紫叶苔(Pleurozia purpurea)线粒体基因组密码子偏好性分析

为了丰富对紫叶苔属遗传基础的认识,探究了紫叶苔([WTBX]Pleurozia purpurea)线粒体基因组密码子使用的偏好性及其影响因素。首先,基于完整紫叶苔线粒体基因组筛选蛋白编码序列;然后,利用CodonW、EMBOSS explorer计算各基因的GC含量、有效密码子数(ENC)、密码子适应指数(CAI)和同义密码子相对使用度(RSCU)等指标;最后,采用R软件进行参数间的相关性分析并绘图。结果表明:1)从紫叶苔线粒体基因组中筛选出31条蛋白编码序列,平均GC含量和密码子第3位碱基的GC含量分别为38.17%和32.28%,ENC和CAI平均值为50.29、0.17,表明密码子偏好使用以A/U结尾的碱基,且密码子偏好性较弱;2)ENC分别与GCall、GC3以及基因编码氨基酸的长度呈极显著正相关,表明GC含量和基因的长度对密码子的偏好有显著影响;3)中性绘图、ENC-plot和PR2-plot分析表明,突变压力和自然选择等因素共同塑造了密码子的使用偏好性,其中突变是最主要的影响因素;4)筛选出23个最优密码子,多数以A/U结尾。因此,紫叶苔线粒体基因组密码子偏好性较弱,除了主要受突变压力影响外,还受到自然选择、碱基组成和基因长度等因素的影响,研究结果可为苔藓植物线粒体基因组的进化研究提供参考。

Studies on Distribution and Chemical Characteristics of Copper in Cells of Setcreasea purpurea Boom

[Objective] The aim of this study was to reveal the mechanism of Cu2+ hyper-accumulator in Setcreasea purpurea Boom from the angle of distribution characteristics and binding form of Cu2+ in tissue cells.[Method]The distribution characteristics of Cu2+ in subcells of Setcreasea purpurea Boom was studied by the technique of differential centrifugation,and the binding form of Cu2+ in roots and leaves of Setcreasea purpurea Boom was also investigated by the sequential chemical extraction method and the enzymolysis method.[Result]Cu2+ in roots mainly distributed in the cell wall which is accounting for one third of the Total Cu2+ in roots,while Cu2+ in leaves mainly distributed in the chloroplast which is accounting for a quarter to the Total Cu2+ in leaves.Under the high concentration of Cu2+ or the extended treatment duration,the translocation of Cu2+ in root cells into the cell wall increased but the translocation of Cu2+ in root cells into the plastid decreased,while the translocation of Cu2+ in leaf cells into the chloroplast increased but the translocation of Cu2+ in leaf cells into the cell wall decreased.Cu2+ in leaves was mainly combined with amino acid,small molecular polymeric pigments,protein and polysaccharide,while Cu2+ in roots was mainly combined with cell wall substances such as cellulose and membrane-bound protein.[Conclusion]The distribution characteristics and binding form of Cu2+ in cells is possibly one of the dominant mechanisms for Cu2+ hyper-accumulator in Setcreasea purpurea Boom.

Analysis of Gene Differential Expression of Setcreasea purpurea Boom under Copper Stress by cDNA-AFLP

[Objective] The aim was to explore biological function of related genes in Setcreasea purpurea Boom under copper stress and to study the mechanism of its copper-resistance from standpoint of molecular biology in order to solve the problem of copper pollution. [Method] Setcreasea purpurea Boom was taken as experimental material which enjoys enrichment ability to cupric ion. About 90 fragments of differential expression were obtained by cDNA-AFLP and silver staining technique, among which, 17 fragments were amplified. After purification and identification, sequences of 6 differential fragments were got and used for BLAST X contrast. [Result] Six differential expressed fragments may play roles when Setcreasea purpurea Boom was under copper stress. The homology achieved 49% between differential sequences of E5MG-3 and of Arabidopsis thaliana mRNA （accession numbers： AAM62956.1）, homology was 53% between sequences of E4MB-2 and Solanum tuberosum mRNA （accession numbers： A5A717.1）, and 65% between sequences of E6MG-1 and Colocasia esculenta （L.） Schott mRNA （accession numbers： AAO62313.1）. It can be concluded that differential expressed genes are related to cell signaling, antioxidation, metabolism and protein modification. [Conclusion] The study has laid foundation for further exploration of regulatory network about response of Setcreasea purpurea Boom to copper stress.

Specific Protein Properties of Setcreasea pupurea Boom under Copper Stress

[Objective] This study was to investigate the expression of the specific protein in Setcreasea purpurea Boom under copper stress, with the aim to clarify the copper tolerance mechanism of S. purpurea. [Method] Methods of water culture, elec- trophoresis and chromatography were used to analyze the molecular weight of the specific protein in the copper hyperaccumulator S. purpurea, as well as its expression time and the minimum copper concentration for the expression. And the specific protein was isolated and purified. [Result] Under copper stress, the minimum concentra- tion of copper to induce the expression of the specific protein from S. purpurea was 50 umol/L, and the expression time of the protein was in the 4th week with the molecular weight of 89.4 kDa. [Conclusion] The results show that the copper tolerance of S. purpurea is closely related with the expression of the specific protein.

脱氧胆酸钠对麦角菌 Claviceps purpurea 94002 (EKP_N94002)产生 α-麦角隐亭的刺激作用

脱氧胆酸钠对麦角菌Ｃｌａｖｉｃｅｐｓｐｕｒｐｕｒｅａ９４００２（ＥＫＰＮ９４００２）产生α麦角隐亭的刺激作用朱平何惠霞陈世智（中国医学科学院、中国协和医科大学药物研究所，卫生部天然药物生物合成重点实验室，北京１０００５０）麦角菌属（Ｃｌａｖｉｃｅｐ...

影响绛红色小单孢菌(Micromonospora Purpurea)S-1212孢子萌发的几个主要因子的研究

影响绛红色小单孢菌孢子同步萌发的因子有营养、p H、温度、密度、溶氧等 ,试验结果表明 :p H6 .7～ 6 .8、温度 36℃、孢子密度以 1× 1 0 9个 / ml为宜 ,溶氧及一些生长因子 ,特别是甘氨酸、缬氨酸、丝氨酸 ,对该菌孢子萌发也是相当重要的因子 .

免疫增强剂紫锥菊(Echinacea purpurea)提取物对大菱鲆(Scophthalmus maximus)头肾内非特异性免疫基因表达量的影响

通过给大菱鲆(Scophthalmus maximus)注射不同浓度的紫锥菊(Echinacea purpurea)提取物,测定其非特异性免疫指标的变化水平,研究其对大菱鲆的头肾内非特异性免疫分子基因相对表达量的影响,进而为其在大菱鲆养殖生产过程中推广应用提供科学依据。试验选取体重(250±20)g的大菱鲆作为试验动物,分别注射10、20、40mg/m L浓度的紫锥菊提取物,试验共进行28d,注射试验结束后,分别从高、中、低剂量组及空白对照组选出6尾大菱鲆,分别取出大菱鲆的头肾并提取总RNA。采用?Ct法进行目标基因的荧光定量分析,再应用SPSS17.0软件对获得的实验数据进行单因素方差分析。研究结果表明,紫锥菊提取物能够不同程度地提高大菱鲆头肾内非特异性免疫分子基因相对表达量。注射紫锥菊提取物28d后对大菱鲆头肾中Lysozyme基因相对表达量的影响显著(P<0.05),对C3补体基因相对表达量的影响极显著(P<0.01),对Transferrin基因相对表达量的影响显著(P<0.05),对TGF-β1基因相对表达量的影响极显著(P<0.01),对IL-1h基因相对表达量的影响极显著(P<0.01),本试验研究表明,紫锥菊提取物能够显著提高大菱鲆头肾内非特异性免疫分子基因的相对表达量。

Effects of Echinacea purpurea Polysaccharide on IEC-6 Cell Proliferation

[Objective] This study was conducted to investigate the effects of Echi-nacea purpurea polysaccharides （EPS） on proliferation of rat intestinal epithelial cel IEC-6. [Method] The proliferation rate of IEC-6 cel s cultured in EPS at different concentrations and for different time was measured by MTT assay and analyzed by statistic methods. [Result] The proliferation rate of IEC-6 cel s cultured in EPS at al the concentrations and for different time was improved by different extents in com-parison with the control. In detail, 50 and 200 μg/ml EPS greatly improved the IEC-6 cel proliferation after 24 h of culture; then, the cel proliferation rate in the two treatments increased from 24 to 48 h, and declined from 48 to 72 h. The cel pro-liferation was also significantly improved by culturing in 100 μg/ml EPS for 72 h and in 500 μg/ml EPS for 48 h. After 48 h of culture, the proliferation rate of IEC-6 cel increased in a EPS dose-dependent manner. [Conclusion] EPS can promote IEC-6 cel proliferation, and thus improve the intestinal mucosal absorption and immune function of rat.

内生真菌对根寄生逆境下紫花针茅根系形态的影响

甘肃马先蒿是一种根寄生杂草,利用吸器在紫花针茅根部建立营养摄取通道,并掠夺性获取紫花针茅体内营养物质,从而抑制紫花针茅生长。研究表明,Epichloë内生真菌与紫花针茅共生能缓解甘肃马先蒿寄生对紫花针茅的生长抑制作用,这可能与内生真菌调控紫花针茅根系形态特征有关。本研究以带菌(E+)、不带菌(E-)紫花针茅为研究对象开展盆栽试验,探究甘肃马先蒿寄生密度对紫花针茅根系形态的影响。结果表明:1)甘肃马先蒿寄生显著降低了紫花针茅地上和根系生物量,抑制了紫花针茅根系的生长发育,且随寄生密度增加,紫花针茅生物量和根系形态指数持续降低,但E+植株的生物量和根系形态优于E-植株。2)通过结构方程模型分析发现,甘肃马先蒿寄生能抑制紫花针茅根长、根表面积和根系拓扑指数,导致紫花针茅生物量降低,且随寄生密度的增大,甘肃马先蒿对紫花针茅生长的抑制作用不断增强。内生真菌通过改变紫花针茅根长、根表面积和外部连接数等根系形态特征对甘肃马先蒿寄生作出正向的积极响应,进而增加紫花针茅生物量。由此可见,内生真菌能通过调控根系形态特征来增强紫花针茅根系和地上生物量,进而缓解甘肃马先蒿对紫花针茅的根寄生危害,这将为利用禾草内生真菌共生体这一特殊资源对根寄生杂草防控提供新思路。

紫花亚菊(Ajania purpurea)愈伤组织诱导及植株再生体系的建立

以紫花亚菊(Ajania purpurea)组培苗茎段、叶片为外植体,在附加不同浓度植物生长调节剂6-BA和NAA的MS培养基上诱导培养,获得再生植株。对试验结果进行观察分析筛选出合适的配方:愈伤组织诱导培养基为MS+1.0 mg·L-16-BA+0.1 mg·L-1NAA,诱导率达95.6%;芽诱导培养基为:MS+1.0 mg·L-16-BA+0.2 mg·L-1NAA,诱导率达82%;不定根最适诱导培养基为:1/2 MS+0.15 mg·L-1IBA,生根率达90%以上,再生植株移栽成活率达90%。

Study on Flavonoids from the Methanol Extracts of Tephrosia purpurea Leaves

[Objective] The aim was to determine flavonoids from the MeOH extracts of Tephrosia purpurea leaves and their cytotoxicitives against the ovarian cells from Sprodenia litura （SL cells）.[Method] The compounds were isolated with column chromatography and their structures were established on the basis of various spectroscopic analysis （including UV,1D and 2D NMR analyses as well as HR-ESRMS）.The cytotoxicity against the SL cells was evaluated by using MTT method.[Result] Six known flavonoids,6-methoxykaempferol （1）,6-methoxykaempferol 7-O-α-rhamnopyranoside （2）,6-methoxykaempferol 3-O-α-rhamnopyranosyl（1→2）[α-rhamno-pyranosyl（1→6）]-β-galactopyranoside （3）,6-methoxykaempferol 3-O-α-rhamnopyranosyl （1 →2）[α-rhamnopyranosyl （1 →6）]-β-galactopyranoside-7-O-α-rhamnopyranoside （4）,pongachin （5）,5,7-dimethoxy-8-（3-hydroxy-3-methylbut-1Z-enyl） flavanone （6） were isolated and determined.Except compound 5,the others were isolated from T.purpurea for the first time.For the cytotoxicity compound 5 had significant activity with the IC50 value of 4.4 mg/L while compound 1 and 3,whose cytotoxicity exceeded rotenone,also showed moderate activity.[Conclusion] Of all the compounds from T.purpurea leaves,the content of 6-methoxykaempferol compounds was considerable.The profiles of these compounds against SL cells suggested that compounds 1,3 and 5,whose cytotoxicity exceeded rotenone,were worth further research.

Effect of Echinacea purpurea Polysaccharide on IL-6 m RNA Expression Level in IEC-6 Cell after LPS Injury

[Objective] The aim was to investigate the effect of Echinacea purpurea polysaccharide （EPS） on IL-6 mRNA expression level in IEC-6 cell after lipopolysac- charide （LPS） injury. [Method] Total RNA of IEC-6 cell was extracted with TRIzon reagent and amplified by R-r-PCR. The amplification products were examined by a- garose gel electrophoresis and graphed for analysis. [Result] After stimulation by LPS, the IL-6 mRNA expression level in iEC-6 cell increased. However, EPS could inhibit this effect, and the inhibitory effect was dose-dependent. At the concentration of 50 μg/ml, EPS could partially inhibit the IL-6 mRNA expression in IEC-6 cell after LPS stimulation; in the concentration range of 100-500 μg/ml, the inhibitory effect of EPS on IL-6 mRNA expression in iEC-6 cell increased with the increase of con- centration. When the IEC-6 cell was pre-treated with EPS （50, 100, 200 and 500 μg/ml） for 24 h and then stimulated with LPS （10 μg/ml） for 1 and 4 h, respectively, it was found that the LPS-induced mRNA expression of IL-6 in IEC-6 cell was in- hibited by EPS, and this kind of inhibitory effect was time-dependent. [Conclusion] After small intestinal epithelial cells were stimulated by LPS, the IL-6 mRNA expres- sion level increased. However, EPS could inhibit the LPS-induced mRNA expression of IL-6, thus protecting the intestinal mucosa. In addition, this kind of inhibitory effect showed time and concentration dependence.

南海海绵Spirastrella purpurea Lamarck的化学成分研究

从中国南海海绵 Spirastrella purpurea Lamarck中分离得到 5个有机化合物。通过现代波谱技术并结合X-衍射分析,确定其结构分别为胆甾醇(Ⅰ)、麦角甾-6,22-二烯-5α,8α-环二氧-3α-醇(Ⅱ)、对羟基苯乙酸胺(Ⅲ)、腺嘌呤(Ⅳ)和(±)-尿囊素(Ⅴ)。本文是对该种海绵化学成分的首次报道。

灰毛豆Tephrosia purpurea中查尔酮类化合物的分离鉴定

【目的】为探讨灰毛豆属植物Tephrosia purpurea化学成分中的查尔酮类化合物,对其枝、叶部位进行化学成分分离及结构解析。【方法】采用乙醇提取、溶剂萃取、柱层析分离方法得到多种灰毛豆属T.purpurea化学成分,并用波谱分析手段对其进行结构表征。【结果】从灰毛豆属T.purpurea中共分离鉴定出11个查尔酮类化合物,分别为elatadihydrochalcone(1),spinochalcone A(2),3′,5′-diisopentenyl-2′,4′-dihydroxychalcone(3),candidachalcone(4),O-methylpongamol(5),(+)-tephrosone(6),spinochalcone C(7),epoxyobovatachalcone(8),tunicatachalcone(9),crassichalone(10),(S)-elatadihydrochalcone(11)。【结论】以上化合物均为查尔酮,且均属首次从该植物中分离得到。

紫锥菊中烷基酰胺抗神经炎症活性研究

为了探究天然植物紫锥菊中烷基酰胺(AADs)对帕金森病(Parkinson’s disease,PD)神经炎症小鼠的保护作用,文章以紫锥菊醇水粗提物为原料,通过石油醚萃取、硅胶柱层析、葡聚糖凝胶LH-20分离纯化获得AADs;采用腹腔注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)1周构建PD神经炎症小鼠模型,研究灌胃给予AADs(0.75、3.00 mg/kg)或多巴胺片(3.00 mg/kg,阳性药)3周后对小鼠3种行为学、血清多巴胺和炎症因子质量浓度、纹状体内酪氨酸羟化酶(tyrosine hydroxylase,TH)阳性细胞比率的影响。研究结果表明:纯化物中的AADs质量比较醇水粗提物提高了36.3倍,达到(5.30±0.25)mg/g;AADs给药(3.00 mg/kg)7 d使PD神经炎症小鼠爬杆时长缩短、悬挂和游泳评分得到明显改善(P<0.01);与给药7 d相比,延长给药周期(21 d)可进一步改善小鼠行为学指标;AADs给药(3.00 mg/kg)21 d,小鼠血清中多巴胺质量浓度提高了43.9%;而一氧化氮合酶(inducible nitric oxide synthase,iNOS)、环氧合酶-2(cyclooxygenase-2,COX-2)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白细胞介素-6(interleukin-6,IL-6)4种炎症因子水平分别降低了12.3%、10.9%、15.3%、17.8%;纹状体内TH阳性细胞比率增加了94.1%。总之,AADs能够显著改善PD神经炎症小鼠的运动灵敏性、握力和协调性,该保护作用可能通过抑制炎症因子释放、维持多巴胺正常水平、增加TH阳性细胞比率而实现。

Anticancer activity of Tephrosia purpurea and Ficus religiosa using MCF 7 cell lines

Objective:To investigate anticancer activity of different fractions of Tephrosia purpurea[TP] （Sharapunkha,Fabaceae） and Ficus religiosa[FR]（Peepal,Moraceae）.Methods:The fractions of TP and FR were prepared and tested for in vitro anticancer activity using human MCF 7 cell line by trypan blue exclusion method.Results:The result showed that among all these fractions of TPI.TPIII.FRI and FRIII showed better anticancer activity compared to other fractions.The IC50 value for TPI（152.4μM）,TPIII（158.71μM）.FRI（160.3μM） and for FRIII（222.7μM） was observed.Conclusions:The present study shows anticancer potential of TP and FR fractions in MCF 7 cell line.

Evaluation of antiinflammatory activity of Tephrosia purpurea in rats

Objective:To evaluate the antiinflammatory activity of orally administered ethanolic extract of Tephrosia purpurea in acute and subacute inflammation in rats.Methods:An ethanolic extract of Tephrosia purpurea was prepared.Carrageenan induced paw edema and cotton pellet granuloma were the models for acute and subacute inflammation respectively.Four groups of rats in each model were treated orally with 2%gum acacia,100 mg /kg of aspirin,500 mg/kg and 1 000 mg/kg of ethanolic extract of Tephrosia purpurea respectively.In carrageenan induced paw edema model, subplantar injection of 1%carrageenan was made into the hind paw of the rats sixty minutes after the administration of the respective drugs.The paw volume was measured immediately after injection of carrageenan,at 3 hours and at 6 hours.Then percentage inhibition of edema was calculated.In the cotton pellet granuloma model,animals were administered drugs for six days after placing cotton pellets in the axilla on each side.On the 7th day,dry weight of granuloma was calculated.Results:The rats treated with Tephrosia purpurea did not exhibit any significant decrease in paw volume and serum ceruloplasmin levels as compared to the control and aspirin treated groups in the acute inflammation model;while,there was a significant(P 【 0.01) decrease in the weight of granuloma in Tephrosia purpurea and aspirin treated groups as compared to control in subacute inflammation.Conclusions:The ethanolic extract of orally administered Tephrosia purpurea shows significant antiinflammatory effect in subacute inflammation but not in acute inflammation in rats.

Trait complementarity between fine roots of Stipa purpurea and their associated arbuscular mycorrhizal fungi along a precipitation gradient in Tibetan alpine steppe

Development of fine roots and formation of symbiosis with arbuscular mycorrhizal(AM) fungi represent two strategies for plants to acquire nutrient and water from soil. Here, we elucidated how fine root development and symbolized mycorrhizal fungi with Stipa purpurea responded to the precipitation change in Tibetan alpine steppe ecosystem across a precipitation gradient from 50 mm to 400 mm. As precipitation increased, the proportion of thinner fine roots(diameter < 0.4 mm) in total roots increased significantly; while the mycorrhizal colonization percentage, either associated with thinner or thicker roots, decreased. This phenomenon indicated that fine root development and symbolized mycorrhizal fungi are likely alternative, and plant preferred to develop fine root rather than build a symbiotic relationship with mycorrhizal fungi in more benign niches with higher precipitation. Also, root diameter was negatively correlated with specific root length(SRL), but positively correlated with AM fungal colonization percentage, indicating thicker-root species rely more on mycorrhizal fungi in alpine steppe. The complementarity between fine root and mycorrhizal fungi of S. purpurea is mediated by precipitation in Tibetan alpine steppe.

The Synthesis and Storage Sites of Phenolic Compounds in the Root and Rhizome of <i>Echinacea purpurea</i>

Cichoric acid is the main phenolic compound in the root and rhizome of the medicinal part, Echinacea purpurea that is known for possessing immune enhancing characteristics. In this study, we analysis the the synthesis and storage sites of phenolic compound in E. purpurea. We used fluorescent microscopy, transmission electron microscopy, cytochemical and immunocytochemical localization to observe the distribution of phenolic compounds. Our results show that the phenolic compounds were mostly distributed in the cortex parenchyma cells, vascular parenchyma cells and pith parenchyma cells in the root and rhizome, and mainly present in the vacuoles, large intercellular spaces and their surrounding cell walls. No phenolic compounds were observed in the cytoplasm and the organelles. We concluded that the phenolic compounds were synthetized in the cortex parenchyma cells, vascular parenchyma cells and pith parenchyma cells in the root and rhizome, and stored in the vacuoles of parenchyma cells. The above results provided significantly cytological information for further approaching the metabolic regulation and transfer pathways of phenolic compounds in biochemistry and molecular biology.

A Monoterpene Glycoside from Ehinacea purpurea

Aim To separate and identify chemical constituents of Ehinacea purpurea . Methods Five compounds were isolated from the plant using chromatography. Their structures were elucidated by spectroscopy. Results Five compounds were isolated and their structures were identified as 2, 6 dimethyl 7 octene 2, 3, 6 triol 2 O β D glucopyranoside (1), 7, 8 furocoumarin (2), 6 methoxy 7 hydroxycoumarin (3), caffeic acid (4), methyl caffeate (5), and ethyl caffeate (6). Conclusion All these compounds were obtained from the plant for the first time.