Origanum vulgare L.leaf extract alleviates finasteride-induced oxidative stress in mouse liver and kidney

Objective:To investigate the hepatorenoprotective effects of Origanum vulgare L.against finasteride-induced oxidative injury in the liver and kidney of mice.Methods:Liquid chromatography-electrospray ionization-tandem mass spectrometry(LC-ESI/MS)analysis was utilized to yield a fingerprint of Origanum vulgare polyphenolic constituents.Thirty BALB/c mice received 0.5 mL/day distilled water,finasteride(25 mg/kg/day for 10 d),and 100,200,or 400 mg/kg/day finasteride+Origanum vulgare extract with 6 mice per group for five weeks.On day 36,liver and kidney function as well as pro-and antiinflammatory(IFN-γ,IL-12,IL-6,TNF-α,IL-1β,and IL-10)cytokines were measured.The total antioxidant status,nitric oxide(NO),and malondialdehyde levels as well as the activities of NO synthase and catalase were also evaluated.Histopathological study was conducted to assess the effect of Origanum vulgare extract on finasteride-induced renal and hepatic toxicities.Results:Twenty-five major polyphenolic compounds were identified in the Origanum vulgare extract by LC-ESI/MS.Origanum vulgare extract,especially at 200 and 400 mg/kg/day doses,significantly improved liver and kidney biochemical indices,decreased inflammatory cytokines,increased total antioxidant status and NO synthase and catalase activities,as well as decreased plasma NO and malondialdehyde levels in a dose-dependent manner as compared to the finasteride group.Histopathological results further confirmed the protective effect of Origanum vulgare extract.Conclusions:Origanum vulgare extract ameliorates finasterideinduced hepatic and renal biochemical and histopathological alterations,and restores antioxidant/oxidant balance.

Influence of Abiotic Elicitors on Accumulation of Thymol in Callus Cultures of Origanum vulgare L.

Callus cultures of Origanum vulgare L. were established from leaf discus on Murashige and Skoog （MS） medium containing different levels of growth regulators, i.e., 2,4-Dichlorophenoxyacetic acid （2,4-D）, Naphthalene acetic acid （NAA）, Benzyl Adenine （BA） and Kinetin （Kn） and incubated under dark condition. Callus tissues were employed to study the influence of abiotic elicitors on the production of thymol. Constant weights of callus （300 mg） were cultured on accumulation medium treated separately with each one of elicitors used （50 g/L sucrose, 200 mg/L NaC1 and 50 or 100 mg/L proline）. The fresh and dry weights of callus were recorded after six weeks. The result indicated that maximum production of fresh and dry callus weight were 1,014 mg and 46.20 mg respectively achieved at 0.5 mg/L 2,4-D and 3 mg/L BA adding to the medium. Dry callus tissues were extracted with 70% methanol and analyzed by HPLC to determine the concentrations of thymol. The addition of abiotic elicitors to MS medium caused significant reduction in fresh weight of callus compared with control treatment. The concentration of thymol in the callus cultured on control treatment was 146.6 ppm. The data showed that 50 or 100 mg/L proline produced the highest yield of thymol 181.48 ppm and 174.58 ppm respectively, followed by sucrose 162.9 ppm, whereas the treatment with NaCI caused reduction in thymol concentration to percentage of 50.56% compared with the control.

Micropropagation and Acclimatization of Common Oregano (Origanum vulgare L. Subsp. vulgare) by Shoot Tip Culture

Origanum vulgare L. is a commercially valued species with remarkable biological properties. It is subject to over-exploitation practices that seriously threaten its sustainability for future generations. Thus, micropropagation serves as a tool for the protection and domestication of this species. In this study, we established an in vitro vegetative propagation protocol for Origanum vulgare. This is done through the axillary bud technique by carrying out various tests. Six culture media (MS, MSm, N30K, SD, SH and B5) were tested. Therefore, SD was chosen for the following experiments. Seven cytokinins (adenine (Ad), N6-(2-isopentenyl) (2ip), zeatin (Zeat), kinetin (Kin), benzyladenine (BAP), 1,3-diphenylurea (DPU) and thidiazuron (TDZ) at 5 concentrations (0.44, 1.33, 2.22, 3.11 and 4.44 μM/L) were evaluated. Thus, Kin at 3.11 μM allowed high regeneration of vitroplants, optimal elongation, total rooting of explants, maximum bud multiplication, and absence of hyperhydric explants. In fact, the integration of auxins (indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), and 1-naphthaleneacetic acid (NAA)) into the culture medium and their combinations with 3.11 μM Kinetin contributed to the optimization of the root part. Thus, it was improved in particular in the case of 3.11 μM Kin and 6.27 μM IBA. Three polyamines (Putrescine, Spermidine and Spermine) at different concentrations (1.134, 3.402, 5.67, 7.938 and 11.34 μM/L) combined at 3.11 μM Kin and 6.27 μM IBA were tested. In fact, 1.304 μM putrescine was considered to be the most suitable for in vitro culture of explants, since it allowed optimal propagation of buds and roots, also a high rate of regeneration and rhizogenesis. GA3 at 1.15 μM combined with 3.11 μM Kin and 6.27 μM IBA permitted maximum bud multiplication. The acclimatization was carried out successfully using vitroplants showing good foliar and root development. Thus, three months after acclimatization, the seedlings were transferred into large pots under natural light and temperature conditions. Almost all acclimatized plants developed flowers in the first year between May and July.

Chemical Composition and Antiviral Effect of Extracts of Origanum vulgare

This study aimed determine the activity of aqueous and ethanolic extracts of Origanum vulgare against some viruses of veterinary importance (bovine viral diarrhea virus (BVDV), equine arteritis virus (EAV), equine influenza virus (EIV), feline calicivirus (FCV), canine distemper virus (CDV), canine adenovirus (CAV), and canine cororavirus (CCoV) by evaluating the possibility of inhibition of viral particles production. The aqueous extract from 1600 μg/mL did not show cytotoxicity for all cellular lineages evaluated, Madin Darby bovine kidney cells (MDBK), Rabbit kidney cells (RK 13), Madin Darby canine kidney cells (MDCK) and Crandell feline kidney cells (CRFK), and the ethanolic extract of Origanum vulgare was not toxic at 600 μg/mL. The addition of aqueous extract of Origanum vulgare in media resulted in a significant reduction of the EAV titer from 105.42 infecting dose for cellular culture at 50% (TCID50) to 102.09 TCID50/100 μL while in the presence of the ethanolic extract of Origanum vulgare in media resulted in a significant reduction of the EAV titer from 105.42 TCID50 to 100.79 TCID50/100 μL. To CDV the addition of aqueous extract resulted in a reduction from 102.00 TCID50 to 100.00 TCID50/100 μL while in the presence of the ethanolic extract titers were reduced from 102.00 TCID50 to 101.50 TCID50/100 μL. No significant differences in titers regarding the others analyzed viruses were detected. With respect to chemical analysis of the extracts of Origanum vulgare, were identified in the ethanol extract phenolics rosmarinic acid, caffeic acid, carnosol, p-coumaric acid, carnosic acid, luteolin, apigenin, kaempferol and quercetin. In aqueous extracts of Origanum vulgare were detected rosmarinic acid, p-coumaric acid carnosic acid, luteolin, apigenin, kaempferol and quercetin. The data obtained stimulate other biological assays in order to determine which compounds are responsible for the antiviral activity as well as which are the mechanisms involved. The results presented and the considerations we were able to draw from them allowed us to conclude that the ethanolic extract of Origanum vulgare demonstrated lower cell viability than the aqueous extract and has significant antiviral activity against EAV and the both aqueous and ethanolic extracts have antiviral action against CDV.

Allelopathy of Cold Water Extracts from Origanum vulgare ssp. vulgare L.

Secondary metabolites in medicinal plants could lead to discovery of new classes of herbicides. Recently aromatic plants have gained interest as a source of allelopathic secondary metabolites. Origanum vulgare ssp. vulgare L. infusions in hot water are used in folk medicine and possess proved beneficial biological activity. Plant-to-plant variability of metabolites due to genetic heterogeneity is established in Lamiaceae family. From this point of view, studies on plants from different geographic regions might reveal important sources of variability. The objective of this study was to evaluate allelopathic activity of cold water extracts made from the aerial parts of O. vulgare ssp. vulgare growing wild in Northeast Bulgaria in laboratory conditions. The allelopathic effect was evaluated using root elongation test and Allium cepa-test. Oregano extracts (17.5 g/l, 52.5 g/l) significantly decreased root length of Triticum aestivum L. (P ≤ 0.001). The root growth reduction could serve as a sign for presence of water soluble allelopathic secondary metabolites in the plant tested. Oregano (3.5 g/l) inhibited cell division in Allium root meristematic cells. The decline of the mitotic index indicates the occurrence of a cytotoxic effect. Oregano induced abnormalities in mitotic and interphase cells, so can be also considered as genotoxic. The observed macroscopic and microscopic effects of tested extracts indicated presence of water soluble allelochemicals in O. vulgare ssp. vulgare. This characteristic could be further studied as a possibility to be used in weed management programs.

Effect of Wild Marjoram (<i>Origanum vulgare</i>) Plant Extracts on Capacitation of Sheep Spermatozoa <i>in Vitro</i>

The purpose of this study was to evaluate the effect of the addition of Origanum vulgare extract to in vitro capacitation sperm medium (IVCSM). This study investigated the antioxidant and antimicrobial effects of O. vulgare extracts at different concentrations (0.3, 0.6, 1.2 μg/ml and 25.0, 50.0, 100.0 μg/ml, respectively) in IVCSM. Significant enhancements in semen quality parameters such as total motility, live and live capacitated sperm were found when O. vulgare extract was added as an antioxidant source (1.2 μg/ml). The treatment of spermatozoa with O. vulgare extract at the highest concentration (100 μg/ml) for 2 hrs without antibiotics improved sperm characteristics. In conclusion, incubation of sperm with O. vulgare extract in capacitation medium had beneficial effects on the characteristics of ram sperm.

<i>In Vitro </i>Antibacterial and Antifungal Activity of Methanol, Chloroform and Aqueous Extracts of <i>Origanum vulgare</i>and Their Comparative Analysis

The work reports antibacterial and antifungal activity of different solvent extracts of Origanum vulgare. The antimicrobial activity of methanol, chloroform and aqueous extracts were determined against nine different gram negative and gram positive bacterial strains and three fungal stains. The bacterial strains were Bacillus subtilis (ATCC 6633), Staphylococcus aureus (ATCC 29213), Micrococcus luteus (ATCC 9341), Pseudo-monas aeruginosa (ATCC 33347), Escherichia coli (ATCC 25922), Salmonella typhi (ATCC 19430), Shig-ella flexneri (ATCC 25929), Salmonella para typhi A (ATCC 9150) and Proteus mirabilis (ATCC 49565) and fungal strains were Aspergillus flavus, Aspergillus nigar and Aspergillus pterus. Agar well diffusion method was followed in this study. The comparative analysis of antibacterial activity reflects that among these three extracts, chloroform and methanol extracts shows promising result by exhibiting maximum anti-bacterial activity, whereas aqueous extract is not active against most of these strains. The analysis of anti-fungal activity reveals chloroform extract as most efficacious unlike methanol and Aqueous extracts.

Genetic Diversity of SSR Markers in Cultivated Hordeum vulgare L.in Qinghai Province

[Objective]The aim was to analyze genetic diversity of SSR markers in Hordeum vulgare L.in Qinghai Province and lay a foundation for screening and protecting some excellent H.vulgare cultivars.[Method]SSR markers were used to evaluate the genetic diversity of 42 cultivated H.vulgare from Qinghai Province.[Result]42 H.vulgare showed polymorphism in 7 SSR markers locus.A total of 24 alleles were identified,and the number of alleles per locus ranged from 1 to 6,with an average of 3.0.According to SSR markers polymorphism,42 H.vulgare could be divided into 4 groups,namely I,II,III and IV.[Result]The study indicated that cultivated H.vulgare from Qinghai Province is rich in genetic diversity,which will provide reference for selecting parent of H.vulgare breeding.

利用聚丙烯酰胺凝胶电泳和高效液相色谱法对Scarlett和Prestige大麦（大麦属vulgare L）的麦芽、麦汗以及啤酒中蛋白组分的比较研究

本研究利用聚丙烯酰胺凝胶电泳（SDS－PAGE）和反相高效液相色谱法（RP－HPLC），对Scarlett和Prestige大麦麦芽的蛋白组分（大麦醇溶蛋白、清蛋白和其他可溶性蛋白）进行了研究，同时跟踪分析了由这两种麦芽制成的麦芽汁以及啤酒中的蛋白质变化情况。此外，对这两个大麦品种进行了工业酿造规模的研究。 通过对反相高效液相色谱数据进行统计学分析，发现大麦醇溶蛋白在发芽期间存在一个水解过程，导致其含量降低，同时产生较少的疏水性缩氨酸。相比之下，清蛋白和其他可溶性蛋白质在发芽过程中含量会增加。一些麦芽水溶性蛋白会由大麦醇溶蛋白质水解产生。我们还观察分析了这两个品种麦芽蛋白组分数量上的差别。 聚丙烯酰胺凝胶电泳图显示出麦芽汁中的绝大部分组分在成品啤酒中也可以观测到。然而，高效液相色谱数据的化学计量分析显示Scarlett和Prestige大麦麦汁蛋白组分间存在数量上的差异。Scarlett大麦麦汁比Prestige麦汁中含有更多的蛋白质。与两种麦芽汁相比，两种成品啤酒样品中的蛋白相似性更强。 试验将最优方法成功用于两种大麦麦芽蛋白质组分的研究中，同时跟踪分析了发芽期间蛋白质组分的变化情况以及麦芽汁和成品啤酒中蛋白质组分的变化。

QTL analysis of flag leaf in barley (Hordeum vulgare L.) for morphological traits and chlorophyll content

To understand genetic patterns of the morphological and physiological traits in flag leaf of barley, a double haploid （DH） population derived from the parents Yerong and Franklin was used to determine quantitative trait loci （QTL） controlling length, width, length/width, and chlorophyll content of flag leaves. A total of 9 QTLs showing significantly additive effect were detected in 8 intervals on 5 chromosomes. The variation of individual QTL ranged from 1.9% to 20.2%. For chlorophyll content expressed as SPAD value, 4 QTLs were identified on chromosomes 2H, 3H and 6H; for leaf length and width, 2 QTLs located on chromosomes 5H and 7H, and 2 QTLs located on chromosome 5H were detected; and for length/width, I QTL was detected on chromosome 7H. The identification of these QTLs associated with the properties of flag leaf is useful for barley improvement in breeding programs.

Differential Responses of Antioxidative System to Soil Water Shortage in Barley (<i>Hordeum vulgare</i>L.) Genotypes

Drought is one of the major factors limiting the yield and quality of crops in the world. The activity of antioxidative system to tolerate the drought stress is significant in plants. In the present study, the activities and isoform profiles of catalase (CАТ), ascorbate peroxidase (APX), glutathione reductase (GR), and superoxide dismutase (SOD) were analyzed in four barley genotypes grown under soil water restriction. Drought stress caused increase in the activities of CАТ and SOD in all studied genotypes, while APX activity decreased. The total GR activity increased substantially in genotypes K 2778 and St.Garabag 7 and decreased in No. 77 local and St.Pallidum 596 genotypes under conditions of severe water stress. No detectable differences were observed in the isoenzyme pattern (the appearance of a new isoenzymes and disappearance of another one) between control plants and those subjected to soil drought. However, intensification of corresponding isoforms in electrophoretic spectra was observed in stressed barley leaves relative to watered ones. The obtained results possibly suggest that antioxidant protection in barley plants under drought conditions could be attributed mainly to SOD and CAT.

Anti-yeast activities of Origanum oil against human pathogenic yeasts

Outbreak of autoimmune diseases by pathogenic yeasts has led to a serious medical threat. As these organisms evolve resistance to existing antifungal drugs, the concern could be further compounded. The realm of plant derived products offers a wide spectrum of potentially valuable alternatives to the existing synthetic fungicides. Essential oils from sev-eral medicinal plants have been shown to exhibit pharmacological attributes. In the present study, anti-yeast properties of Oregano essential oil (OEO) were examined in vitro against four human patho-genic yeasts i.e., Candida albicans, Cryptococcus al-bidus, Cryptococcus neoformans and Rhodotorula ru-brum. OEO concentration of 200 μg/mL was found to be growth inhibitory against all four yeasts examined, thereby showing its potential to function as a natural anti-yeast agent.

Analysis and Identification of Flavanoids and Phenolcarbonic Acid in Extract Plant of <i>Clinopodium vulgare</i>

Among the plants used by Bulgarian plants traditional medicine is Clinopodium vulgare L. (Lamiaceae). The aim of this study is to identify natural substances like some flavanoids and phenolcarbonic acids discovered in the plant with another technic because of Phenol carbonic acids. Instrumentation: The chromatographic studies were performed on a liquid chromatography. The identification of the components was carried out according to the spectral modifications after addition of the reagents of standard shifts. The results presented in this report seemed appropriate for the identification of flavonoids and phenol carbonic acids in extracts of Clinopodium vulgare.

普通大麦中spontaneum和vulgare亚种染色体N带带型比较研究

本文研究了普通大麦（ＨｏｒｄｅｕｍｖｕｇａｒｅＬ．）中ｓｐｏｎｔａｎｅｕｍ亚种的Ｎ带带型，并与ｖｕｌｇａｒｅ亚种中的栽培大麦进行了比较，发现西亚种的Ｎ带带型存在一定的差异，这种差异主要表现在染色体长短臂中间带数目和位置的不同，而着丝粒带是一致的．因此两亚种Ｎ带带型差异是染色体种内多态性的表现，Ｎ带带型总体上的一致性一定程度上反映了它们间杂交的可亲和性．

大麦HvLEC1基因的克隆及其表达特征分析

植物LEC蛋白是NF-Y转录因子的一类B亚基,在植物胚状体形成过程中起重要作用。为了研究大麦小孢子体外培养形成胚状体的机理,本研究利用RACE技术在大麦中克隆了一个新的LEC基因,该基因cDNA全长为1004 bp,开放阅读框全长为597 bp,编码198个氨基酸,其蛋白1~59位氨基酸含有LEC结构域,命名为HvLEC1。HvLEC1在大麦的根、茎、叶和小孢子培养过程中均能表达,其中小孢子培养7 d时表达量最高,且HvLEC1在大麦品系BI04中的表达量比基19高,BI04愈伤产量也比基19高,表明HvLEC1表达量和愈伤产量有相关性,受盐胁迫后HvLEC1在大麦的根中快速上调表达,提示HvLEC1可能不仅参与小孢子胚状体发生,而且参与盐胁迫响应。

大麦cDNA-AFLP技术体系的优化及其应用

为构建适合分析大麦杂种优势的cDNA-AFLP技术体系,以4个大麦不育系、2个大麦恢复系以及按4×2配制的8个杂交种为材料,对影响大麦cDNA-AFLP技术体系的几个关键因素进行了研究。结果表明,大麦cDNA-AFLP技术按以下程序可得到多态性好的清晰条带:使用百泰克公司的TRIpure Reagent总RNA提取试剂可提取到高质量的叶片总RNA;用TaKaRa的M-MLV RTase反转录合成双链cDNA;用MseⅠ和EcoRⅠ酶切6h,再用T4连接酶15℃连接接头18h;预扩PCR反应的dNTP终浓度为0.2mmol.L-1,扩增20循环效果最好;预扩产物稀释20倍后进行选择性扩增,最终用6%的变性聚丙烯酰胺凝胶进行电泳分离条带。利用该cDNA-AFLP体系对参试杂交种及其亲本的谱带类型进行了分析,亲本与杂种之间共扩增出7种类型的谱带,分别是共同表达型(P1P2F1)、P1表达型(P1)、P2表达型(P2)、F1表达型(F1)、P2F1表达型(P2F1)、P1F1表达型(P1F1)和P1P2表达型(P1P2)。其中差异谱带类型可分为四种表达模式:(1)单亲显性表达型,包括P2F1表达(P2F1)和P1F1表达(P1F1);(2)单亲沉默表达型,包括仅P1表达(P1)和仅P2表达(P2);(3)杂种上调表达型,即仅F1表达(F1);(4)杂种下调表达型,即仅P1P2表达(P1P2)。

维生素C前体2-酮基-L-古龙酸二步混菌发酵研究新进展

维生素C(Vc)二步混菌发酵是我国首创具有自主知识产权的唯一应用于工业化生产Vc的微生物转化方法,该方法利用混合菌发酵L-山梨糖生产Vc前体物质-2-酮基-L-古龙酸,再经化学转化合成Vc;具有简化工艺,减少污染,降低能耗等优点。本文主要从产酸菌代谢关键酶、伴生菌胞外物质、组学以及外源添加物等方面综述Vc二步混菌发酵的最新研究进展,并提出进一步研究和探索的方向。

大麦种子萌发lncRNA-miRNA-mRNA差异表达分析及ceRNA调控网络构建

为解析长链非编码RNA(lncRNA)、microRNA(miRNA)和mRNA如何在ceRNA互作模式下调控大麦种子萌发,揭示大麦(Hordeum vulgare L.)种子萌发转录水平分子调控机制,基于大麦种子萌发相关的RNA-Seq数据,共鉴定出6447个差异表达的mRNA,661个差异表达的lncRNA,310个差异表达的miRNA;对差异表达的mRNA和lncRNA进行加权基因共表达网络(WGCNA)分析,各鉴定到1个与大麦种子萌发高度相关的共表达模块。模块内相关基因涉及水分响应、胚发育等生物学过程,以及MAPK信号转导、植物激素信号转导等信号通路。通过对lncRNA/mRNA和miRNA进行靶基因预测,得到168个miRNA-mRNA靶基因对以及310个miRNA-lncRNA靶基因对,进而构建了包含2个子网络的ceRNA调控网络,发现有16个miRNA、38个lncRNA以及18个mRNA可能具有ceRNA功能。最后,利用qRT-PCR验证了2个关系对:HORVU0Hr1G039470-miR2611-MSTRG.21252以及HORVU6Hr1G031480-miR1858-MSTRG.23227。

NaCl对大麦硝态氮吸收动力学特征的影响

采用营养液培养方法研究了不同盐分和氮水平对大麦‘鉴四’(Hordeum vulgare L.)生长及硝态氮(NO3?-N)吸收动力学参数特征的影响。结果表明:不同浓度NO3?-N与Na Cl预处理后,大麦对NO3?吸收符合离子吸收动力学模型,其吸收动力学参数表现为NO3?预处理浓度增加后,Vmax增大,Km值增加,但增加的幅度不一致。对高亲和力系统来说,所有预处理大麦的NO3?-N吸收曲线均符合Michaelis-Menten方程的描述。1 mmol?L?1NO3?-N预处理中,120 mmol?L?1 Na Cl预处理比1 mmol?L?1 Na Cl预处理显著提高了NO3?-N的吸收速率;10 mmol?L?1NO3?-N预处理时,120 mmol?L?1 Na Cl预处理与1 mmol?L?1 Na Cl预处理无显著性差异。表明低氮环境下生长的大麦,其根系NO3?-N高亲和力系统受Na Cl影响较大。对低亲和力系统来说,所有预处理大麦的NO3?-N吸收曲线均符合Michaelis-Menten方程的描述。在1 mmol?L?1 NO3?-N预处理中,与1 mmol?L?1 Na Cl预处理相比,120 mmol?L?1 Na Cl胁迫的大麦NO3?-N吸收速率显著提高;10 mmol?L?1 NO3?-N预处理中,120 mmol?L?1 Na Cl与1 mmol?L?1 Na Cl处理相比,降低了大麦的NO3?-N吸收速率。表明低氮环境下生长的大麦,盐胁迫对其根系低亲和力系统有促进作用,而高氮环境下生长的大麦,盐胁迫对其低亲和力系统的NO3?-N吸收没提高作用。

噬菌体对2-酮基-L-古龙酸生产的影响

研究了噬菌体对2-酮基-L-古龙酸混菌发酵的影响,在混菌培养12小时后分别接种2×105pfu/ml巨大芽孢杆菌的两种噬菌体BS601和BS801,发现:(1)噬菌体BS601虽能裂解B.megaterium,但能通过释放胞内活性物质促进K.vulgare生长,从而促进2-KLG的合成并缩短发酵周期;(2)添加噬菌体BS801能同时裂解B.megaterium和K.vulgar,导致代谢停止,无法合成2-KLG。基于上述研究结果,通过在不同发酵周期添加一定浓度的噬菌体BS601,显著提高了2-KLG的产量和生产强度。