Essential oil constituents of Valeriana italica and Valeriana tuberosa. Stereochemical and conformational study of 15-acetoxyvaleranone

The chemical composition of the essential oils obtained by hydrodistillation of roots, stems with leaves and inflorescences of Valeriana italica Lam. and Valeriana tuberosa L. were studied by GC/MS. Seventy-three and forty-one constituents were identified from each plant, respectively. The major constituent of the oil obtained from the roots of V. italica was isolated and identified as 15-acetoxyvaleranone. Its stereochemistry and conformation has been studied using NMR spectroscopy and molecular modelling. The oils obtained from V. tuberosa completely lacked the characteristic valerane or kessane sesquiterpenes.     

UHPLC-Q-Orbitrap HR-MS-Based Metabolomics for Profiling the Sida rhombifolia Metabolites with Different Plant Organs and Cultivation Ages

Plant organs and cultivation ages can result in different compositions and concentration levels of plant metabolites. The metabolite profile of plants can be determined using liquid chromatography. This study determined the metabolite profiles of leaves, stems, and roots of Sida rhombifolia at different cultivation ages at 3, 4, and 5 months post-planting (MPP) using liquid chromatography-mass spectrometry/mass spectrometry (LC/MS/MS). The results identified that 41 metabolites in S. rhombifolia extract for all plant organs and cultivation ages. We successfully identified approximately 36 (leaves), 22 (stems), and 18 (roots) compounds in all extract. Using principal component analysis (PCA) with peak area as the variable, we clustered all sample extracts based on plant organs and cultivation ages. As a result of PCA, S. rhombifolia extracts were grouped according to plant organs and cultivation ages. In conclusion, a clear difference in the composition and concentration levels of metabolites was observed in the leaves, stems, and roots of S. rhombifolia harvested at 3-, 4-, and 5-MPP.     

4种鼠尾草属植物根、茎、叶中脂溶性成分的分布差异性

为了说明4种鼠尾草属植物脂溶性成分在根、茎、叶中的分布情况,本文采用了组织化学定位和HPLC图谱分析相结合的方法,对4种鼠尾草属植物不同部位的脂溶性成分进行了分析.结果表明:4种鼠尾草属植物的根、茎、叶中脂溶性成分的分布有差异,根中脂溶性成分分布于周皮,茎、叶中脂溶性成分分布于表皮.丹参和白花丹参根中的脂溶性成分主要是丹参酮ⅡA、丹参酮Ⅰ和二氢丹参酮Ⅰ,鼠尾草和药用鼠尾草根中仅含丹参酮ⅡA,不含丹参酮Ⅰ和二氢丹参酮Ⅰ.茎、叶中的脂溶性成分不是丹参酮ⅡA、丹参酮Ⅰ和二氢丹参酮Ⅰ.脂溶性成分的种类和含量随品种和引种地的不同而产生差异.这一结果明确了脂溶性成分在4种鼠尾草属植物不同部位的确切分布,为准确用药提供了依据.     

蚜虫寄主植物与取食部位的多样性

在中国科学院动物研究所已有标本采集记录和国内外文献资料的基础上,分别从蚜虫类Aphidina科级和属级阶元系统研究世界范围内蚜虫寄主植物与取食部位的多样性.蚜虫的寄主植物种类繁多,涉及267科2120属,尤其以菊科、禾本科、豆科、伞形科、唇形科、蔷薇科、茜草科、兰科、壳斗科、杨柳科、胡桃科等植物为主.13科蚜虫的寄主植物差别很大:在科级水平,球蚜科Adelgidae、纩蚜科Mindaridae和平翅绵蚜科Phloeomyzidae的寄主植物类群比较单一,蚜科Aphididae和瘿绵蚜科Pemphigidae的寄主植物范围最为广泛;在属级水平,各属蚜虫间寄主植物也有明显差异,有143属蚜虫的寄主植物多于2科,其中蚜科的属占多数,蚜属Aphis、瘤蚜属Myzus、长管蚜属Macrosiphum、粗额蚜属Aulacorthum和声蚜属Toxoptera的寄主植物最多,各超过100科290属.在不同的寄主植物上寄生着不同种类的蚜虫,其中16科47属寄主植物上寄生的蚜虫多于14属.蚜虫在寄主植物上的取食部位丰富多样,可分为7类,分别为叶片、嫩梢、嫩枝、茎、花、根部、果实等.以叶片、茎、嫩枝和嫩梢为蚜虫主要取食部位.平翅绵蚜科主要取食嫩枝、茎和根部,短痣蚜科Anoeciidae主要取食叶片、嫩梢和根部,球蚜科、群蚜科Thelaxidae和毛管蚜科Greenideidae取食叶片、嫩梢、嫩枝和茎等部位,纩蚜科取食叶片、嫩梢、茎和花等部位,大蚜科Lachnidae和斑蚜科Drepanosiphidae除取食叶片、嫩梢、嫩枝、茎等部位外,前者还在根部取食,后者还寄生在果实上,根瘤蚜科Phylloxeridae在除花以外的其它6个部位取食,蚜科、瘿绵蚜科和毛蚜科Chaitophoridae的取食部位最为多样,它们在上述7个部位均可取食.还初步讨论了在不同寄主植物上蚜虫物种的分化,以及蚜虫与寄主植物之间的对应关系.     

Tea prepared from Anastatica hirerochuntica seeds contains a diversity of antioxidant flavonoids, chlorogenic acids and phenolic compounds

HPLC-PDA-MS(2) was used to identify phenolic and polyphenolic compounds in an herbal tea made from seeds of Anastatica hirerochuntica, a plant found in the Sahara-Arabian deserts and used to treat a variety of ailments. Twenty compounds comprising a series of flavone C- and O-linked glycosides, phenolic acids, chlorogenic acids and flavonols were identified or partially identified on the basis of co-chromatography with reference compounds and MS(2) and MS(3) fragmentation patterns. The flavones were the principal components, occurring as luteolin, apigenin and diosmetin conjugates. The antioxidant potential of individual compounds in Anastatica was assessed using HPLC with an on-line ABTS·(+) detection system. This revealed that 14 compounds exhibited antioxidant activity. The highest contribution to the antioxidant capacity of the tea, 56%, came from 3,4-dihydroxybenzoic acid and caffeoyl- and dicaffeoylquinic acids while 6-C-glucosides of luteolin and apigenin contributed 41%.     

干旱胁迫对苗期大豆异黄酮合成的影响

以不同耐旱性的2个大豆品种(高耐旱JP-6、低耐旱JP-16)为研究材料,采用高效液相色谱和实时荧光定量PCR技术,分析不同时间持续干旱胁迫下,大豆叶片和根系中异黄酮的积累变化及关键酶基因的表达情况.结果表明:大豆根部异黄酮含量显著高于叶部,而异黄酮关键酶基因的表达量则在叶片中更高,耐旱品种JP-6根部的异黄酮积累量更大.随着干旱胁迫持续时间的增加,不同耐旱品种的异黄酮合成与积累变化规律存在显著差异:强耐旱品种JP-6的根和叶中,异黄酮积累量均呈现先下降后升高的趋势;而弱耐旱品种JP-16则相反,异黄酮积累量在不同部位中均呈现先上升后降低的趋势;除JP-6叶中C4H、4CL、IFS2等异黄酮合成上游基因外,其他不同品种、不同部位的关键酶基因表达量均随着干旱胁迫持续时间的增加,呈现先下降后上升的趋势.大豆叶片是异黄酮的主要合成部位,大豆根部也存在少量的异黄酮合成.弱耐旱大豆根部的异黄酮合成和最终积累量均较低,强耐旱品种则较高.根部异黄酮积累量高的大豆品种,其耐旱性更强.     

光照对大豆幼苗组织中异黄酮含量和分布的影响

利用高效液相色谱（ＨＰＬＣ）测定了不同光照处理的大豆（Ｇｌｙｃｉｎｅｍａｘ（Ｌ．）Ｍｅｒｒｉ．）幼苗不同组织的异黄酮类含量。子叶中最高,叶片和根部相对较少。子叶的异黄酮以大豆甙和染料木甙及其丙二酰基结合体为主,且在光照条件下,异黄酮含量随光照时间的增加而显著升高;相反,黑暗中的异黄酮含量随苗龄的增加呈下降趋势;当子叶由黑暗转为光照处理以后,异黄酮含量同样随光照时间的增加而升高。在叶片和根部异黄酮含量和种类也因光照条件的不同而有很大差异。光照条件下,叶片中以染料木甙及其丙二酰结合体和黄酮芦丁为主,且随时间增加呈上升趋势;黑暗中的黄化叶片,则以大豆甙和丙二酰结合体为主,但随时间的变化不明显。在幼苗根部,黑暗条件下几乎检测不出异黄酮的存在;光照条件下,则可检测到５种异黄酮,其中以大豆甙元及其衍生物占主要部分。实验证实了光照对大豆异黄酮的积累有明显的促进作用     

Light Effect on the Tissue Contents and Distribution of Isoflavones in the Developing Seedling of Soybean

The contents of isoflavones in the developing soybean seedling tissues under different light conditions were examined by high-performance liquid chromatography (HPLC). The results indicated that the total contents of isoflavones in soybean seedling tissues were remarkably different, being the highest in the cotyledons and lower in the leaves and roots. In the cotyledons, the isoflavones like daidzin, genistin and their malonyl conjugates were the major metabolites and were increased markedly with the time of light exposure. In contrast, there was a decrease in the content of isoflavones in the dark. When the dark grown seedlings were tranferred to the light, an increase of the isoflavone content was observed. In the leaves and roots, there was also a marked difference in the contents and types of isoflavones due to various light conditions. Leaves contained mainly the isoflavones such as genistin, malonyl gensistin and flavones rutin, which were increased gradually under light condition, but in the dark, daidzin and malonyl daidzin were the major metabolites and did not change markedly with time. In the roots, no isoflavone was detected in the dark, while five isoflavones were found in the light-grown seedlings, and all root sections were predominated by daidzein and its conjugates. These results provided the evidence that light strongly stimulates the accumulation of isoflavones in soybean seedling tissues.     

Profiling changes in metabolism of isoflavonoids and their conjugates in Lupinus albus treated with biotic elicitor

Liquid chromatography with ultraviolet and mass spectrometric detection was applied to monitor changes in profiles of isoflavonoid glycosides and free isoflavonoid aglycones in Lupinus albus L. Four isoflavonoid aglycones, fourteen isoflavonoid glycosides, four flavonol glycosides and flavone glycoside were identified in lupin tissue after LC/ESI/MS analyses. An elicitor preparation from purified yeast cell wall was used to inject the shoots of 3-week old seedlings or to infiltrate the cut lupin leaves. Qualitative and quantitative changes of isoflavonoids were measured at different time points after elicitation. In elicited lupin seedlings increased amounts of prenylated isoflavone aglycones were identified. The concentrations of glycosidic conjugates of isoflavones present in plant tissue were less affected.     

一氧化氮参与盐胁迫下长春花酚类代谢的调控研究

为研究外源一氧化氮(NO)调控盐胁迫下长春花中酚类化合物的响应,采用液相色谱—质谱联用(LCMS)技术靶向分析梯度浓度硝普纳(SNP)处理对盐胁迫下长春花幼苗根、茎、花、叶4个部位中酚类化合物组分及含量水平的变化。结果共鉴定出L-苯丙氨酸和18种酚类物质,C6C1类5种、C6C3类5种、C6C3C6类8种,其中原儿茶酸、绿原酸、槲皮素在长春花根、茎、花、叶4个部位中均存在;不同浓度SNP处理后长春花不同部位酚类化合物响应积累明显不同,其中C6C1和C6C3小分子酚酸类化合物主要积累在根和茎中,C3C6C3类主要富集在花和叶中;L-苯丙氨酸在茎、叶中相对含量较高,盐胁迫下茎中含量显著升高,且随外源NO浓度增大呈下降趋势。外源NO影响盐胁迫下植物器官中酚类化合物的积累和变化,其中根和茎响应敏感,从种类和相对含量的角度,茎和叶更适合检测酚类化合物。     

UHPLC/HR‐ESI‐MS/MS Profiling of Phenolics from Tunisian Lycium arabicum Boiss. Antioxidant and Anti‐lipase Activities’ Evaluation

This study was performed in the aim to evaluate nine different extracts from Tunisian Lycium arabicum for their total phenolic and total flavonoid contents, phytochemical analyses as well as their antioxidant and anti‐lipase activities. The in vitro antioxidant property was investigated using three complementary methods (DPPH, ferric reducing antioxidant power (FRAP), and β‐carotene‐linoleic acid bleaching assays) while anti‐lipase activity was evaluated using 4‐methylumbelliferyl oleate method. From all of the tested extracts the most potent found to be the polar MeOH extracts especially those of stems and leaves. In order to investigate the chemical composition of these extracts and possible correlation of their constituents with the observed activities, an UHPLC/HR‐ESI‐MS/MS analysis was performed. Several compounds belonging to different chemical classes were tentatively identified such as rutin and kampferol rutinoside, the major constituents of the leaves, and N‐caffeoyltyramine, lyciumide A, N‐dihydrocaffeoyltyramine as well as fatty acids: trihydroxyoctadecadienoic acid and hydroxyoctadecadienoic acid isomers were detected abundantly in the stems. These results showed that the MeOH extracts of stems and leaves of L. arabicum can be considered as a potential source of biological active compounds.     

Profiling of phenolic glycosidic conjugates in leaves of Arabidopsis thaliana using LC/MS

Profiling of plant secondary metabolites is still a very difficult task. Liquid chromatography (LC) or capillary electrophoresis hyphenated with different kinds of detectors are methods of choice for analysis of polar, thermo labile compounds with high molecular masses. We demonstrate the applicability of LC combined with UV diode array or/and mass spectrometric detectors for the unambiguous identification and quantification of flavonoid conjugates isolated from Arabidopsis thaliana leaves of different genotypes and grown in different environmental conditions. During LC/UV/MS/MS analyses we were able to identify tetra-, tri-, and di-glycosides of kaempferol, quercetin and isorhamnetin. Based on our results we can conclude that due to the co-elution of different chemical compounds in reversed phase HPLC systems the application of UV detectors does not allow to precisely profile all flavonoid conjugates existing in A. thaliana genotypes. Using MS detection it was possible to unambiguously recognize the glycosylation patterns of the aglycones. However, from the mass spectra we could not conclude neither the anomeric form of the C-1 carbon atoms of sugar moieties in glycosidic bonds between sugars or sugar and aglycone nor the position of the second carbon involved in disaccharides. The applicability of collision induced dissociation techniques (CID MS/MS) for structural analyses of the studied group of plant secondary metabolites with two types of analyzers (triple quadrupole or ion trap) was demonstrated.     

Distribution of chromenes and benzofurans in Encelia californica

Two populations of Encelia californica (Asteraceae) were analysed for chromenes and benzofurans on an organ specific basis using HPLC. Both classes of compounds were present in all parts of the plant studied including roots, stems, leaves, capitula and achenes. The distribution patterns were less complex in roots and achenes when compared to stems, leaves and capitula which yielded three chromenes and two benzofurans. In addition to the chromenes and benzofurans the capitula afforded a sesquiterpene lactone of the eudesmanolide type which was absent in the other parts of the plant.     

Secondary metabolites in in vitro cultured plants of the genus Drosera

Extracts from plantlets of different species of the genus Drosera, grown as in vitro cultures, were evaluated for the level of phenolic secondary metabolites from the group of naphthoquinones and flavonols. The profiles of natural products in the extracts obtained from different species were monitored by HPLC with UV detection at 260 and 330 nm. On the basis of the data obtained, Drosera binata, the species with the highest amount of plumbagin, was selected for further studies. The most effective method of extraction of quinones was established and the composition of phenolic secondary metabolites in the tissues was determined. For the identification of phenolic compounds, HPLC-UV and HPLC-ESI/MS were applied.     

LC/MS profiling of flavonoid glycoconjugates isolated from hairy roots, suspension root cell cultures and seedling roots of Medicago truncatula

Hairy roots and suspension cell cultures are commonly used in deciphering different problems related to the biochemistry and physiology of plant secondary metabolites. Here, we address about the issue of possible differences in the profiles of flavonoid compounds and their glycoconjugates derived from various plant materials grown in a standard culture media. We compared profiles of flavonoids isolated from seedling roots, hairy roots, and suspension root cell cultures of a model legume plant, Medicago truncatula. The analyses were conducted with plant isolates as well as the media. The LC/MS profiles of target natural products obtained from M. truncatula seedling roots, hairy roots, and suspension root cell cultures differed substantially. The most abundant compounds in seedlings roots were mono- and diglucuronides of isoflavones and/or flavones. This type of glycosylation was not observed in hairy roots or suspension root cell cultures. The only recognized glycoconjugates in the latter samples were glucose derivatives of isoflavones. Application of a high-resolution mass spectrometer helped evaluate the elemental composition of protonated molecules, such as [M + H]⁺. Comparison of collision-induced dissociation MS/MS spectra registered with a quadrupole time-of-flight analyzer for tissue extracts and standards allowed us to estimate the aglycone structure on the basis of the pseudo-MS³ experiment. Structures of these natural products were described according to the registered mass spectra and literature data. The analyses conducted represent an overview of flavonoids and their conjugates in different types of plant material representing the model legume, M. truncatula.     

Simultaneous quantification of phytohormones in fermentation extracts of Botryodiplodia theobromae by liquid chromatography–electrospray tandem mass spectrometry

Fermentation broth and biomass from three strains of Botryodiplodia theobromae were characterized by high performance liquid chromatography–electrospray tandem mass spectrometry (HPLC–ESI–MS/MS) method, in order to quantify different phytohormones and to identify amino acid conjugates of jasmonic acid (JA) present in fermentation broths. A liquid–liquid extraction with ethyl acetate was used as sample preparation. The separation was carried out on a C18 reversed-phase HPLC column followed by analysis via ESI–MS/MS. The multiple reaction monitoring mode was used for quantitative measurement. For the first time, indole-3-acetic acid, indole-3-propionic acid, indole-3-butyric acid and JA were identified and quantified in the ethyl acetate extracts from the biomass, after the separation of mycelium from supernatant. The fermentation broths showed significantly higher levels of JA in relation to the other phytohormones. This is the first report of the presence of gibberellic acid, abscisic acid, salicylic acid and the cytokinins zeatin, and zeatin riboside in fermentation broths of Botryodiplodia sp. The presence of JA-serine and JA-threonine conjugates in fermentation broth was confirmed using HPLC-ESI tandem mass spectrometry in negative ionization mode, while the occurrence of JA-glycine and JA-isoleucine conjugates was evidenced with the same technique but with positive ionization. The results demonstrated that the used HPLC–ESI–MS/MS method was effective for analysing phytohormones in fermentation samples.     

基于广泛靶向代谢组学分析黑老虎不同部位成分差异

为提高黑老虎(Kadsura coccinea)资源的综合利用率，采用广泛靶向代谢组学技术鉴定并分析了根、茎、叶代谢组分差异及高度富集成分。结果表明，在根、茎和叶中分别鉴定出642、650和619个代谢物，以酚酸、脂质、类黄酮和有机酸为主；叶与根、茎与根的共有成分分别为566和650个，显著差异成分有442和393个，主要为酚酸、类黄酮和脂质，差异代谢物在苯丙烷生物合成、黄酮与黄酮醇生物合成通路中显著富集。代谢物总丰度和次生代谢物丰度均表现为叶>根>茎，叶中酚酸、类黄酮和脂质及茎中酚酸积累量显著高于根，而氨基酸及其衍生物、萜类、木脂素、香豆素、生物碱的丰度在根中显著上调。因此，黑老虎根、茎、叶有大量共有成分，叶和茎中酚酸、叶中类黄酮和脂质高度富集，含有新绿原酸、绿原酸、槲皮素等多个丰度较高且具有重要生物活性化合物，具有较高利用价值。     

Variations in the essential oil composition from different parts of Coriandrum sativum L. cultivated in Tunisia

Essential oils obtained by hydrodistillation from different organs (flowers, leaves, stems and roots) of Tunisian coriander (Coriandrum sativum L.) was analyzed. GC and GC-MS analysis enabled us to identify 64 compounds and revealed great qualitative and quantitative differences between the analyzed parts. In all organs, the main compound was (E)-2-dodecenal, followed by (E)-2-tridecenal, gamma-cadinene, (Z)-myroxide, neryl acetate and eugenol. Multivariate analysis (PCA) revealed a high similarity in the essential oils composition between upper leaves and flowers in one hand and basal leaves, roots and stems on the other hand. Further, it has been reported an organ-dependant distribution of essential oil compounds.