Aucuba japonica Thunb. is an evergreen understory shrub that grows naturally at a mine site. The mine soil contains high concentrations of heavy metals, and A. japonica appears to maintain ...detoxification mechanisms against heavy metals in the study site's understory. This study aimed to investigate the heavy metal tolerance mechanisms in A. japonica, considering the possible roles of arbuscular mycorrhizal and root-endophytic fungi. We conducted fieldwork in summer (canopy-foliation season) and winter (canopy-defoliation season) to measure the heavy metal concentrations in leaves, branches, and roots and analyze possible detoxicants in the roots. The infection rates of arbuscular mycorrhizal and root-endophytic fungi were evaluated via microscopic observation, and heavy metal (Zn) localization in A. japonica roots was observed using confocal laser scanning microscopy. Field analysis showed that A. japonica accumulated excessive Zn and produced aucubin and citric acid in the roots in both summer and winter. Zn localization observations clarified that Zn was distributed in thickened epidermal and cortical cell walls, suggesting that the cell walls functioned as Zn deposition sites, reducing Zn toxicity. It was further clarified that Zn was contained within cortical cells, indicating that Zn might be detoxified by aucubin and citric acid. Arbuscular mycorrhizal and root-endophytic fungi within cortical cells adsorbed Zn on fungal cell walls, indicating that these fungi would reduce Zn content within root cells and might alleviate Zn toxicity. Our results indicated that A. japonica would maintain Zn tolerance in both summer and winter via Zn immobilization in the cell walls and production of aucubin and citric acid, and that arbuscular mycorrhizal and root-endophytic fungi might play important roles in the Zn tolerance of A. japonica.
Clethra barbinervis Sieb. et Zucc. is a tree species that grows naturally at several mine sites and seems to be tolerant of high concentrations of heavy metals, such as Cu, Zn, and Pb. The purpose of ...this study is to clarify the mechanism(s) underlying this species' ability to tolerate the sites' severe heavy-metal pollution by considering C. barbinervis interaction with root fungal endophytes. We measured the heavy metal concentrations of root-zone soil, leaves, branches, and fine roots collected from mature C. barbinervis at Hitachi mine. We isolated fungal endophytes from surface-sterilized root segments, and we examined the growth, and heavy metal and nutrient absorption of C. barbinervis seedlings growing in sterilized mine soil with or without root fungal endophytes. Field analyses showed that C. barbinervis contained considerably high amounts of Cu, Zn, and Pb in fine roots and Zn in leaves. The fungi, Phialocephala fortinii, Rhizodermea veluwensis, and Rhizoscyphus sp. were frequently isolated as dominant fungal endophyte species. Inoculation of these root fungal endophytes to C. barbinervis seedlings growing in sterilized mine soil indicated that these fungi significantly enhanced the growth of C. barbinervis seedlings, increased K uptake in shoots and reduced the concentrations of Cu, Ni, Zn, Cd, and Pb in roots. Without root fungal endophytes, C. barbinervis could hardly grow under the heavy-metal contaminated condition, showing chlorosis, a symptom of heavy-metal toxicity. Our results indicate that the tree C. barbinervis can tolerate high heavy-metal concentrations due to the support of root fungal endophytes including P. fortinii, R. veluwensis, and Rhizoscyphus sp. via growth enhancement, K uptake promotion and decrease of heavy metal concentrations.
At a sedimentary site in an old mine site, Miscanthus sinensis formed patches, where Pinus densiflora seedlings could grow better compared with those outside the patches, indicating that M. sinensis ...would improve P. densiflora seedling establishment. The purpose of this study was to understand the mechanisms by which M. sinensis facilitates the survival of P. densiflora seedlings by considering the soil properties, heavy metal tolerance, and root endophytes in P. densiflora seedlings at the sedimentary site. The sedimentary site, which is a bare ground, contained high concentrations of Fe, indicating that plants should be exposed to Fe and high soil temperature stresses. Measurement of soil temperature revealed that M. sinensis suppressed sharp increases and alternation of soil temperature, resulting in reducing high soil temperature stress in P. densiflora seedlings. To adapt to the Fe stress environment, P. densiflora outside and inside the patches produced Fe detoxicants, including catechin, condensed tannin, and malic acid. Ceratobasidium bicorne and Aquapteridospora sp. were commonly isolated from P. densiflora seedlings outside and inside the patches as root endophytes, which might enhance Fe tolerance in the seedlings. Aquapteridospora sp., which is considered as a dark-septate endophyte (DSE), was also isolated from the roots of M. sinensis, suggesting that M. sinensis might play a source of a root endophyte to P. densiflora seedlings. Ceratobasidium bicorne could be classified into root endophytes showing symbiosis and weak pathogenicity to host plants. Therefore, high soil temperature stress would weaken P. densiflora seedlings, causing root endophytic C. bicorne to appear pathogenic. We suggested that P. densiflora could adapt to the Fe stress environment via producing Fe detoxicants, and M. sinensis would facilitate the establishment of P. densiflora seedlings in the sedimentary site by providing a DSE, Aquapteridospora sp., and maintaining symbiosis of C. bicorne from high soil temperature stress.
To date, studies on terrestrial plant ecology and evolution have focused primarily on the trade-off patterns in the allocation of metabolic production to roots and shoots in individual plants and the ...scaling of whole-plant respiration. However, few empirical studies have investigated the root : shoot ratio by considering scaling whole-plant respiration at various sizes throughout ontogeny.
Here, using a whole-plant chamber system, we measured the respiration rates, fresh mass and surface area of entire roots and shoots from 377 Fagus crenata individuals, from germinating seeds to mature trees, collected from five different Japanese provenances. Non-linear regression analysis was performed for scaling of root and shoot respiration, fresh mass and surface area with body size.
Whole-plant respiration increased rapidly in germinating seeds. In the seedling to mature tree size range, the scaling of whole-plant respiration to whole-plant fresh mass was expressed as a linear trend on the log-log coordinates (exponent slightly greater than 0.75). In the same body size range, root and shoot respiration vs. whole-plant fresh mass were modelled by upward-convex (exponent decreased from 2.35 to 0.638) and downward-convex trends (exponent increased from -0.918 to 0.864), respectively. The root fraction in whole-plant respiration, fresh mass and surface area shifted continuously throughout ontogeny, increasing in smaller seedlings during early growth stages and decreasing in larger trees.
Our results suggest a gradual shift in allocation priorities of metabolic energy from roots in seedlings to shoots in mature trees, providing insights into how roots contribute to shoot and whole-plant growth during ontogeny. The models of root : shoot ratio in relation to whole-plant physiology could be applied in tree growth modelling, and in linking the different levels of ecological phenomena, from individuals to ecosystems.
Metallophores are low-molecular-weight compounds capable of chelating heavy metals, which have recently been reported to alleviate heavy metal stress in plants. We isolated two undescribed compounds ...as Zn-chelating metallophores from the culture broth of the root endophytic Pezicula ericae w12-25, which was collected from a Zn-accumulating plant, Aucuba japonica Thunb. These two compounds were determined to be (3aS,4S,6aR)-3a-hydroxy-3-methylene-4-octyldihydrofuro3,4-bfuran-2,6(3H,4H)-dione and (3S,3aS,4S,6aR)-3a-hydroxy-3-(hydroxymethyl)-4-octyldihydrofuro3,4-bfuran-2,6(3H,4H)-dione using spectroscopic methods (HRMS, 1H and 13C NMR, and 2D NMR) and X-ray crystallography, respectively. The two compounds, classified as furofurandiones, were named isoavenaciol and 7-hydroxy-isoavenaciol. After the hydrolysis of the lactone moiety, isoavenaciol would release the carboxyl group to show Zn-chelating activity. Their antifungal activities were confirmed using Cladosporium herbarum (AHU9262).
Root endophytic Pezicula ericae, isolated from Zn-accumulating plant, Aucuba japonica, produced two undescribed metallophores showing Zn chelating activity. Display omitted
•Pezicula ericae were isolated from Zn-accumulating plant, Aucuba japonoica.•P. ericae produced two undescribed furofurandiones as Zn-chelating metallophores.•Two compounds were named isoavenaciol and 7-hydroxy-avenaciol.•Hydrolysis of lactone moiety would increase the Zn-chelating activity.•Their antifungal activities were evaluated using Cladosporium herbarum.
Miscanthus sinensis Andersson naturally grows at mining sites and accumulates heavy metals, suggesting heavy metal tolerance. This study aimed to elucidate the heavy metal tolerance mechanisms of M. ...sinensis, which naturally grew at a mine site, considering root endophytes. Elemental analysis by inductively coupled plasma-optical emission spectroscopy showed that M. sinensis highly accumulated zinc and lead in adventitious roots and dead root bark, exhibiting tolerance via these elemental removals in dead root bark. Phenolic analysis by high-performance liquid chromatography revealed chlorogenic acid production in adventitious roots. Phialocephala fortinii, which showed the highest appearance, and Talaromyces verruculosus, which produced gluconic acid as a zinc detoxicant, were isolated from adventitious roots of M. sinensis. In inoculation tests, P. fortinii and T. verruculosus significantly enhanced seedling growth by increasing potassium uptake and possibly indole-3-acetic acid production. Our results provide fundamental knowledge for using M. sinensis and root endophytes for revegetation at mining sites.
Miscanthus sinensis Andersson is a pioneer plant species that grows naturally at mining sites. Miscanthus sinensis can detoxify aluminium (Al) by producing phytosiderophores, such as chlorogenic ...acid, citric acid, and malic acid, and localizing Al in cell walls. Root-endophytic Chaetomium cupreum, which produces microbial siderophores, enhances Al tolerance in M. sinensis. However, we could not determine whether the siderophores produced by C. cupreum actually enhance Al tolerance in M. sinensis, because the microbial siderophores have not yet been identified in previous research. The purpose of this study was to clarify how C. cupreum chemically increases Al tolerance in M. sinensis under acidic mining site conditions, especially considering siderophores. Using instrumental analyses, the siderophore produced by C. cupreum was identified as oosporein. Comparison of the stability constant between Al and phytosiderophores and oosporein indicated that oosporein could detoxify Al similarly to chlorogenic acid, which shows higher stability constant than citric acid and malic acid. Inoculation test of C. cupreum onto M. sinensis in acidic mine soil showed that C. cupreum promoted seedling growth, and enhanced Al tolerance via inducing chlorogenic-acid production and producing oosporein. These results suggested that C. cupreum could chemically enhance Al tolerance and might promote growth via reducing excessive Al in cell walls, the main site of Al accumulation. In addition, the chemical enhancement of Al tolerance by C. cupreum might be important for M. sinensis to adapt to acidic mining sites.
For growing plants at mine sites, plant species that accumulate metals in tissues and are tolerant to high metal concentrations should be selected from the perspective of phytostabilization. However, ...the eco-chemical or elemental information of the plant species at the mine sites is limited. The purpose of this study was to identify plants that can adapt to natural growth at mine sites, via: (1) vegetation survey, (2) elemental analysis in soil and plants, and (3) detoxicant detection in plant cells. Our vegetation survey indicated that plants growing at our study site are consistent with plant species confirmed at other mine sites in previous reports. A. indica var. maximowiczii and F. sachalinensis, present at the mine site, highly accumulated Fe, Al, and Cu in the roots, indicating their metal tolerance. Furthermore, A. indica var. maximowiczii produced detoxicants such as chlorogenic acid and 3,5-dicaffeoylquinic acid in the roots, which exhibited high antioxidative activity that would play an important role in metal tolerance in A. indica var. maximowiczii. This study will be effective in providing fundamental information on phytostabilization at mine sites.
, a root endophyte in
, enhances Al tolerance in
by changing aluminum (Al) localization and the production of a siderophore, oosporein, which chelates Al for detoxification. Oosporein has various ...functions, including insecticidal activity, phytotoxicity, antifungal activity, and a siderophore. In our study, we focused on the detoxification effect of oosporein as a siderophore and on the growth of
under Al exposure. In addition, the phytotoxicity of oosporein to
was confirmed to compare with those in
and
as control plants. Under Al stress, oosporein promoted plant growth in
seedlings at 10 ppm, which was the same concentration as that detected in
roots infected with
in our previous study. Oosporein also showed low phytotoxicity to
compared with
at even high concentrations of oosporein. These results suggest that the concentration of oosporein in
roots would be maintained at the appropriate concentration to detoxify Al and would promote
growth under Al stress, although oosporein would show low phytotoxicity to the natural host plant,
, compared with the non-host plant,
.
Mine drainage is a vital water problem in the mining industry worldwide because of the heavy metal elements and low pH. Rhizofiltration using wetland plants is an appropriate method to remove heavy ...metals from the water via accumulation in the rhizosphere. Phragmites australis is one of the candidate plants for this method because of metal accumulation, forming iron plaque around the roots. At the study site, which was the mill tailings pond in the Ningyo-toge uranium mine, P. australis has been naturally growing since 1998. The results showed that P. australis accumulated Fe, Mn, and 238U in the nodal roots without/with iron plaque compared with other plant tissues. Among the 837 bacterial colonies isolated from nodal roots, 88.6% showed siderophore production activities. Considering iron plaque formation around P. australis roots, we hypothesized that microbial siderophores might influence iron plaque formation because bacterial siderophores have catechol-like functional groups. The complex of catechol or other phenolics with Fe was precipitated due to the networks between Fe and phenolic derivatives. The experiment using bacterial products of root endophytes, such as Pseudomonas spp. and Rhizobium spp., showed precipitation with Fe ions, and we confirmed that several Pseudomonas spp. and Rhizobium spp. produced unidentified phenolic compounds. In conclusion, root-endophytic bacteria such as Pseudomonas spp. and Rhizobium spp., isolated from metal-accumulating roots of P. australis, might influence iron plaque formation as the metal accumulation site. Iron plaque formation is related to tolerance in P. australis, and Pseudomonas spp. and Rhizobium spp. might indirectly contribute to tolerance. Although there are many issues to be resolved in this research, we hope that the fundamental analysis of plant-microbe interactions would be helpful for phytoremediation at mine sites.
PDF
