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Lipoxygenase (LOX, EC1.13.11.12) catalyzes the oxygenation of polyunsaturated fatty acids and further generates oxylipins via enzymatic or non-enzymatic pathways, thus participating in the regulation of growth and development, stress response, and other processes. The objective of this research was to characterize the function and evaluate the mechanism of lipoxygenase gene AfLOX4 ’s role in enhancing drought tolerance. Transcriptomic data were generated and analyzed for the LOX gene, under different concentrations of drought (polyethylene glycol-6000) (PEG6000) and abscisic acid (ABA) stress treatment, to determine their effects on the expression of the AfLOX4 gene and its role in plant physiology. The localization of the AfLOX4 protein was detected through protoplast transformation. The expression of AfLOX4 in Amorpha fruticosa tissues was determined by quantitative real-time PCR (qRT-PCR). Six LOX genes of A. fruticosa that showed an upregulated pattern under different concentrations of drought (PEG6000) stress were found via transcriptomic analysis and were named AfLOX1  ~ 6. The AfLOX4 protein was localized in the cytoplasm. AfLOX4 was expressed in the stems, roots, flowers, and leaves of Amorpha fruticosa L., and its expression level was the highest in leaves. The expression of AfLOX4 increased with the prolongation of stress time under different stresses. After ABA treatment, the germination rate of tobacco lines overexpressing AfLOX4 was significantly increased, while under NaHCO 3 stress, the tolerance of tobacco plants overexpressing AfLOX4 was higher than that of wild-type tobacco. The tobacco with overexpression of the AfLOX4 gene detected by the natural drought method had a higher survival rate in the drought stress recovery experiment, and improved the drought tolerance of tobacco. This study reveals the function of the AfLOX4 gene in resisting drought stress and lays a solid theoretical foundation for the genetic improvement of Amorpha fruticosa L. plant varieties, as well as improvement of resource utilization.

The studied introduction population of the alien North American species Amorpha fruticosa L. was formed in the Main Botanical Garden of the Russian Academy of Sciences (Moscow) 80 years ago from seeds of different geographical origin. Currently, this population consists of individuals of the second and third generations, which arose as a result of the spontaneous pollination of parental plants. It is the northernmost site of A. fruticosa growth in the secondary distribution range; in southern regions, it usually behaves like an aggressive invasive species and invades natural plant communities. A. fruticosa is known to contain a number of bioactive compounds with valuable pharmacological properties, and the aim of this study was to evaluate the biochemical composition of leaves and fruits at the northern limit of the species’ growth, since stress conditions promote active accumulation of secondary metabolites. The data on the composition of secondary metabolites, on the mineral composition, phenolic compounds, and flavonoids in the leaves and fruits of A. fruticosa, and on the amount and composition of essential oil in the extract from fruits are presented. High levels of adonitol, which is used as a sweetener in products for diabetic patients, have been reported in the fruits of A. fruticosa. α-Copaene, α-pinene, δ-cadinene, α-muurolene, and β- and α-caryophyllene predominate in the essential oil of the fruit, providing its antimicrobial activity. The phytochemical analysis of A. fruticosa from the secondary distribution range confirms the potential of this species as a valuable resource for the pharmacopoeia industry.

L. is a deciduous shrub that is native to North America and has been introduced to China as an ornamental plant. In order to clarify the drought resistance characteristics of L. and excavate the related genes involved in drought resistance regulation pathway, the mechanism of drought resistance stress of L. was revealed by the changes of transcriptome of L. under drought stress.Through the changes of the transcriptome of L. under drought stress, the mechanism of anti-stress of L. could be revealed. Different concentrations of polyethylene glycol-6000 (PEG-6000) was used to simulate drought stress, and transcriptomic analysis was used to reveal the changes of gene expression patterns in L. seedlings. Results showed that L. seedlings were seriously affected by PEG-6000. As for the differently expressed genes (DEGs), most of them were up-regulated. The additional Go and KEGG analysis results showed that DEGs were functionally enriched in cell wall, signal transduction and hormonal regulation related pathways. DEGs like AfSOD, AfHSP, AfTGA, AfbZIP and AfGRX play roles in response to drought stress. In conclusion, L. seedlings were sensitive to drought, which was different from L. tree, and the genes functions in drought stress responses via ABA-independent pathways. The up-regulation of Salicylic acid signal related DEGs (AfTGA and AfPR-1) indicated that Salicylic acid play a key role in response to drought stress in L.

To combat the decline in North American grasslands and prairies, innovative strategies to establish new native grass and forb plantings must be considered. Integrated vegetation management entails the use of many practices to cultivate desirable vegetation along roadsides, including mowing, applying herbicides, burning, and replanting. Currently, only a limited selection of postemergence herbicides are available to improve native plant establishment along roadsides. A greenhouse herbicide screen that included four postemergence herbicides registered for use on Conservation Reserve Program (CRP) acres and rights-of-way was conducted to test their safety for use on four native grasses (big bluestem, buffalograss, sideoats grama, and switchgrass) and seven forb species (ashy sunflower, black-eyed Susan, butterfly milkweed, desert false indigo, Illinois bundleflower, Mexican hat plant, and purple coneflower). Clopyralid (689 g ae ha–1), metsulfuron (4.18 g ai ha–1), and quinclorac (418 g ai ha–1) applied at labeled rates caused no injury to the native grass species or butterfly milkweed. However, florpyrauxifen-benzyl (38.4 g ai ha–1) caused significant injury to buffalograss and switchgrass. None of the herbicides tested were universally safe to use on all forb species evaluated in this trial, with each herbicide causing unacceptable injury (≥25%) to one or more forb species. None of the herbicides studied here would be completely safe for use on mixed stands of native grasses and native forbs at the seedling growth stage, indicating that prairie establishment must use alternative chemistries, plant mixes with fewer species, or avoid postemergence applications shortly after emergence of native forbs. Nomenclature: Clopyralid; florpyrauxifen-benzyl; metsulfuron; quinclorac; ashy sunflower, Helianthus mollis Lam.; big bluestem, Andropogon gerardii Vitman; black-eyed Susan, Rudbeckia hirta L.; buffalograss, Buchloe dactyloides [Nutt.] J.T. Columbus; butterfly milkweed, Asclepias tuberosa L.; desert false indigo, Amorpha fruticosa L.; Illinois bundleflower, Desmanthus illinoensis (Michx.) MacMill. ex B.L. Rob. & Fernald; Mexican hat plant, Ratibida columnifera (Nutt.) Wooton & Standl.; purple coneflower, Echinacea purpurea (L.) Moench; sideoats grama, Bouteloua curtipendula [Michx.] Torr.; switchgrass, Panicum virgatum L.

A first assessment of genetic diversity and expansion of the invasive species Amorpha fruticosa L. was estimated in the 91E0* (Annex I Dir. 92/43/EEC) priority habitat of Nestos. A. fruticosa is a perennial deciduous shrub belonging to the family Fabaceae and is native to the south-western part of North America, which recently was introduced to Europe as an ornamental plant. Its expansion through the cross-border river of Nestos has significant negative effects on the 91E0* habitat (a habitat with Alnus glutinosa, Populus alba, and Ulmus minor). Herein, we studied the genetic diversity with amplified fragment length polymorphisms fragments of eastern and western populations in order to see if there is one genetic pool for the species. Furthermore, we estimated the regeneration index (IR) in order to see the potential expansion of this invasive species. Results showed that eastern and western populations have almost equal heterozygosity (He) and with a slightly higher value for the western population, which was in agreement with the results of the IR. This is an innovative study that discusses the implications of A. fruticosa of 91E0* habitat in Nestos and proposes management measures to eliminate its expansion and improve the resilience of the habitat.

In this paper, the ultrasonic-assisted extraction process of flavonoid compounds from leaves of Amorpha fruticosa is optimized. In single factor experiments, solid/liquid ratios, ultrasonic power, ethanol concentrations and extraction cycles were experimental factors. Box–Behnken central composite design and RSM analyzed the effects of the four factors on the yield of total flavonoids. The optimal extraction parameters were solid/liquid ratio 1:50 g/mL, ultrasonic power 316 W, ethanol concentration 50%, 4 extraction cycles. In the optimized condition, the estimated value of the regression model was 66.6456 mg/g while the measured value was 66.4329 mg/g.

Amorpha fruticosa L. (false indigo bush) is an invasive shrub species from Fabaceae family originating from North America. Nowadays this species creates great problems because it is invading floodplains along the rivers from Europe, even the Danube River floodplain reaching to the Danube Delta. The floodplain area of the river Mureş is highly affected by this invasion, this species occupying great surfaces of grassland and arable land. The researches were developed on the lowland area alongside the river Mureş (Arad County, western Romania) in the perimeter of the locality Pecica, respectively belonging to Mures Floodplain Natural Park. The purpose of the work was to characterize different relationships between the features of the grassland vegetation with soil features in three different situations, respectively permanent grassland, grassland totally invaded with A. fruticosa, and transitional grassland partially invaded with A. fruticosa. The soil samples were collected from 0 - 25 cm depth. The researches were investigated the influence of the false indigo bush invaded grassland on the soil features as pH, nitrogen index, total nitrogen, humus content, phosphorus, potassium and carbonates. The considered grassland vegetation features related with the soil features were: floristic composition (main functional groups of plants, some biodiversity indexes and pastoral value. The obtained results showed the existence of complex relationships among the analysed vegetation and soil features.

Amorpha fruticosa L. is a Chinese folk medicine and rich in polyphenols. Fifteen known compounds were isolated and identified from the leaves of A. fruticosa L. They are tephrosin (1), 6a,12a-dehydrodeguelin (2), vitexin (3), afrormosin-7-O-β-d-glucopyranoside (4), 2″-O-α-l-rhamnopyranosyl isovitexin (5), rutin (6), chrysoeriol (7), 7-O-methylluteolin (8), trans-p-coumaric acid (9), 2-benzyl-4,6-dihydroxybenzoic acid-4-O-β-d-glucopyranoside (10), formononetin (11), quercetin (12), apigenin (13), β-sitosterol (14), and β-daucosterol (15). Compounds 3, 4, 5, and 7–9 were isolated from A. fruticosa L. for the first time. Cytotoxicity of individual compounds 3–10 and 90% ethanol extract against human cancer cell lines HCT116 and HepG2 were reported. The results suggested that compounds 7 and 8, and the crude extract exhibited inhibitory effects on human cancer cell line HCT116, at concentrations of 100 μg/mL, 5 μg/mL, and 25 μg/mL at <60% of cell viability rate, respectively. In addition, a valid high-performance liquid chromatography diode array detector method was established to quantitatively analyze compounds 1–12 in the leaves of A. fruticosa L., which was harvested at three different stages of maturity from May 20 to August 10, 2014. The results demonstrated that contents were greatly influenced by the maturity. Total amounts of the analytical constituents gradually increased from May 20 to August 10, with the values ranging from 10.86 mg/g to 18.84 mg/g, whereas bioactive compounds 7 and 8 presented the opposite variation trend. The results of this study may provide data for further study and comprehensive utilization of A. fruticosa L. resource. [Display omitted] •Fifteen compounds were isolated and identified.•Six compounds were isolated from A. fruticosa L. for the first time.•Cytotoxicity of eight compounds and 90% ethanol extract against human cancer cell lines HCT116 and HepG2 was reported.•A valid HPLC-DAD method was established to quantitatively analyze 12 compounds in the leaves of A. fruticosa L.

The supercritical CO 2 fluid extraction process was used to obtain seed oils. The compositional analyses of the seed oils of Amorpha fruticosa L., Caragana microphylla Lam and Elaeagnus angustifolia L. were performed by GC–MS, from which 18–22 fatty acid compositions were identified, and their main components were linoleic acid (49.10–66.39 %) and oleic acid (11.95–41.10 %). The seed oils were rich in unsaturated fatty acids, which accounted for 79.75–91.19 %. The in vitro anti-oxidant activities of the seed oils were measured by the DPPH, ABTS and phosphomolybdenum complex assays. The IC 50 s of the seed oils were 6.5110–12.6599 mg/mL tested by DPPH assay, and the IC 50 s were in the range of 0.6872–1.9310 mg/mL tested by ABTS assay. The anti-oxidant activities of seed oils decreased in the order of A. fruticosa  >  C. microphylla  >  E. angustifolia (A) >  E. angustifolia (D). In vitro antimicrobial activities of seed oils against five bacteria were tested by microdilution method. The MIC values of the oils against Bacillus subtilis , E. coli , S. aureus , P. solanacearum and B. thuringiensis were in the range of 26–36, 30–38, 58, 56–58 and 56–58 mg/mL, respectively, while the MBC values were in the range of 30–40, 34–42, 58, 56–58 and 58 mg/mL, respectively. In vitro antifungal properties of the seed oils against four plant pathogenic fungi were tested by the mycelial growth rate method. The EC 50 s of seed oils against V. mali , C. gloeosporioides , F. graminearum and B. cinerea were in the range of 17.51–19.25, 15.58–19.12, 8.00–17.12 and 18.76–19.14 mg/mL, respectively. The seed oils showed moderate anti-oxidant and antimicrobial activities.

The present study focuses on the chemical and biological analysis of the ethanolic 70% v/v extracts obtained from Amorpha fruticosa leaves, branches and fruits. The total phenolic content determined by Folin-Ciocalteu method was the highest in leaves extract (159.5 mg/g). The HPLC-MS analysis indicated the presence of ferulic acid, luteolin and rutoside in all three extracts. The antioxidant activity was tested by the DPPH method and the results indicated a good scavenging activity (EC50 = 18.8 ± 0.2 µg/mL) for the fruit extract, followed by leaf extract (EC50 = 38.03 ± 0.75 µg/mL) and branch extract (EC50 = 221.16 ± 1.7 µg/mL). The antimicrobial activity evaluated by agar disc diffusion method indicated the inhibition of growth for Staphylococcus aureus and Sarcina lutea for all samples.