Physiological responses of low- and high-cadmium accumulating Robinia pseudoacacia-rhizobium symbioses to cadmium stress.

The role of rhizobia in alleviating cadmium (Cd) stress in woody legumes is still unclear. Therefore, two types of black locust (Robinia pseudoacacia L.) with high and low Cd accumulation abilities were selected from 11 genotypes in China, and the effects of rhizobium (Mesorhizobium huakuii GP1T11) inoculation on the growth, CO2 and H2O gas exchange parameters, Cd accumulation, and the absorption of mineral elements of the high (SX) and low Cd-accumulator (HB) were compared. The results showed that rhizobium-inoculation significantly increased biomass, shoot Cd contents, Cd accumulation, root-to-shoot translocation factor (TF) and the absorption and accumulation of mineral elements in both SX and HB. Rhizobium-inoculation increased chlorophyll a and carotenoid contents, and the intercellular carbon dioxide concentrations in HB plants. Under Cd exposure, the high-accumulator SX exhibited a significant decrease in photosynthetic CO2 fixation (Pn) and an enhanced accumulation of Cd in leaves, but coped with Cd exposure by increasing chlorophyll synthesis, regulating stomatal aperture (Gs), controlling transpiration (Tr), and increasing the absorption and accumulation of mineral elements. In contrast, the low-accumulator HB was more sensitive to Cd exposure despite preferential accumulation of Cd in roots, with decreased chlorophyll and carotenoid contents, but significantly increased root biomass. Compared to the low-accumulator HB, non-inoculated Cd-exposed SX plants had higher chlorophyll contents, and rhizobium-inoculated Cd-exposed SX plants had higher Pn, Tr, and Gs as well as higher levels of P, K, Fe, Ca, Zn, and Cu. In conclusion, the high- and low-Cd-accumulator exhibited different physiological responses to Cd exposure. Overall, rhizobium-inoculation of black locust promoted the growth and heavy metal absorption, providing an effective strategy for the phytoremediation of heavy metal-contaminated soils by this woody legume.

Exogenous chelating agents influence growth, physiological characteristics and cell ultrastructure of Robinia pseudoacacia seedlings under lead-cadmium stress.

Heavy metal pollution of soil, especially by lead (Pb) and cadmium (Cd), is a serious problem worldwide. The application of safe chelating agents, combined with the growing of tolerant trees, constitutes an approach for phytoremediation of heavy-metal-contaminated soil. This study aimed to determine whether the two safe chelators, tetrasodium glutamate diacetate (GLDA) and citric acid (CA), could improve the phytoremediation capacity of black locust (Robinia pseudoacacia L.) in a Pb-Cd-contaminated soil and to find the key factors affecting the biomass accumulation of stressed black locust. In Pb- and Cd-stressed black locust plants, medium- and high-concentration GLDA treatment inhibited the growth, chlorophyll synthesis and maximum photochemical efficiency (Fv/Fm), promoted the absorption of Pb and Cd ions and resulted in the shrinkage of chloroplasts and starch grains when compared with those in Pb- and Cd-stressed plants that were not treated with GLDA. The effects of CA on plant growth, ion absorption, chlorophyll content, chlorophyll fluorescence and organelle size were significantly weaker than those of GLDA. The effect of both agents on Cd absorption was greater than that on Pb absorption in all treatments. The levels of chlorophyll a and plant tissue Cd and rates of starch metabolism were identified as the key factors affecting plant biomass accumulation in GLDA and CA treatments. In the future, GLDA can be combined with functional bacteria and/or growth promoters to promote the growth of Pb- and Cd-stressed plants and to further improve the soil restoration efficiency following pollution by heavy metals. Application of CA combined with the growing of black locust plants has great potential for restoring the Cd-polluted soil. These findings also provide insights into the practical use of GLDA and CA in phytoremediation by R. pseudoacacia and the tolerant mechanisms of R. pseudoacacia to Pb-Cd-contaminated soil.

The Rpf107 gene, a homolog of LOR, is required for the symbiotic nodulation of Robinia pseudoacacia.

MAIN CONCLUSION: Rpf107 is involved in the infection process of rhizobia and the maintenance of symbiotic nitrogen fixation in black locust root nodules. The LURP-one related (LOR) protein family plays a pivotal role in mediating plant defense responses against both biotic and abiotic stresses. However, our understanding of its function in the symbiotic interaction between legumes and rhizobia remains limited. Here, Rpf107, a homolog of LOR, was identified in Robinia pseudoacacia (black locust). The subcellular localization of Rpf107 was analyzed, and its function was investigated using RNA interference (RNAi) and overexpression techniques. The subcellular localization assay revealed that Rpf107 was mainly distributed in the plasma membrane and nucleus. Rpf107 silencing prevented rhizobial infection and hampered plant growth. The number of infected cells in the nitrogen fixation zone of the Rpf107-RNAi nodules was also noticeably lower than that in the control nodules. Notably, Rpf107 silencing resulted in bacteroid degradation and the premature aging of nodules. In contrast, the overexpression of Rpf107 delayed the senescence of nodules and prolonged the nitrogen-fixing ability of nodules. These results demonstrate that Rpf107 was involved in the infection of rhizobia and the maintenance of symbiotic nitrogen fixation in black locust root nodules. The findings reveal that a member of the LOR protein family plays a role in leguminous root nodule symbiosis, which is helpful to clarify the functions of plant LOR protein family and fully understand the molecular mechanisms underlying legume-rhizobium symbiosis.

Invasive Alien Species as a Potential Source of Phytopharmaceuticals: Phenolic Composition and Antimicrobial and Cytotoxic Activity of Robinia pseudoacacia L. Leaf and Flower Extracts.

Black locust (Robinia pseudoacacia L.), an invasive tree in Europe, commonly known for its negative impact on biodiversity, is a rich source of phenolic compounds recognized in traditional medicine. Since the metabolite profile depends on the environment and climate, this study aimed to provide the first LC-MS phytochemical screening of the black locust from the Istria region (Croatia). The compounds were extracted from leaves and flowers with 70% ethanol and 80% methanol. Total phenolics (TP) and flavonoids (TF), as well as antioxidant capacity (AC) measured by ABTS (17.49-146.41 mg TE/g DW), DPPH (24.67-118.49 mg TE/g DW), and FRAP (7.38-77.53 mg TE/g DW) assays, were higher in leaf than in flower extracts. Higher TP and total non-flavonoid (TNF) values were displayed in ethanolic than in methanolic extracts. In total, 64 compounds were identified, of which flavonols (20) and hydroxycinnamic acid derivatives (15) were the most represented. Flavanols such as catechin dominated in leaf extracts, followed by flavonols, with kaempferol glucuronyl rhamnosyl hexosides as the main compound, respectively. Flower extracts had the highest share of flavones, followed by ellagitannins, with luteolin dirhamnosyl hexosides and vescalagin, respectively, being predominant. The extracts had good quorum sensing, biofilm formation prevention, and eradicating capacity. The results provided new insights into the phytochemical properties of R. pseudoacacia as the first step toward its potential pharmaceutical use.

Temporal shifts in root exudates driven by vegetation restoration alter rhizosphere microbiota in Robinia pseudoacacia plantations.

Plant-soil-microbiota interactions mediated by root exudates regulate plant growth and drive rhizosphere microbial feedbacks. It remains unknown how root exudates affect rhizosphere microbiota and soil functions in the course of forest plantation restoration. The metabolic profiles of tree root exudates are expected to shift with stand age, leading to variation in rhizosphere microbiota structure, and in turn, potentially altering soil functions. To unravel the effects of root exudates, a multi-omics study was conducted using untargeted metabonomic profiling, high-throughput microbiome sequencing and functional gene array. The interactions among root exudates, rhizosphere microbiota and nutrient cycling-related functional genes were explored under 15- to 45-year-old Robinia pseudoacacia plantations in the Loess Plateau region of China. Root exudate metabolic profiles, rather than chemodiversity, markedly changed with an increase in stand age. A total of 138 age-related metabolites were extracted from a key module of root exudates. The relative contents of six biomarker metabolites, such as glucose-1-phosphate, gluconic acid and N-acetylneuraminic acid, increased distinctly over time. The biomarker taxa (16 classes) of rhizosphere microbiota varied in a time-sensitive manner, which played potential roles in nutrient cycling and plant health. Nitrospira, Alphaproteobacteria and Acidobacteria were enriched in the rhizosphere of older stands. Key root exudates influenced functional gene abundances in the rhizosphere via direct effects or indirectly through biomarker microbial taxa (e.g., Nitrososphaeria). Overall, root exudates and rhizosphere microbiota are essential for soil function maintenance in R. pseudoacacia plantation restoration.

Nitrogen and light regulate symbiotic nitrogen fixation by a temperate forest tree.

Symbiotic nitrogen fixation (SNF) is a critical mechanism of ecosystem recovery, and in forests of the eastern United States, the most common tree species that supports SNF is black locust (Robinia pseudoacacia L.). Despite its prevalence, black locust's fixation strategy-whether it maintains fixation at a constant rate (obligate fixation) or reduces its fixation rate (facultative fixation)-is unknown. Here, we examined how nitrogen and light control SNF by black locust, by growing seedlings under two nitrogen levels and across four levels of light. Seedlings were harvested after 12 weeks to determine biomass changes, nodule activity, and photosynthetic rates. Black locust seedlings increased biomass growth with increasing light, but only in the absence of nitrogen addition, while seedling root:shoot (biomass) modestly declined with increasing light regardless of nitrogen level. We found that black locust behaved like a facultative fixer, and regulated fixation by excising or maintaining nodules, and by controlling nodule biomass and activity. Specifically, nitrogen addition reduced seedling investment in nodule biomass (g g-1) by 63%, and reduced seedling allocation to nitrogen fixation (µmol C2H4 g-1 h-1) by 66%. In contrast, light affected nitrogen fixation through two indirect pathways. First, light increased plant growth, and hence nitrogen demands, which caused an increase in nitrogen fixation proportional to biomass. Second, light increasd photosynthetic activity, which was positively associated with nodule activity, but only in the absence of nitrogen addition. Our findings for how black locust regulates SNF can improve predictions of ecosystem SNF under the changing environmental conditions.

Degradation of Chemical Components of Thermally Modified Robinia pseudoacacia L. Wood and Its Effect on the Change in Mechanical Properties.

Currently, emphasis is placed on using environmentally friendly materials both from a structural point of view and the application of protective means. For this reason, it is advisable to deal with the thermal modification of wood, which does not require the application of protective substances, to increase its service life. The main reason for the thermal modification of black locust is that although black locust grows abundantly in our country, it has no industrial use. It is mainly used outdoors, where thermal modification could increase its resistance. This article deals with the thermal modification of black locust wood (Robinia pseudoacacia L.) and the impact of this modification on the chemical components of the wood with an overlap in the change in mechanical properties compared to untreated wood. Static (LOP, MOR, and MOE) and dynamic (IBS) bending properties were evaluated as representative mechanical properties. At the same time, the impact of thermal modification on the representation of chemical components of wood (cellulose, lignin, hemicellulose) was also tested. As a result of the heat treatment, the mechanical properties gradually decreased as the temperature increased. The highest decrease in mechanical values found at 210 °C was 43.7% for LOP and 45.1% for MOR. Thermal modification caused a decrease in the content of wood polysaccharides (the decrease in hemicelluloses content was 33.2% and the drop in cellulose was about 29.9% in samples treated at 210 °C), but the relative amount of lignin in the wood subjected to increased temperature was higher due to autocondensation, and mainly because of polysaccharide loss. Based on the correlations between chemical and mechanical changes caused by thermal modification, it is possible to observe the effects of reducing the proportions of chemical components and changes in their characteristic properties (DP, TCI) on the reduction in mechanical properties. The results of this research serve to better understand the behavior of black locust wood during thermal modification, which can primarily be used to increase its application use.

Genome-wide high-resolution mapping of DNA methylation reveals epigenetic variation in the offspring of sexual and asexual propagation in Robinia pseudoacacia.

KEY MESSAGE: We detected the genome-wide pattern of DNA methylation and its association with gene expression in sexual and asexual progenies of mature Robinia pseudoacacia trees. DNA methylation plays an important role in plant reproduction and development. Although some studies on sexual reproduction have been carried out in model plants, little is known about the dynamic changes in DNA methylation and their effect on gene expression in sexual and asexual progeny of woody plants. Here, through whole-genome bisulfite sequencing, we revealed DNA methylation patterns in the sexual and asexual progenies of mature Robinia pseudoacacia to understand the regulation of gene expression by DNA methylation in juvenile seedlings. An average of 53% CG, 34% CHG and 5% CHH contexts was methylated in the leaves of mature and juvenile individuals. The CHH methylation level of asexually propagated seedlings was significantly lower than that of seed-derived seedlings and mature trees. The intergenic regions had the highest methylation level. Analysis of differentially methylated regions (DMRs) showed that most of them were hypermethylated and located in the gene upstream and introns. A total of 24, 108 and 162 differentially expressed genes containing DMRs were identified in root sprouts (RSs), root cuttings (RCs) and seed-derived seedlings (SSs), respectively, and a large proportion of them showed hypermethylation. In addition, DMRs were enriched within GO subcategories including catalytic activity, metabolic process and cellular process. The results reveal widespread DNA methylation changes between mature plants and their progenies through sexual/asexual reproduction, which provides novel insights into DNA methylation reprogramming and the regulation of gene expression in woody plants.

The seeds of invasion: enhanced germination in invasive European populations of black locust (Robinia pseudoacacia L.) compared to native American populations.

Local adaptation and the evolution of phenotypic plasticity may facilitate biological invasions. Both processes can enhance germination and seedling recruitment, which are crucial life-history traits for plants. The rate, timing and speed of germination have recently been documented as playing a major role during the invasion process. Black locust (Robinia pseudoacacia L.) is a North American tree, which has spread widely throughout Europe. A recent study demonstrated that a few populations are the source of European black locust. Thus, invasive populations can be compared to native ones in order to identify genetic-based phenotypic differentiation and the role of phenotypic plasticity can thereby be assessed. A quantitative genetics experiment was performed to evaluate 13 juvenile traits of both native and invasive black locust populations (3000 seeds, 20 populations) subjected to three different thermal treatments (18 °C, 22 °C and 31 °C). The results revealed European populations to have a higher germination rate than the native American populations (88% versus 60%), and even when genetic distance between populations was considered. Moreover, this trait showed lower plasticity to temperature in the invasive range than in the native one. Conversely, other studied traits showed high plasticity to temperature, but they responded in a similar way to temperature increase: the warmer the temperature, the higher the growth rate or germination traits values. The demonstrated genetic differentiation between native and invasive populations testifies to a shift between ranges for the maximum germination percentage. This pattern could be due to human-mediated introduction of black locust.

Rapid nitrogen fixation contributes to a similar growth and photosynthetic rate of Robinia pseudoacacia supplied with different levels of nitrogen.

Robinia pseudoacacia L. is a legume species that is widely used in afforestation, which has high N2 fixation capacity and rapid growth rate. Both nitrogen (N) supply and phenology affect plant growth, photosynthesis and leaf senescence. The aim of this study was to determine how N supply affects N2 fixation, leaf photosynthesis and senescence of R. pseudoacacia at different phenological stages. Seedlings of R. pseudoacacia were supplied with different levels of 15N-labelled NH4NO3 solution, with seedlings of Sophora japonica Linn. as reference plants to calculate the percentage of N derived from the atmospheric N2 (%Ndfa). Compared with plants supplied with a high N level, those with a low N supply had a higher %Ndfa at an early developmental stage. Nitrogen fixation compensated the effect of a low N supply on plant growth in R. pseudoacacia. A high N supply decreased biomass allocation to lateral roots and nodules, and increased the relative growth rate of plant height as well as specific leaf area. The eighth mature compound leaf of R. pseudoacacia tended to have a higher net photosynthetic rate than the fourth leaf, and the leaves still maintained a moderate photosynthetic rate in early autumn. Plants tended to allocate more biomass to leaves at an early developmental stage and to stems and roots at a later developmental stage (3 months old). The N level did not affect leaf photosynthesis at different phenological stages, primarily due to (i) a high %Ndfa under low N supply at early growing stage, and a similar high %Ndfa under all N supplies at a late growing stage, and (ii) the delayed greening phenotype of expanding leaves to save nutrients for mature leaves.

Quantitative and Qualitative Identification of Bioactive Compounds in Edible Flowers of Black and Bristly Locust and Their Antioxidant Activity.

Black and bristly locust flowers are an excellent source of polyphenols, including flavonols, phenolic acids, and anthocyanins. In the present literature, there is a lack of studies showing the quantity and quality of phenolic compounds from different locust flowers. There are a few studies on the status of polyphenols in black locust flowers and their products but not bristly locusts. The aims of this study were to analyze and compare the concentrations of bioactive compounds from Robinia pseudoacacia and Robinia hispida flowers over two years. These two species of plants from six independent locations (parks and green areas) located in Warsaw were assessed in this study. The dry matter and polyphenol contents of the flowers were determined. Black locust flower samples contained significantly more myricetin and luteolin. Only bristly locust flowers contained anthocyanins. Five individual anthocyanins were identified in the pink-colored bristly locust flowers. Pelargonidin-3-O-glucoside and cyanidin-3-O-glucoside were the predominant forms in the pool of total anthocyanins.

A consecutive 4-year elevated air temperature shaped soil bacterial community structure and metabolic functional groups in the rhizosphere of black locust seedlings exposed to lead pollution.

Global warming may influence the bioavailability and mobility of heavy metals by stimulating or inhibiting plant growth, thereby influencing rhizosphere soil chemistry and microbial characteristics. Black locust has been widely planted in China as a promising species for afforestation programs, farmland shelterbelt projects, and soil restoration in mined areas because of its rapid growth and adaptability to environmental stressors. Here, we examined soil bacterial community structure and predicted bacterial metabolic function in the rhizosphere of black locust exposed to elevated temperature (+1.99 °C) and Pb for 4 years. Elevated temperature significantly (p < 0.05) reduced total carbon (TC), total nitrogen (TN), and total sulfur (TS) contents in above-ground parts but increased TC and TN contents in roots and seedling height under Pb exposure. Elevated temperature significantly (p < 0.05) increased Pb availability and raised pH, TC, TN, TS and water-soluble organic carbon (WSOC) contents, and the C:H ratio in rhizosphere soils under Pb exposure. The interactive effects between Pb and temperature on pH, TC, TH, TS, WSOC, and the C:H ratio were significant (p < 0.05). Elevated temperature significantly (p < 0.05) reduced the diversity and the richness of bacterial community, altered genus-level bacterial community composition, and improved (p < 0.05) the relative abundances of some bacteria involving in terpenoids and polyketides and xenobiotics biodegradation metabolism under Pb exposure. Canonical correspondence analysis indicated that pH, WSOC, C:N ratio, and soluble Pb were significant (p < 0.05) factors on the relative abundance of bacterial genera, such as Ochrobactrum and Sphingomnas. Overall, long-term elevated temperature resulted in changes in rhizosphere soil characteristics and Pb availability, thus affecting the bacterial community structure and metabolic functional groups. The conclusion helps us understand the response mechanism of soil bacteria in the rhizosphere to heavy metals under global warming scenarios.

Robinia pseudoacacia-dominated vegetation types of Southern Europe: Species composition, history, distribution and management.

Knowledge of the species composition of invaded vegetation helps to evaluate an ecological impact of aliens and design an optimal management strategy. We link a new vegetation analysis of a large dataset to the invasion history, ecology and management of Robinia pseudoacacia stands across Southern Europe and provide a map illustrating Robinia distribution. Finally, we compare detected relationships with Central Europe. We show that regional differences in Robinia invasion, distribution, habitats and management are driven both by local natural conditions (climate and soil properties, low competitive ability with native trees) and socioeconomic factors (traditional land-use). Based on the classification of 467 phytosociological relevés we distinguished five broad vegetation types reflecting an oceanity-continentality gradient. The stands were heterogeneous and included 824 taxa, with only 5.8% occurring in more than 10% of samples, representing mainly hemerobic generalists of mesophilous, nutrient-rich and semi-shady habitats. The most common were dry ruderal stands invading human-made habitats. Among native communities, disturbed mesic and alluvial forests were often invaded throughout the area, while dry forests and scrub dominated in Balkan countries. Continuous, long-term and large-scale cultivation represent a crucial factor driving Robinia invasions in natural habitats. Its invasion should be mitigated by suitable management taking into account adjacent habitats and changing cultivation practices to select for native species. Robinia invasion has a comparable pattern in Central and Southern Europe, but there is a substantial difference in management and utilization causing heterogeneity of many South-European stands.

The Rpf84 gene, encoding a ribosomal large subunit protein, RPL22, regulates symbiotic nodulation in Robinia pseudoacacia.

MAIN CONCLUSION: A homologue of the ribosomal protein L22e, Rpf84, regulates root nodule symbiosis by mediating the infection process of rhizobia and preventing bacteroids from degradation in Robinia pseudoacacia. Ribosomal proteins (RPs) are known to have extraribosomal functions, including developmental regulation and stress responses; however, the effects of RPs on symbiotic nodulation of legumes are still unclear. Ribosomal protein 22 of the large 60S subunit (RPL22), a non-typical RP that is only found in eukaryotes, has been shown to function as a tumour suppressor in animals. Here, a homologue of RPL22, Rpf84, was identified from the leguminous tree R. pseudoacacia. Subcellular localization assays showed that Rpf84 was expressed in the cytoplasm and nucleus. Knockdown of Rpf84 by RNA interference (RNAi) technology impaired the infection process and nodule development. Compared with the control, root and stem length, dry weight and nodule number per plant were drastically decreased in Rpf84-RNAi plants. The numbers of root hair curlings, infection threads and nodule primordia were also significantly reduced. Ultrastructure analyses showed that Rpf84-RNAi nodules contained fewer infected cells with fewer bacteria. In particular, remarkable deformation of bacteroids and fusion of multiple symbiosomes occurred in infected cells. By contrast, overexpression of Rpf84 promoted nodulation, and the overexpression nodules maintained a larger infection/differentiation region and had more infected cells filled with bacteroids than the control at 45 days post inoculation, suggesting a retarded ageing process in nodules. These results indicate for the first time that RP regulates the symbiotic nodulation of legumes and that RPL22 may function in initiating the invasion of rhizobia and preventing bacteroids from degradation in R. pseudoacacia.

Comparative analysis of chloroplast genomes of five Robinia species: Genome comparative and evolution analysis.

The black locust (Robinia pseudoacacia) is widely distributed, and has strong drought resistance and salt tolerance. These characteristics make it the best type of tree for landscaping and resource conservation in China. In this study, the chloroplast genomes of five black locusts were identified de novo and the evolutionary relationship among these black locusts and their taxonomic statuses in leguminous plants were determined. The chloroplast structures of the five black locusts were typical and had four parts, including two single copy regions (large and small single copy sections) and a pair of inverted repeats (IRs). Genome sizes were between 155,364 bp and 155,655 bp; the genome of R. pseudoacacia var. decaisneana was the smallest, while that of R. pseudoacacia var. tortuosa was the largest. The genomes contained 124-130 protein-coding genes; R. pseudoacacia var. tortuosa had the fewest, while R. hispida and R. pseudoacacia var. decaisneana had the most. In this study, eight to ten genes from chloroplast genomes contained introns. Nine genes from the chloroplast genomes of R. pseudoacacia and R. pseudoacacia f. unifolia contained introns that had lost the trnL-CAA gene via evolution, while eight chloroplast genes of R. pseudoacacia var. tortuosa contained introns that had lost the trnL-CAA and psaA genes. Among them, the rpoC1 gene had the longest introns at 2828 bp, and rps12+ had the smallest introns at only 533 bp. There were various amplification phenomena in the IR region among the five black locusts. Most of the protein-coding genes of the five black locusts had a high degree of codon preference. To determine the phylogenetic positions of the five black locusts, we conducted a systematic evolutionary analysis using common protein-coding genes in chloroplast sequences from 34 species of leguminous plants and 12 other species. The results showed that the relationship between Robinia and Acacia ligulata was the most distant among those of the leguminous plants, and the relationship between Robinia and Lotus japonicus was the closest. The chloroplast protein-coding genes in different black locusts were relatively conservative by evolutionary selection pressure analysis standards. These results are important for our understanding of their photosynthetic mechanisms and evolution, and the transgenic engineering of their chloroplasts.

From Robinia pseudoacacia L. nectar to Acacia monofloral honey: biochemical changes and variation of biological properties.

BACKGROUND: Robinia pseudoacacia L. nectar and its derivative monofloral honey were systematically compared in this study, to understand how much the starting solution reflected the final product, after re-elaboration by Apis mellifera ligustica Spinola. RESULTS: Subjected to dehydration in the hive, nectar changed in its water and sugar content when transformed into honey, as physicochemical and gas chromatographic-mass spectrometric analyses revealed. Spectrophotometric measurements and characterization by high-performance liquid chromatography-diode array detection of 18 plant molecules demonstrated honey to be richer than nectar in secondary metabolites. For the first time, the hypothesis of the existence of a nectar redox cycle in R. pseudoacacia was reported, as previously described for Nicotiana sp., based on 1D-protein profiles, western blot analysis and detection of H2 O2 and ascorbate. The bioactivity of both matrices was also investigated. Antiradical in vitro tests showed that Acacia honey was more antioxidant than nectar, which was even able to induce oxidative stress directly in a eukaryotic cell system. Antimicrobial assays demonstrated that nectar was bacteriostatic, due to H2 O2 activity, whereas honey was even bactericidal. CONCLUSION: All these data support the ecological role of nectar and honey in nature: protection of the gynoecium from pathogens and preservation from degradative processes, respectively. © 2018 Society of Chemical Industry.

How ecosystems change following invasion by Robinia pseudoacacia: Insights from soil chemical properties and soil microbial, nematode, microarthropod and plant communities.

Biological invasions are a global threat to biodiversity. Since the spread of invasive alien plants may have many impacts, an integrated approach, assessing effects across various ecosystem components, is needed for a correct understanding of the invasion process and its consequences. The nitrogen-fixing tree Robinia pseudoacacia (black locust) is a major invasive species worldwide and is used in forestry production. While its effects on plant communities and soils are well known, there have been few studies on soil fauna and microbes. We investigated the impacts of the tree on several ecosystem components, using a multi-trophic approach to combine evidence of soil chemical properties and soil microbial, nematode, microarthropod and plant communities. We sampled soil and vegetation in managed forests, comparing those dominated by black locust with native deciduous oak stands. We found qualitative and quantitative changes in all components analysed, such as the well-known soil nitrification and acidification in stands invaded by black locust. Bacterial richness was the only component favoured by the invasion. On the contrary, abundance and richness of microarthropods, richness of nematodes, and richness and diversity of plant communities decreased significantly in invaded stands. The invasion process caused a compositional shift in all studied biotic communities and in relationships between the different ecosystem components. We obtained clear insights into the effects of invasion of managed native forests by black locust. Our data confirms that the alien species transforms several ecosystem components, modifying the plant-soil community and affecting biodiversity at different levels. Correct management of this aggressive invader in temperate forests is urgently required.

Disturbance induced dynamics of a tritrophic novel ecosystem.

Novel ecosystems formed by invasive plants provide a good opportunity to get insight into early dynamics and pattern formation of these ecosystems. The invasive black locust as host plant, Bruchophagus robiniae as host-specific seed predator and its parasitoids were the components of the studied tritrophic system. To investigate disturbance-driven dynamics of this system we created seed-vacated host plant patches in a field experiment. We removed all pods from selected patches of black locust resulting in an induced local extinction of seed predators and their parasitoids. We hypothesized that disturbance enhances top-down control by parasitoids; this enhanced top-down control decreases seed predation, facilitating the host plant's spread. We found that disturbance modified only parasitism after controlling with year effect: in vacated patches median parasitism was higher than in control patches. Seed predation exceeded its initial level in vacated patches in the third year after the disturbance, but in the fourth year it dropped again presumably due to the strong top-down control. Our findings also suggested that the seed predator was also affected by the bottom-up control of its host plant's density. We found that in the studied new ecosystem the top-down control was strengthened by the disturbance. Since the host plant of the tritrophic system is an invasive species, partial habitat disturbance of such species may increase the severity of parasitoid top-down control, which may reduce seed predation by the herbivores.

Comparative mitochondrial proteomic, physiological, biochemical and ultrastructural profiling reveal factors underpinning salt tolerance in tetraploid black locust (Robinia pseudoacacia L.).

BACKGROUND: Polyploidy is an important phenomenon in plants because of its roles in agricultural and forestry production as well as in plant tolerance to environmental stresses. Tetraploid black locust (Robinia pseudoacacia L.) is a polyploid plant and a pioneer tree species due to its wide ranging adaptability to adverse environments. To evaluate the ploidy-dependent differences in leaf mitochondria between diploid and tetraploid black locust under salinity stress, we conducted comparative proteomic, physiological, biochemical and ultrastructural profiling of mitochondria from leaves. RESULTS: Mitochondrial proteomic analysis was performed with 2-DE and MALDI-TOF-MS, and the ultrastructure of leaf mitochondria was observed by transmission electron microscopy. According to 2-DE analysis, 66 proteins that responded to salinity stress significantly were identified from diploid and/or tetraploid plants and classified into 9 functional categories. Assays of physiological characters indicated that tetraploids were more tolerant to salinity stress than diploids. The mitochondrial ultrastructure of diploids was damaged more severely under salinity stress than that of tetraploids. CONCLUSIONS: Tetraploid black locust possessed more tolerance of, and ability to acclimate to, salinity stress than diploids, which may be attributable to the ability to maintain mitochondrial structure and to trigger different expression patterns of mitochondrial proteins during salinity stress.

Tolerance or avoidance: drought frequency determines the response of an N2 -fixing tree.

Climate change is increasing drought frequency, which may affect symbiotic N2 fixation (SNF), a process that facilitates ecosystem recovery from disturbance. Here, we assessed the effect of drought frequency on the ecophysiology and SNF rate of a common N2 -fixing tree in eastern US forests. We grew Robinia pseudoacacia seedlings under the same mean soil moisture, but with different drought frequency caused by wet-dry cycles of varying periodicity. We found no effect of drought frequency on final biomass or mean SNF rate. However, seedlings responded differently to wet and dry phases depending on drought frequency. Under low-frequency droughts, plants fixed carbon (C) and nitrogen (N) at similar rates during wet and dry phases. Conversely, under high-frequency droughts, plants fixed C and N at low rates during dry phases and at high rates during wet phases. Our findings suggest that R. pseudoacacia growth is resistant to increased drought frequency because it employs two strategies - drought tolerance or drought avoidance, followed by compensation. SNF may play a role in both by supplying N to leaf tissues for acclimation and by facilitating compensatory growth following drought. Our findings point to SNF as a mechanism for plants and ecosystems to cope with drought.