Chemical constituents,biological functions and pharmacological effects for comprehensive utilization of Eucommia ulmoides Oliver

Eucommia ulmoides Oliver is a native plant and valuable tonic Chinese medicine in China with a long history,great economic value and comprehensive development potential.Traditionally,the comprehensive utilization rate of E.ulmoides Oliv.is still very low,only bark has been used as medicine and other parts of Eucommia ulmoides Oliv.cannot be fully utilized,even the leaves have been well utilized in food products in Japan in the past decades.In order to improve the comprehensive utilization efficiency of E.ulmoides Oliv.,in this review,we summarized the varieties and contents of main active compounds,biological functions and pharmacological effects in different parts of E.ulmoides Oliv.The findings suggest that other parts of E.ulmoides Oliv.could replace the bark of E.ulmoides Oliv.to some extent besides of their respective applications.The unique and extensive physiological functions between different parts of E.ulmoides Oliv.indicate that the comprehensive utilization of E.ulmoides Oliv.has a wide space to develop,which is also an effective way to protect E.ulmoides Oliv.resources and improve its the utilization rate.

Comparative analysis of chemical components between barks and leaves of Eucommia ulmoides Oliver

The chemical components of the essential oils in the barks and leaves of Eucommia ulmoides Oliver were analyzed and compared by chromatograms and mass spectra technique, heuristic evolving latent projections (HELP), alternative moving window factor analysis (AMWFA) algorithms and normalization method based on the peak areas; the flavones in the barks and leaves of Eucommia ulmoides Oliver were separated on an ODS column by gradient elution carried out with the flow phase consisting of water, methanol and phosphoric acid (0.1%), and their contents were quantitatively determined by standard curve method and diode array detection (DAD) at 362 nm. The results show that 68 and 73 compounds respectively from essential oils of the barks and leaves of Eucommia ulmoides Oliver are identified, and there are 33 mutual compounds among 108 compounds determined. The total contents of these volatile components of the two samples possess 92.9% and 97.75% of the gross of the relevant essential oils, respectively; the contents of the rutin, quercetin and kaempferol in the barks and leaves of Eucommia ulmoides Oliver are 0.016 9, 0.003 6, 0.002 1 and 0.064 4, 0.030 2, 0.010 0 mg/g, respectively, and the determination recoveries are 95.2%-106.2%. The comparative analysis shows that for the barks and leaves of Eucommia ulmoides Oliver, there are significant differences in their components of the relevant essential oils and flavones.

The Extraction of Eucommia Ulmoides Oliv Polysaccharides and Its Protective Effect on Liver of Clophasphamidecy Injured Mice

Objective:To study the influenceof eucommia polysaccharide on the mice' liver damaged by clophasphamidecy (CY).Methods:Injecting CY build mice liver damage model,eucommia polysaccharide given different doses, measured blood serum ALT,AST and the liver's SOD,MDA. Results:After the injection CY,blood serum ALT,AST and the MDA of liver rise and the SOD of liver reduce comparedwith the blank group. The eucommia polysaccharide can improve these index.Conclusion:The Eucommia polysaccharide may protect the mice' liver damaged by CY.

Isolation of a novel antifungal peptide from the bark of Eucommia ulmoides Oliv

A novel Eucommia antifungal peptide,named EAFP3,was isolated from the bark of Eucommia ul-too/des by NaC1 extract,gel filtration and reverse phase high performanee liquid ehromatography.Themolecular mass of EAFP3 is 4157.3 Da,and its partial amino acid sequenee is-.LYQQLIAGITLNK.-.EAFP3 exerts an inhibitory activity against Candida albicans in vitro and the drug concentration requiredfor 50% growth inhibition(IC50)is 31.25μg/mL.

Cloning and Characterization of EuGID1 in Eucommia ulmoides Oliver

Gibberellic acid controlled the key developmental processes of the life cycle of landing plants,and regulated the growth and development of plants.In this study,a novel gibberellin receptor gene EuGID1 was obtained from Eucommia ulmoides Oliver.The cDNA of EuGID1 was 1556 bp,and the open reading frame was 1029 bp,which encoded 343 amino acids.EuGID1 had the homology sequence with the hormone-sensitive lipase family.Amino acid sequence alignment confirmed EuGID1 protein had the highest homology with the GID1 protein of Manihot esculenta.EuGID1 was located in the nucleus and cell membrane and had expression in four plant organs.Overexpression of EuGID1 in transgenic Arabidopsis plants promoted plant elongation and increased siliques yield.

Distribution Prediction of Suitable Growth Area for Eucommia ulmoides in China under Climatic Change Background

[Objective]The research aimed to study distribution prediction of suitable growth area for Eucommia ulmoides in China under climatic change background.[Method]By using the maximum entropy model and many kinds of climate change scenarios,we predicted current and future distribution patterns of suitable growth area for Eucommia ulmoides in China and its change process.[Result]At present,highly suitable growth area of E.ulmoides mainly distributed in Sichuan,Shaanxi and Chongqing.Under climate change background,total suitable growth areas in future three decades all drastically reduced when compared with that at present.It was noteworthy that moderately and highly suitable growth areas of wild E.ulmoides all disappeared,and junction between Shaanxi and Gansu and Taibai Mountain would be stable suitable growth area of wild E.ulmoides.[Conclusion]The research could provide useful reference data for investigation,protection and sustainable development of the wild E.ulmoides resources.

Research Progress of Cell Wall Breaking Technology and Its Application in Eucommia ulmoides Male Flower Tea

Eucommia ulmoides male flowers are rich in secondary metabolite,which have anti-tumor,sedative hypnotic,hypotensive,hypolipidemic,anti-fatigue,bacteriostatic,antioxidant and anti-aging effects.The conventional processing of E.ulmoides male flowers leads to the loss of nutrients and active ingredients.In recent years,cell wall breaking technology has been developed and utilized in various fields such as traditional Chinese medicine,good,chemical industry and biology.In order to promote the development of the E.ulmoides industry,the cell wall breaking technology and its characteristics are reviewed,and the application advantages of the cell wall breaking technology in the male flowers of E.ulmoides are discussed,and the prospect of the cell wall breaking of E.ulmoides is proposed in this article.

Ectopic Overexpression of EuCHIT30.7 Improves Nicotiana tabacum Resistance to Powdery Mildew

Various strains of powdery mildew(PM),a notorious plant fungal disease,are prevalent and pose a significant threat to plant health.To control PM,transgenic technology can be used to cultivate more resistant plant varieties.In the present study,we utilized the rapid amplification of cDNA ends(RACE)technique to clone the full-length cDNA sequence of the EuCHIT30.7 gene to explore plant genes with disease resistance functions.Bioinformatics analysis revealed that this gene belongs to the GH18 family and is classified as a class III chitinase.The EuCHIT30.7 gene is expressed throughout the Eucommia ulmoides plant,with the most abundant expression in male flowers.Subcellular localization analysis indicated that the protein encoded by this gene was detected within both the cell membrane and cytoplasm.Upon PM inoculation,overexpression of EuCHIT30.7 in tobacco plants led to a significantly reduced relative lesion area and a decreased spore count compared to both wild-type and empty vector control plants.Activities of the protective enzymes,namely,peroxidase(POD),superoxide dismutase(SOD),catalase(CAT),and phenylalaninammo-nialyase(PAL),in tobacco plants overexpressing EuCHIT30.7 were significantly greater than those in wild-type and empty vector tobacco plants.Furthermore,the rate of increase in malondialdehyde(MDA)content was significantly lower in tobacco plants expressing EuCHIT30.7 compared to control tobacco plants.In EuCHIT30.7 transgenic tobacco,the expression of pathogen-related protein genes,namely,PR2,PR5,PR1a,PDF1.2,and MLP423,along with the tobacco PM negative regulatory gene,MLO2,were significantly higher compared to control tobacco plants.These findings suggested that EuCHIT30.7 significantly enhances the resistance of tobacco to PM.

杜仲胶合成相关基因EuFPS的克隆及序列分析

本文采用逆转录—聚合酶链式反应 (RT PCR)方法 ,以杜仲树叶总RNA为模板扩增出杜仲法呢基焦磷酸合酶基因EuFPS ,克隆至质粒 pUCm T中。序列分析表明 ,所克隆的cDNA序列全长 12 71bp ,开放阅读框共编码 32 0个氨基酸残基 ,其核酸序列与拟南芥菜、银胶菊和巴西橡胶树FPP合酶的序列同源性分别为 81% ,87% ,82 %。

杜仲对黄曲霉的抑制及在养殖中的有益作用

饲料中黄曲霉污染严重制约了饲料业和养殖业的高质量发展,在我国目前饲料端全面禁抗背景下,天然植物原料作为理想的抗生素替代品被推崇。杜仲是我国传统的药食两用植物资源,将其提取物或原粉添加到饲料中可有效遏制黄曲霉污染,并提高畜禽品质,是理想的饲料替抗物。文章阐述了杜仲提取物及其活性成分对黄曲霉的抑制作用以及在养殖中的有益作用,旨在为其在饲料行业中的应用推广提供参考。

SFE-CO_2法与SD法提取杜仲叶挥发油成分的比较

分别采用超临界CO2流体萃取法(SFE-CO2法)与水蒸气蒸馏法(SD法)从杜仲(Eucommia ulmoids Oliv.)叶中提取挥发油,并结合气相色谱-质谱仪(GC-MS)对其进行检测。从SFE-CO2法提取的杜仲叶挥发油中共鉴定出59种成分,占挥发油总量的82.57%;从SD法提取的杜仲叶挥发油中共鉴定出39种成分,占挥发油总量的84.48%。两种提取方法得到的挥发油组分及其含量差异较大。SFE-CO2法比SD法能更真实、全面的反映杜仲叶药材的挥发油成分。

EuFPS基因表达载体构建及对杜仲遗传转化的研究

本实验用EcoRI和BamHI双酶切植物表达载体pSH737和含有目的基因的pUC—FPS，定向连接得到重组质粒pSH-FPS，将其导入农杆菌EHA105。采用农杆菌介导法对杜仲进行遗传转化，研究了卡那霉素（kanamycin，Kin）浓度、预培养时间、菌液浓度及侵染时间、乙酰丁香酮（acetosyringone，AS）浓度、共培养时间等对杜仲遗传转化效率的影响。结果表明，选择无菌苗苗龄15d的杜仲下胚轴，卡那霉素浓度50mg/L，农杆菌浓度OD600值0．3～0．6，侵染时间8min，侵染时菌体重悬液中添加50μmol／L乙酰丁香酮，共培养时间3d，抗性芽的获得率最高。对再生植株进行GUS检测发现有45％的植株呈阳性。

杜仲抗菌肽EuCHIT1对白色念珠菌的抑菌作用及机制研究

目的:研究杜仲抗菌肽EuCHIT1对白色念珠菌的抗菌活性及机制。方法:通过聚合酶链式反应(PCR)扩增EuCHIT1蛋白的编码区,构建原核表达质粒并转化到大肠杆菌Escherichia coli BL21(DE3),经异丙基硫代半乳糖苷(IPTG)诱导,采用镍亲和色谱法纯化得到可溶性的EuCHIT1;采用二倍最小抑菌浓度(MIC)法稀释EuCHIT1并测定其对白色念珠菌的抑菌圈直径;采用荧光分光光度计和酶标仪分别检测EuCHIT1对白色念珠菌细胞膜的去极化作用及对细胞膜通透性的影响;通过鲎试验动态浊度法和高效液相色谱法分别测定白色念珠菌中(1,3)-β-D-葡聚糖和麦角甾醇含量;酶联免疫吸附法(ELISA)检测EuCHIT1对白色念珠菌感染的SD大鼠肺上皮WTRL1细胞中白细胞介素-1β(IL-1β)、IL-6及肿瘤坏死因子-α(TNF-α)表达的影响。结果:纯化得到的杜仲抗菌肽EuCHIT1相对分子质量为77210,对白色念珠菌MIC值为32μg·mL^-1,EuCHIT1质量浓度与抑菌活性之间具有量效关系;EuCHIT1可增加白色念珠菌细胞壁的去极化及通透性,降低白色念珠菌细胞壁(1,3)-β-D-葡聚糖及麦角甾醇的含量,同时EuCHIT1可降低白色念珠菌感染的WTRL1细胞中IL-6及TNF-α水平(P<0.05)。结论:EuCHIT1对白色念珠菌具有抑菌作用,主要机制可能是破坏细胞壁,导致细胞膜去极化、通透性增加,并降低白色念珠菌感染的肺上皮细胞IL-6及TNF-α水平。

杜仲翅果果实形态和含胶量与产地环境相关性分析

该研究选择我国7个杜仲产地的杜仲为研究对象,采用随机取样的方法,利用SPSS软件进行统计分析。结果表明:不同产地果实形态和含胶量存在显著差异,贵州剑河最高为13.16%,河北保定最低为9.34%,含胶量与所在环境气候条件密切相关,纬度越高,年日照时数越长,翅果含胶量越低;无霜期越长,翅果含胶率越高。

杜仲EuHMGRS基因家族的鉴定及生物信息学分析

萜类物质合成关键酶3-羟基-3-甲基戊二酰辅酶A还原酶(HMGRS)是甲羟戊酸途径(mevalonate pathway,MVA)的第1个限速酶,对萜类物质的合成具有重要的作用。为了探明EuHMGRS家族基因对杜仲萜类物质合成的调控机制,对EuHMGRS家族基因进行生物信息学分析,结果表明:杜仲EuHMGRS基因中存在HMGRS特有的保守的活性位点,其中包括2个HMG-CoA结合位点(EMPVGYVQIP、TTEGCLVA)和2个NADP(H)结合位点(DAMGMNM、GTVGGGT),因此可以确定为杜仲HMGRS基因家族,在杜仲中该家族成员至少包含3个基因,记为EuHMGR12、EuHMGR18、EuHMGR19。杜仲HMGRS与其它植物的HMGRS蛋白相比在中间区域和C端保守性高,在N端的序列相似性较低,预示不同的HMGRS基因功能上具有高度的保守性,但是对于在亚细胞定位可能会具有多样性。

杜仲叶提取物对MC3T3-E1细胞增殖作用的实验研究

目的研究杜仲叶提取物（Euec）对小鼠成骨样细胞株MC3T3-E1细胞生长、增殖的影响及其分子机制。方法采用Euec与MC3T3-E1细胞体外共同培养，MTT法检测不同浓度Euec处理MC3T3-E1细胞24、48、72h后的细胞增殖情况，流式细胞仪检测细胞周期分布。结果4．0～125．0μg／ml的Euec能促进MC3T3-E1细胞增殖，其中7.8μg／ml的Euec对MC3T3-E1细胞增殖的促进作用最强。流式细胞仪检测显示，6.25μg／ml的Euec作用MC3T3-E1细胞48h后，G0/G1期成骨样细胞显著减少，S期成骨样细胞增多，G2/M期成骨样细胞显著增多。结论Euec可促进MC3T3-E1细胞向G2／M期转换，使G1期的成骨样细胞减少，G2期的成骨样细胞显著增加，从而促进细胞的有丝分裂和细胞的增殖。

杜仲叶酸性多糖EOP-1的分离纯化、结构解析

采用热水提取、用Savage法除蛋白,过氧化氢法脱色得到杜仲叶粗多糖(crude polysaccharide from leaves of eucommia ulmoides oliver)。经Sepharose CL-6B分离纯化得到杜仲多糖的一个组分EOP-1,通过HPSEC分析其纯度及测定分子量,表明该多糖约为6.0×105 D。部分酸水解和甲基化分析,结合GC-MS测试手段,对杜仲多糖EOP-1的糖链结构进行了研究。结果显示,EOP-1的糖残基主要由D-GalpA、D-Glcp、D-Galp、L-Araf和L-Rhap组成,主链由1,4-D-GalpA连接,为半乳糖醛酸聚糖,侧链主要由1,4-D-Galp、1,6-D-Galp、1,5-L-Araf、1,2-L-Rhap构成,其相对摩尔百分比为45.11∶35.9∶0.9∶10.5∶6.9。

杜仲提取组分对MC3T3-E1 Subclone 14成骨细胞的影响

目的:研究杜仲中各成分对MC3T3-E1 Subclone 14成骨细胞增殖、成骨细胞保护素(DPG)与核因子κB受体活化因子配体(RANKL)比值的影响。方法:对杜仲皮进行组分提取,得到A组分(水洗脱液)、b组分(20%乙醇洗脱液)和C组分(40%乙醇洗脱液);对照组MC3T3-E1 Subclone 14成骨细胞加入DMEM/F-2培养基(Hyclon)培养,给药组分别加入浓度为200、100、50、25和12.5 mg/L含A、B、C组分的培养基培养;采用MTT法检测MC3T3-E1 Subclone 14成骨细胞增殖情况;ELISA法检测MC3T3-E1 Subclone 14成骨细胞OPG及RANKL活性,紫外分光光度法检测MC3T3-E1 Subclone 14成骨细胞碱性磷酸酶(ALP)活性。结果:与对照组比较,给药24 h后,100和200 mg/L C组分给药组的OD值明显增高(P<0.05);与对照组比较,给药48 h后,200 mg/LC组ALP的活性以及OPG与RANKL比值明显增高,差异具有统计学意义(P<0.05)。结论:杜仲40%乙醇提取组分能明显上调OPG与RANKL比值,并能促进MC3T3-E1 Subclone 14成骨细胞的增殖。

杜仲EuREF1基因表达水平与橡胶积累的关系

[目的]为了探明EuREF1与胶分子量和含胶率的关系,深入探究杜仲胶合成机制。[方法]在杜仲新枝和叶片生长较快的4月中旬到6月中旬,利用RT-PCR技术分析杜仲EuREF1基因在雌雄株叶片和茎皮中的表达水平,以索氏提取法和GPC/SEC分别检测叶片和树皮中橡胶的含量和分子量。[结果]在4月至6月期间,雌雄株叶片、茎皮EuREF1表达水平分别与这些器官中胶含量及胶分子量呈显著或极显著正相关(R=0.8982~0.9880)。在雌株叶片、茎皮、果皮和雄株叶片、茎皮中,分子量在1.0×10^6~5.0×10^6 Da所占比例最多,即长度在1.5×10^4~7.4×10^4个异戊二烯单体的橡胶最多。[结论]结果推测EuREF1基因产物与杜仲胶积累程度有紧密的关系,可能在橡胶链的延伸中发挥重要作用。

杜仲糖基转移酶基因EuCGT1克隆及序列分析

本研究以杜仲(Eucommia ulmoides)7月雌株新生幼枝的树皮为材料,根据杜仲转录组测序数据中筛选的一个糖基转移酶基因(CGT)片段信息设计引物,采用SMART RACE技术,克隆了5’-端1432bp和3’-端535 bp cDNA片段,测序拼接后获得全长1 583 bp的杜仲CGT的cDNA序列。分析结果表明,该cDNA开放阅读框长1 464 bp,编码487个氨基酸,命名为EuGT基因。EuGT在NCBI中经blastn比对,显示序列与甜橙和草莓的UDP-葡萄糖基转移酶基因同源性分别为72%和67%;推测的氨基酸序列经blastp和CDD比对,显示序列与UDP-葡萄糖类黄酮3-O-糖基转移酶-6同源,其中含有UDP-葡萄糖基转移酶的保守域(pfam00201),初步推测EuCGT1蛋白属于糖基转移酶家族。本研究克隆的EuGT基因是首次从杜仲中克隆的糖基转移酶家族基因。以EuCGT1基因构建的植物表达载体pSH-35S-EuCGT1采用农杆菌介导法遗传转化烟草,获得了含EuCGT1基因的转基因烟株,移栽生长1个月的转基因烟草植株在表型上与野生型烟草无明显差异。