Masanori Nonaka

Arum- and Paris-type arbuscular mycorrhizas in a mixed pine forest on sand dune soil in Niigata Prefecture, central Honshu, Japan

Arbuscular mycorrhizas (AM) are the most widespread mycorrhiza in nature and form two morphologies, Arum- and Paris-type. The determining factors defining the two different morphologies are not well understood. In this study, the distribution of Arum- and Paris-type AM was determined in a mixed pine forest. A total of 35 plant species belonging to 20 families and 32 genera were identified and examined for AM colonization and morphological types. AM morphological types in 14 families were confirmed as follows: Arum-type in Rosaceae, Oleaceae, Lauraceae, Vitaceae and Compositae, Paris-type in Aquifoliaceae, Ulmaceae, Araliaceae, Theaceae, Magnoliaceae, Rubiaceae and Dioscoraceae, and both and/or intermediate types in Caprifoliaceae and Gramineae. Plant families whose AM morphological status was previously unknown were clarified as follows: Polygonaceae and Commelinaceae showed Arum-type morphology; Celastraceae, Menispermaceae and Elaeagnaceae had typical Paris-type morphology. The proportion of Arum-type to Paris-type species decreased in the following order: annuals > perennials > deciduous species > evergreen species, and pioneer group > early successional group > late successional group. Evergreen plants had a higher tendency to form Paris-type AM than annuals, perennials and deciduous plants. The results indicate that environmental changes, such as shade during plant succession, control the distribution of plant growth forms in mixed pine forest and may also play a part in the distribution of Arum- and Paris-type morphology. The identity of the plant seems to strongly influence AM morphology, though control by the fungal genome cannot be ruled out.

Cooccurring plants forming distinct arbuscular mycorrhizal morphologies harbor similar AM fungal species

Arbuscular mycorrhizal (AM) fungal spores were isolated from field transplants and rhizosphere soil of Hedera rhombea (Miq) Bean and Rubus parvifolius L., which form Paris-type and Arum-type AM, respectively. DNA from the spore isolates was used to generate molecular markers based on partial large subunit (LSU) ribosomal RNA (rDNA) sequences to determine AM fungi colonizing field-collected roots of the two plant species. Species that were isolated as spores and identified morphologically and molecularly were Gigaspora rosea and Scutellospora erythropa from H. rhombea, Acaulospora longula and Glomus etunicatum from R. parvifolius, and Glomus claroideum from both plants. The composition of the AM fungal communities with respect to plant trap cultures was highly divergent between plant species. Analysis of partial LSU rDNA sequences amplified from field-collected roots of the two plant species with PCR primers designed for the AM fungi indicated that both plants were colonized by G. claroideum, G. etunicatum, A. longula, and S. erythropa. G. rosea was not detected in the field-collected roots of either plant species. Four other AM fungal genotypes, which were not isolated as spores in trap cultures from the two plant species, were also found in the roots of both plant species; two were closely related to Glomus intraradices and Glomus clarum. One genotype, which was most closely related to Glomus microaggregatum, was confined to R. parvifolius, whereas an uncultured Glomeromycotan fungus occurred only in roots of H. rhombea. S. erythropa was the most dominant fungus found in the roots of H. rhombea. The detection of the same AM fungal species in field-collected roots of H. rhombea and R. parvifolius, which form Paris- and Arum-type AM, respectively, shows that AM morphology in these plants is strongly influenced by the host plant genotypes as appears to be the case in many plant species in natural ecosystems, although there are preferential associations between the hosts and colonizing AM fungi in this study.

137Cs in irrigation water and its effect on paddy fields in Japan after the Fukushima nuclear accident

There is concern that radiocesium deposited in the environment after the accident at the Fukushima Daiichi Nuclear Power Plant (FDNPP) in March 2011 will migrate to paddy fields through hydrological pathways and cause serious and long-lasting damage to the agricultural activities. This study was conducted in the Towa region of Nihonmatsu in the northern part of Fukushima Prefecture, Japan, (1) to quantify (137)Cs in stream water used to irrigate paddy fields by separating the dissolved and particulate components in water samples and then fractionating the particulate components bonded in different ways using a sequential extraction procedure, and (2) to determine the amounts of radiocesium newly added to paddy fields in irrigation water relative to the amounts of radiocesium already present in the fields from the deposition of atmospheric fallout immediately after the FDNPP accident. Three catchments were studied, and the (137)Cs activity concentrations in stream water samples were 79-198 mBq L(-1) under stable runoff conditions and 702-13,400 Bq L(-1) under storm runoff conditions. The residual fraction (F4, considered to be non-bioavailable) was dominant, accounting for 59.5-82.6% of the total (137)Cs activity under stable runoff conditions and 69.4-95.1% under storm runoff conditions. The (137)Cs newly added to paddy fields in irrigation water only contributed 0.03-0.05% of the amount already present in the soil (201-348 kBq m(-2)). This indicates that the (137)Cs inflow load in irrigation water is negligible compared with that already in the soil. However, the contribution from the potentially bioavailable fractions (F1+F2+F3) was one order of magnitude larger, accounting for 0.20-0.59%. The increase in the dissolved and soluble radiocesium fraction (F1) was especially large (3.0% to infinity), suggesting that radiocesium migration in irrigation water is increasing the accumulation of radiocesium in rice.

Soil radiocesium distribution in rice fields disturbed by farming process after the Fukushima Dai-ichi Nuclear Power Plant accident

A magnitude 9.0 earthquake and subsequent large tsunami hit the northeastern coast of Japan on March 11, 2011. This resulted in serious damage to the reactors of the Fukushima Dai-ichi Nuclear Power Plant (FDNPP), operated by the Tokyo Electric Power Company. Large amounts of radionuclides were released from the FDNPP, a proportion of which were deposited onto the ground. In this study, we investigated soil radiocesium contamination of rice fields in Aga and Minamiuonuma, Niigata, ~130 and 200 km away from the FDNPP, respectively, as Niigata is one of the largest rice growing regions in Japan. Soil samples were collected from the plow layer of five rice fields in August and September, 5-6 months after the FDNPP accident. Results showed that radiocesium concentrations (the sum of Cs-134 and Cs-137) in the rice soil samples were ~300 Bq (kg dry soil)(-1). All samples contained a Cs-134/Cs-137 activity ratio of 0.68-0.96 after correction to March 11, 2011, showing that the radiocesium released from the FDNPP were deposited on these areas. Although the rice fields had been disturbed by farming processes after the FDNPP accident, the depth distribution of radiocesium concentrations in the plow layers showed higher concentrations in the upper soil layers. This suggests that spring tillage, flooding and puddling performed before rice transplantation may not disperse radiocesium deposited on the surface through the whole plow layer. In addition, the planar distribution of radiocesium concentrations was examined near the water inlet in one of the rice fields. Highest activities were found aligned with the direction of irrigation water discharge, indicating that radioactivity levels in rice fields may be elevated by an influx of additional radionuclides, probably in irrigation water, during farming.

Radiocesium immobilization to leaf litter by fungi during first-year decomposition in a deciduous forest in Fukushima.

Vast forest areas in eastern Japan have been contaminated with radio-isotopes by the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident. Radiocesium (radioCs) is known to remain bioavailable in forest ecosystems for a long time, and it is necessary to terminate the cycling process to decontaminate the forest ecosystem. We observed radiocesium concentrations of leaf litter during decomposition on a forest floor where radiocesium ((137)Cs) contamination was ∼155 kBq/m(2). Litter bag experiments were conducted with newly fallen mixed deciduous leaf litter in a deciduous forest (alt. 610 m) about 50 km from the FDNPP. Litter bags were retrieved in April, June, August, October, and December 2012. Fresh litter (137)Cs concentration was ∼3000 Bq/kg in December 2011. During the decomposition process on the forest floor, litter (137)Cs concentration increased rapidly and exceeded 25,000 Bq/kg after 6 months, whereas potassium (K) concentration in the litter was rather stable, indicating that radiocesium and K showed contrasting dynamics during the early decomposition phase. Nitrogen, phosphorus, and (137)Cs contents were positively correlated to fungal biomass, evaluated by phospholipid fatty acids in the litter during decomposition. The increase of radiocesium concentration mainly occurred during from April to October, when fungal growth peaked. Therefore, this suggests fungal translocation of nutrients from outside the litter substrate (immobilization) is the mechanism to increase radiocesium in the decomposing litter. The amount of (137)Cs contained in the 1-year-old decomposed leaf litter was estimated to be 4% per area of the soil-contaminated (137)Cs.

Enhanced transformation of diphenylarsinic acid in soil under sulfate-reducing conditions

Diphenylarsinic acid (DPAA) is known to be the major contaminant in soils where diphenylchloroarsine and diphenylcyanoarsine were abandoned after World Wars I and II. In this study, experimental model studies were performed to elucidate key factors regulating the transformation of DPAA under anaerobic soil conditions. The elimination of DPAA in Gleysol soils (Qiqihar and Shindori soils) was more rapid than in Mollisol and Regosol soils (Heihe and Ikarashi soils, respectively) during a 5-week incubation. No clear relationship between decreasing rates of DPAA concentrations and soil Eh values was found. The Ikarashi soil showed the slowest decrease in DPAA concentrations among the four soils, but the transformation of DPAA was notably enhanced by addition of exogenous sulfate together with acetate, cellulose or rice straw. Addition of molybdate, a specific inhibitor of sulfate reduction, resulted in the stagnation of DPAA transformation, suggesting that indigenous sulfate reducers play a role in DPAA transformation under anaerobic conditions. Arsenate, phenylarsonic acid, phenylmethylarsinic acid, diphenylmethylarsine oxide and three unknown compounds were detected as metabolites of DPAA. This is the first study to reveal enhancement of DPAA transformation under sulfate-reducing conditions.

Host-related variability in arbuscular mycorrhizal fungal structures in roots of Hedera rhombea , Rubus parvifolius , and Rosa multiflora under controlled conditions

The arbuscular mycorrhizal (AM) morphology of three host plant species inoculated with single and mixed fungal culture and the distribution of AM fungal species in roots of the hosts treated with a mixed culture of AM fungi were determined. The aim was to investigate the effect of host plants and AM fungi on AM morphology of coexisting plant species. Noncolonized rooted cuttings of Hedera rhombea (Miq) Bean, Rubus parvifolius L., and Rosa multiflora Thunb. were inoculated with five fungal species as single and mixed culture inocula. The fungal species used were Gigaspora rosea and Scutellospora erythropa, previously isolated from H. rhombea; Acaulospora longula and Glomus etunicatum from R. parvifolius; and Glomus claroideum from both plant species. A few hyphal and arbusculate coils were seen in the mixed culture-inoculated roots of R. parvifolius; all fungal treatments produced this Paris-type AM in H. rhombea and Arum-type AM in R. parvifolius, and R. multiflora indicates that AM morphology is strongly controlled by the identity of the host plants used in this study. AM fungal rDNA was extracted separately from roots of each replicate plant species inoculated with the mixed fungal culture, amplified, cloned, sequenced, and analyzed to determine the AM fungal species and their respective proportions in roots of each plant species. Glomus etunicatum and G. claroideum of the family Glomaceae generally occurred more frequently in R. parvifolius and R. multiflora, which form Arum-types, whereas S. erythropa, of the family Gigasporaceae, was the most frequently detected species in H. rhombea, which produced Paris-type AM. Although the genotype of the plant species used appears to determine the AM morphologies formed, there was preferential association between the hosts and AM fungal inoculants.

First report of white mould caused by Sclerotinia sclerotiorum on jackfruit

Symptoms of white mould were first observed on jackfruit (Artocarpus heterophyllus) in Rangpur, Bangladesh, during February 2012. Fluffy, white mycelia developed on the fruit surface, along with large irregular black sclerotia. Morphological characteristics and the internal transcribed spacer sequences of ribosomal DNA identified the fungus as Sclerotinia sclerotiorum.

Native Trichoderma strains isolated from Bangladesh with broad spectrum antifungal action against fungal phytopathogens

Nineteen Trichoderma isolates, collected from different locations in Bangladesh, were characterised through phenotypic, biochemical and molecular means. Besides, they were assessed for their antifungal action in vitro. The isolates were divided into three groups: T. asperellum, T. virens and T. harzianum. A dual culture assay and a culture filtrate assay against 6 phytopathogens revealed that 9 of the 19 isolates showed significant antifungal activities. The isolate T. harzianum TR05 showed the highest inhibition against Fusarium oxysporum, Rhizoctonia solani, Fusarium circinatum and Phomopsis vexans, followed by T. asperellum TR08 and T. virens TR06. TR08 had the highest inhibition against Sclerotium rolfsii and Pythium aphanidermatum, followed by TR05 and TR06. These findings were in agreement with their activities of extracellular hydrolytic enzymes, including chitinase, β-1,3-glucanase, and proteinase. Our results suggest that isolates TR05, TR06 and TR08 have the potential to be effective biocontrol agents against the phytopathogenic fungi.

Formation of diphenylthioarsinic acid from diphenylarsinic acid under anaerobic sulfate-reducing soil conditions

Diphenylarsinic acid (DPAA) is a toxic phenylarsenical compound often found around sites contaminated with phenylarsenic chemical warfare agents, diphenylcyanoarsine or diphenylchloroarsine, which were buried in soil after the World Wars. This research concerns the elucidation of the chemical structure of an arsenic metabolite transformed from DPAA under anaerobic sulfate-reducing soil conditions. In LC/ICP-MS analysis, the retention time of the metabolite was identical to that of a major phenylarsenical compound synthesized by chemical reaction of DPAA and hydrogen sulfide. Moreover the mass spectra for the two compounds measured using LC/TOF-MS were similar. Subsequent high resolution mass spectral analysis indicated that two major ions at m/z 261 and 279, observed on both mass spectra, were attributable to C12H10AsS and C12H12AsSO, respectively. These findings strongly suggest that the latter ion is the molecular-related ion ([M+H](+)) of diphenylthioarsinic acid (DPTA; (C6H5)2AsS(OH)) and the former ion is its dehydrated fragment. Thus, our results reveal that DPAA can be transformed to DPTA, as a major metabolite, under sulfate-reducing soil conditions. Moreover, formation of diphenyldithioarsinic acid and subsequent dimerization were predicted by the chemical reaction analysis of DPAA with hydrogen sulfide. This is the first report to elucidate the occurrence of DPAA-thionation in an anaerobic soil.