater fish such as eel can be achieved using a closed recirculating system without emission.     

Differences in oxygen and carbon stable isotope ratios between hatchery and wild pink salmon fry

We examined differences in otolith oxygen (δ¹⁸O_otolith) and carbon (δ¹³C_otolith) stable isotope ratios between hatchery and wild pink salmon fry. The δ¹⁸O_otolith values of hatchery and wild fry were ?7.7 ± 0.2 ‰ and ?8.3 ± 0.3 ‰ (1σ), respectively. This difference reflected differences in temperature conditions experienced by each fry. The δ¹³C_otolith values of hatchery and wild fry were ?19.2 ± 0.3 ‰ and ?11.1 ± 1.8 ‰ (1σ), respectively. The lower δ¹³C_otolith values of hatchery fry were probably related to their intake of artificial diets. Discriminant analysis of δ¹⁸O_otolith and δ¹³C_otolith values demonstrated a highly significant difference between hatchery and wild fry with 95.8 % classification accuracy. Therefore, analysis of δ¹⁸O_otolith and δ¹³C_otolith precipitated in the fry stage may be useful for discriminating the origin of returning adult pink salmon. The discrimination method for returning adult fish would provide important information for evaluating the effect of hatchery release and their impact on the wild population.     

Identification of ommastrephid squid paralarvae collected in northern Hawaiian waters and phylogenetic implications for the family Ommastrephidae using mtDNA analysis

ABSTRACT:   A total of 110 adult individuals from four ommastrephid (family Ommastrephidae) squid species ( Ommastrephes bartramii, Sthenoteuthis oualaniensis, Eucleoteuthis luminosa, and Hyaloteuthis pelagica ) were used to obtain diagnostic DNA markers for species identification. Restriction fragment length polymorphism (RFLP) analysis of a partial segment (855 bp) of the mitochondrial DNA cytochrome oxidase I (COI) gene amplified by polymerase chain reaction (PCR) revealed that the restriction profiles of two endonucleases ( Alu  I and Tsp 509 I) were diagnostic for species identification. The restriction assay partially supplemented with nucleotide sequence analysis successfully assigned 69 damaged and morphologically equivocal ommastrephid paralarvae collected in northern Hawaiian waters, identifying 60 O. bartramii , eight S. oualaniensis , and one E. luminosa . The family Ommastrephidae appears to be monophyletic. Although the phylogenetic relationships among genera were not resolved well due to apparent homoplasy and large genetic divergence between species, COI sequence data without transitions provided support for subfamily level relationships.     

Genetic variability and stock structure of red tilefish Branchiostegus japonicus inferred from mtDNA sequence analysis

Sequence analysis based on the anterior part of the mitochondrial DNA control region was carried out to reveal the genetic diversity, stock structure, and historical demography of the red tilefish. Nucleotide sequences of 388 bp in length were determined for 280 individuals taken from eight localities. The molecular diversity, haplotype diversity, and nucleotide diversity were relatively high (average h = 0.929 ± 0.011; average π = 0.008 ± 0.005). The mismatch distribution was not significantly different from the expected distribution for a rapidly expanding population (P = 0.453). The minimum spanning network connecting with full-sequence haplotypes contained star-like topologies derived from multiple ancient lineages, supporting the mismatch distribution analysis. No significant genetic differentiation was observed among eight localities (maximum pairwise F _ST = 0.023, reduced-sequence data set). Our results suggest a large panmictic population of the red tilefish along the coast of Honshu to the East China Sea.     

Characterization of ethylene biosynthesis and its regulation during fruit ripening in kiwifruit,Actinidia chinensis ‘Sanuki Gold’

Ethylene biosynthesis in kiwifruit, Actinidia chinensis ‘Sanuki Gold’ was characterized using propylene, an ethylene analog, and 1-methylcyclopropene (1-MCP), an inhibitor of ethylene perception. In fruit harvested between a young stage (66 days after pollination) (DAP) and an early commercial harvesting stage (143 DAP), 2 days of exposure to propylene were sufficient to initiate ethylene biosynthesis while in fruit harvested at commercial harvesting stage (154 DAP), 4 days of propylene treatment were required. This observation suggests that response of ethylene biosynthesis to propylene treatment in kiwifruit declined with fruit maturity. Propylene treatment resulted in up-regulated expression of AC-ACO1, AC-ACO2, AC-SAM1 and AC-SAM2, prior to the induction of AC-ACS1 and ethylene production, confirming that AC-ACS1 is the rate limiting step in ethylene biosynthesis in kiwifruit. Treatment of fruit with more than 5 μL L^?1 of 1-MCP after the induction of ethylene production subsequently suppressed ethylene production and expression of ethylene biosynthesis genes. Treatment of fruit with 1-MCP at harvest followed with propylene treatment delayed the induction of ethylene production and AC-ACS1 expression for 5 days. These observations suggest that in ripening kiwifruit, ethylene biosynthesis is regulated by positive feedback mechanism and that 1-MCP treatment at harvest effectively delays ethylene production by 5 days.