Chebulic acid derivatives from Balakata baccata and their antineuroinflammatory and antioxidant activities.

Sixteen chebulic acid derivatives, including nine new (1-9) and seven known (10-16) ones, were isolated from an ethanol extract of the branches and leaves of Balakata baccata. The structures of the new compounds were elucidated by their UV, IR, HRESIMS, NMR, electronic circular dichroism (ECD) and single-crystal X-ray diffraction data. The effects of all the isolates on antineuroinflammatory and antioxidant activities were evaluated. Compared with the positive control minocycline (IC50 = 1.21 ± 0.71 µM), compounds 1-16 with IC50 values being greater than 50 µM, displayed almost no effects on the inhibition of NO production in LPS-induced BV-2 microglial cells, however, the results of antioxidant activity for compounds 1-16 showed significant DPPH-radical scavenging abilities with EC50 value ranging from 3.98 to 14.24 µM, while the EC50 value of positive control vitamin C was 14.31 µM. At last, the results of PCR (qRT-PCR) analysis showed that compound 1 could enhance the expression of antioxidases (HO-1, GCLC, and NQO1) at the mRNA levels.

Two new ent-atisane diterpenes from Sapium sebiferum.

Two new ent-atisane diterpenes, sebiferumnins K and L (1 and 2), were isolated from the branches of Sapium sebiferum, together with four known compounds, ent-3-oxoatis-16α, 17-acetonide (3), ent-l6α, 17-dihydroxyatisan-3-one (4), ent-atisane-3ß, 16α, 17-triol (5), ent-3α-hydroxyatis-16α,17-acetonide (6). The structures of these two ent-atisane diterpenes were elucidated by extensive analyses of the spectroscopic data. [Formula: see text].

Overexpression of the Stress-Inducible SsMAX2 Promotes Drought and Salt Resistance via the Regulation of Redox Homeostasis in Arabidopsis.

Recent studies have demonstrated that strigolactones (SLs) also participate in the regulation of stress adaptation; however, the regulatory mechanism remains elusive. In this study, the homolog of More Axillary Branches 2, which encodes a key component in SL signaling, in the perennial oil plant Sapium sebiferum was identified and functionally characterized in Arabidopsis. The results showed that the expression of SsMAX2 in S. sebiferum seedlings was stress-responsive, and SsMAX2 overexpression (OE) in Arabidopsis significantly promoted resistance to drought, osmotic, and salt stresses. Moreover, SsMAX2 OE lines exhibited decreased chlorophyll degradation, increased soluble sugar and proline accumulation, and lower water loss ratio in response to the stresses. Importantly, anthocyanin biosynthesis and the activities of several antioxidant enzymes, such as superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX), were enhanced in the SsMAX2 OE lines, which further led to a significant reduction in hydrogen peroxide levels. Additionally, the SsMAX2 OE lines exhibited higher expression level of several abscisic acid (ABA) biosynthesis genes, suggesting potential interactions between SL and ABA in the regulation of stress adaptation. Overall, we provide physiological and biochemical evidence demonstrating the pivotal role of SsMAX2 in the regulation of osmotic, drought, and salt stress resistance and show that MAX2 can be a genetic target to improve stress tolerance.

Accumulation of uranium and heavy metals in the soil-plant system in Xiazhuang uranium ore field, Guangdong Province, China.

Plants that have grown for many years in the special environmental conditions prevailing in mining areas are naturally screened and show strong capacity to adapt to their environment. The present study investigated the enrichment characteristics of U and other heavy metals (As, Cu, Pb, Mn, Mo, Zn, Cd, Co, and Ni) in the soil-plant system in Xiazhuang uranium mine. Four dominant plants (Castanopsis carlesii, Rhus chinensis, Liriodendron chinense, and Sapium discolor) and soil samples were collected from the mined areas, unmined areas, and background areas away from the ore field. U, As, Cu, Pb, Mn, Mo, Zn, Cd, Co, and Ni concentrations were analyzed by ICP-MS. The results demonstrate that (1) The highest concentrations of U (4.1-206.9 mg/kg) and Pb (43.3-126.0 mg/kg) with the geoaccumulation index (Igeo) greater than 1 show that they are the main soil pollutants in the research area. (2) The biological accumulation coefficient (LBAC) values for Cd, Mn, and Cu are greater than zero in S. discolor, L. chinense, and C. carlesii and these three plants indicate that they can be used for remediation of the soil in the ore field. (3) R. chinensis inhibits the accumulation of heavy metals and shows sensitive pigment responses to the accumulation of U in the leaves. L. chinense has the strongest enrichment effect on heavy metals but exhibits weak biochemical responses under U stress. C. carlesii demonstrates strong adaptation to U and can maintain healthy pigment characteristics in case of high U enrichment. (4) S. discolor, L. chinense, C. carlesii and R. chinensis have strong tolerance to U toxicity and different biochemical responses.

[Study on chemical constituents of stems and leaves of Sapium discolor].

Seventeen compounds were isolated from the 95% ethanol extract of the stems and leaves of Sapium discolor by using various chromatographic techniques,including silica gel,Sephadex LH-20,MCI,ODS,and semi-preparative HPLC. Their structures were elucidated as sapiumin F( 1),kadsulignan C( 2),ciwujiatone( 3),ethylbrevifolin carboxylate( 4),7-hydroxy-8-methoxycoumarin( 5),fraxetin( 6),fraxidin( 7),isofraxidin( 8),6,7,8-trimethoxycoumarin( 9),5,6,7,8-tetramethoxycoumarin( 10),8-hydroxy-5,6,7-trimethoxycoumarin( 11),3,3'-di-O-methylellagic acid( 12),3,3',4'-tri-O-methylellagic acid( 13),3'-methoxyellagic acid 4'-O-α-rhamnopyranoside( 14),4,5-didehydro-chebulic acid triethyl ester( 15),ent-kaurane-3-oxo-16α,17-diol( 16),and abscisic acid( 17) by spectroscopic data. Compound 1 is a new compound. Except for compounds 4,11,and 13,the remaining compounds were isolated from this plant for the first time. All the isolates were evaluated for their in vitro antineuroinflammatory activities,and the results showed that compounds 6 and 15 significantly inhibited nitric oxide production in lipopolysaccharide-induced BV-2 microglial cells,with IC50 values of 6. 06 and 6. 05 µmol·L-1,respectively.

Comparative transcriptome analysis reveals the regulatory networks of cytokinin in promoting the floral feminization in the oil plant Sapium sebiferum.

BACKGROUND: Sapium sebiferum, whose seeds contain high level of fatty acids, has been considered as one of the most important oil plants. However, the high male to female flower ratio limited the seed yield improvement and its industrial potentials. Thus, the study of the sex determination in S. sebiferum is of significant importance in increasing the seed yield. RESULTS: In this study, we demonstrated that in S. sebiferum, cytokinin (CK) had strong feminization effects on the floral development. Exogenous application with 6-benzylaminopurine (6-BA) or thidiazuron (TDZ) significantly induced the development of female flowers and increased the fruit number. Interestingly, the feminization effects of cytokinin were also detected on the androecious genotype of S. sebiferum which only produce male flowers. To further investigate the mechanism underlying the role of cytokinin in the flower development and sex differentiation, we performed the comparative transcriptome analysis of the floral buds of the androecious plants subjected to 6-BA. The results showed that there were separately 129, 352 and 642 genes differentially expressed at 6 h, 12 h and 24 h after 6-BA treatment. Functional analysis of the differentially expressed genes (DEGs) showed that many genes are related to the hormonal biosynthesis and signaling, nutrients translocation and cell cycle. Moreover, there were twenty one flowering-related genes identified to be differentially regulated by 6-BA treatment. Specifically, the gynoecium development-related genes SPATULA (SPT), KANADI 2 (KAN2), JAGGED (JAG) and Cytochrome P450 78A9 (CYP79A9) were significantly up-regulated, whereas the expression of PISTILLATA (PI), TATA Box Associated Factor II 59 (TAFII59) and MYB Domain Protein 108 (MYB108) that were important for male organ development was down-regulated in response to 6-BA treatment, demonstrating that cytokinin could directly target the floral organ identity genes to regulate the flower sex. CONCLUSIONS: Our work demonstrated that cytokinin is a potential regulator in female flower development in S. sebiferum. The transcriptome analysis of the floral sex transition from androecious to monoecious in response to cytokinin treatment on the androecious S. sebiferum provided valuable information related to the mechanism of sex determination in the perennial woody plants.

Coumarinolignoids and Taraxerane Triterpenoids from Sapium discolor and Their Inhibitory Potential on Microglial Nitric Oxide Production.

Seventeen compounds, including three new pairs of coumarinolignoid enantiomers, (7' S,8' S)-sapiumins A-C (1a-3a) and (7' R,8' R)-sapiumins A-C (1b-3b), six new taraxerane triterpenoids, sapiumic acids A-F (4-9), and five known taraxerane triterpenoids (10-14), were isolated from an ethanol extract prepared from the stems and leaves of Sapium discolor. The structures of 1-9 and their relative configurations were determined by spectroscopic data analysis, and the absolute configurations of the coumarinolignoids 1a/1b-3a/3b and triterpenoids 6-9 were assigned using experimental and calculated ECD data. Compounds 1a/1b-3a/3b are the first coumarinolignoids to be reported from the genus Sapium. Among all the isolates, compounds 1b, 2a/2b, 3a/3b, and 6-9 inhibited nitric oxide production in lipopolysaccharide-stimulated BV-2 microglial cells, with IC50 values of 1.7-15.3 µM.

Anti-diabetic potential of Sapium ellipticum (Hochst) Pax leaf extract in Streptozotocin(STZ)-induced diabetic Wistar rats.

BACKGOUND: Ethnobatanical survey associates Sapium ellipticum (SE) with antidiabetic usage among other medicinal functions in different parts of Africa. More importantly, previous studies on the plant extract in our laboratory showed that SE has significant effects on the activities of carbohydrate metabolizing enzymes such as glucokinase, glucose-6-phosphatase, α-amylase and α-glucosidase. In view of these, the anti-diabetic potential of the plant leaf extract in streptozotocin (STZ)-induced diabetes rat model (Wistar strain) was examined. METHODS: Diabetes was induced in experimental animals via single intraperitoneal dose (55 mg/kg BW) of freshly prepared STZ. SE was evaluated at 400 and 800 mg kg-1 of body weight (BW), against metformin (12 mgkg-1 BW). Treatments were done orally (p.o), twice daily at 8 h interval for a period of 21 days. RESULTS: SE significantly reduced fasting blood glucose (FBG) level by 46.5 and 44.4% (400 and 800 mg dosage respectively) compared to initial diabetic values. However, the effects were significantly lower than 72.6% glucose reduction produced by metformin. Hepatic and skeletal muscle glycogens were observed to increase by 27.06 and 12.55% respectively in SE-treated rats (800 mg dosage) compared to their corresponding values in diabetic control animals. Plasma and pancreatic insulin contents were also improved (31.77 and 52.34% respectively) by SE administration. The histopathological examination of the pancreas indicates beta cells regeneration in the treated animals, particularly in diabetic rats treated with 800 mg dosage of the extract compared to the diabetic control animals and metformin group. The presence of phenolic compounds namely amentoflavone, lupeol and luteolin-7-O-glucoside in SE as characterized and reported in our previous study is likely responsibly for the antidiabetic effects of the plant extract noted in the present study. CONCLUSION: The outcome of this study provides scientific basis in support of the medicinal relevance of SE and lend credence to its utilization in folk medicine for the treatment of diabetes and other oxidative stress-related ailments.

BRIEF-REPORT New set of microsatellites for Chinese tallow tree, Triadica sebifera.

Chinese tallow (Triadica sebifera) is an important crop and ornamental tree. After it was introduced into the USA, it gradually became a noxious invasive tree in south-eastern America since the middle of the 1900s. Because only six microsatellites were reported previously in T. sebifera, to better understand the genetic diversity and population dynamics of such species, we reported here 28 new microsatellite markers. For these 28 microsatellites, the number of alleles per locus ranged from 2-16. The expected heterozygosity and the expected heterozygosity corrected for sample size varied from 0.0796 to 0.9081 and from 0.0805 to 0.9176, respectively. These microsatellites will provide additional choice to investigate the genetic diversity and structure in T. sebifera.

Identification, characterization and structure analysis of a type I ribosome-inactivating protein from Sapium sebiferum (Euphorbiaceae).

Ribosome-inactivating proteins (RIPs) are N-glycosidases (EC3.2.2.22) that universally inactivate the ribosome, thereby inhibiting protein biosynthesis. In this study, a novel type I RIPs named SEBIN was identified in Sapium sebiferum. Nuclear acid depurine experiment showed that SEBIN had rRNA N-Glycosidase activity. Further experiment indicated that SEBIN significantly inhibited Caenorhabditis elegans development as well as resulted in worm cell apoptosis. This is the first report to evaluate RIPs toxicity using C. elegans. We proposed that SEBIN may impaire C. elegans reproduction in a DNA-damage manner besides traditional protein synthesis inhibition approach. The predicted 3D structure was modeled using threading and ab initio modeling, and the r-RNA binding residue of SEBIN was identified through the protein-ligand docking approach. It showed the amino acid residues, Glu195, Asn81, Ala82, Tyr83, Glu164, Ser163, Ile159 and Arg167, played critical roles in catalytic process. Our results provided the theoretical foundation of structure-function relationships between enzymatic properties, toxicity and structural characterization of SEBIN.

New diterpenoids from Sapium discolor.

Chemical fractionation of the ethanolic extract generated from the twigs and leaves of Sapium discolor led to the isolation and identification of four new macrocyclic diterpenoids including three members of the rare casbane family, sapidisins A-C (1-3), and an analog of the cembrane class, sapidisin D (4), a new 3,4-seco ent-kaurane diterpenoid (5), and 18 known phenolic compounds. Their structures were elucidated by comprehensive spectroscopic analyses especially 1D NMR (1)H-(1)H couplings and 2D NMR ROESY data. The discovery of 1-4 from S. discolor provides a clue for further study on the biogenetic evolution of the widely existent tigliane-type diterpenoids in the Sapium species.

[Chemical constituents of Sapium sebiferum leaves].

Chemical constituents of ethyl acetate extract of Sapium sebiferum leaves were isolated and purified by various chromatographic methods, including column chromatographies over silica gel, macroporous adsorption resin, and Sephadex LH-20, as well as preparative TLC and semi preparative HPLC. As a results, 15 compounds were separated from Sapium sebiferum leaves and their structures were examined by spectral analysis including NMR and MS data and identified as( + )-(7R,7'R,7"S,7'"S,8S,8'S,8"S,8'"S)-4", 4"'-dihydroxy-3,3',3",3',5,5'-hexamethoxy-7,9';7',9-diepoxy-4,8";4',8'"-bisoxy-8,8'-dineo-lignan-7",7"',9",9"'-tetraol(1) ,1-(4'- hydroxy-3'-methoxyphenyl)-2-[4"-(3-hydroxypropyl) -2", 6"-dimethoxyphenoxy] propane-1, 3-diol (2), Thero-2, 3-bis-(4-hydroxy-3- methoxypheyl)-3-methoxy-propanol(3) , threo-5-hydroxy-3,7-dimethoxyphenyl propane-8,9-diol (4), boropinol B (5), threo-8S-7-methoxysyringylglycerol(6), 5-hydroxymethylfurfural(7), 5-( methoxy-methyl)-1H-pyrrole-2-carbaldehyde (8), quercetin (9) , kaempferol (10), ethyl gallate(11), coniferaldehyde(12), vanillin(13), 7-hydroxy-6-methoxy-2H-1-henzopyran-2-one(14),and 1-heptacosanol (15). All compounds except for compounds 9-11,14 were separated from this plant for the first time.

Loss of specificity: native but not invasive populations of Triadica sebifera vary in tolerance to different herbivores.

During introduction, invasive plants can be released from specialist herbivores, but may retain generalist herbivores and encounter novel enemies. For fast-growing invasive plants, tolerance of herbivory via compensatory regrowth may be an important defense against generalist herbivory, but it is unclear whether tolerance responses are specifically induced by different herbivores and whether specificity differs among native and invasive plant populations. We conducted a greenhouse experiment to examine the variation among native and invasive populations of Chinese tallow tree, Triadica sebifera, in their specificity of tolerance responses to herbivores by exposing plants to herbivory from either one of two generalist caterpillars occurring in the introduced range of Triadica. Simultaneously, we measured the specificity of another defensive trait, extrafloral nectar (EFN) production, to detect potential tradeoffs between resistance and tolerance of herbivores. Invasive populations had higher aboveground biomass tolerance than native populations, and responded non-specifically to either herbivore, while native populations had significantly different and specific aboveground biomass responses to the two herbivores. Both caterpillar species similarly induced EFN in native and invasive populations. Plant tolerance and EFN were positively correlated or had no relationship and biomass in control and herbivore-damaged plants was positively correlated, suggesting little costs of tolerance. Relationships among these vegetative traits depended on herbivore type, suggesting that some defense traits may have positive associations with growth-related processes that are differently induced by herbivores. Importantly, loss of specificity in invasive populations indicates subtle evolutionary changes in defenses in invasive plants that may relate to and enhance their invasive success.

Phenological documentation of an invasive species, Sapium sebiferum (L.) Roxb.

Phenological documentation and plant invasion are key contemporary issues in the biological sciences. Recognizing this, the present paper describes the phenological stages of an invasive species, Sapium sebiferum (L.) Roxb. A tree of the species was marked, and daily visits were made for recording its phenology. The Biologische Bundesanstalt, Bundessortenamt and CHemical industry (BBCH) scale was used for documenting the same. Of the total ten principal stages, eight were observed in the species. These stages started with bud development (stage 01) in the month of March and ended with leaf fall (stage 97) in the month of December. Descriptions of the phenological stages and their dates are provided in the text. It is hoped that such a phenological scale will be helpful in managing plant invasion.

Glycosides with galloyl groups from Balakata baccata and their antineuroinflammatory activities.

Seven new glycosides (1 - 7) with galloyl groups and two known kaempferol glycosides (8 and 9) were obtained from the overground parts of Balakata baccata. The structures of the new compounds were determined by comprehensive spectroscopic analyses. The rarely seen allene moiety in compounds 6 and 7 were described by detailed analysis of 1D and 2D NMR data. The antineuroinflammatory effect of all the isolates was assessed through inhibiting nitric oxide (NO) production in lipopolysaccharide (LPS)-induced BV-2 microglial cells. Compounds 1, 2, 6, and 7 showed potent inhibitory activities with IC50 values of 25.7, 17.2, 15.5 and 24.4 µM, respectively, compared with the positive control minocycline (IC50 = 16.1 µM).

Hemiptera-induced galls of Sapium glandulosum have histological and cytological compartmentalization created with a large amount of carbohydrate.

Gall formation impacts the development of plant species by altering the structure and mobilization of reserves, and the functional and physiological patterns of the host organ. The current study aimed to evaluate the impact generated by the Neolithus fasciatus galling insect (Hemiptera: Triozidae) in Sapium glandulosum leaves (Euphorbiaceae) at the cytological, histological, histochemical, and biochemical levels. Non-galled leaves and galls in the young, mature, and senescent stages were evaluated. The non-galled leaf has a uniseriate epidermis, stomata only on the abaxial side, a dorsiventral mesophyll, and parenchyma cells with thin primary walls containing chloroplasts with plastoglobules. The gall has a parenchymatous compartmentalized cortex. The young and mature galls already have a dense cytoplasm, especially in the inner cells of the cortex, with chloroplasts, mitochondria, Golgi complex, and large and evident nuclei. In senescent galls, there are signs of organelle degradation and cell digestion. Carbohydrates occur in greater amounts in the mature gall, mainly in the starch grain form, while proteins and lipids predominate in non-galled leaves. Secondary metabolites occur mainly in the young gall and may be related to its protection and to the signaling of its development. Sapium glandulosum galls have histological and cytological compartmentalization of the cortex with a large amount of carbohydrates, which supply energy to maintain the development of the structure.

Isolation of high-quality total RNA from lipid-rich seeds.

Obtaining high-quality RNA from lipid-rich seeds has been a challenge, since lipids can interfere with tissue disruption and block cells from extraction buffer, and polysaccharides, polyphenols, and other secondary metabolites can bind or coprecipitate with RNA. Using an improved CTAB-based extraction buffer and modified grinding and incubation method, we developed a protocol that is able to minimize the negative effects caused by lipids and other compounds. Using this protocol, we obtained high-quality and high-quantity RNA from six species of lipid-rich seeds within 3 hours. The isolated RNA was demonstrated to be suitable for downstream applications.

Dielectric characterization of the seeds of invasive Chinese tallow tree.

Biofuels have the potential to replace a significant portion of the transportation needs of the USA and the world. Low-cost lipid feedstock (i.e. tallow tree seeds) can be used for production of biodiesel, but these seeds need to be dried. Microwave drying is utilized to dry various seeds and grain, but for this process to be most efficient the dielectric properties of the materials need to be known. This study presents, for the first time, the dielectric properties of the seeds of the Chinese tallow tree. The dielectric constant and dielectric loss factor were determined using a modified free-space method for different moisture contents at frequency ranges centered around 915 and 2450 MHz respectively. The dielectric constant decreased with frequency in those respective ranges, and increased linearly with increasing moisture content, from 1.96 at 0% m.c. to 2.3 at 7% m.c. at 915 MHz, and from 2.08 at 0% m.c. to 2.42 at 7.6% m.c. at 2450 MHz. The dielectric loss factor generally decreased with frequency in both frequency ranges, and increased linearly with moisture content from a low of 0.16 at 0% m.c. to 0.21 at 7.56% at 915 MHz, respectively from a low of 0.13 at 0% m.c. to 0.2 at 7.6% m.c. at 2450 MHz. These results can be used to design microwave processing operations and systems for drying of tallow tree seeds, as well as for non-destructive determination of theses seeds' moisture content.

Thermophysical characterization of the seeds of invasive Chinese tallow tree: importance for biofuel production.

The limited supply of traditional fossil based fuels, and increased concern about their environmental impact has driven the interest in the utilization of biomass based energy sources, including those that are underutilized or otherwise nuisance species such as Chinese tallow trees (Triadica sebifera [L.]). This species is a prolific seeds producer, and this paper shows that they contain more than 50% lipids by mass that are suitable for conversion into biodiesel. We present here, for the first time, the seeds' thermophysical properties important for biofuel production. The seeds were characterized using thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and ultimate analysis; their thermal conductivity, thermal diffusivity, and specific heat were determined. The characterization results were correlated to fatty acid composition and lipid content for whole seeds and individual layers, as well as to the protein, hemicellulose, cellulose, and lignin content. The TGA analysis indicated the presence, in addition to lipids, of hemicellulose, cellulose, lignin, and proteins, depending on the layer analyzed. Thermal conductivity and specific heat were, respectively 0.14 ± 0.007 W/mK and 3843.5 ± 171.16 J/kgK for wax, 0.20 ± 0.002 W/mK and 2018.7 ± 5.18 J/kgK for shells, 0.13 ± 0.0 W/mK and 1237 ± 3.15 J/kgK for internal kernel, and 0.13 ± 0.000 W/mK and 2833.9 ± 104.11 J/kgK for whole seeds. These properties and characterization method can be further used in engineering analysis used to determine the most optimum processing method for production of biofuels from this feedstock.

Interactive effects of herbivory and competition intensity determine invasive plant performance.

Herbivory can reduce plant fitness, and its effects can be increased by competition. Though numerous studies have examined the joint effects of herbivores and competitors on plant performance, these interactive effects are seldom considered in the context of plant invasions. Here, we examined variation in plant performance within a competitive environment in response to both specialist and generalist herbivores using Chinese tallow as a model species. We combined tallow plants from native and invasive populations to form all possible pairwise combinations, and designated invasive populations as stronger neighbours and native populations as weaker neighbours. We found that when no herbivory was imposed, invasive populations always had higher total biomass than natives, regardless of their neighbours, which is consistent with our assumption of increased competitive ability. Defoliation by either generalist or specialist herbivores suppressed plant growth but the effects of specialists were generally stronger for invasive populations. Invasive populations had their lowest biomass when fed upon by specialists while simultaneously competing with stronger neighbours. The root/shoot ratios of invasive populations were lower than those of native populations under almost all conditions, and invasive plants were taller than native plants overall, especially when herbivores were present, suggesting that invasive populations may adopt an "aboveground first" strategy to cope with herbivory and competition. These results suggest that release from herbivores, especially specialists, improves an invader's performance and helps to increase its competitive ability. Therefore, increasing interspecific competition intensity by planting a stronger neighbour while simultaneously releasing a specialist herbivore may be an especially effective method of managing invasive plants.