Citrus aurantium L. extract alleviate depression by inhibiting gut microbiota-mediated inflammation in mice

Gut microbiota plays a crucial role in the pathophysiology of depression.This study aimed to explore the antidepressant effect of mature whole Citrus aurantium fruit extract(FEMC)in the chronic unpredictable mild stress(CUMS)model.The behavioral tests were applied to assess antidepressant effect and 16S rRNA sequencing was used to analyze the changes of gut microbiota.The results showed that the major components of FEMC were naringin and neohesperidin and significantly increased the sucrose preference index of the mice.FEMC also could reduce the feeding latency in an open field test and the rest time in a novelty suppressed feeding test.In addition,FEMC could increase CUMS-induced reduction in the levels of BDNF,PSD95,and SYN in the hippocampus.Moreover,FEMC intervention slightly decreased the ratio of Firmicutes to Bacteroidota.Meanwhile,FEMC reduced the abundance of the Prevotellaceae_Ga6A1_group,[Ruminococcus]_torques_group,which have been reported to be closely related to inflammation.Bioinformatics analysis revealed that mitogen-activated protein kinase(MAPK)signaling pathway and lipopolysaccharide biosynthesis were involved in the anti-inflammatory effect of FEMC in the CUMS animal model.Finally,the ELISA results showed that FEMC could significantly reduce the expression of pro-inflammatory cytokines IL-6 and TNF-αin the serum of depressive mice.Our results suggest FEMC can am eliorate depressive behavior by i nhibiting gut microbiota-mediated inflammation in mice.

柚子(Citrus grandis Osbeck)S-like核酸酶基因CgSL1的分离与特征分析

从柚子 (Citrus grandis Osbeck) 2个自交不亲和品种溪蜜柚和度尾蜜柚的花柱中克隆出一个类似 S核酸酶基因 Cg SL1(C.grandis S- like RNase) ,它的 c DNA序列全长 10 74 bp,编码 2 97个氨基酸。通过与其它植物S- like核酸酶和 S核酸酶氨基酸序列进行比较 ,发现 Cg SL1类似于 S- like核酸酶 ,与拟南芥中的 RNS2一致性为6 2 .5 %。对 Cg SL1的表达分析表明该基因在花柱、花药、叶片不同器官以及花柱的不同发育阶段均有表达 ,且在花柱中的表达随衰老增强 ,由此推测它可能与衰老有关。

Effects of Different Fertilization Treatments on Soil, Leaf Nutrient and Fruit Quality of Citrus grandis var. longanyou

The five-year-old “Longanyou” trees were used as the experimental material to study the effects of different fertilization treatments. The nutrient contents in soil and leaves, fruit yield and quality were determined, and then the correlations were analyzed. The results showed that: 1) The soil nutrient contents of 0 - 20 cm depth were more than the 20 - 40 cm, and the trends of nutrient contents of the 0 - 20 cm soil layers were as follows: treatment 2 (T2) > treatment 3 (T3) > treatment 4 (T4) > treatment 1(T1) > control (CK). However, the 20 - 40 cm depth had not significant difference between different treatments, but T2, T4 and T3 were higher than T1 and CK. It indicated that the soil effective nutrient content increased in T2 and T3. 2) Compared with the control, the content of K and B elements was improved obviously in leaves with the increase of organic manure application. The contents of P (1.60 g·kg-1), B (26.00 mg·kg-1) and Mg (1.18 g·kg-1) were the highest, and other nutrients contents were also higher, indicating that T2 could effectively improve the leaves’ nutrient contents. 3) The fruit yield per plant was the highest in T2 (95.40 kg plant-1), and the single fruit weight, total sugar, sugar and acid ratio, vitamin C were also the highest, but titratable acid was lower. It indicated that T2 effectively improved fruit yield and quality. 4) There were positive correlations between multiple factors of soil nutrients and the quality index, such as fruit peel thickness, total sugar, solid acid ratio, sugar and acid ratio, Vc content and single yield etc. There was significant correlation between K, B, Zn, Fe contents and fruit yield and quality index, and the contents of B, Zn and Fe in leaves were significantly correlated with soil nutrient, indicating that the contents of K, B, Zn, Fe in soil and leaf were closely related to fruit yield and quality. In sum, the T2 was the best fertilization scheme for orchard management practice of “Longanyou”.

基于UPLC特征图谱和多成分定量评价不同采收时间枸橘药材质量

目的:基于不同采收时间枸橘药材UPLC特征图谱,结合黄酮类和香豆素类成分的含量测定,明确枸橘成分积累规律,为枸橘的采收和质量控制提供依据。方法:采用UPLC法构建30批枸橘特征图谱,采用相似度评价、聚类分析、主成分分析和正交偏最小二乘判别分析对数据进行处理。采用HPLC法测定其中6个成分的含量。结果:30批样品UPLC特征图谱共标定25个共有峰,相似度为0.913~0.993,指认了7个成分,其中6个成分芸香柚皮苷、柚皮苷、枸橘苷、欧前胡素、异欧前胡素、葡萄内酯的含量分别为0.62~2.40 mg/g、5.75~27.52 mg/g、7.00~53.86 mg/g、0.63~2.94 mg/g、0.11~0.45 mg/g、3.69~9.29 mg/g;聚类分析可将样品分为3类,正交偏最小二乘判别分析筛选出5个质量差异标志性成分。不同采收时间枸橘药材可以通过果皮颜色区分,枸橘苷、柚皮苷、欧前胡素等是导致枸橘药材质量产生差异的主要标志物。结论:果皮颜色可作为枸橘药材的采收依据;枸橘苷、柚皮苷和欧前胡素等是枸橘药材质量差异标志物。

Study on Infrared Spectra of Citrus grandis Peels from Guangxi Zhuang Autonomous Region

[ Objective ] This study aimed to establish the infrared （IR） spectra fingerprints for peels of Citrus grandis from Guangxi Zhuang Autonomous Region. [ Method ] The fingerprint was established by fourier transform infrared spectroscopy （ FT-IR）, and then were analyzed by Omic software. [ Result ] IR spectra fin- gerprint of C. grand/s were successfully established. There were six common peaks in the characteristic spectra of 12 samples. [ Conclusion] The specific, rapid and stable method can be applied to quality control of C. grandis.

Optimizing Culture System of Ri T-DNA Transformed Roots for Citrus grandis cv. Changshou Shatian You

Genetic transformation experiments of the different explants from Citrus grandis cv. Changshou Shatian You infected with Agrobacterium rhizogenes were carried out in darkness or in light. The optimizing culture system of Ri T-DNA transformed roots for C. grandis cv. Changshou Shatian You was constructed as follows： After the ventral wounded striations on the single activation cotyledon were inoculated by A. rhizogenes A4 （logarithmic period）, they were cocultured at （25 ±2）℃ in darkness for 25-30 days; some transformed roots were generated from wounded striations of most cotyledons. The genetically transformed ratio is （83 ± 11）%. Axenic Ri T-DNA transformed roots （hairy roots） were harvested after five subcultures. Explants were activated on MT medium. The MS medium was used for subculture of transformed roots. Mass Ri T-DNA transformed roots in which the hormone was produced independently were harvested from this optimizing culture system. White, fresh Ri T-DNA transformed roots were （1.14 ±0.07） cm long, （0.73 ±0.04） mm wide, and the growth direction of transformed roots was negative geotropism.

Identification and Control of HLB Disease in Citrus grandis

HLB disease has been endangering citrus production,and it is an important task in Citrus grandis production to identify and control HLB disease. Diaphorina citri and Trioza erytreae are the main vectors for the spreading of HLB disease. Scientific and proper release of predatory natural enemies such as ladybugs,combined with chemical control can effectively control psyllid. For suspected HLB disease strains,a simple " HLB disease detection reagent" can be used for detection. This method is simple,cheap and accurate,and it is an economical and feasible identification method for ordinary growers.

High-yielding Fertilization Technology for Citrus grandis

As of April 2017,the planting area of Citrus grandis in Ruili City was 594. 3 ha. Among it,the area of fruiting trees was 267. 2 ha,and they were mainly owned by small growers. In view of the serious fragmentation of pomelos,the large investment in fertilizers,and the unsatisfactory production efficiency of C. grandis in Ruili,the fertilization technology for C. grandis was discussed. The problems in the fertilization process,as well as the principles and modes of fertilization were analyzed briefly. The fertilization methods for saplings and fruiting trees were introduced,in order to promote the improvement of the fertilization and management level of local pomelo growers.

柚树(Citrus grandis)叶片光合作用对补增UV-B辐射的响应

生长在人工光照 4 0 0μmol m- 2 s- 1 下的柚树幼树光合速率的最大值为 1 0 .2± 0 .5μmol m- 2 s- 1 ;而补增UV-B辐射 ( 3.8-4 .2μW cm- 2 ,2 4 5～ 2 97nm,4 5d)的叶片则为 6.4± 0 .8μmol m- 2 s- 1 ,较对照植株降低37.2 %。对照植物的表观量子产率 (固定 mol CO2 mol- 1量子 )为 0 .0 75± 0 .0 1 2 ,而经 UV-B辐射处理植株则为0 .0 4 1± 0 .0 0 8,明显较对照植株低。UV-B辐射处理使植株叶片的光呼吸和不包括光呼吸的 CO2 补偿点增高。对照植株叶片的最大值的 CO2 羧化速率 (μmol m- 2 s- 1 )为 57.1± 1 .5μmol m- 2 s- 1 ,较 UV-B辐射处理的高30 .9% ,而 UV-B辐射处理的植株的光合电子传递速率较对照低 30 %。同时 UV-B辐射植株叶片有较低的光能转化效率 ,其较对照低 39.1 % ,叶片亦含有较低的叶绿素含量。结果表明 ,UV-B辐射明显抑制叶片光合羧化速率和光合电子传递速率 ,UV-B辐射可能抑制包括 Rubisco羧化作用在内的多个光合生理过程 ,降低叶片光合速率。柚树叶片对 UV-B辐射敏感 ,选育抗 UV-B辐射的柚树品种势在必行。

Identification of Genetic Variation in Citrus sinensis from Hunan Based on Start Codon Targeted Polymorphism

[Objective] The aim was to identify genetic variation in Citrus sinensis （sweet orange） germplasm from Hunan Province according to the Start Codon Targeted （SCoT） Polymorphism. [Method] The reaction system for SCoT amplification from sweet orange was first optimized, and then the SCoT fragments were amplified from 24 sweet orange cultivars collected in Hunan Province and sequenced for genetic variation analysis. [Result] The optimum reaction system for SCoT markers amplification was 2.0 μl containing 80 ng of template DNA, 0.3 mmol/L dNTPs, 0.2 μmol/L primer, 1.6 mmol/L Mg2＋, 1.6 U of Taq DNA polymerase and 10×PCR buffer. By using this reaction system, the PCR products from the sweet orange cultivars produced clear and reproducible bands at 100-2 000 bp through electrophoresis. The SCoT fragments of the 24 sweet orange cultivars were 1 090-1 091 bp, with the homology of 99.84% and nucleotide deletion and substitution. After being sequenced, the SCoT polymorphisms could distinguish 12 sweet orange cultivars. In addition, the BLAST result showed that part of the SCoT fragments coding region shared high homology with ribosomal protein S3 N superfamily. [Conclusion] This study will provide a theoretical basis for breeding sweet orange cultivars.

热带柚木(Tectona grandis L. f.)人工林中不同邻体干扰强度下林木CO2气体交换特征

研究以海南岛尖峰岭21a生的珍贵树种柚木人工林为对象,以邻体干扰指数为干扰强度评价指标,用Li-6400(Li-cor,Inc.,USA)便携式光合作用测定系统研究了邻体干扰强度对柚木个体CO2气体交换的日动态和季节变化特性,光合-光响应特性及光合-CO2响应特性的影响。结果表明:邻体干扰并未改变个体叶片光合作用日进程,但对日净光合速率(Pn)的影响差异较大,表现为弱、中、强和极强度干扰下日Pn时段均值比为2.5∶2.3∶1.7∶1.0,弱干扰下日最大净光合速率为极强干扰下的2.8倍;随着干扰强度的增加,柚木叶片的蒸腾速率、气孔导度等特征参数也出现不同程度的下降。邻体干扰对光合-光响应和光合-CO2响应特征参数的影响明显,弱干扰下柚木叶Asat、Qsat、αA、CE、Vcmax和Jmax分别是极强干扰下的2.7、1.3、1.4、2.7、1.9和2.8倍,Qcomp和Rd则受邻体干扰的影响不明显。干扰强度对光合光响应和光合CO2响应特征的影响随光环境和CO2浓度的改变而有所差异,光环境的改变对弱干扰下柚木个体的影响明显要强于强度以上干扰的个体,而CO2浓度的改变对弱干扰个体的影响则明显要弱于强度干扰以上个体的影响。因此,在林分结构相对单一、经营管理措施基本一致的人工林内,邻体干扰是个体竞争的主要影响因素之一,干扰强度的不同将影响种内个体光合碳同化能力出现差别,导致个体生长差异。可以基于邻体干扰指数评价柚木人工林内种内竞争程度的大小,并以此为依据合理调整柚木人工林的林分结构,达到对环境资源的最合理利用。

红肉脐橙(Citrus sinensis L.)果肉中特征色素提取方法探索

红肉脐橙是目前为止惟一果肉呈粉红色或红色的脐橙,其呈色色素尚未见报道。为深入研究其果肉中特征色素的构成及在果实生长发育和贮存过程中色素含量的变化,本研究在对该色素初步定性的基础上对提取方法进行了探索。在分析红肉脐橙果肉中色素萃取液的紫外可见吸收光谱时发现,该色素有类胡萝卜素的特征吸收峰,将萃取液进行薄层层析则进一步证实其内含有番茄红素。在色素提取的前处理方式上,本研究认为冷冻干燥样品的浸提效果较真空干燥的样品稳定(变异系数小),且浸提率相差无几,所以选择冷冻干燥作为合适的样品前处理方式。色素浸提条件的正交实验结果表明,最佳浸提条件为50℃、1.5h内浸提4次,浸提的料液比为1:60。此外,浸提温度、浸提时间、浸提料液比和浸提次数对浸提率都有显著影响,影响最大的因素是浸提温度,最佳温度为50℃,随温度降低浸提率显著降低。其次是侵提时间的长短,最佳浸提时间为1.5h,延长或缩短0.5h都会使浸提率下降。浸提次数对浸提率的影响是最小的。

柚(Citrus grandis Osbeck cv.)MADS box基因的表达分析

以De novo转录组学方法分析柚(Citrus grandis Osbeck cv.)MADS box基因家族的基因.以特早熟龙柚(变异型)和琯溪蜜柚(参照型)嫩果(不超过5天)和新叶为材料,经过RNA提取、反转录、测序、组装得到Unigene序列.Unigene序列在Nr、Swiss-Prot、KEGG和COG四大数据库中进行比对,获得MADS box转录因子家族基因注释结果.采用数字基因表达标签(digital gene expression tag,DGE)技术,对样本中基因转录丰度以RPKM值表示,比较处理组和对照组的RPKM值,以倍性变化≥2确定差异表达基因(differentially expressed genes,DEGs).从特早熟龙柚和琯溪蜜柚嫩果和叶片中,总共注释到28个MADS box基因家族基因.在龙柚嫩果(FE)和蜜柚嫩果(FL)中表达的有25个,其中有6个属于差异表达基因.在龙柚叶片(LE)和蜜柚叶片(LL)中表达的有27个,其中有7个属于差异表达基因.注释到的28个MADS box基因家族基因在克莱门特柚(Citrus clementina)和中国甜橙(Citrus sinensis)报道过的分别有18个和20个,其中有5个基因既在中国甜橙和克莱门特柚都没有报道过,也没有发现在其它Citrus物种报道过.

Phenolic and flavonoid contents of mandarin (Citrus reticulata Blanco) fruit tissues and their antioxidant capacity as evaluated by DPPH and ABTS methods

The total phenolic and flavonoid contents in the fruit tissues （peels, pulp residues, seeds, and juices） of 19 citrus genotypes belonged to Citrus reticulata Blanco were evaluated and their antioxidant capacity was tested by 2,2-diphenyl-l-picrylhydra- zyl radicals （DPPH） method and 2,2＇-azino-bis（3-ethylbenzthiozoline-6）-sulphonic acid （ABTS） method. The total phenolic and flavonoid contents, and their antioxidant capacity varied in different citrus fruit tissues. Generally, the peel had both the highest average of total phenolics （27.18 mg gallic acid equivalent （GAE） g^-1 DW） and total flavonoids （38.97 mg rutin equivalent （RE） g^-1 DW）. The highest antioxidant capacity was also the average of DPPH value （21.92 mg vitamin C equiv- alent antioxidant capacity （VCEAC） g^-1 DW） and average of ABTS value （78.70 mg VCEAC g-1 DW） in peel. The correlation coefficient between the total phenolics and their antioxidant capacity of different citrus fruits tissues ranged from 0.079 to 0.792, and from -0.150 to 0.664 for the total flavonoids. The antioxidant capacity of fruit tissues were correlated with the total phenoilc content and flavonoid content except in case of the peel. In addition, the total phenolic content and antioxidant capacity varied in different citrus genotypes. Manju and Karamandarin were better genotypes with higher antioxidation and the phenolic content, however Shagan was the poorest genotype with lower antioxidation and the phenolic content.

Effect of <i>Citrus aurantium</i>L. Essential Oil and Its Interaction with Fluoxetine on Anxiety in Male Mice

Anxiety is a very common mental disorder among neurological diseases. Some herbs have soothing effects and play an important role in reducing anxiety. The purpose of this study is to investigate the effect of Citrus aurantium L. essential oil on anxiety and its interference with serotonergic pathway. Sixty male mice were assigned into control, sham (saline and olive oil), and experimental groups. Intraperitoneal injection of Citrus aurantium L. essential oil was applied at doses of 0.5, 2.5, and 5 percent for 5 days. In another set of experiments, after intraperitoneal injection of Citrus aurantium L. essential oil at doses of 0.5, 2.5, and 5 percent for 5 days, on the 5th day, 30 minutes before applying essential oil, fluoxetine (2 mg/kg) was injected. Then, the anxiety-related behavior was assessed using elevated plus maze test. The results revealed that injection of essential oil of Citrus aurantium L. alone or along with fluoxetine led to increasing the number of entries into the open arms and the time spent in open arms that was significantly different compared with control and sham groups (P?< 0.001). Besides, further effects revealed when fluoxetine added to essential oils, however no more effects obtained when compared to fluoxetine alone. It is concluded that Citrus aurantium L. essential oil can reduce the anxiety in male mice and due to fluoxetin potentiation and maximum response observed, the herb may express its anxiolytic effects in part, via serotonergic system.

An efficient method for clonal propagation and in vitro establishment of softwood shoots from epicormic buds of teak(Tectona grandis L.)

Softwood shoots were produced from 40 cm long stem segments placed horizontally in flat trays containing sterilized sand under natural light or shade conditions for subsequent rooting and micropropagation studies in teak （Tectona grand＆ L.）. Higher number of shoots （6.17） per log was produced under natural light as compared to shade conditions. Forcing was also better in natural light as compared to shade in terms of shoot length, number of nodes or leaves. For rooting, 2-4 cm long softwood shoots were excised and treated with either indole-3-butyric acid （IBA） or α-naphthyl acetic acid （NAA） at 0, 1000, 2000 or 3000 μmol.L^-1 each or with combinations （1000 ＋ 1000, 2000 ＋ 2000 or 3000 ＋ 3000 μmol.L^-1） and then placed in flat trays containing autoclaved sand at 25 ± 2℃ in 16 h photoperiod at 35 μmol.m^-2.s^-1. After 28 days, softwood cuttings treated with IBA ＋ NAA （3000 ＋ 3000 μmol.L^-1） had highest rooting percentage （89.3%） with 5.5 mean roots. Shoot apex and nodal explants of softwood cuttings were pretreated with 0.1% （w/v） ascorbic acid, boric acid, activated charcoal, citric acid, glutamine or polyvinylpolypyrollidone （PVP） for 24 h to remove phenolic compounds before surface disinfestation. Glutamine （G1） and PVP were equally effective resulting in 60% establishment of shoot apices on MS medium supplemented with 10 μmol.L^-1 6-benzylaminopurine （BAP） ＋ 5 μmol.L^-1 NAA. Using shoot apices, highest （42.80） number of multiple shoots with 54.33 mm shoot length were obtained on MS ＋ BAP （8.8 p.mol.L 1） ＋ IBA （2 μmol.L^-1） after 45 days. Shoots were successfully rooted and acclimatized to greenhouse conditions.

Integrated multi-omics analysis of developing‘Newhall’orange and its glossy mutant provide insights into citrus fragrance formation

Sesquiterpene valencene is dominant in flavedo tissues of sweet oranges and imparts a unique woody aroma.However,the interaction between the biosynthetic pathways of valencene and other nutritional compounds is less studied.Sesquiterpenoids were significantly accumulated in a previously reported glossy mutant of orange(MT)than the wild type(WT),especially valencene and caryophyllene.In addition,we identified several other pathways with variations at both the transcriptional and metabolic levels in MT.It’s interesting to found those upregulated metabolites in MT,such as eukaryotic lipids,kaempferol and proline also showed strong positive correlation with valencene along with fruit maturation while those down-regulated metabolites,such as phenylpropanoid coumarins and most of the modified flavonoids exhibited negative correlation.We then categorized these shifted pathways into the‘sesquitepenoid-identical shunt’and the sesquitepenoid-opposite shunt’and confirmed the classification result at transcriptional level.Our results provide important insights into the connections between various fruit quality-related properties.

A Study of the Effect of <i>Citrus aurantium</i>L. Essential Oil on Anxiety and Its Interaction with GABAergic Pathways in Male Mice

Anxiety is among the most common disorders affecting a great number of people. Different neurotransmitter systems, such as GABAergic system, play an important role in emergence of anxiety-related behaviors. In this study, we have investigated the effect of Citrus aurantium L. essential oil on anxiety and its interaction with GABAergic pathways. Male mice were assigned into control, sham, and experimental groups. Intraperitoneal injection of Citrus aurantium L. essential oil was applied at doses of 0.5, 2.5, and 5 percent for 5 days. Diazepam (0.1 mg/kg) was injected on the fifth day, thirty minutes before applying Citrus aurantium L. essential oil. The anxiety-related behavior of mice was then assessed using elevated plus-maze test. In groups receiving Citrus aurantium L. essential oil at doses of 2.5 and 5 percent, the percent time spent in the open arms increased significantly (P Citrus aurantium L. essential oil resulted in increasing in the number of entries to the open arms and the percent time spent in the open arms (P Citrus aurantium L. essential oil can reduce anxiety-related behaviors in male mice thah may act via GABAergic system.

<i>In-Vitro</i>Regeneration of <i>Citrus sinensis</i>(L.) Osbeck from Mature Seed Derived Embryogenic Callus on Different Solid Basal Media

In-vitro callus induction and regeneration method was developed using different plant growth regulators (PGRs), and basal media (Murashige and Skoog (MS), CHU (N6) and Gamborg (B5) media) of Citrus sinensis (L.) Osbeck. Observations of the effect of PGRs were carried out using different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D),1-naphthalene acetic acid (NAA) and combinations of 2,4-D and NAA using different basal media. This study found Citrus sinensis (L.) Osbeck exhibited a high frequency of callus induction on MS medium supplemented with 3 mg/L 2,4-D and callus induction frequency was 86.7% ± 3.4% whereas N6 and B5 showed lower callus induction frequency of 83.3% ± 8.8% and 82.2% ± 1.9% respectively compared to that of MS media with supplementation of the same hormone. Among the induced calli, the morphological analysis showed only 40% - 50% was embryogenic calli. Regeneration of plantlets from calli was done using different concentrations and combinations of auxin and cytokinin. The study showed that 3 mg/L 6-benzylaminopurine (BAP) supplemented medium has the maximum potential to promote regeneration of Citrus sinensis (L.) Osbeck from embryogenic calli with the frequency of 89.3% ± 8.8% but no regeneration occurred from the non-embryogenic calli. The regenerated plantlets were rooted on MS medium with supplementation of 5 mg/l NAA. These observations in Citrus sinensis (L.) Osbeck regeneration will be helpful for genetic improvement with desired traits.

Effect of <i>Citrus aurantium</i>L. Essential Oil and Haloperidol on Anxiety in Male Mice

Relationship between sociability and the amount of brain’s dopamine is very well known. In this study, we have examined the effect of Citrus aurantium L. essential oil on anxiety and its interaction with dopaminergic pathways. 70 male mice were assigned into experimental, control, and sham groups. Essential oil of Citrus aurantium L. was injected intraperitonealy at doses of 0.5%, 2.5% and 5% for 5 days. Subcutaneous injection of haloperidol was administered on the fifth day, 30 minutes before the injection of the essential oil. The anxiety-related behavior of mice was then assessed by elevated plus-maze test. The result of this study showed that the injection of Citrus aurantium L. essential oil at doses of 2.5% and 5% increased significantly the time spent in the open arms (OAT) (p < 0.001), also there was a significant increase in the number of entries into the open arms (OAE). Injection of different doses of the essential oil along with haloperidol significantly increased OAT (p < 0.001(. The results demonstrate that the essential oil of Citrus aurantium L. along with haloperidol medication reduces anxiety-related behaviors.