Guaiane-type sesquiterpenoids from Alisma orientalis

Two guaiane-type sesquiterpenoids named orientalol E (1) and orientalol F (3) were isolated from the rhizome of Alisma orientalis (SAM) JUZEP together with two known guaiane-type sesquiterpenoids alismol (2) and alismoxide (4). Their relative stereostructures were elucidated by spectroscopic methods, whereas absolute stereostructures were determined on the basis of chemical correlation.     

Saponin-rich fraction from Clematis chinensis Osbeck roots protects rabbit chondrocytes against nitric oxide-induced apoptosis via preventing mitochondria impairment and caspase-3 activation

Our previous study reported that the saponin-rich fraction from Clematis chinensis Osbeck roots (SFC) could effectively alleviate experimental osteoarthritis induced by monosodium iodoacetate in rats through protecting articular cartilage and inhibiting local inflammation. The present study was performed to investigate the preventive effects of SFC on articular chondrocyte, and explore the underlying mechanisms. Primary rabbit chondrocytes were cultured and exposed to sodium nitroprusside (SNP), a NO donor. After treatment with different concentrations of SFC (30, 100, 300, 1,000 μg/ml) for 24 h, nucleic morphology, apoptotic rate, mitochondrial function and caspase-3 activity of chondrocytes were examined. The results showed that SNP induced remarkable apoptosis of rabbit chondrocytes evidenced by Hoechst 33258 staining and flow cytometry analysis, and SFC prevented the apoptosis in a concentration-dependent manner. Further studies indicated that SFC could prevent the depolarization of mitochondrial membrane potential (∆ψm) in SNP-treated chondrocytes and suppress the activation of caspase-3. It can be concluded that the protection of SFC on articular chondrocytes is associated with the anti-apoptosis effects via inhibiting the mitochondrion impairment and caspase-3 activation.     

中华猕猴桃基因组可变剪接事件鉴定及分析

可变剪接使一个基因能产生多种m RNA成熟体,极大地增加蛋白多样性.采用中华猕猴桃基因组数据做参考数据,利用中华猕猴桃叶片和果实3个不同发育时期(未成熟、半成熟和成熟期)的转录组数据,从中华猕猴桃基因组(39040个基因)中共鉴定出11651个基因(占总基因数的29%)对应的32180个可变剪接事件.在可变剪接不同类型中,内含子保留类型的发生频率最高,占50%以上;3′可变位点类型频率约为5′端可变类型的2倍.GO富集分析结果表明,可变剪接的基因主要富集于酶调控及核苷酸结合相关功能的GO类别中,而组织特有可变剪接基因功能富集热点与组织的重要功能关联,叶片多为肌动蛋白及微管相关;未成熟果实与双组分信号系统相关;半成熟果实多与磷脂合成过程相关;成熟果实多与信号传递过程相关.另外,55.6%的维生素合成相关基因发生可变剪接事件,显著高于基因组水平的29.6%,暗示着可变剪接参与维生素合成相关基因代谢过程中的重要作用.通过对中华猕猴桃全基因组可变剪接的分析,为解析中华猕猴桃基因组及进一步开展相关分子育种工作提供依据.     

A compound isolated from Schisandra chinensis induces apoptosis

Schizandra chinensis has been known to have five predominant tastes: salty, sweet, sour, astringent, and bitter. It has also been shown to have various effects on the cardiovascular system, gastrointestinal system, anti-inflammatory, central nervous system, endocrine system, and stress protect. However, its anti-cancer activity on colon carcinoma HCT-116 cells has not been yet been examined. Thus, in this study, we attempted to isolate a compound from Schisandra chinensis that induced apoptosis in HCT-116 cells. An active compound was found and identified to be Gomisin A. It displayed apoptotic activity through caspase-7 cleavage in colon carcinoma HCT-116 cells. In addition, we further assessed the effects of this compound using long-term survival clonogenic assay with HCT116 cells.     

Stereumin A-E, sesquiterpenoids from the fungus Stereum sp. CCTCC AF 207024

Five cadinane sesquiterpenoids, named stereumin A (1), B (2), C (3), D (4) and E (5) were isolated from the CHCl(3) extract of the culture broth of the fungal strain CCTCC AF 207024. Based on the sequences at the internal transcribed spacer (ITS) region and partial 28S rDNA, this fungus was identified as a Stereum sp. The structures of the five compounds were elucidated using spectroscopic data from 1D, 2D NMR and HRESIMS experiments, and the structures of 1 and 2 were further confirmed by single-crystal X-ray diffraction analysis. Compounds 1-5 showed nematicidal activities against the nematode Panagrellus redivivus at 400 mg l(-1). Among these five compounds, compounds 3 and 4 killed 84.4% and 94.9% of P. redivivus, respectively in 48 h.     

Eryvarins F and G,two 3-phenoxychromones from the roots of Erythrina variegata

Two 3-phenoxychromones, eryvarins F and G, were isolated from the roots of Erythrina variegata. Their structures were established to be 3-(2,4-dihydroxyphenoxy)-7-hydroxy-6,8-di(3,3-dimethylallyl)chromen-4-one and 3-(2,4-dihydroxyphenoxy)-8-(3,3-dimethylallyl)-2,2-dimethylpyrano[5,6:6,7]chromen-4-one on the basis of spectroscopic and chemical evidence. Eryvarins F and G are unusual 3-phenoxychromone derivatives with two isoprenoid groups.     

Synaptonemal complex analysis in spermatocytes of diploid and triploid Chinese shrimp Fenneropenaeus chinensis

A modified surface spreading technique for synaptonemal complex (SC) analysis was tested to assess the process of chromosome synapsis in spermatocytes of diploid and induced triploid Fenneropenaeus chinensis. Spermatocytes of diploid shrimp showed typical morphological characteristics of eukaryote SC, with complete synapsis of bivalents. No recognizable bivalent associated with sex chromosomes was observed in spermatocytes of diploid shrimp. However, differences in morphology of SC, including unsynapsed univalents, bivalents, totally paired trivalents with non-homologous synapsis, partner switches and triple synapsis were identified at early pachytene stage of triploid spermatocytes. Triple synapsis was especially common at late pachytene stage in spermatocytes of triploid shrimp. The observed abnormal synapsis behavior of chromosomes in spermatocytes indicated that triploid male shrimp may find it difficult to develop normal haploid sperm.     

Irregular sesquiterpenoids from Ligusticum grayi roots

Root oil of Ligusticum grayi (Apiaceae) contains numerous irregular sesquiterpenoids. In addition to the known acyclic sesquilavandulol and a sesquilavandulyl aldehyde, two thapsanes, one epithapsane, and 14 sesquiterpenoids representing eight hitherto unknown carbon skeletons were found. These skeletons are: prethapsane, i.e. 1,1,2,3a,7,7-hexamethyloctahydro-1H-indene; isothapsane, i.e. 1,2,3a,6,7,7a-hexamethyloctahydro-1H-indene; ligustigrane, i.e. 1,1,2,7,7,7a-hexamethyloctahydro-1H-indene; isoligustigrane, i.e. 1,1,2,6,7,7a-hexamethyloctahydro-1H-indene; preisothapsane, i.e. 1,1,2,3a,6,7-hexamethyloctahydro-1H-indene; isoprethapsane, i.e. 1,1,2,4,7,7-hexamethyloctahydro-1H-indene; allothapsane, i.e. 1,1,2,3a,7,7a-hexamethyloctahydro-1H-indene; and oshalagrane, i.e. 1,1,2,4,6,6-hexamethylspiro[4.4]nonane.The bicyclic sesquiterpenoids are presumably biosynthesized by head-to-head coupling of geranyl diphosphate and dimethylallyl diphosphate, followed by a cyclization sequence leading to a hydroindane skeleton with six one-carbon substituents. Subsequent rearrangements—primarily methyl migrations—account for the remarkable variety of structures represented in L. grayi root oil.     

Purification and characterization of hatching enzyme from shrimp Penaeus chinensis

By using Penaeus chorion as a specific substrate, the hatching enzyme (HE) from Penaeus chinensis was purified by gel-filtration and ion-exchange chromatography, and characterized in terms of its molecular weight and enzymatic properties in this study. It was found that the molecular weight of Penaeus HE is about 43.0 kDa in SDS-PAGE. The Penaeus HE had obvious choriolytic activity, which was optimal at pH 6.0 and temperature of 40 degrees C, respectively. The Km value of the HE for casein was 7.47 mg ml(-1). The HE activity was almost completely inhibited by SBTI, p-APMSF, bestatin, and NEM, greatly inhibited by ovomucoid, TLCK, IAM, chymostatin, and PMSF, and slightly inhibited by pepstatin A, TPCK, LBTI, and leupeptin. These results indicate that the HE is most probably a trypsin-type serine protease. Besides of these, the HE was extremely sensitive to EDTA, Zn2+, Ca2+, Mg2+, and Cu2+. Combined with the results that the EDTA-pretreated HE activity could be perfectly recovered by Zn2+, it is indicated that shrimp HE is most probably a kind of Zn-metalloprotease.     

Structure of a highly substituted beta-xylan of the gum exudate of the palm Livistona chinensis (Chinese fan)

Structural features of the acidic, highly substituted glycanoxylan (LCP; 87% yield) from the gum exudate of the palm, Livistona chinensis, family Arecaceae, were determined. It had [alpha]D -30 degrees, Mw 1.9x10(5) and a polydispersity ratio Mw/Mn of approximately 1.0. Acid hydrolysis gave rise to Rha, Fuc, Ara, Xyl, and Gal, in a 1:6:46:44:3 molar ratio, and 12% of uronic acid was present. LCP had a highly branched structure with side-chains containing nonreducing end-units (% values are approximate) of Araf (15%), Fucp (4%), Xylp (7%), GlcpA, and 4-Me-GlcpA, and internal 2-O- (5%) and 3-O-substituted Araf (8%), and 2-O-substituted Xylp (14%) units. The (1-->4)-linked beta-Xylp main-chain units of LCP were substituted at O-3 (4%), O-2 (17%), and O-2,3 (16%). Partial acid hydrolysis gave 4-Me-alpha-GlcpA-(1-->2)-[beta-Xylp-(1-->4)](0-2)-Xyl, identified by showing that the uronic acids were single-unit side-chain substituents on O-2. Milder hydrolysis conditions removed from O-3 other side-chains containing Fucp and Araf nonreducing end-units and internal Arap, and 2-O- and 3-O-substituted Araf units. Carboxyl-reduced LCP contained 4-O-methylglucose and glucose in a 3.2:1 molar ratio, arising from GlcpA and 4-OMe-GlcpA nonreducing end-units, respectively. The gum contained small amounts of free alpha-Fucp-(1-->2)-Ara, which corresponds to structures in the polysaccharide. Free myo- and D- or L-chiro-inositol were present in a 9:1 ratio.     

Laboratory Evaluation of Seven Pakistani Strains of Entomopathogenic Nematodes Against a Stored Grain Insect Pest, Pulse beetle Callosobruchus chinensis (L.)

Seven Pakistani strains of entomopathogenic nematodes belonging to the genera Steinernema and Heterorhabditis were tested against last instar and adult stages of the pulse beetle, Callosobruchus chinensis (L.). These nematodes included Steinernema pakistanense Shahina, Anis, Reid and Maqbool (Ham 10 strain); S. asiaticum Anis, Shahina, Reid and Rowe (211 strain); S. abbasi Elawad, Ahmad and Reid (507 strain); S. siamkayai Stock, Somsook and Reid (157 strain); S. feltiae Filipjev (A05 strains); Heterorhabditis bacteriophora Poinar (1743 strain); and H. indica Poinar, Karunakar and David (HAM-64 strain). Activity of all strains was determined at four different nematode densities in Petri dishes and in concrete containers. A significant nematode density effect was detected for all nematode species tested. Overall, Heterorhabditis bacteriophora, S. siamkayai, and S. pakistanense were among those that showed the highest virulence to pulse beetle larvae and adults. For all nematode species, the last larval stage of the pulse beetle seems to be more susceptible than the adult. LC(50) values in Petri dish and concrete containers were 14-340 IJs/larvae and 41-441 IJs/larvae, respectively, and 59-1376 IJs/adult and 170-684/adult, respectively.     

Cytotoxic sesquiterpenoids from Winteraceae of Caledonian rainforest

One secobutanolide, two butanolides and six drimane sesquiterpenoids were isolated from the bark and leaves of Zygogynum pancheri and Zygogynum acsmithii (Winteraceae) along with six known drimanes, isodrimanial, 1β-O-p-methoxy-E-cinnamoyl-bemadienolide, 7-ketoisodrimenin, drimenin, polygodial and 1β-E-cinnamoyl-6α-hydroxypolygodial. Their structures were elucidated through analysis of spectroscopic data. Drimane sesquiterpenoids with a dialdehyde function exhibited significant inhibitory activities in the in vitro cytotoxic assays against KB, HL60 and HCT116 cancer cell lines.     

Costs of mating and egg production in female Callosobruchus chinensis

Costs of reproduction include the costs of mating and egg production. Specific techniques such as irradiation or genetic mutation have been used to divide the expense into costs of mating and egg production in previous studies. We tried to divide the costs in the adzuki bean beetle, Callosobruchus chinensis (Coleoptera: Bruchidae), which needs some kinds of bean as an oviposition substrate. Mated females that were not allowed to lay eggs had a shorter life span than virgin females, but they had a longer life span than mated females that were allowed to lay eggs. The results showed two independent significant costs, mating and egg production, on the life span in C. chinensis. Costs of mating, however, include the costs of sexual harassment by males and copulation itself, and we need further studies to divide the costs. The present method for dividing the cost of reproduction into costs of mating and egg production can be applied to a broad taxonomic range of insect species, and thus it will be a useful model system for inter-specific comparisons of costs of mating and egg production.     

Nardonaphthalenones A and B from the roots and rhizomes of Nardostachys chinensis Batal.

Nardonaphthalenones A and B (1–2), one new apo-α-carotenone (3) and four new monoterpenoids (4, 8–9 and 11), along with six known compounds (5–7, 10, 12–13) were isolated from the dried roots and rhizomes of Nardostachys chinensis Batal. Their structures were elucidated by analysis of the spectroscopic data including NMR, HRESIMS and circular dichroism data. Furthermore, the serotonin transporter (SERT)-regulating activities of these isolates were evaluated, among them compound 3 showed the strongest enhancement activity while compound 12 showed a moderate inhibition activity on SERT.     

Alginate-encapsulation of shoot tips of jojoba [Simmondsia chinensis (Link) Schneider] for germplasm exchange and distribution

Shoot tips excised from in vitro proliferated shoots derived from nodal explants of jojoba [Simmondsia chinensis (Link) Schneider] were encapsulated in calcium alginate beads for germplasm exchange and distribution. A gelling matrix of 3 % sodium alginate and 100 mM calcium chloride was found most suitable for formation of ideal calcium alginate beads. Best response for shoot sprouting from encapsulated shoot tips was recorded on 0.8 % agar-solidified full-strength MS medium. Rooting was induced upon transfer of sprouted shoots to 0.8 % agar-solidified MS medium containing 1 mg l⁻¹ IBA. About 70 % of encapsulated shoot tips were rooted and converted into plantlets. Plants regenerated from encapsulated shoot tips were acclimatized successfully. The present encapsulation approach could also be applied as an alternative method of propagation of desirable elite genotype of jojoba.     

Constituents of Chrysothamnus viscidiflorus

A phytochemical investigation of the aerial parts of Chrysothamnus viscidiflorus var. viscidiflorus afforded three new [chrysothol (1), 2 and 4] and seven known compounds, including five sesquiterpenes, two cinnamic acid derivatives, two ketoalcohol derivatives and one coumarin glucoside. The structures of two previously reported compounds, 1b and 1c, were revised on the basis of chemical reaction. Structures of the compounds were determined by extensive NMR studies, including DEPT, COSY, NOE, HMQC, HMBC and X-ray analysis. The unpublished X-ray data of the known compounds 6 and 7 are reported. Compounds chrysothol (1), and 8-10 showed anti-cancer activity against human breast cancer cells.     

An efficient and reproducible indirect shoot regeneration from female leaf explants of Simmondsia chinensis,a liquid-wax producing shrub

Simmondsia chinensis (Link) Schneider is a perennial, dioecious, drought resistant and multipurpose seed oil crop grown in arid and semi-arid conditions throughout the world. A reproducible and more efficient method for indirect shoot organogenesis from female leaf explants has been standardized. The leaf explants cultured on Murashige and Skoog (MS) medium with 1.0 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) alone produced the highest frequency of callus compared with 1.5 mg l⁻¹ IBA. Maximum proliferation of callus was observed on MS medium containing a combination of 1.0 mg l⁻¹ 2,4-D with 0.5 mg l⁻¹ BAP. For shoot differentiation, the proliferated callus was subcultured on MS medium supplemented with 6-benzylaminopurine (BAP) (1.0–4.0 mg l⁻¹) along with 40 mg l⁻¹ adenine sulphate as additive or in combination with α-naphthalene acetic acid (NAA) or Indole-3-butyric acid (IBA). Optimum shoots differentiated from callus was obtained on MS medium supplemented with 2.0 mg l⁻¹ BAP and 0.2 mg l⁻¹ NAA. On this medium, 100 % cultures were responded with an average number of 14.44 shoots per explant with their mean length of 4.78 cm. In vitro rooting (6.22 roots per explant) was achieved on half strength MS medium containing 2 % sucrose with 3.0 mg l⁻¹ IBA and 300 mg l⁻¹ activated charcoal (AC). Rooted plantlets were successfully hardened under control conditions and acclimatized under field conditions with 90 % success rate. The present protocol is highly efficient, reproducible and economically viable for large scale production of female plants.     

不同增温处理对夏蜡梅光合特性和叶绿素荧光参数的影响

利用红外线辐射器(Electric radiant infrared heater)设置3个不同增温处理:(1)晚上增温2℃(18:00—翌日6:00);(2)白天增温2℃(6:00—18:00);(3)全天增温2℃;以不做增温处理作为对照,模拟气候变暖对夏蜡梅(Sinocalycanthus chinensis)的光合特性和叶绿素荧光参数的影响。3个季节中,3种增温处理下的夏蜡梅净光合速率(Anet)和最大净光合速率(Amax)均高于对照。表明3种增温处理均可以提高夏蜡梅的光合作用能力。增温处理提高了气孔导度(Gs)、暗呼吸速率(Rd),降低了胞间CO2浓度(Ci)。3种增温处理中,夏蜡梅在3个生长季节的Anet和Amax均以白天增温最大,表明白天增温下夏蜡梅具有最高的光合作用能力。白天增温处理下的表观量子效率(AQY)、光饱和点(LSP)显著高于全天增温、晚上增温以及对照。白天增温和晚上增温使夏蜡梅的相对叶绿素含量显著增加,且白天增温叶绿素相对含量显著高于晚上增温、全天增温及对照。白天增温、晚上增温与全天增温处理下的夏蜡梅的PSⅡ最大光化学效率(Fv/Fm)均显著低于对照,在12:00和14:00时,三者之间存在显著性差异,以白天增温最高,晚上增温次之,全天增温最低。总之,3种增温处理夏蜡梅的光合能力总体上均有不同程度的增强,且在白天增温处理下,夏蜡梅的Anet、Amax值最高,表明白天增温下夏蜡梅具有最高的光合能力,这与白天增温引起的相对叶绿素含量增高、叶片对强光的适应能力加强、较低程度的光胁迫等因素有关。本研究说明未来增温条件下,将提高夏蜡梅的光合生理活性,促进夏蜡梅的生长。     

祁连圆柏和圆柏色素含量及其花青苷合成酶活性的季节性变化

以祁连圆柏(Sabina przewalskii)和圆柏(S. chinensis)为材料, 测定2种植物花青苷、类黄酮、紫松果黄素、叶绿素和 类胡萝卜素的含量及花青苷合成过程中关键酶苯丙氨酸解氨酶(PAL)和类黄酮糖基转移酶(UFGT)的活性, 并分析了各值的季节性变化。结果表明, 祁连圆柏和圆柏叶片中PAL及UFGT的活性、花青苷、类黄酮﹑紫松果黄素以及类胡萝卜素的含量在低温季节均明显高于其它季节; 叶绿素含量在低温季节低于其它季节; 并且祁连圆柏中花青苷含量及其合成酶PAL和UFGT的活性以及类黄酮、紫松果黄素和叶绿素含量始终高于圆柏。结果说明花青苷是圆柏属植物中具有抗冻特性的重要次生代谢物,是抵御低温和辐射胁迫的一种重要保护物质; 紫松果黄素等色素对圆柏属植物抵抗低温诱导的光抑制起重要作用。