Polar constituents from the aerial parts of Origanum vulgare L. Ssp. hirtum growing wild in Greece

From the polar extracts of Origanum vulgare L. ssp. hirtum 19 compounds have been isolated. The structures and relative stereochemistry have been elucidated by spectroscopic analysis and determined as apigenin, luteolin, chrysoeriol, diosmetin, quercetin, eriodictyol, cosmoside, vicenin-2, caffeic acid, p-menth-3-ene-1,2-diol 1-O-beta-glucopyranoside, thymoquinol 2-O-beta-glucopyranoside, thymoquinol 5-O-beta-glucopyranoside, thymoquinol 2,5-O-beta-diglucopyranoside, 12-hydroxyjasmonic acid, 12-hydroxyjasmonic acid 12-O-beta-glucopyranoside, lithospermic acid B, rosmarinic acid, 10-epi-lithospermic acid, and epi-lithospermic acid B. The three latter products display unusual stereochemistry of the 3,4-hydroxyphenyllactic acid unit(s), which to the authors' best knowledge has never been reported before in similar compounds. Moreover, lithospermic acid B (and its stereoisomers), p-menth-3-ene-1,2-diol 1-O-beta-glucopyranoside, 12-hydroxyjasmonic acid, and 12-hydroxyjasmonic acid 12-O-beta-glucopyranoside were isolated for the first time from Origanum species.     

Evaluation of barley landraces and selections from natural outcrosses ofH. vulgare ssp.spontaneum with ssp.vulgare for breeding in semi-arid areas

Summary Landraces of barley and tough rachis genotypes selected from natural crosses of ssp.spontaneum withssp. vulgare were evaluated in Cyprus under rainfed conditions with the best commercially grown grain varieties. Landraces, eliminated from cultivation around 40 years ago, gave grain yields comparable to those to the best commercially grown varieties, Athenais and Kantara. Also, selected genotypes from the natural hybrids gave grain yields approaching those of Athenais and Kantara, in spite of the fact that human interference was limited only to visual individual plant selection in natural habitats, followed by the evaluation of 44 genotypes in the present studies. Locally adapted landraces and selected genotypes from the natural hybrids expressed valuable measurable traits, such as 1000-grain weight, crude protein content, volume weight, lodging and disease resistance, which could be used in breeding programmes. The heading date of these 44 selected tough rachis genotypes was within a narrow range, which lies within the optimum range determined by long term studies for Cyprus. Furthermore, non-visible gene complexes for adaptation to moisture, heat and other stresses present in land races and natural hybrids could be exploited for breeding in semi-arid areas. These natural hybrids apparently contributed towards maintaining genes present in landraces which became extinct long time ago.     

Effects of genotype and weed control on the nutrient composition of winter spelt (Triticum aestivum ssp. spelta L.) and common wheat (Triticum aestivum ssp. vulgare)

The renaissance of spelt is associated with the development of alternative farming and a trend to maintain biodiversity. The present study carried out a comparison between two methods of weeding (mechanical and chemical) as well as a cereal cultivar influence on chemical composition of spelt and wheat. The research material was taken from field experiments carried out during three years. In order to assess the chemical composition, the following cereal cultivars were used: lines of winter spelt – STH 8 and STH 11 and a winter common wheat cv. Tonacja. In all cases, the overall protein content in spelt was significantly higher than in common wheat. Spelt grain is also richer in fats than common wheat. A visible tendency to higher concentration of raw fibre was observed when mechanical weeding was applied. Also, mechanical weeding as well as the variety significantly influenced β-glucan content. The study of the impact of a weeding method and a variety does not clearly show the direction of the amino acid content changes. The research indicates that there are ample opportunities to shape the level of nutrients, which decide about spelt grain application in the food industry through agronomic factors (methods of weeding and cultivar).     

Chemical compositions and antibacterial effects of essential oils of Turkish oregano (Origanum minutiflorum), bay laurel (Laurus nobilis), Spanish lavender (Lavandula stoechas L.), and fennel (Foeniculum vulgare) on common foodborne pathogens

Chemical compositions and inhibitory effects of essential oils of Turkish oregano (Origanum minutiflorum O. Schwarz & P. H. Davis), bay laurel (Laurus nobilis L.), Spanish lavender (Lavandula stoechas subsp. stoechas L.), and fennel (Foeniculum vulgare Mill.) on Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, and Staphylococcus aureus were determined. After the essential oils were applied on the foodborne pathogens at doses of 0 (control), 5, 10, 20, 30, 40, 50, and 80 microL/mL, the resultant numbers of cells surviving were counted. Results revealed that all essential oils exhibited a very strong antibacterial activity against the tested bacteria (P < 0.05). Gas chromatography-mass spectrophotometry analyses revealed that carvacrol (68.23%), 1,8-cineole (60.72%), fenchone (55.79%), and trans-anethole (85.63%) were the predominant constituents in Turkish oregano, bay laurel, Spanish lavender, and fennel essential oils, respectively.     

Biotransformation and pharmacokinetics of 4-(3,4-dihydroxybenzoyloxymethyl)phenyl-O-beta-D-glucopyranoside, an antioxidant isolated from Origanum vulgare

4-(3,4-Dihydroxybenzoyloxymethyl)phenyl- O-beta-D-glucopyranoside (OV-16) is a polyphenolic glycoside isolated from oregano (Origanum vulgare L.), which is a popular Chinese herb and a common spice in Western diet. To understand the biotransformation and pharmacokinetics of OV-16, rats were orally administered OV-16 and oregano decoction. Blood samples were withdrawn at specific time points. The presence of OV-16 and its metabolites protocatechuic acid (PCA) and p-hydroxybenzyl alcohol (HBA) in serum were determined by HPLC method, whereas their conjugated metabolites were assayed indirectly through hydrolysis with beta-glucuronidase and sulfatase. Our results showed that when OV-16 was orally administered, free forms of OV-16, PCA, and HBA were not present in blood and the major metabolites were the glucuronides/sulfates of PCA and HBA sulfate. The serum metabolites of OV-16 exhibited free radical scavenging activity. When oregano decoction was given, the glucuronides and sulfates of PCA were the major metabolites in blood.     

Exploring the Serbian GenBank barley (Hordeum vulgare L. subsp. vulgare) collection for powdery mildew resistance

Unlocking resistance genes in genbank collections are of prime importance for securing sustainable crop production. In this regard, the Serbian GenBank barley collection, comprising 93 local landraces and 36 commercial cultivars and elite barley breeding lines, was screened for novel resistances to powdery mildew (Blumeria graminis f. sp. hordei) using a set of 28 isolates with a wide spectrum of virulences/avirulences. No line was resistant to all the isolates, but one and three accessions showed resistance to 27 and 26 isolates, respectively. Twenty landraces (21.51 %) and ten cultivars (27.78 %) exhibited resistance to 50 % of the isolates. Infection type 2 was most frequent among resistant accessions. Nine B. graminis isolates were sufficient for gene postulation in 73 barley lines. In total, thirty-five different resistance spectra were recorded and the following known resistance genes were postulated namely, Mlra,Mlh,Mla12, Mla7(Mlu), Mlg, MlLa, Mla6, Mla7, Mlt, Mla22, Mlat, Mla1, Mlk. The majority of resistance profiles was constituted by only one line. Unidentified genes alone or in combination were proposed for twenty landraces and six cultivars. This report demonstrated that the barley collection held at the Serbian GenBank could be exploited as a new source for powdery mildew resistance.     

小麦-大麦大穗大粒渐渗系WB13的分子细胞学鉴定

小麦新种质WB13是普通小麦(Triticum aestivum L.)品种7182与农家二棱大麦(Hordeum vulgare ssp.distichon Hsü.)杂交后回交多代衍生而来的大穗大粒材料。为了明确WB13的遗传基础,本研究利用形态学、细胞学、分子标记及特异片段回收测序等技术对其进行了鉴定。结果表明,采用小麦不同同源群简单重复序列(simple sequence repeat,SSR)标记对WB13和小麦7182进行遗传背景分析发现,两者遗传相似系数(genetic similarity coefficient,GS)达97.0%,田间表现为大穗、大粒,综合农艺性状较好;根尖细胞染色体数目为2n=42,以大麦基因组DNA为探针进行基因组原位杂交(genomic in situ hybridization,GISH)未出现杂交信号;利用大麦特异序列标签位点(sequence-tagged site,STS)标记对WB13和农家二棱大麦进行扩增,发现ABG054(4H)和ABC305B(7H)两个标记在WB13中扩增出了大麦特征条带(分别记为WS1和WS3),经测序及序列比对,发现其与扩增到的大麦序列分别具有100%和98%的相似性,与EMBL数据库中的序列比对,与两者有相似性的全为大麦的序列。利用大麦4H和7H染色体上的35对SSR引物对WB13及其亲本进行扩增,发现与千粒重相关的标记MGB396(4H)在WB13中扩增出了大麦的特异条带。综合以上结果确定WB13含有大麦4H和7H染色体的遗传物质,为小麦-大麦渐渗系材料,且4H染色体渗入片段中可能携带与千粒重相关的有益基因。本研究确定了WB13为小麦-大麦杂种后代,此材料的育成丰富了小麦-大麦中间材料和大穗大粒材料的种质资源。同时本研究在分子水平上初步揭示了WB13大穗大粒特性的成因,为后续构建遗传分析群体进行相关数量性状(quantitative trait loci,QTL)定位及推动WB13的研究利用积累了基础资料。     

Antibacterial activity of Coriandrum sativum L. and Foeniculum vulgare Miller Var. vulgare (Miller) essential oils

Essential oils were extracted from the fruits of Coriandrum sativum L. and Foeniculum vulgare Miller var. vulgare (Miller) and assayed in vitro for antibacterial activity to Escherichia coli and Bacillus megaterium, bacteria routinely used for comparison in the antimicrobial assays, and 27 phytopathogenic bacterial species and two mycopathogenic ones responsible for cultivated mushroom diseases. A significant antibacterial activity, as determined with the agar diffusion method, was shown by C. sativum essential oil whereas a much reduced effect was observed for F. vulgare var. vulgare oil. C. sativum and F. vulgare var. vulgare essential oils may be useful natural bactericides for the control of bacterial diseases of plants and for seed treatment, in particular, in organic agriculture. The significant antibacterial activity of essential oils to the bacterial pathogens of mushrooms appears promising.     

Kinetics of carotenoids degradation during the storage of einkorn (Triticum monococcum L. ssp. monococcum) and bread wheat (Triticum aestivum L. ssp. aestivum) flours

To evaluate the effect of storage temperature, the degradation kinetics of carotenoids in wholemeal and white flour of einkorn cv. Monlis and bread wheat cv. Serio, stored at -20, 5, 20, 30, and 38 degrees C, was assessed by normal-phase high-performance liquid chromatography. In Monlis, the carotenoids content (8.1 and 9.8 mg/kg for wholemeal and white flour, respectively) was 8-fold higher than in Serio (1.0 and 1.1 mg/kg). Only lutein and zeaxanthin were detected in bread wheat, while significant quantities of (alpha and beta)-carotene and beta-cryptoxanthin were observed in einkorn. Carotenoids degradation was influenced by temperature and time, following first-order kinetics. The degradation rate was similar in wholemeal and white flour; however, loss of lutein and total carotenoids was faster in Serio than in Monlis. The activation energy E(a) ranged from 35.2 to 52.5 kJ/mol. Temperatures not exceeding 20 degrees C better preserve carotenoids content and are recommended for long-term storage.     

Leaf photosynthesis,chlorophyll fluorescence and ion content of barley (Hordeum vulgare) in response to salinity

Eight barley genotypes contrasting in their salinity tolerance were assessed for their chlorophyll fluorescence, photosynthetic performance, lipid peroxidation level and ionic content. A pot experiment was conducted in Borj-Cedria (Tunisia), in a wire house with a glass roof to avoid rainfall. The assay consisted of three treatments (0, 100 and 200 mM NaCl) with eight barley genotypes following a completely randomized design. Each pot was considered as one replicate and nine replicates were used for each genotype and treatment. The salt-tolerant genotypes Kerkna and Tichedrett recorded the highest tolerance for photosynthesis and potassium accumulation, whereas the susceptible genotypes were mostly affected at severe salinity. Contrasting behavior was noted for fluorescence attributes, while PSII yield was unchanged reflecting good protection against photodamage. Photosynthetic performance, enhanced water use efficiency, maintained leaf K⁺ and oxidative defense remain the key components for tolerance mechanisms. Salt-tolerant barley could be suitable for management of salt-affected soils.     

Investigation of the Origanum onites L. essential oil using the chorioallantoic membrane (CAM) assay

The in vivo test on the chorioallantoic membrane of the fertilized hen's egg (CAM assay) is a current method to determine antiangiogenic, antiinflammatory activity and toxic effects of individual compounds or complex plant extracts. The method is used for testing natural compounds in small amounts for revealing various modes of action and the complex mechanisms related to angiogenesis and inflammation. Furthermore, possible side effects such as membrane irritation, toxic, and anticoagulant properties of the investigated material in question can be detected. For the evaluation, the essential oil obtained by hydrodistillation of the aerial parts of Origanum onites L., a common spice and medicinal plant, was tested for its effect in the chorioallantoic membrane (CAM) assay. The essential oil composition was revealed by means of gas chromatography-mass spectrometry (GC-MS). Eighty three components were identified, representing 99.1% of the total oil. Carvacrol, thymol, p-cymene, and gamma-terpinene were found as major components and were also individually tested in the CAM assay. Along with the monoterpenes carvacrol and thymol, their methyl ether derivatives were also examined for comparison of their physiological action. Neither the essential oil nor its components showed any pronounced antiinflammatory or antiangiogenic property in the CAM assay, at 10-250 microg/pellet. However, the irritant effect of the essential oil was linked to thymol in a dose-response fashion, up to 10 microg/pellet, where it was still showing irritation.     

A case study of seed exchange networks and gene flow for barley (Hordeum vulgare subsp. vulgare) in Morocco

Local patterns of seed regeneration and trade that occur outside the formal breeding sector (seed exchange networks) can have a strong influence on the genetic diversity and evolution of traditional crop varieties. Despite this, little is known about the extent to which seed exchange networks influence gene flow and genetic structure in traditional crop varieties. Here we study barley (Hordeum vulgare subsp. vulgare) in rural communes of Northern Morocco in 2008 and 2009. We quantified seed regeneration and exchange by farmers within the seed exchange network using structured interviews. Using SSR markers, we also quantified the neutral genetic diversity and structure of a complex of traditional varieties referred to as Beldi that is managed in this exchange network. The majority of farmers (>88 %) report cultivating Beldi. Most seeds of Beldi (70–90 %) are maintained on-farm, while the remainder of seeds are obtained from local markets within the commune. Beldi has high genetic diversity and there is weak but significant genetic structure between communes (F_ST = 0.031). From SSR marker data there is evidence of a high level of gene flow between communes not reported in interviews. Seeds purchased in local markets likely represent seeds from a larger geographic region, leading to lower genetic structure among communes than expected based on the reported level of on-farm seed regeneration and local sourcing of seed. We discuss the implications of this seed exchange network for the conservation of traditional barley varieties in the study region.     

Identification of three novel polyphenolic compounds, origanine A-C, with unique skeleton from Origanum vulgare L. using the hyphenated LC-DAD-SPE-NMR/MS methods

Identification of new compounds especially those with new skeletons from plant kingdom has long been a vital aspect for understanding phytochemistry, plant metabolisms and discovering new bioactive compounds. In this study, we identified and isolated three novel polyphenolic compounds, origanine A-C, from a well-researched plant Origanum vulgare L. using the hyphenated LC-DAD-SPE-NMR/MS methods. Based on the combined information from UV-visible, accurate mass and 2D NMR spectra together with computational calculations, we found that these compounds all had a novel skeleton of cyclohexenetetracarboxylic acids attached with some well-known bioactive moieties including 3,4-dihydroxyphenyl, 4-(β-d-glucopyranosyloxy)benzyl alcohol (gastrodin), and 3-(3,4-dihydroxyphenyl)lactic acid (danshensu) residues. These findings provided crucial information to fill the gaps in our knowledge in terms of the plant secondary metabolism. This study also indicated the necessity for further research in plant secondary metabolism for even well-studied plants and demonstrated the powerfulness of the hyphenated LC-DAD-SPE-NMR/MS methods for comprehensive analysis of plant metabolites in particular for discovering new natural compounds.     

Study of metallopeptidase isozymes from malted barley (Hordeum vulgare cv. Morex)

It has been reported that germinated barley contains peptidases that are sensitive to metal-chelating agents; however, none of these enzymes have been isolated, nor have their roles in germinated barley been investigated. Anion-exchange chromatography and chromatofocusing have been used to isolate a group of peptidases from barley (Hordeum vulgare cv. Morex) green malt that are sensitive to metal-chelating agents. Their activities were studied using one- and two-dimensional polyacrylamide gel electrophoresis. When analyzed on two-dimensional PAGE gels that contained gelatin as substrate, the enzymes separated into three major and approximately six minor activity spots with acidic pI values. The enzymes were optimally active against the gelatin substrate at pH 8.0 and were completely inhibited by 1,10-phenanthroline and EDTA, indicating that they belonged to the metallopeptidase class (EC 3.4.24.x). After the enzymes were inhibited with EDTA, the activities were recovered in the presence of low concentrations of metal ions. The hydrolysis of gelatin substrate was also impaired by the presence of reducing agents. The metallopeptidases readily digested, in vitro, the barley prolamine D hordein, indicating that they may be involved in degrading storage proteins during barley germination.     

HPLC analysis of flavonoids and secoiridoids in leaves of Ligustrum vulgare L. (Oleaceae)

Identification and quantification of flavonol glycosides and secoiridoids was carried out on leaves of Ligustrum vulgare L. (Oleaceae) by means of HPLC-DAD and HPLC-MS analysis. In addition to previously reported secoiridoids (oleuropein, ligustaloside A, ligustaloside B, and ligstroside) four kaempferol glycosides (kaempferol 3-O-glucoside 7-O-rhamnoside, kaempferol 3, 7-O-dirhamnoside, kaempferol 3-O-rhamnoside, and kaempferol 3-O-glucoside) and two quercetin glycosides (quercetin 3-O-glucoside 7-O-rhamnoside and quercetin 3,7-O-dirhamnoside) were present in leaves of L. vulgare L. Although secoiridoids accounted for nearly the 76% of the total leaf polyphenols content (with ligustaloside A as the main component), kaempferol glycosides were also accumulated in the leaves of L. vulgare L. to a relatively high extent (23%). Contribution of quercetin derivatives was minor under our experimental conditions. Our findings suggest that flavonol glycosides may have a central role in both the ecology and the biology of L. vulgare L.     

大白菜表达序列标签SSR标记分析*

对大白菜（Brassica campestris L．ssppekinensis）13007个EST序列进行拼接共得到7714个片段重叠群（contig），其中10．4％（890个）非冗余EST含有SSR标记（EST-SSR），标记间平均间隔距离为3．9kb，单、双和三核苷酸重复序列大约各占1／3左右。CDS所包含的三核苷酸重复序列最多，而且AAG／CTT重复序列的含量最高。通过同源性比较分析，获得功能已知的SSR-ESTs774个，共计含有846个SSR，其中50．2％位于5＇-UTR，31．4％位于3＇-UTR，18．4％位于CDS。实验选取123个SSR座位设计引物，利用芜菁花叶病毒（Turnip mosaic virus，TuMV）的高抗和高感大白菜F6代自交系材料各2份，分析EST-SSR标记多态性。结果表明，其中37个标记可进行有效扩增，但是没有表现出多态性，23个标记（18．7％）至少在1份材料中具有多态性。对这些EST-SSR标记在大白菜和其它芸薹属植物的系统发育关系分析、遗传作图和基因定位的应用进行了讨论。     

小白菜硝酸还原酶基因的克隆与初步鉴定

蔬菜是一种易于富集硝酸盐的植物性食物，人体摄入的硝酸盐大部分（70％～80％）来自蔬菜。然而，我国蔬菜生产中氮肥用量迅速提高，造成了蔬菜，特别是喜氮的叶菜类蔬菜硝酸盐含量过高。不同蔬菜硝酸盐含量存在很大差异，同一种蔬菜不同品种体内硝酸盐含量也存在差异。不同器官累积硝酸盐的能力不同，硝酸盐累积取决于硝态氮的吸收与同化的结果。     

rDNA polymorphism of foxtail millet (Setaria italica ssp. italica) landraces in northern Pakistan and Afghanistan and in its wild ancestor (S. italica ssp. viridis)

Ribosomal DNA (rDNA) spacer length polymorphism was studied in foxtail millet (Setaria italica ssp. italica) landraces from Pakistan and Afghanistan and in its wild ancestor (S. italica ssp. viridis) from Pakistan by PCR-based methods. Sequence polymorphism was also investigated for accessions selected based on the observed length polymorphism. The PCR-based length polymorphism and sequence polymorphism of rDNA intergenic spacer (IGS) clearly demonstrated genetic differentiation between cultivated and wild forms in the region. Genetic differentiation was observed between different areas to some extent in the cultivated form, and between different regions in the wild form of northern Pakistan. Based on the results, we discuss the genetic differentiation of foxtail millet and wild ancestor in this region and possible utility of rDNA markers to trace the dispersal of this crop in the region.     

Epithiospecifier protein from broccoli (Brassica oleracea L. ssp. italica) inhibits formation of the anticancer agent sulforaphane

In some cruciferous plants, epithiospecifier protein (ESP) directs myrosinase (EC 3.2.3.1)-catalyzed hydrolysis of alkenyl glucosinolates toward epithionitrile formation. Here, for the first time, we show that ESP activity is negatively correlated with the extent of formation of the health-promoting phytochemical sulforaphane in broccoli (Brassica oleracea L. ssp. italica). A 43 kDa protein with ESP activity and sequence homology to the ESP of Arabidopsis thaliana was cloned from the broccoli cv. Packman and expressed in Escherichia coli. In a model system, the recombinant protein not only directed myrosinase-dependent metabolism of the alkenyl glucosinolate epi-progoitrin [(2S)-2-hydroxy-3-butenyl glucosinolate] toward formation of an epithionitrile but also directed myrosinase-dependent hydrolysis of the glucosinolate glucoraphanin [4-(methylsulfinyl)butyl glucosinolate] to form sulforaphane nitrile, in place of the isothiocyanate sulforaphane. The importance of this finding is that, whereas sulforaphane has been shown to have anticarcinogenic properties, sulforaphane nitrile has not. Genetic manipulation designed to attenuate or eliminate expression of ESP in broccoli could increase the fractional conversion of glucoraphanin to sulforaphane, enhancing potential health benefits.     

结球白菜结球前期基因表达序列标签（EST）分析

以结球白菜(Brassica rapa L.ssp．pekinensis)结球前期心叶为材料，构建了结球前期球叶cDNA文库，随机挑选克隆单向单次测序后，共得到1162条峰图良好和插入片段长度大于150bp的可用序列。BLASTX及BLASTN序列比对分析表明，94．8％(1102／1162)的表达序列标签(EST)可在蛋白质或核苷酸水平上找到同源类似物。同源性最大的蛋白质按物种来源分析后发现，大约77％的功能已知蛋白质来自拟南芥(Arabidopsis thaliana)，另外还有60个EST与已发表的植物基因没有同源性，这一部分EST对于研究结球白菜独特的生理和形态发育途径以及开展基因组分子作图具有重要的意义。同源蛋白质按其功能进行分类表明，参与蛋白质合成过程的酶或蛋白质的EST数量最多，其次是参与能量代谢过程(包括光合作用)的EST序列。在核苷酸水平上，全部EST中只有51％的同源物来自拟南芥。对上述1162条EST进行片段重叠群分析(contig analysis)共获得895个非冗余片段重叠群，其中723个仅由一个EST组成(即singletons)，表明结球白菜结球过程中表达的基因种类繁多。大量EST的获得为进一步了解结球白菜结球机制及获取相关基因提供了重要的序列信息。