A passive integrated transponder tag implanted by a new alternative surgical method: effects on the oriental weather loach (Misgurnus anguillicaudatus) and application in a small irrigation system

We validated the effects of a passive integrated transponder (PIT) tagging process on the oriental weather loach Misgurnus anguillicaudatus. Laboratory experiments were conducted to assess the effects of PIT tagging on fish survival, growth, wound healing, and tag omission. Two tagging protocols, standard syringe injection versus insertion through a small hole pierced by a fine needle-shaped awl, were compared using a 12.5 × 2.07 mm² tag. A control group was also included. In comparison with the awl technique, syringe injection heightened the mortality of the loach and delayed healing of the wound caused by tag insertion. No effects of either PIT tagging method were detected on the growth of surviving loach. We also field-tested similarly tagged populations within a river-based irrigation system of Sado Island, Japan. Two different sized tags (long, 12.5 × 2.07 mm²; short, 8.5 × 2.12 mm²) were compared by using antenna loggers which detected fish movement through gates and automatically logged tagged fish’s tag IDs and timestamps. By comparing logged data and actual fish collection surveys both below and above the gates, 77% and 30% of actual loach movements were confirmed to have been successfully logged for the long and short tags, respectively. The awl insertion technique with the longer tag is therefore recommended for use in similar studies of smaller fish species.     

The patch mosaic of an old-growth warm-temperate forest: patch-level descriptions of 40-year gap-forming processes and community structures

Old-growth forests consist of various types of small patches that reflect their own gap-forming process, which includes changes in environmental conditions occurring over several decades. We reconstructed the gap-forming processes that had occurred during a 40-year period for eight representative patches of an old-growth evergreen broad-leaved forest in Japan, and examined the current community structure. The selected patches were based on (1) changes in canopy heights estimated from aerial photographs taken in four different years, (2) long-term ecological research (LTER) monitoring records, and (3) a recent field survey, so that they sufficiently covered characteristic gap-forming processes such as a new gap, an old gap and consistently closed canopy. Height and diameter at breast height (DBH) were measured on all living trees taller than 1.3 m. In their height distributions, currently almost closed patches that were open in 1966 show a rotated sigmoid, whereas their DBH distributions are an inverse J-shape. In contrast, patches that have been consistently under a closed canopy exhibit gentle inverse J-shapes for both distributions. For species composition, there are no clear contrasts associated with the past gap-forming processes except for the existence of fast-growing deciduous species in large currently open patches. Our results suggested that the variation in several decades of gap-forming processes played a central role in the high patch diversity and the complex patch mosaic of the forest. Diverse gap-forming processes created micro-environmental heterogeneity both vertically and horizontally, and contributed to the maintenance of the species-rich, warm-temperate old-growth forest.     

Construction of a LukS-PV mutant of a staphylococcal Panton-Valentine leukocidin component having a high LukS-like function

A 2-residue (D12I13) segment of LukS of a staphylococcal leukocidin component is an essential region for the hemolytic function of LukS towards rabbit erythrocytes in the presence of LukF. Here, we report that insertion of D, I, or AA residue(s) between A11 and E12 residues of LukS-PV, in which the 2-residue D12I13 segment in LukS was absent, confers the full LukS function on LukS-PV, which has only 4% hemolytic activity of that of LukS towards rabbit erythrocytes.     

Functional reconstitution and characterization of the Arabidopsis Mg2+ transporter AtMRS2-10 in proteoliposomes

Magnesium (Mg²⁺) plays critical role in many physiological processes. The mechanism of Mg²⁺ transport has been well documented in bacteria; however, less is known about Mg²⁺ transporters in eukaryotes. The AtMRS2 family, which consists of 10 Arabidopsis genes, belongs to a eukaryotic subset of the CorA superfamily proteins. Proteins in this superfamily have been identified by a universally conserved GlyMetAsn motif and have been characterized as Mg²⁺ transporters. Some members of the AtMRS2 family, including AtMRS2-10, may complement bacterial mutants or yeast mutants that lack Mg²⁺ transport capabilities. Here, we report the purification and functional reconstitution of AtMRS2-10 into liposomes. AtMRS2-10, which contains an N-terminal His-tag, was expressed in Escherichia coli and solubilized with sarcosyl. The purified AtMRS2-10 protein was reconstituted into liposomes. AtMRS2-10 was inserted into liposomes in a unidirectional orientation. Direct measurement of Mg²⁺ uptake into proteoliposomes revealed that reconstituted AtMRS2-10 transported Mg²⁺ without any accessory proteins. Mutation in the GMN motif, M400 to I, inactivated Mg²⁺ uptake. The AtMRS2-10-mediated Mg²⁺ influx was blocked by Co(III)hexamine, and was independent of the external pH from 5 to 9. The activity of AtMRS2-10 was inhibited by Co²⁺ and Ni²⁺; however, it was not inhibited by Ca²⁺, Fe²⁺, or Fe³⁺. While these results indicate that AtMRS2-10 has similar properties to the bacterial CorA proteins, unlike bacterial CorA proteins, AtMRS2-10 was potently inhibited by Al³⁺. These studies demonstrate the functional capability of the AtMRS2 proteins in proteoliposomes to study structure–function relationships.     

A study of seed dispersal by flood flow in an artificially restored floodplain

Riverine floodplains play many important roles in river ecosystems. However, many floodplains have suffered degradation or loss of ecological function due to excessive river improvements or through changes in agricultural systems. As a result, many floodplain restoration projects are being conducted worldwide. One of the many methods being implemented to restore floodplain vegetation is flood water seed dispersal. In this technique, precisely estimating the effect of seed dispersal by flood water is important in order to achieve successful floodplain revegetation. Here, we focus our attention on sediment transport by flood water into the Azamenose Swamp, a restored floodplain. We attempt to estimate the function of seed deposition in the restored floodplain and explain how the seeds are deposited in the floodplain by flood water. The result suggests that the restored floodplain functions as a more appropriate deposition site for seeds than the riverbanks of the main river. It was also found that the distance from the inflow site and the weight of the sediment were related to seed deposition.     

Describing size-related mortality and size distribution by nonparametric estimation and model selection using the Akaike Bayesian Information Criterion

When we calculate mortality along a gradient such as size, dividing into size classes and calculating rates for every class often involves a trade-off: fine class intervals produce fluctuating rates along the gradient, whereas broad ones may miss some trends within an interval. The same trade-off occurs when we want to illustrate size distribution by a histogram. This paper introduces nonparametric methods, published in a statistical journal, into forest ecology, in which the fine-class strategy is used in an extreme way: (1) a smoothly changing pattern is approximated by a fine step function, (2) the goodness-of-fit to the data and the smoothness along the gradient are formulated as a weighting sum within a Bayesian framework, (3) the Akaike Bayesian Information Criterion (ABIC) selects the weighting system that most appropriately balances the two demands, and (4) the values of the step function are optimized by the maximum likelihood method. The nonparametric estimates enable us to represent various patterns visually and, unlike parametric modeling, calculations do not demand the determination of a functional form. Mortality and size distribution analyses were conducted on 12-year forest tree monitoring data from a 4 ha permanent plot in an old-growth warm–temperate evergreen broad-leaved forest in Japan. From trees of 11 evergreen species with a diameter at breast height (DBH) greater than 5 cm, we found three types of trend with increasing DBH: decreasing, ladle-shaped and constant mortality. These patterns reflect variations in life history particular to each species.     

Fat-tailed gene flow in the dioecious canopy tree species Fraxinus mandshurica var. japonica revealed by microsatellites

Pollen flow, seed dispersal and individual reproductive success can be simultaneously estimated from the genotypes of adults and offspring using stochastic models. Using four polymorphic microsatellite loci, gene flow of the wind-pollinated and wind-seed-dispersed dioecious tree species, Fraxinus mandshurica var. japonica, was quantified in a riparian forest, in northern Japan. In a 10.5-ha plot, 74 female adults, 76 male adults and 292 current-year seedlings were mapped and genotyped, together with 200 seeds. To estimate dispersal kernels of pollen and seeds, we applied normal, exponential power, Weibull, bivariate t-distribution kernels, and two-component models consisting of two normal distribution functions, one with a small and one with a large variance. A two-component pollen flow model with a small contribution (26.1%) from short-distance dispersal (sigma = 7.2 m), and the rest from long-distance flow (sigma = 209.9 m), was chosen for the best-fitting model. The average distance that integrated pollen flows inside and outside the study plot was estimated to be 196.8 m. Tree size and flowering intensity affected reproduction, and there appeared to be critical values that distinguished reproductively successful and unsuccessful adults. In contrast, the gene flow model that estimated both pollen and seed dispersal from established seedlings resulted in extensive seed dispersal, and the expected spatial genetic structures did not satisfactorily fit with the observations, even for the selected model. Our results advanced small-scale individual-based parentage analysis for quantifying fat-tailed gene flow in wind-mediated species, but also clarified its limitations and suggested future possibilities for gene flow studies.     

Glutathione reductase/glutathione is responsible for cytotoxic elemental sulfur tolerance via polysulfide shuttle in fungi

Fungi that can reduce elemental sulfur to sulfide are widely distributed, but the mechanism and physiological significance of the reaction have been poorly characterized. Here, we purified elemental sulfur-reductase (SR) and cloned its gene from the elemental sulfur-reducing fungus Fusarium oxysporum. We found that NADPH-glutathione reductase (GR) reduces elemental sulfur via glutathione as an intermediate. A loss-of-function mutant of the SR/GR gene generated less sulfide from elemental sulfur than the wild-type strain. Its growth was hypersensitive to elemental sulfur, and it accumulated higher levels of oxidized glutathione, indicating that the GR/glutathione system confers tolerance to cytotoxic elemental sulfur by reducing it to less harmful sulfide. The SR/GR reduced polysulfide as efficiently as elemental sulfur, which implies that soluble polysulfide shuttles reducing equivalents to exocellular insoluble elemental sulfur and generates sulfide. The ubiquitous distribution of the GR/glutathione system together with our findings that GR-deficient mutants derived from Saccharomyces cerevisiae and Aspergillus nidulans reduced less sulfur and that their growth was hypersensitive to elemental sulfur indicated a wide distribution of the system among fungi. These results indicate a novel biological function of the GR/glutathione system in elemental sulfur reduction, which is distinguishable from bacterial and archaeal mechanisms of glutathione- independent sulfur reduction.