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Highlights: [•] cis-Cinnamic acid analogues was synthesized having substituents on the aromatic ring, and their inhibitory activities on the elongation of lettuce roots were evaluated. [•] Para- and ortho-substitution decreased potency. [•] meta-Substituted cis-cinnamic acid analogues were more likely to be potent. [•] m-Iodo, m-methoxy and m-trifluoromethyl substituents induced higher potency. [Copyright &y& Elsevier]

Highlights: [•] The citrate content in the roots was increased with Al treatments. [•] The activities of aconitase and NADP+-ICDH were decreased with Al treatments. [•] The activity of citrate synthase was not affected by Al. [•] Al–citrate complexes were a major chemical form in the cell sap of root tips. [Copyright &y& Elsevier]

Abstract: 1-O-cis-cinnamoyl-β-d-glucopyranose is one of the most potent allelochemicals isolated from Spiraea thunbergii Sieb. It is suggested that it derives its strong inhibitory activity from cis-cinnamic acid, which is crucial for phytotoxicity. It was synthesized to confirm its structure and bioactivity, and also a series of cis-cinnamic acid analogues were prepared to elucidate the key features of cis-cinnamic acid for lettuce root growth inhibition. The cis-cyclopropyl analogue showed potent inhibitory activity while the saturated and alkyne analogues proved to be inactive, demonstrating the importance of the cis-double bond. Moreover, the aromatic ring could not be replaced with a saturated ring. However, the 1,3-dienylcyclohexene analogue showed strong activity. These results suggest that the geometry of the C–C double bond between the carboxyl group and the aromatic ring is essential for potent inhibitory activity. In addition, using several light sources, the photostability of the cinnamic acid derivatives and the role of the C–C double bond were also investigated. [Copyright &y& Elsevier]

The mycotoxin deoxynivalenol (DON) causes serious problems worldwide in the production of crops such as wheat and barley because of its toxicity toward humans and livestock. A bacterial culture capable of degrading DON was obtained from soil samples collected in wheat fields using an enrichment culture procedure. The isolated bacterium, designated strain WSN05-2, completely removed 1,000 μg/mL of DON from the culture medium after incubation for 10 days. On the basis of phylogenetic studies, WSN05-2 was classified as a bacterium belonging to the genus Nocardioides. WSN05-2 showed significant growth in culture medium with DON as the sole carbon source. High-performance liquid chromatography analysis indicated the presence of a major initial metabolite of DON in the culture supernatant. The metabolite was identified as 3- epi-deoxynivalenol (3- epi-DON) by mass spectrometry and H and C nuclear magnetic resonance analysis. The amount of DON on wheat grain was reduced by about 90% at 7 days after inoculation with WSN05-2. This is the first report of a Nocardioides sp. strain able to degrade DON and of the yet unknown 3- epi-DON as an intermediate in the degradation of DON by a microorganism. [ABSTRACT FROM AUTHOR]

Abstract: Ogasawara Islands are important ecosystems sustaining many indigenous spices. To clarify the indigenous soil environments of Ogasawara Islands, we studied the chemistry of the soils. Many surface soils were low in bio-available P (0 to 0.55g P2O5 kg−1, average: 0.04g P2O5 kg−1 as Bray II P, n =22), but several soils were found to contain extremely large amounts of bio-available P (1.36 to 6.98g P2O5 kg−1, average: 2.93g P2O5 kg−1, n =5). From soil profile analyses, the authors concluded that the extremely large amount of bio-available P could not be explained by the effects of parent materials with high P contents nor the effect of fertilizations by human activity, but the effects of natural seabird activities in the past could be the cause. The soil profiles with large amounts of bio-available P indicate deep migration of soil materials from A horizons, which could be a result of intensive mixing of upper horizons by seabird activities. The intensive mixing was supported by the low mechanical impedance of the horizons for the P-accumulating soils (8.17±2.54kgcm−2, n =8) than those for the non-P-accumulating soils (17.46±3.52kgcm−2, n =36). It is likely that in the past seabirds, such as shearwaters, made burrows in the soils for nesting and propagating and inadvertently transported a large amount of P from the sea to the soils, resulting in the extremely large amounts of bio-available P in the present soils. [ABSTRACT FROM AUTHOR]

Abstract: To quantitatively clarify the organic carbon accumulation processes on the forest floor during an early stage of humification (3 years), solid-state 13C cross polarization magic angle spinning nuclear magnetic resonance (CPMAS NMR) signals were monitored for phased–humified beech and oak litters and soil surface horizons in the northern Kanto District, Japan. The mass loss rate of the carbon components during the humification for both litters was in the following order: O-alkyl > aromatic > aliphatic > carbonyl carbons. This result indicates that the labile O-alkyl carbons, probably dominated by holocellulose were selectively degraded compared to the other components. 44% of O-alkyl carbon mass for beech and 38% for oak lost throughout 3 years of incubation. Inversely, the mass of aliphatic carbons, which is mainly composed of saturated hydrocarbons, decreased quite slowly from 20 to 10% with humification, probably because a large proportion of the aliphatic carbons are secondary products of microorganisms. The aromatic carbon mass, which would be derived from lignin/tannin and their metabolites, also decreased gradually from 17 to 6% over 3 years. While, the carbonyl carbon mass was quite stable at around 2% throughout the incubation period, probably because the hydrolysis reactions of organic carbon would contribute to the formation of the carbonyl carbons. According to an exponential model, the total carbon stocks on the forest floor converged at 4.2 Mg C ha−1 for the first few years at the studying site. The carbon compositions converged to intermediate levels between those of the F and A 1 horizons. The simulation in the present study is able to represent the carbon accumulation process on the forest floor including a part of the mineral. [Copyright &y& Elsevier]

The article offers information on the adaptive aluminum resistance in Rhodotorula glutinis through mitochondrial regulation. It examines the microbial acclimation to increasing aluminum stress, as well as the mechanisms responsible for adaptive aluminum resistance. Wild-type and aluminum resistant cells were compared to determine the mechanism underlying the acquired aluminum resistance of Rhodotorula glutinis IFO1125. A greater number of mitochondria was found in resistant cells. It is revealed that increased number of mitochondria and increased mitochondrial DNA content is directly related to increased aluminum resistance.

Abstract: Cyanamide (NH2CN) has recently been proven to be a natural product, although it has been synthesized for over 100 years for agricultural and industrial purposes. The distribution of natural cyanamide appears to be limited, as indicated by our previous investigation of 101 weed species. In the present study, to investigate the distribution of natural cyanamide in Vicia species, we monitored the cyanamide contents in V. villosa subsp. varia, V. cracca, and V. amoena during their pre-flowering and flowering seasons. It was confirmed that V. cracca was superior to V. villosa subsp. varia in accumulating natural cyanamide, and that V. amoena was unable to biosynthesize this compound under laboratory condition examined. The localization of cyanamide in the leaves of V. villosa subsp. varia seedlings was also clarified. In a screening study to find cyanamide-biosynthesizing plants, only Robinia pseudo-acacia was found to contain cyanamide among 452 species of higher plants. We have investigated 553 species to date, but have so far found the ability to biosynthesize cyanamide in only three species, V. villosa subsp. varia, V. cracca and R. pseudo-acacia. [Copyright &y& Elsevier]

Abstract: To determine the mechanism of aluminum (Al) detoxification in the roots of tea plants (Camellia sinensis (L.) Kuntze), the amounts of Al and Al-chelating compounds (fluoride (F), organic acids and catechins) were measured and the chemical forms of Al in root cell extracts were identified by the application of 27Al-nuclear magnetic resonance (NMR) spectroscopy. Tea plants were cultivated in nutrient solutions containing 0, 4, 1.0 and 4.0mM of Al at pH 4.2 for approximately 10 weeks. The levels of soluble Al, water-soluble oxalate and citrate, but not F, malate or catechins in young roots increased with an increase in the concentration of Al in the treatment solution. The 27Al NMR spectra of root tips and cell sap extracted from root tips that had been treated with Al were almost identical and had four signals, with two (11 and 16ppm) apparently corresponding to the known chemical shifts of Al–oxalate complexes. In the spectra of cell sap, the resonances at 11 and 16ppm increased with an increase in the Al contents. These results suggest that the levels of Al–oxalate complexes increased in response to an increase in the Al level, implying that oxalate is a key Al-chelating compound in the mechanism of Al detoxification in the tea root. [Copyright &y& Elsevier]

The production of quorum-sensing-related signal molecules (QSRMs) among culturable bacteria comprising the community on wheat heads was investigated. The taxonomic position of 186 bacterial isolates obtained from ten heads was inferred based on 16S rRNA gene sequences, and their QSRM production was determined using two bioreporter strains of N-acylhomoserine lactones. Approximately 33% of isolates produced QSRMs, though the proportion of QSRM-producing isolates on a wheat head was significantly negatively correlated with population size. Most of the producing isolates were Pantoea species, most commonly Pantoea ananatis. Furthermore, the proportion of Pantoea ananatis that produced QSRMs was significantly negatively correlated with the number of bacterial genera (community richness) on each head. Finally, community richness was positively correlated with population size. Qualitative analysis using thin-layer-chromatography revealed that the QSRMs of Pantoea isolates were composed of at least two compounds. This is the first report indicating that Pantoea ananatis isolates inhabiting wheat heads are capable of producing QSRMs. QSRM production by Pantoea spp. may contribute to the predominance of this genus on wheat heads, particularly at relatively low population densities and community diversity. [ABSTRACT FROM AUTHOR]

• Hydrophilic fulvic acid fractions had received much biological transformations. • Humic acid fractions had formed at early stage of soil formation with less biological transformation. • Fire event could be a possible formation mechanism of HA fractions. • Mobility of soil organic carbon fractions was limited to <4 mm per century. • The thick humic horizon reflects the successive up-building accumulation of soil particles containing soil organic carbon. A buried humic horizon (14C age between 5.4 and 6.8 kyr BP) of a volcanic ash soil in Aomori, Japan, which was collected from the depth between 147 and 187 cm at 5 cm-interval (total eight sub-horizon samples), was investigated to clarify the degree of biological transformation and mobility of soil organic carbon (SOC) fractions. The SOC fractions were prepared from each sub-horizon sample by extraction and precipitation procedures with controlling pH of the extracted solution, resulting in humin, humic acid (HA), and four fulvic acid (FA) fractions (two hydrophilic FA fractions: FA 1 and FA 2 , and two hydrophobic FA fractions: FA 3 and FA IHSS). The prepared SOC fractions were characterized by 14C age and stable isotopic ratios of 13C (δ13C) and 15N (δ15N). The hydrophilic FA fractions showed the highest δ13C and δ15N values, indicating that these SOC fractions had been most enriched with 13C and 15N by biological metabolic processes. On the other hand, the HA fraction showed the lowest δ13C and δ15N values, therefore this fraction would have been less-metabolized, although HA fraction has been regarded as well-processed in general. The 14C age of the HA fraction was almost same as the deposition age of the corresponding sub-horizon, indicating that the C in the HA fraction would have been fixed in situ right after photosynthesis by plants at the early stage of soil formation and chemically stabilized at soil surface, by fire event, etc. The average rates of vertical translocation of the SOC fractions were low (humin and HA fractions: <1 mm per century, FA fractions: 1–4 mm per century), implying that the vertical translocation of SOC would not be the main mechanisms for forming thick humic horizons. The present study showed strong evidence of the in situ formation of SOC on soil surface, and successive up-building accumulation of soil particles containing SOC would contribute to the formation of the thick humic horizons. [ABSTRACT FROM AUTHOR]

• We simulated the eradication of invasive goats and rats on an oceanic island. • Simultaneous eradication both goats and rats was favorable for ecosystem restoration. • The simultaneous eradication made the ecosystem be bi-stable. • In rare cases, the ecosystem was crashed after the eradication. The decline of the natural ecosystem on the Ogasawara Islands caused by invasive species has been an important environmental problem and projects for eradicating invasive species are currently underway. Some invasive species have proliferated in the host environment and are affecting the material cycle in the ecosystem. If the invasive species were removed from the environment, the material cycle of the ecosystem may be altered and native species that should be protected may be negatively affected. After the introduction of invasive alien species, namely goats and rats, the vegetation declined on Nakoudojima Island, an island that belongs to the Ogasawara Islands. Therefore, in this study, a mathematical model was developed to simulate and analyze the ecosystem changes that occurred following the eradication of goats and rats. When only rats were eradicated, the island's vegetation changed to grassland but revegetation was delayed and the biomass of the native animal species did not recover significantly. When only goats were eradicated, the vegetation recovered and the biomass of the native animal species increased; however, the island became forested and many native herbaceous plant species became extinct. When goats and rats were eradicated simultaneously, the vegetation recovered and the biomass of the native animal species increased; however, the ecosystem became very unstable and two extreme results were obtained – the vegetation of the entire island changed to either forest or grassland. The results of this study suggest that instead of simply eradicating invasive species, it is necessary to monitor post-eradication changes in the ecosystem and take adaptive measures as required. [ABSTRACT FROM AUTHOR]

Disturbances caused by invasive ungulates alter soil environments markedly and can prevent ecosystem recovery even after eradication of the ungulates. On oceanic islands, overgrazing and trampling by feral goats has caused vegetation degradation and soil erosion, which can alter soil chemistry. To understand the effects of the changes on plant performance, we conducted a laboratory experiment to assess herbaceous species growth under various soil conditions with phosphorous, nutrients, and acidity. Subsoil was collected from Nakodo-jima in the northwest Pacific. Six herbaceous species dominating the island were grown in soils with three levels of added CaCO 3 and P 2 O 5 and two levels of added KNO 3 . After 4 weeks of growth, the total dry plant weight was significantly lower with no added P 2 O 5 , regardless of the addition of KNO 3 . Three species weighed more under P 2 O 5 and KNO 3 addition in high-pH soil, whereas the remaining three weighed less. Our results indicated that herbaceous species growth is limited primarily by phosphorous availability; the limitation is dependent on soil pH, and the trend of dependency differs among species. This implies that ecosystems with extreme disturbances cannot recover without improving the soil chemistry., (© 2023. The Author(s).)

Restoration of species-rich semi-natural grassland requires not only a seed source but also appropriate soil properties. In Europe, approximately 10 years are required for the properties of fertilized soils to reach suitable conditions and be considered successfully restored. However, restoration may require additional time in Japan because heavier precipitation causes leaching of basic cations from soils, resulting in soil acidification; volcanic ejecta also forms active Al and Fe hydroxides with high phosphate sorption. Within this context, we aimed to answer the following questions: i) whether and how the impacts of fertilization remain in the soil properties after half a century in Japan; and ii) how fertilization affects the restoration of semi-natural grasslands in Japan. We investigated the vegetation and soil properties of a Zoysia japonica pasture improved half a century ago with a single application of fertilizer and an adjacent semi-natural grassland (native pasture) in Japan, and found the following: (1) the two pastures had similar dominance of Z. japonica, but differed in the species composition; (2) the improved pasture exhibited lower species richness than the native pasture; (3) soil nutrients, including N, P, K, Mg, and Ca, were higher in the improved pasture than in the native pasture; and (4) many chemical properties of the soils were associated with species composition; namely, the vegetation on nutrient-rich soil had more alien species and fewer native species. We conclude that a single dose of fertilization can affect soil properties in semi-natural grasslands over half a century in Japan, leading to species loss and changing the species composition. We suggest that fertilized soils under grazing in Japan may require more than half a century to restore the nutrients to suitable levels for the establishment of a species-diverse grassland., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2022 Tsutsumi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Leaf ionome indicates plant phylogenetic evolution and responses to environmental stress, which is a critical influential factor to the structure of species populations in local edaphic sites. However, little is known about leaf ionomic responses of local plant species to natural edaphic mineral variations. In the present study, all plant species and soil samples from a total of 80 soil sites in Shiozuka Highland were collected for multi-elemental analysis. Ioniomic data of species were used for statistical analysis, representing 24 species and 10 families. Specific preferences to ionomic accumulation in plants were obviously affected by the phylogeny, whereas edaphic impacts were also strong but limited within the phylogenetic preset. Correlations among elements resulted from not only elemental synergy and competition but also the adaptive evolution to withstand environmental stresses. Furthermore, ionomic differences of plant families were mainly derived from non-essential elements. The majority of variations in leaf ionome is undoubtedly regulated by evolutionary factors, but externalities, especially environmental stresses also have an important regulating function for landscape formation, determining that the contributions of each factor to ionomic variations of plant species for adaptation to environmental stress provides a new insight for further research on ionomic responses of ecological speciation to environmental perturbations and their corresponding adaptive evolutions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Zhang, Hiradate, Kusumoto, Morita, Koyanagi, Chu and Watanabe.)

Selective removal of lignin and other recalcitrant compounds, collectively registered as acid-unhyrolyzable residue (AUR), results in bleaching of leaf litter, but the importance of bleaching in decomposition processes on forest soil has not been fully evaluated. The aims of this study were to elucidate the occurrence of bleached area in decomposing leaf litter and to compare chemical composition between bleached and nonbleached portions in a subtropical forest in Japan. Field incubation of leaf litter was performed over an 18-month period with the litterbag method. The decomposition processes during the first 9 month were characterized by the relatively rapid mass loss and increase of bleached area, whereas the mass loss was slowed down and the bleached area decreased thereafter. Mass loss of leaf tissues was faster and AUR content was lower in bleached than in nonbleached portions, indicating the acceleration of mass loss in bleached leaf tissues by the selective decomposition of recalcitrant compounds. The decrease in carbonyl-C in the bleached portions was associated with the increase of extractable nitrogen. The results suggest that the bleaching plays a dominant role in the transformation and turnover of organic compounds and nitrogen in decomposing leaf litter.

Microbes inhabiting the phyllosphere encounter harmful ultraviolet rays, and must develop adaptive strategies against this irradiation. In this study, we screened bacterial isolates originating from the phyllosphere of various plants which harbored absorbers of ultraviolet A (UVA), a wavelength range which is recognized as harmful to human skin. Of the 200 phyllosphere bacterial isolates we screened, methanol extracts from bacterial cells of seventeen isolates absorbed wavelengths in the range of 315-400nm. All of the UVA-absorbing strains belonged to Methylobacterium species based on 16S ribosomal RNA gene sequences, suggesting that cells of this bacterial genus contain specific UVA-absorbing compounds. When cells of a representative Methylobacterium strain were extracted using various solvents, UVA absorption was observed in the extracts obtained using several aqueous solvents, indicating that the UVA-absorbing compounds were highly polar. A compound was purified using solid columns and HPLC separation, and comparative analysis revealed that the absorption strength and spectrum of the compound were similar to those of the known UVA filter, avobenzone. The compound was also verified to be stable under UVA exposure for at least 480min. Based on these results, the UVA-absorbing compound harbored by Methylobacterium has potential to be used as a novel sunscreen ingredient., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The relationship between degradation speed of soil-buried biodegradable polyester film in a farmland and the characteristics of the predominant polyester-degrading soil microorganisms and enzymes were investigated to determine the BP-degrading ability of cultivated soils through characterization of the basal microbial activities and their transition in soils during BP film degradation. Degradation of poly(butylene succinate-co-adipate) (PBSA) film was evaluated in soil samples from different cultivated fields in Japan for 4 weeks. Both the degradation speed of the PBSA film and the esterase activity were found to be correlated with the ratio of colonies that produced clear zone on fungal minimum medium-agarose plate with emulsified PBSA to the total number colonies counted. Time-dependent change in viable counts of the PBSA-degrading fungi and esterase activities were monitored in soils where buried films showed the most and the least degree of degradation. During the degradation of PBSA film, the viable counts of the PBSA-degrading fungi and the esterase activities in soils, which adhered to the PBSA film, increased with time. The soil, where the film was degraded the fastest, recorded large PBSA-degrading fungal population and showed high esterase activity compared with the other soil samples throughout the incubation period. Meanwhile, esterase activity and viable counts of PBSA-degrading fungi were found to be stable in soils without PBSA film. These results suggest that the higher the distribution ratio of native PBSA-degrading fungi in the soil, the faster the film degradation is. This could be due to the rapid accumulation of secreted esterases in these soils.

To improve the biodegradation of biodegradable plastic (BP) mulch films, 1227 fungal strains were isolated from plant surface (phylloplane) and evaluated for BP-degrading ability. Among them, B47-9 a strain isolated from the leaf surface of barley showed the strongest ability to degrade poly-(butylene succinate-co-butylene adipate) (PBSA) and poly-(butylene succinate) (PBS) films. The strain grew on the surface of soil-mounted BP films, produced breaks along the direction of hyphal growth indicated that it secreted a BP-degrading enzyme, and has directly contributing to accelerating the degradation of film. Treatment with the culture filtrate decomposed 91.2 wt%, 23.7 wt%, and 14.6 wt% of PBSA, PBS, and commercially available BP polymer blended mulch film, respectively, on unsterlized soil within 6 days. The PCR-DGGE analysis of the transition of soil microbial community during film degradation revealed that the process was accompanied with drastic changes in the population of soil fungi and Acantamoeba spp., as well as the growth of inoculated strain B47-9. It has a potential for application in the development of an effective method for accelerating degradation of used plastics under actual field conditions.

Mealybugs, which include several agricultural pests, are small sap feeders covered with a powdery wax. They exhibit clear sexual dimorphism; males are winged but fragile and short lived, whereas females are windless and less mobile. Thus, sex pheromones emitted by females facilitate copulation and reproduction by serving as a key navigation tool for males. Although the structures of the hitherto known mealybug pheromones vary among species, they have a common structural motif; they are carboxylic esters of monoterpene alcohols with irregular non-head-to-tail linkages. However, in the present study, we isolated from the Matsumoto mealybug, Crisicoccus matsumotoi (Siraiwa), a pheromone with a completely different structure. Using gas chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy, we identified the pheromone as 3-methyl-3-butenyl 5-methylhexanoate. Its attractiveness to males was confirmed in a series of field trapping experiments involving comparison between the isolated natural product and a synthetic sample. This is the first report of a hemiterpene mealybug pheromone. In addition, the acid moiety (5-methylhexanoate) appears to be rare in insect pheromones.

Velvetbean (Mucuna pruriens) plants impede the growth of neighboring plants. One compound, 3-(3',4'-dihydroxyphenyl)-L-alanine (L-DOPA), is responsible for the allelopathic capacity of velvetbean. This compound is an active allelochemical that decreases root growth of several plant species. In mammals, L-DOPA is a well-known therapeutic agent for the symptomatic relief of Parkinson's disease. However, its mode of action in plants is still not well understood. To address such issues, gene expression in Arabidopsis thaliana plants, which had been exposed to L-DOPA, was analyzed using DNA microarrays. After 6 h of L-DOPA exposure, the expression of 110 genes was significantly upregulated, and the expression of 69 genes was significantly downregulated. These induced genes can be divided into different functional categories, mainly on the basis of subcellular localization, metabolism, and proteins with a binding function or cofactor requirement. Based on these results, we suggest that L-DOPA acts by two mechanisms: it influences amino acid metabolism and deregulates metal homeostasis, especially that of iron, which is required for the fundamental biological processes of all organisms.

Plant roots exude viscous polysaccharides, called mucilage. One of the suggested roles of mucilage is immobilization of toxic metal cations, including aluminum (Al), in the rhizosphere. Mucilage exuded from roots of Melastoma malabathricum (Al accumulator) was characterized in comparison with that of Zea mays (maize; Al nonaccumulator). Removal of mucilage significantly reduced Al accumulation in M. malabathricum. The cation adsorption affinity of M. malabathricum mucilage was higher for Al and lanthanum (La) than for barium (Ba), whereas that of maize mucilage was in the order Ba > La > Al. A (27)Al nuclear magnetic resonance (NMR) spectrum of the Al-adsorbed mucilage and bioassay with alfalfa seedlings indicated that the concentrated Al in the mucilage of M. malabathricum, unlike that of maize, bound very weakly to cation exchange sites of mucilage. The higher charge density in M. malabathricum mucilage, derived from unmethylated uronic acid, is inferred to be related to preferential adsorption of trivalent cation. Not only a higher degree of methylation in the uronic acid (glucuronic acid) but also H(+) release from roots to the mucilage appears to be responsible for the loose binding of Al in M. malabathricum mucilage. These characteristics of mucilage may help Al hyperaccumulation in M. malabathricum.

3-(3',4'-Dihydroxyphenyl)-L-alanine (L-DOPA), which is synthesized in velvet bean (Mucuna pruriens), inhibits plant growth. The concentration of L-DOPA in soil is reduced by adsorption and transformation reactions, which can result in the reduction of its plant-growth-inhibitory activity. To determine which part of the L-DOPA structure is involved in the adsorption and soil transformation reactions, we compared the kinetics of L-DOPA disappearance in a volcanic ash soil with that of L-phenylalanine (3-phenyl-L-alanine) and L-tyrosine (3-(4'-hydroxyphenyl)-L-alanine), compounds that are similar in structure to L-DOPA but do not have a catechol (o-dihydroxybenzene) moiety. L-Phenylalanine and L-tyrosine were not adsorbed and transformed in the soil at equilibrium pH values between 4 and 7. These results suggest that the adsorption and transformation reactions of L-DOPA in the soil involve the catechol moiety and not the amino and carboxylic acid groups, which are common to all three compounds. Like L-DOPA, (+)-catechin, another allelochemical that contains a catechol moiety, underwent adsorption and soil transformation reactions. Thus, we concluded that the concentrations of allelochemicals bearing a catechol moiety in soils will decrease rapidly owing to adsorption and transformation reactions, and this decrease will be faster in soils with a high pH value or high adsorption ability. Owing to this decrease in concentration, allelopathic phenomena may not occur.

Cyanamide is a multifunctional agrochemical used, for example, as a pesticide, herbicide, and fertilizer. Recent research has revealed that cyanamide is a natural product biosynthesized in a leguminous plant, hairy vetch (Vicia villosa). In the present study, gas chromatography-mass spectrometry (GC-MS) equipped with a capillary column for amines was used for direct quantitative determination of cyanamide. Quantitative signals for ((14)N(2))cyanamide, ((15)N(2))cyanamide (internal standard for stable isotope dilution method), and m-(trifluoromethyl)benzonitrile (internal standard for correcting errors in GC-MS analysis) were recorded as peak areas on mass chromatograms at m/z 42 (A(42)), 44 (A(44)), and 171 (A(IS)), respectively. Total cyanamide content, ((14)N(2))cyanamide plus ((15)N(2))cyanamide, was determined as a function of (A(42)+A(44))/A(IS). Contents of ((14)N(2))cyanamide and ((15)N(2))cyanamide were then calculated by multiplying the total cyanamide content by A(42)/(A(42)+A(44)) and A(44)/(A(42)+A(44)), respectively. The limit of detection for the total cyanamide content by the GC-MS analysis was around 1ng. The molar ratio of ((14)N(2))cyanamide to ((15)N(2))cyanamide in the injected sample was equal to the observed A(42)/A(44) value in the range from 0.1 to 5. It was, therefore, possible to use the stable isotope dilution method to quantify the natural cyanamide content in samples; i.e., the natural cyanamide content was derived by subtracting the A(42)/A(44) ratio of the internal standard from the A(42)/A(44) ratio of sample spiked with internal standard, and then multiplying the resulting difference by the amount of added ((15)N(2))cyanamide (SID-GC-MS method). This method successfully gave a reasonable value for the natural cyanamide content in hairy vetch, concurring with the value obtained by a conventional method in which cyanamide was derivatized to a photometrically active compound 4-cyanimido-1,2-naphthoquinone and analyzed with reversed-phase HPLC (CNQ-HPLC method). The determination range of cyanamide in the SID-GC-MS method was almost the same as that in the CNQ-HPLC method; however, the SID-GC-MS method was much simpler than the CNQ-HPLC method.

Robinia pseudo-acacia L. (black locust) is a nonindigenous species currently invading the central part of Japanese grasslands. Several allelochemicals were identified and characterized from the leaf tissue. The growth of both radicle and hypocotyl in the tested species (barnyard grass, white clover, lettuce, and Chinese cabbage) was reduced when grown in soil mixed with the leaves of R. pseudo-acacia at various concentrations. Aqueous leaf extracts, when bioassayed, exhibited a significant suppression of radicle growth. Chromatographic separation of an ethanolic extract of R. pseudo-acacia leaves resulted in isolation of three compounds, identified as robinetin (1), myricetin (2), and quercetin (3) by nuclear magnetic resonance and mass spectroscopy. All inhibited root and shoot growth of lettuce. Robinetin, found in a large amount, caused 50% suppression of the root and shoot growth of lettuce at 100 ppm. The presence of these bioactive substances in leaf tissue suggests a potential role for flavonoids in R. pseudo-acacia invasion in introduced habitats.

Spiraea thunbergii Sieb. contains 1-O-cis-cinnamoyl-beta-D-glucopyranose (CG) and 6-O-(4'-hydroxy-2'-methylene-butyroyl)-1-O-cis-cinnamoyl-beta-D-glucopyranose (BCG) as major plant growth inhibiting constituents. In the present study, we determined the inhibitory activity of CG and BCG on root elongation of germinated seedlings of lettuce (Lactuca sativa), pigweed (Amaranthus retroflexus), red clover (Trifolium pratense), timothy (Phleum pratense), and bok choy (Brassica rapa var chinensis) in comparison with that of two well-known growth inhibitors, 2,4-dichlorophenoxyacetic acid (2,4-D) and (+)-2-cis-4-trans-abscisic acid (cis-ABA), as well as two related chemicals of CG and BCG, cis-cinnamic acid (cis-CA) and trans-cinnamic acid (trans-CA). The EC50 values for CG and BCG on lettuce were roughly one-half to one-quarter of the value for cis-ABA. cis-Cinnamic acid, which is a component of CG and BCG, possessed almost the same inhibitory activity of CG and BCG, suggesting that the essential chemical structure responsible for the inhibitory activity of CG and BCG is cis-CA. The cis-stereochemistry of the methylene moiety is apparently needed for high inhibitory activity, as trans-CA had an EC50 value roughly 100 times that of CG, BCG, and cis-CA. Growth inhibition by CG, BCG, and cis-CA was influenced by the nature of the soil in the growing medium: alluvial soil preserved the bioactivity, whereas volcanic ash and calcareous soils inhibited bioactivity. These findings indicate a potential role of cis-CA and its glucosides as allelochemicals for use as plant growth regulators in agricultural fields.

A sex pheromone component of Pseudococcus cryptus has been isolated and identified. The crude pheromone extract obtained by airborne collection was fractionated by liquid chromatography (LC) on Florisil, and further purified by high performance liquid chromatography and preparative Gas Chromatography (GC). The pheromone component was shown to be an ester, the alcohol part of which was identical to the known alcohol moiety of the pheromone of Planococcus citri. The chemical structure was determined to be 3-isopropenyl-2,2-dimethylcyclobutylmethyl 3-methyl-3-butenoate by MS and 1H NMR analyses. The absolute configuration of the pheromone was assigned as (1R,3R) by comparison of the retention time of the alcohol derived from the P. cryptus pheromone with those of the alcohol derived from P. citri pheromone, and a synthetic sample of alcohol enriched in the (1R,3R)-enantiomer, using a chiral GC stationary phase. The structure of the pheromone was confirmed by synthesis, and by bioassays in a glasshouse.

The formation and presence of aluminum tridecamer (Al(13), [AlO(4)Al(12)(OH)(24)(H(2)O)(12)](7+)) in solution was strongly inhibited by soil humic acids (HA), as shown by liquid-state 27Al-nuclear magnetic resonance (NMR) analyses. It was also observed that Al(13) coprecipitated with HA in the form of Al(13)-HA complexes, as shown by solid-state magic angle spinning (MAS) 27Al-NMR analyses. Tetrahedral Al in the Al(13)-HA complexes was gradually converted to octahedral Al, indicating that Al(13) in Al(13)-HA complexes was decomposed into Al-HA complexes with reaction time. When HA was present in the acidic Al solution before the partial neutralization, the precipitates formed after the partial neutralization did not contain any tetrahedral Al, indicating the absence of Al(13) and that HA is a strong inhibitor of Al(13) formation. These results indicate that Al(13) is not formed from Al complexed with soil HA after partial neutralization. If Al(13) was added to soils, it would preferentially form precipitates of non-phytotoxic Al(13)-HA complex in the presence of excess HA (i.e. a COOH/Al(13) molar ratio of more than 9) and gradually be converted to an octahedral Al-HA complex. Therefore, Al(13) is not a major plant-growth inhibitor in humus-rich acidic soils.

Cyanamide was isolated from the leaves and stems of hairy vetch (Vicia villosa), guided by plant growth inhibitory activity against lettuce (Lectuca sativa) seedlings. A large proportion of the inhibitory activity in the crude extract was explained by the presence of cyanamide, suggesting it to be a possible allelochemical in this species. The amount in a 9-day-old seedling, which had been grown without nutrients, reached approx. 40 times that of a nongerminated seed, demonstrating cyanamide biosynthesis in the seedlings. This is the first report on the isolation of a possible allelochemical from hairy vetch and also of the finding of cyanamide as a natural product.