Development of CAPS markers and their application in breeding for mango, Mangifera indica L.

To develop cleaved amplified polymorphic sequence (CAPS) markers for the identification of true hybrids in F1 progeny, we cloned sucrose synthase (SS) and sucrose phosphate synthase (SPS) genes from 19 mango cultivars. Through comparison of amino acid sequences in a BLASTX search, cloned SS and SPS genes were found to be homologs of Citrus unshiu CitSUS1 (AB022092) and CitSPS1 (O22060), respectively. Therefore, we designated the SS and SPS genes as MiSUS1 and MiSPS1, respectively. There were 11 genotypes in MiSUS1 and 10 in MiSPS1 sequences. In the MiSUS1 nucleotide sequences, 46 SNPs and 5 in/dels were detected. Twenty-six out of the 46 SNPs mapped on the restriction enzyme recognition sites were successfully converted to 25 CAPS markers. In the MiSPS1 nucleotide sequences, 99 SNPs and 6 in/dels were detected. Thirty out of 99 SNPs and 2 out of 6 in/dels mapped on the recognition sites were also converted to 32 CAPS markers. By using two CAPS markers, SS-2798 and SPS-5745, we identified 37 true hybrids that had different genotypes to ‘Irwin’ from 82 F1 progeny obtained through a cross between ‘Irwin’, the maternal parent, and paternal parents by using flies as pollinators. The method developed here to identify true hybrids is a simple, rapid and highly reliable tool for efficient mango breeding.     

Cesium-137 discharge into the freshwater fishery ground of grazing fish, ayu Plecoglossus altivelis after the March 2011 Fukushima nuclear accident

The Fukushima nuclear accident led to deposition of radionuclides into the freshwater fishery ground of ayu Plecoglossus altivelis. For the purpose of more comprehensive understanding of the radioactive contamination, cesium-137 (¹³⁷Cs) was analyzed in running water, muddy sediment, attached algae, and ayu using samples from three river systems with different levels of contamination within Fukushima Prefecture in summer and autumn 2011. The amount of ¹³⁷Cs contained in water as well as in sediment declined over an interval of two months, although the concentration in sediment samples was up to 10,000-fold higher than that in water samples. The changes in ¹³⁷Cs concentration of algae showed different courses among the three river systems, the concentration being greater in autumn in the river systems with high and moderate radioactive contamination. ¹³⁷Cs detected within a single stock of ayu appeared to be excreted rather rapidly with a calculated half-life of approximately 39 days, while the concentration ratio ranged from 10³ to 10⁴ l/kg, attaining higher levels than previously reported. These findings indicate that the most important pathway of ¹³⁷Cs into ayu is via intake of algae as food but that the retention time of ¹³⁷Cs inside ayu is relatively short.     

The ex vivo effects of eyestalk peptides on ovarian vitellogenin gene expression in the kuruma prawn Marsupenaeus japonicus

Seven major peptides belonging to the crustacean hyperglycemic hormone family were purified from the sinus gland located in the eyestalk of the kuruma prawn Marsupenaeus japonicus, and their effects on vitellogenin gene expression were examined using the ex vivo ovary incubation system. Six molecular species of crustacean hyperglycemic hormone, Pej-SGP-I, -II, -III, -V, VI, and VII, displayed significant inhibitory effects on vg expression with almost the same efficacies, whereas Pej-SGP-IV (known as molt-inhibiting hormone) did not. Two chromatophorotropic peptides, red pigment-concentrating hormone and pigment-dispersing hormone, which were also present in the sinus glands, did not have a clear effect on the gene expression levels in this incubation system. These results suggest that the six crustacean hyperglycemic hormones are potentially capable of acting as vitellogenesis-inhibiting hormones in M. japonicus.     

Stress hormone responses in ayu Plecoglossus altivelis in reaction to different catching methods: comparisons between electrofishing and cast netting

It is important to select the most appropriate method for fish sampling in streams and rivers; the sampling efficiency as well as any negative effect of the method of fish sampling on fishes such as stress, injury, and mortality must be considered. This study aimed to investigate stress hormone responses in ayu Plecoglossus altivelis in reaction to direct current (DC) electrofishing, alternating current (AC) electrofishing, and cast netting in artificial streams. The mortality, injury rate, and catch efficiency of these catching methods were also compared. It was found that immediately after capture, fish caught using a cast net exhibited significantly higher cortisol levels than did control fish that were caught rapidly by a hand net. Cortisol levels did not differ between electroshocked fish and the controls. Time required to catch 12 fish was shorter when DC was used (20 s) than when AC was used (45 s). The time required to catch fish greatly increased when a cast net was used (840 s). Cortisol levels in DC electroshocked and control treatment groups were at resting levels 24 and 48 h after capture. However, higher cortisol levels were found in fish captured using a cast net at both these time points. Cortisol levels in AC electroshocked fish returned to lower levels at 24 h, but increased again at 48 h. Furthermore, 48 h following capture, the fish caught by AC electrofishing exhibited higher mortality (7.5 %) than those captured by other methods (0 %). Spinal injury was also detected in one of the fish in this group. Considering these findings, DC electrofishing is the most effective and least damaging method for collecting P. altivelis in streams with respect to stress, physical damage, and efficiency.     

Effect of fish in rice-fish culture on the rice yield

Rice-fish culture, which means the simultaneous culture of rice and fish, is one of the best options to increase food production from limited land and is practiced in many countries in the world. Although many researchers and farmers believe that the rice yield is increased by fish farming in paddy fields, this hypothesis has never been fully tested. Here, we report ecological processes leading to higher rice yields in the rice-fish culture using crucian carp (Carassius complex), which have adapted to the paddy field ecosystem in Japan. We compare the rice-fish and rice-only plots in the experimental paddy field for biota, water quality, and rice yield. Coverage of duckweed and densities of zooplankton and benthic invertebrates in the rice-fish plots were lower than those in the rice-only plots, indicating that fish utilized them as food. NO₃–N concentration in the rice-fish plots was higher than that in the rice-only plots, indicating that the increase in NO₃–N concentration results from excretion of unutilized food nutrients by the fish. Consequently, rice yield in the rice-fish plots was 20% higher than that in the rice-only plots. The fertilizing effect of the fish excrement probably increased rice yield.     

Species identification method for marine products of Seriola and related species

The complete nucleotide sequences of mitochondrial DNA (mtDNA) from four Seriola spp. (S. quinqueradiata, S. lalandi, S. dumerili, and S. rivoliana) were determined with the aim of developing a species identification analysis method for discriminating between commercially important Seriola spp. and other related species. In addition, the nucleotide sequences of the mitochondrial cytochrome b gene (Cytb) from five related but less expensive species in terms of market value (Seriolella brama, S. caerulea, S. punctata, Hyperoglyphe japonica, and Rachycentron canadum), which are often used as substitutes for Seriola spp., were determined. Restriction enzyme sites were examined by comparing the nucleotide sequences, and species-specific primers were designed for PCR-based restriction fragment length polymorphism (RFLP) analysis. Based on the results of the PCR amplification studies, the four Seriola spp. and the five related species tested could be categorized into three groups according to their PCR product pattern: a 373-bp product from the four Seriola spp., a 513-bp product from three Seriolella spp. and H. japonica, and a 204-bp product from R. canadum. In addition, RFLP analysis of the PCR products was able to differentiate these fish species.     

Ankyrin repeat proteins comprise a diverse family of bacterial type IV effectors

Specialized secretion systems are used by many bacteria to deliver effector proteins into host cells that can either mimic or disrupt the function of eukaryotic factors. We found that the intracellular pathogens Legionella pneumophila and Coxiella burnetii use a type IV secretion system to deliver into eukaryotic cells a large number of different bacterial proteins containing ankyrin repeat homology domains called Anks. The L. pneumophila AnkX protein prevented microtubule-dependent vesicular transport to interfere with fusion of the L. pneumophila-containing vacuole with late endosomes after infection of macrophages, which demonstrates that Ank proteins have effector functions important for bacterial infection of eukaryotic host cells.     

Feeding territory and variations in behavioural modes of algae‐grazing fish Plecoglossus altivelis ryukyuensis (Ryukyu‐ayu) in subtropical island streams

Abstract – Social and foraging modes in fish often vary between individuals with different body sizes and between populations under different ecological conditions. We studied social and foraging behaviour of algae‐grazing fish Plecoglossus altivelis ryukyuensis (Ryukyu‐ayu) inhabiting Japanese subtropical island streams. Ryukyu‐ayu exhibited four behavioural modes: territorial, schooling, solitary and floating. Their body sizes differed among these modes. Territorial fish predominantly foraged on benthic algae, whereas schooling and solitary individuals frequently consumed drifting materials as well. Schooling fish were smaller than territorial fish, but, unlike some other algae‐grazing fish species, did not use schooling to gain access to food within territories. Territorial fish attacked smaller conspecifics but exhibited lateral display towards larger fish and schooling fish while occasionally attacking grazing gobies. Larger fish maintained larger feeding territories and occupied territories for longer periods than smaller fish did. This suggests that, in Ryukyu‐ayu, intra‐ and interspecific food competitions and relative body size can influence diverse behavioural modes and duration of territory occupation. We also found that Ryukyu‐ayu foraged more often and had larger feeding territories than ayu P. a. altivelis inhabiting temperate streams. We conclude that foraging strategies of Ryukyu‐ayu may have adapted to subtropical island streams, where algal productivity is much lower than that in temperate regions.     

Comparison of distal forelimb conformations between Japanese Black and Holstein-Friesian newborn calves

Flexural and hyperextension deformities are congenital problems in calves. We, therefore, aimed to investigate the distal limb conformation in 1 day- and 28-day-old female Holstein-Friesian (HF) calves (n=21), male Japanese Black (JB) calves (n=15), and female JB calves (n=15). The claw angle of the forelimb dorsal claw wall in a standing position and recorded other parameters, including body weight, withers height, circumference of forelimbs, and flexor tendon thickness in the forelimbs, were measured and compared these between the three groups. At 1 day old, the mean claw angles were 51.1° in female HF calves, 47.0° in male JB calves, and 41.8° in female JB calves; the 95% confidence intervals (CIs) of the claw angles showed large distributions in all three groups. One female HF and one male JB calves showed mild flexural deformity, whereas four JB calves showed hyperextension deformity. At 28 days old, the mean claw angles were 51.7° in female HF calves, 51.2° in male JB calves, and 48.4° in female JB calves; the 95% CIs of the claw angles showed smaller distributions than those at 1 day old in all groups. For all groups, the limb deformities had improved without treatment at 28 days old. As a feature of the breed, female JB calves were apt to show hyperextended deformities inversely proportional to the body weight. These limb deformities healed spontaneously and were thought to be physiological.