Vegetation at the former open-pit Ningyo-toge mine, 36 years after closure treatment: Impact of soil cover on woody plant establishment and dominance of the perennial herb Miscanthus sinensis.

Soil cover is a prevailing method used at mine sites to ensure the safety of hazardous materials and restore ecological functions when the base materials are unfavorable for plant growth. The former open-pit Ningyo-toge Mine was backfilled with overburden and neutralized smelting residues and covered with soil in 1987. After 36 years, the vegetation remained dominated by the perennial herb Miscanthus sinensis, and woody plant establishment did not progress successfully. This study investigated the factors that inhibited woody plant establishment at the site. The soil profile survey revealed that the soil cover formed Bg horizons (pseudogley soil) with cloudy mottling, representative of poorly drained soil. In the Bg horizon, woody plant roots of Pinus densiflora and Weigela hortensis exhibited growth inhibition. Elemental analysis revealed that in the Bg horizon the roots of P. densiflora and W. hortensis accumulated high Fe concentrations exceeding 10,000 mg/kg DW at critical levels. Our results suggested that woody plant roots in the Bg horizon may have suffered from chronic oxygen deficiency accompanied by excessive Fe stress in the soil cover. Topsoil water content (<50 mm) and microtopographic features were not critical factors disrupting woody plant establishment because some individuals were growing in areas with high soil water content, exceeding 60%. Considering that woody plant roots were developed primarily in the shallow A horizon, A horizon formation by M. sinensis is a key step in initiating woody plant establishment by improving the soil structure and physiochemical characteristics of the soil cover, such as carbon content, exchangeable nutrients, and air-filled porosity. For successful mine pollution control and vegetation recovery, implementing an appropriate topsoil system, such as native forest soil, loosely graded and minor infiltration layer above the overburden would be necessary.

Highlighting Potential Physical and Chemical Cues Involved in Conspecific Recognition System in a Predator Nematode, Seinura caverna.

Conspecific recognition is the ability to distinguish and respond to individuals of the same species. In nematodes, this behavior can mediate aggregation, feeding behavior, or mating. Here, we investigated whether and how the predatory nematode Seinura caverna recognizes and avoids conspecifics to prey on. In predation assays, S. caverna did not kill conspecifics, but killed nematodes of three heterospecific species. Interestingly, S. caverna did not kill Ektaphelenchoides spondylis nematodes. Seinura caverna did not eject its stylet when encountering conspecifics or E. spondylis. The characterization of the internal cuticle structure of 13 nematode species suggested that the cuticle may play a role in the preying decision, as E. spondylis and S. caverna exhibited similar, type III, cuticle layers. Chemical extracts from S. caverna further repelled conspecifics. We discuss the potential hierarchical use of physical and chemical cues in S. caverna predation behavior and provide insights into the evolutionary adaptations and behavior of this organism.

Cytoplasmic incompatibility in the semivoltine longicorn beetle Acalolepta fraudatrix (Coleoptera: Cerambycidae) double infected with Wolbachia.

Wolbachia are obligatory endosymbiotic α-proteobacteria found in many arthropods. They are maternally inherited, and can induce reproductive alterations in the hosts. Despite considerable recent progress in studies on the associations between Wolbachia and various taxonomic groups of insects, none of the researches have revealed the effects of Wolbachia on longicorn beetles as the host insect. Acalolepta fraudatrix is a forest longicorn beetle that is distributed in East Asia. In this study, the relationship between Wolbachia and A. fraudatrix was investigated. Out of two populations of A. fraudatrix screened for Wolbachia using the genes ftsZ, wsp, and 16S rRNA, only one of the populations showed detection of all three genes indicating the presence of Wolbachia. Electron microscopy and fluorescent in situ hybridization also confirmed that the A. fraudatrix population was infected with Wolbachia. Sequencing the wsp genes derived from single insects revealed that two strains of Wolbachia coexisted in the insects based on the detection of two different sequences of the wsp gene. We designated these strains as wFra1 and wFra2. The bacterial titers of wFra1 were nearly 2-fold and 3-fold higher than wFra2 in the testes and ovaries, respectively. The two strains of Wolbachia in the insects were completely eliminated by rearing the insects on artificial diets containing 1% concentration of tetracycline for 1 generation. Reciprocal crosses between Wolbachia-infected and Wolbachia-uninfected A. fraudatrix demonstrated that only eggs produced by the crosses between Wolbachia-infected males and Wolbachia-uninfected females did not hatch, indicating that Wolbachia infecting A. fraudatrix causes cytoplasmic incompatibility in the host insect. This is the first report showing the effect of Wolbachia on reproductive function in a longicorn beetle, A. fraudatrix.

A novel approach of preventing Japanese cedar pollen dispersal that is the cause of Japanese cedar pollinosis (JCP) using pollen-specific fungal infection.

In Japan, Japanese cedar pollen dispersal is one of the major causes of pollinosis. Sydowia japonica is an ascomycetous fungus that grows exclusively on the male strobili of Japanese cedar, suggesting a possible mechanism for controlling pollen dispersal. To evaluate this possibility, eleven isolates of S. japonica were collected from around Japan and used as an inoculum to male strobili of Japanese cedar. The treatment demonstrated that the fungus infected only the pollen and prevented pollen dispersal. The fungus did not cause any additional symptoms to other parts of Japanese cedar, such as needles, stems, and buds. All S. japonica isolates collected around Japan could serve to control pollen dispersal. Periodic observation of the fungal pathogenesis with stereomicroscope and scanning electron microscope showed that hyphal fragments and conidia of S. japonica germinated on the surface of male strobili, and the germ tube entered pollen sacs through opening microsporophylls. Within the pollen sacs, the hyphae penetrated pollen gradually, such that all pollen was infected by the fungus by approximately one month before the pollen dispersal season. The infected pollen was destroyed due to the fungal infection and was never released. Our data suggests a novel approach of preventing pollen dispersal using pollen-specific fungal infection.

Effect of light conditions on the resistance of current-year Fagus Crenata seedlings against fungal pathogens causing damping-off in a natural beech forest: fungus isolation and histological and chemical resistance.

Forest gap dynamics affects light intensity on the forest floor, which in turn may influence defense and survival of tree seedlings. Current-year Fagus crenata seedlings show high mortality under the canopy caused by damping-off. In contrast, they survive pathogen attacks in gaps. However, defense mechanisms against damping-off have not been fully understood. In order to determine the resistance factors that affect mortality in current-year seedlings, we compared seedling survival and chemical and histological characteristics of the hypocotyls of seedlings from closed-stand and forest-edge plots. Damping-off occurred in the current-year seedlings mainly from the end of June to July; survival rate of the seedlings was higher in the forest-edge plot than in the closed-stand plot. By performing an inoculation test on the seedling hypocotyls, we identified Colletotrichum dematium and Cylindrocarpon sp. as the causative pathogens under low illumination only. In the beginning of July, only seedling hypocotyls from the forest-edge plot exhibited periderm formation. From mid-June to July, seedling hypocotyls from the forest-edge plot accumulated approximately twice the amount of total phenols as those accumulated by seedling hypocotyls from the closed-stand plot. The ethyl acetate phase of methanol extracts of hypocotyls showed antifungal activity. We conclude that seedlings from the forest-edge plot may resist pathogenic attack via periderm formation and increased phenol synthesis. Plant defense mechanisms that are controlled by light intensity may be important for promoting seedling regeneration in forest gap dynamics.