Essential oils of Origanum compactum increase membrane permeability,disturb cell membrane integrity,and suppress quorum-sensing phenotype in bacteria

The aim of this study was to investigate antibacterial activity of Origanum compactum essential oils collected at three phenological stages on Escherichia coli and Bacillus subtilis. The antibacterial activity was evaluated using the agar-well diffusion assay. The MIC and MBC values were determined using the micro-dilution assay. The investigation of the antibacterial action was carried out by the evaluation of the effect of O. compactum essential oils on the antibacterial kinetic growth, the integrity of cell membrane and permeability of the cell membrane. The anti-quorum sensing activity was tested by the inhibition of the biofilm formation. The findings of this study showed that O. compactum essential oil has potent antibacterial activities against E. coli and B. subtilis. The lowest inhibition value against B. subtilis was obtained with O. compactum essential oil at the post-flowering stage (MIC=MBC=0.0312%(v/v)). The antibacterial mechanisms of O. compactum essential oils are related to the disturbing of the cell membrane integrity and the increasing of the membrane permeability, which leads to the leakage of genetic materials (DNA and RNA). Moreover, O. compactum essential oils inhibited the formation of the biofilms, a phenotype that has been known to be quorum sensing regulated.

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 larvicidal activity of essential oil of Origanum majorana(Lamiaceae) cultivated in Morocco against Culex pipiens(Diptera:Culicidae)

Objective:To evaluate the larvicidal activity of essential oil of Origanum mtijoruna(Lamiaceae)cultivated in Morocco against Culex pipiens(Diptera:Culicidae).Methods:The analysis and the identification of the various constituents of essential oil were carried out by gas chromatography coupled with mass spectrometry.Biological test was performed according to a standard methodology inspired by the World Health Organization protocol with slight modification.Results:This oil mainly consisted of monoterpene and sesquiterpenes.The majority compounds are 4-terpinene(28.96%),y-terpinene(18.57%),α-terpinene(12.72%) and sabinene(8.02%).The lethal concentrations(LC_(50) and LC_(90)) measured for the essential oil Origanum majorana,were respectively of the order of 258.71 mg/L and 580.49 mg/L.Conclusions:The results could be useful in search for newer,safer,and more effective natural larvicidal agents.

Experimental Study on the Antibacterial Effect of Origanum Volatile Oilon Dysentery Bacilli In Vivo and In Vitro

To observe the germistatic and germicidal effects of origanum volatile oil (OVI) on the dysentery bacteria, the abdominal cavity of mice was infected with Shigella sonne (Sh. sonnei) and Shigella flexneri (Sh. flexneri) F 2a. After OVI was given to the mice via gastric lavage, the effects of OVI on the infected mice were observed. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) for dysentery bacteria were determined in vitro. The results showed that origanum volatile oil showed obvious protective effect on mice infected with Sh. sonnei and Sh. flexneri F 2a. and it had germistatic and germicidal effects on dysentry bacteria. We are led to conclude that origanum volatile oil is an effective medicine against the infection of dysentery bacteria.

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.

Efficient Plant Regeneration from Explants of Compact Oregano (Origanum compactum Bentham)

Origanum compactum Benth., a species endemic to Morocco characterized by its biological activities, is overexploited because of its commercial value and threatened with extinction. Accordingly, measures for its conservation are needed. Micropropagation serves as a solution for the protection and the domestication of this species. In this investigation, we established a protocol for vegetative multiplication in vitro of Origanum compactum by the axillary bud technique. Six culture media (SH, SD, N30K, MS, MSm, B5) were tested to determine the most suitable mineral medium for growth and development of explants. Four cytokinins: Kinetin, Zeatin, BAP (6-Benzylaminopurine), 2ip (2-Isopentenyladenine) and three compounds with cytokinin activity: Adénine, 1,3-Diphenylurea (DPU) and Thidiazuron (TDZ) at five concentrations (0.44, 1.33, 2.22, 3.11, 4.44 μM) were tested on budding, growth, hyperhydria and rooting. Then three auxins, Indole-3-acetic acid (IAA), Naphthalene acetic acid (NAA) and Indole butyric acid (IBA) at four concentrations (1.14, 2.85, 4.56, 6.27 μM) in presence of 2.22 μM BAP were evaluated. The combination of Gibberellic acid (0.29, 1.5, 2.60, 2.89 μM GA3) and three polyamines (Putrescine, Spermidine, Spermine) at five concentrations (1.134, 3.402, 5.67, 7.938, 11.34 μM) with cytokinins and auxins were considered. Our results show that Margara medium is the most efficient and BAP at 2.22 μM is the best cytokinin for development of the aerial parts, with a regeneration rate of 88, 90%;rhizogenesis is successful with the combination of 6.27 μM IAA and 2.22 μM BAP. Moreover, the integration of 2.89 μM GA3 with 2.22 μM BAP and 6.27 μM IAA promotes vitroplant growth, bud and shoot multiplication and elongation of the aerial part. The addition of polyamines with 2.22 μM BAP and 6.27 μM IAA does not improve the root part, but Spermine at 5.67 μM promotes bud and shoot multiplication with a high percentage of regeneration, and spermidine at the same concentration gives long explants. Finally, plantlets with good root development were successfully acclimatized to natural conditions and served as a source to establish in vitro culture again.

Interaction Effect of Plant Growth Regulators on Shoot Micropropagation of Aromatic Plant Origanum elongatum (Bonnet) Emberger &Maire

Origanum elongatum (Bonnet) Emb. & Maire, is a medicinal, aromatic and endemic plant of Morocco, characterized by its pharmacological effects, and is commonly used for the production of essential oils and aromas, resulting in high harvest and overexploitation pressure. This is why the present study aims to implement the in vitro micropropagation of Origanum elongatum for optimal vitroplant production. Six macroelements were tested (SH, SD, N30K, MS, MSm and B5) and the SD medium was selected for vegetative propagation of the explants. Seven cytokinins: adenine (Ad), N6-(2-Isopentenyl) adenine, zeatin (Zeat), kinetin (Kin), benzyladenine (BAP), 1,3-diphenylurea (DPU) and thidiazuron (TDZ) were then evaluated at five concentrations (0.44, 1.33, 2.22, 3.11 and 4.44 μM/L) on growth, development, budding, rooting and hyperhydricity. 0.44 μM Kin was selected and combined with three auxins: indole-3-acetic acid (IAA), indole-3-butyric acid (AIB), and 1-naphthaleneacetic acid (NAA) at four concentrations (1.14, 2.85, 4.56 and 6.27 μM/L) to improve rooting and association with 1.14 μM IAA was shown to be efficient for roots development. Different concentrations of gibberellic acid (0.29, 1.5, 2.60 and 2.89 μM/L), combined with 0.44 μM/L Kin and 1.14 μM/L IAA, were tested and 2.60 μM/L GA3 gave maximum buds and shoots. Then, the combination of three polyamines at five concentrations (1.134, 3.402, 5.67, 7.938 and 11.34 μM/L) with 0.44 μM Kin and 1.14 μM/L IAA showed an increase in the number of buds and shoots for 7.938 μM/L putrescine and 3.402 μM/L spermine. Finally, seedlings with good foliar and root development were acclimatized.

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.

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.

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.

Genetic variation of oregano (<i>Origanum vulgare</i>L.) for etheric oil in Albania

In this study are presented the results of studies for oregano in 16 locations (sites) of Albania for the content of the quantity of oils and their fractions.Albaniais the one of the important diversity centers for Oregano because of their adaptation to diverse agro ecological conditions as a result of natural selection. The design of the experiment was based on split plot methods. From the total samples collected, were selected 16 mostly widespread samples of natural populations of origano (Origanum vulgare L. sp. vulgare and sp. hirtum). Essential oil isolation from oregano was performed by hydro distillation. Identification of the components was made by comparing mass spectra of components in essential oils with those from Nist, Wiley and Adams mass spectra libraries. The results show that the total oil content varies from 3.45% to 0.1%. The populations of Oregano in the part of southern Albania have a higher amount of etheric oil compared with those of the North part. The higher content of carvacrol in 55.63%, of the samples was determined in sites of the southern part of Albania, while in the other two investigated area central and north part it was 55.63% and 48.49% respectively. The same indicators were realised also for content of linalool and thymol, while North area has sites with high content of caryophyllen-oxide and β-pinen 3. But, in all sites, the oils of thymol and carvacrolit were dominant.

<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.

Origanum majorana Extracts as Mild Steel Corrosion Green Inhibitors in Aqueous Chloride Medium

We optimized Origanum majorana (OM) extraction for mild steel corrosion inhibition in neutral 0.5 M chloride medium. The inhibition mechanism evolved in presence of the optimal extract was discussed when calculating the activation energy (Ea), the activation enthalpy (ΔaH) as well as the activation entropy (ΔaS). The OM extract molecules were investigated using the density functional theory (DFT) at B3LYP/6-31G(d,p) basis set level. 1-methyl-4-propan-2-ylcyclohexa-1,3-diene alpha-terpinene was predicted exhibiting the most inhibition capabilities.

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.

牛至提取物对肉雏鸡生产性能及其免疫功能的影响试验研究

为研究牛至提取物对肉鸡生产性能及其免疫功能的影响效果。该试验选用210只商品代雏鸡,随机分成3组,A组为空白对照组,B组(30 mg/kg)为中剂量组,C组(60 mg/kg)为高剂量组,4个重复,每个重复10只鸡,用作生产性能测定;另2个重复,每个重复15只鸡,用于不同日龄试验雏鸡免疫器官绝对重量及其指数的研究。结果表明,试验B组、C组促生长率显著高于对照组(P<0.05),B组雏鸡促生长率较高,相反C组促生长率低于B组。B组、C组在3段日龄中分别提高增重11%和6.7%,说明促生长率随日粮添加水平的提高而降低。进一步说明在肉雏鸡日粮中添加牛至提取物可以促进肉雏鸡的生长率,当牛至提取物添加水平为30 mg/kg时,对肉雏鸡的促生长率效果最为显著。为牛至提取物在肉鸡生产上的应用提供理论依据。

云南腾冲牛至精油成分及其抗炎和抗衰老活性特征

本研究利用气相色谱-质谱联用仪对云南腾冲产区的牛至精油(Origanum vulgare L.Essential oil,OvEo)进行了成分分析,并评价了其体内外抗炎和抗衰老作用。结果显示,OvEo含有15种化学成分,主要成分为香芹酚(52.68%)、桉叶油醇(11.4%)和松油烯-4-醇(4.97%)。在100μg/mL的质量浓度下,OvEo显著抑制RAW264.7细胞中TNF-α、IL-6和NO的分泌,同时减少CuSO_(4)诱导的斑马鱼中性粒细胞迁移,具有明显的抗炎作用。此外,OvEo有效清除活性氧,抑制β-半乳糖苷酶的分泌,同时对胶原蛋白酶和弹性蛋白酶活性有良好的抑制效果,其半抑制率质量浓度(Half Maximal Inhibitory Concentration,IC50)分别为6.72 mg/mL和30.7 mg/mL,表现出抗衰老作用。云南腾冲地区的OvEo具备一定的抗炎和抗衰老作用,在未来的抗衰老食品、护肤品和药品领域具有广阔的开发前景。

牛至中药组方汤剂治疗兔球虫病的疗效试验及效果评估

为探讨牛至中药组方对兔球虫病的治疗效果,该试验选用了25只体重相近的45日龄健康无球虫试验兔,并随机分成5组。感染后5 d分别口服2.00、1.00、0.50 g/mL的牛至中药组方汤剂,根据感染程度、存活率、每克粪便卵囊数(OPG)指标判断牛至组方汤剂对兔球虫病的治疗效果。结果表明,阳性对照组的临床症状比各用药组的临床症状严重,阳性对照组平均增重最少,中药组与阳性对照组相比差异极显著(P<0.01)。各试验组在感染后5 d均有卵囊排出,治疗组随着服药天数的延长OPG变为0,阳性对照组OPG明显高于治疗组。牛至中药组方中以高浓度组(2.00 g/mL)治疗兔球虫病效果最佳。说明牛至中药组方治疗兔球虫病有一定效果,且存一定的量效关系,该试验为牛至替代抗生素等药物预防和治疗兔球虫病提供理论依据。

牛至油不同复配组合的体外联合抗菌作用及机理研究

目的探讨牛至油与其他中药挥发油的复配组合对革兰阳性菌、革兰阴性菌的联合抗菌作用及机理。方法采用K-B纸片法、96孔板微量稀释法、棋盘稀释法测定牛至油等单方中药挥发油及其不同复配组合对金黄色葡萄球菌(Staphylococcus aureus,S.aureus)、大肠杆菌(Escherichia coli,E.coli)的抑菌圈直径、最低抑菌浓度(minimum inhibitory concentration,MIC)以及联合抑菌指数(fractional inhibitory concentration index,FICI);通过试剂结合盒酶标仪测定最优复配组合对S.aureus、E.coli的核酸、蛋白质等细胞膜内含物的渗透量,同时采用荧光分光光度计测定一定作用时间的荧光强度。结果①牛至油、丁香油、艾叶油、香薷油、薄荷油、菊花油、荆芥油、姜黄油、肉桂油、藿香油、青蒿油、金银花油、蒲公英油、山苍子油、小茴香油、石菖蒲油对S.aureus、E.coli的抑菌圈直径为6.5~43.8 mm、6.2~26.5 mm,MIC为0.20~50.0μL/mL、0.78~>50.0μL/mL;②牛至油与山苍子油、肉桂油、石菖蒲油、丁香油、金银花油组合对S.aureus的FICI为0.25~0.75,牛至油与肉桂油、丁香油、艾叶油、香薷油、石菖蒲油组合对E.coli的FICI为0.375~1.0;③在牛至油(0.125μL/mL)-肉桂油(0.125μL/mL)、牛至油(0.125μL/mL)-肉桂(0.25μL/mL)组合的MIC浓度作用下,S.aureus、E.coli胞膜内的AKP、核酸、蛋白质外泄量均显著增加(P<0.05),胞内荧光强度均显著降低(P<0.05)。结论牛至油对革兰阳性菌、革兰阴性菌均有显著的抑菌作用,与肉桂油复配后能够起到显著的协同抗菌效果,作用机理可能与破坏菌体细胞壁和细胞膜结构有关。

基于accD基因序列的牛至遗传多样性研究

目的基于acc D基因序列分析不同产地牛至的遗传结构与分化,考察其遗传多样性。方法以8个居群69份样品为材料,试剂盒法提取植物基因组DNA,同源克隆法获得acc D序列,Sanger测序法双向测序,利用DNAsp、Arlequin等软件进行多态性分析,MEGA软件计算遗传距离并构建NJ发育树。结果牛至acc D长度为1432-1436 bp,存在10个单倍型与16个变异位点,单倍型多态性为0.595,核苷酸多样性为0.15750;牛至居群遗传分化较高(Gst=0.36751,Nst=0.46976,Fst=0.67627),基因流较低(Nm=0.43),居群间变异(67.62%)是主要的变异来源。系统发育关系显示10种单倍型形成3大分支,主要以H2为中心形成发散状的系统结构,种群未发生扩张。结论牛至居群存在较高的遗传多样性与遗传分化,可能主要与生境片段化、种群隔离及生活史特征相关,本研究为牛至种质资源的保护和开发利用提供理论基础。