Immunoregulatory polysaccharides from Apocynum venetum L.flowers stimulate phagocytosis and cytokine expression via activating the NF-κB/MAPK signaling pathways in RAW264.7 cells

Two immunomodulatory polysaccharides(Vp2a-Ⅱ and Vp3) were isolated and identified from Apocynum venetum L. flowers, and their innate immune-stimulating functions and working mechanisms were evaluated in RAW264.7 cells. Both the level of released nitric oxide(NO) and expression of inducible nitric oxide synthase(iNOS) m RNA were significantly enhanced in the RAW264.7 macrophages cells treated by Vp2a-Ⅱ and Vp3. Vp2a-Ⅱ(100–800 μg/m L) and Vp3(400 μg/mL) could significantly increase the phagocytic activity of RAW264.7 cells and the secretion and m RNA expression of TNF-α and IL-6 in a concentrationdependent manner through affecting mitogen-activated protein kinase(MAPK) activity and nuclear factor κB(NF-κB) nuclear translocation. Vp2a-Ⅱ might activate the MAPK signaling pathways and induce the nuclear translocation of NF-κB p65, whilst Vp3 likely activated the NF-κB and MAPK signaling pathways without influencing the p38 MAPK route.

Influence on the Quality of Apocynum Venetum with Different Degumming Methods

The biochemical degumming and chemical degumming were employed to process apocynum venetum, and indices values for the residual gum content, fineness, strength of single fiber, average length, moisture regain and whiteness of degummed apocynum venetum fibers are evaluated with fuzzy mathematical method. The results obtained from this investigation show that the general index value of degummed apocynum venetum fibers with biochemical degumming （K1= 0.626 09） is prior to that with chemical degumming（K2=0.421 64）.

Feasibility and Prospect about the Cultivation of Apocynum venetum L. in Coastal Salt-affected Soils

[Objective]The aim of this paper was to analyze the feasibility and prospect of Apocynum venetum L.cultivation in coastal salt-affected soils.[Method]The habitat and distribution of Apocynum venetum L.in coastal area of north China were studied.A method to integrate the charicteristics of Apocynum venetum L.（Luobuma） with the utilization of salt-affected soils in this region was proposed.[Result]The introduction,domestication and growth of plants on coastal salt-affected soils can realize the protection of endangered wild species,and achieve the purpose of improving soil.The characteristics of Apocynum venetum L.can produce optimum economic and ecological benefits.[Conclusion]The implementation will provide references for the protection and exploitation of wild plant,and it is of positive significance for the integration of soil and plant resources.

Apocynum venetum Feed Affects the Serum Metabolism of Tan-sheep through the Amino Acid and Glycolipid Metabolic Pathways

[Objective]The paper was to explore the effects of Apocynum venetum diet on nutritional metabolism of Tan sheep.[Method]Forty Ningxia Tan sheep were randomly divided into 4 groups(A,B,C and D),with 10 sheep in each group.The sheep were fed with different contents of A.venetum granule feedstuff(0%,5%,10%and 15%).Blood samples were collected from jugular vein on the 20th,40th and 60th day of the experimental period.Serum samples were prepared and analyzed for differential metabolites by UPLC-Q-TOF/MS technique and annotated to the KEGG pathway.[Result](1)The differential metabolites increased with the extension of feeding time.The up-regulated metabolites in Tan sheep serum were more than the down-regulated ones,and reached the maximum on the 40th day.The up-regulated differential metabolites included 2 amino acids(valine and alanine),7 organic acids(β-hydroxybutyric acid,fumarate,ethylmalonic acid,hydroxyphenylacetic acid,gentiolic acid,protocatechuic acid,and oxycholic acid),pyrocatechol and taurochenocholate.The down-regulated differential metabolite was only p-chlorophenol.(2)With the increase of A.venetum content in pelleted diet,the differential metabolites in the serum of Tan sheep also increased,and the serum metabolic level gradually stabilized after the 10%concentration level.Similarly,the up-regulated metabolites were far more than the down-regulated metabolites.The up-regulated differential metabolites included 3 organic acids(β-hydroxybutyric acid,oxycholic acid and gentiolic acid),2 amino acids(alanine and valine)and catechol,and the down-regulated differential metabolite was only taurochenocholate.(3)The metabolic pathways involving in differential metabolites were mainly tricarboxylic acid cycle,tyrosine metabolism,taurine and bile acid pathway.[Conclusion]The results will provide a scientific basis for the green breeding of Tan sheep.

Protective effect of Apocynum venetum (罗布麻) against lipid peroxidation damage of erythrocyte membrane

Objective: To investigate the protective effect of Apocynum venetum on lipid peroxidation damage of erythrocyte membrane. Methods: Model of lipid peroxidation of erythrocyte membrane was manufactured by three kinds of radicals generation systems. To set up a normal control group (NC),a model control group (MC),and four Apocynum venetum groups(A.V). Observation was made on the content of malondialdehyde (MDA). Results: Compared with MC group,all of the four Apocynum venetum groups dose-dependently inhibited the increases of MDA content in the membrane induced by xanthine - xanthine oxidase system,H 2O 2 or UV light. Conclusions: Apocynum venetum may protect erythrocyte membrane from the lipid peroxidative damage induced by radicals.

抗盐碱罗布麻DNA导入棉花的研究

利用微注射技术，把抗盐碱罗布麻ＤＮＡ导入鲁棉６号。对其后代经人工配制的盐碱土筛选育成了两个耐盐碱品系９１－１１和９１－１５，在含盐量为０．５１％的滨海盐碱地种植，其皮棉产量分别比受体亲本鲁棉６号增产１９１．７％和２３７．８％。对９１－１１和鲁棉６号在盐胁迫条件下测定其细胞质膜透性和超氧化物歧化酶（ＳＯＤ）活性，结果表明，９１－１１在盐胁迫条件下能够维持较高的ＳＯＤ活性和较稳定的膜透性，而鲁棉６号呈现相反的变化，而且差异显著。据此说明，应用这一技术能够筛选出含有抗性基因的改良品种。

基于RAPD标记的罗布麻野生居群遗传多样性分析

利用RAPD技术对采自我国8个省份的9个罗布麻野生居群进行遗传多样性分析.从80条随机引物中筛选出17条引物,共扩增出157条带,其中多态性带108条.物种水平的多态位点百分率(PPB)为68.79%,Nei’s基因多样性指数H为0.2296,Shannon’s多态性信息指数I为0.3390.居群间的遗传分化系数Gst为0.8841,即11.59%的遗传变异来自于居群内,88.41%的遗传变异来自于居群间,居群间的遗传分化水平远高于居群内.居群间遗传距离与聚类结果显示各居群间的亲缘关系与地理分布不完全相关.针对现有罗布麻的遗传多样性现状,建议加强对现有自然居群的保护.

罗布麻提取物对AngⅡ诱导的大鼠心肌纤维化中ERK通路的影响

目的:探讨罗布麻提取物对血管紧张素Ⅱ(AngⅡ)诱导的大鼠心肌纤维化模型中ERK通路的影响,并初步探索罗布麻提取物抗心肌纤维化机制。方法:采用1×10-6 mol.L-1 AngⅡ诱导大鼠心肌成纤维细胞复制心肌纤维化模型,细胞分为正常对照组、AngⅡ组、缬沙坦治疗组1×10-6 mg.L-1)及罗布麻提取物高剂量组(0.8mol.L-1)、中剂量组(0.4 mol.L-1)、低剂量组(0.2 mol.L-1)。通过对羟脯氨酸及Ⅰ、Ⅲ型胶原(ColⅠ、ColⅢ)的检测鉴定模型的成功复制,RT-PCR及Western blotting方法检测raf、ERK、c-fos mRNA及ERK蛋白在各组细胞中的表达量。结果:成功建立AngⅡ诱导的心肌纤维化模型。与AngⅡ组比较,不同剂量罗布麻提取物组羟脯氨酸及ColⅠ、ColⅢ的表达量均下调(P<0.05或P<0.01);RT-PCR及Western blotting检测发现,罗布麻提取物不同剂量组的raf、ERK、c-fos mRNA及ERK蛋白表达量较AngⅡ组降低(P<0.05或P<0.01)。结论:ERK信号通路在罗布麻提取物调控心肌纤维化发生和发展的过程中发挥重要作用。

基于DNA非编码区序列探讨罗布麻的分类问题(英文)

通过测定中国产罗布红麻、大叶白麻和白麻的ITSt、rnL内含子和trnL-F非编码区序列,并与其它相关植物的相应序列比较,探讨“罗布麻”的系统分类。结果显示:罗布红麻、大叶白麻和白麻3种植物的ITS序列完全一致,而与罗布麻属加拿大麻的ITS序列相差较大;在trnL内含子和trnL-F非编码区,大叶白麻和白麻的完全一致,与罗布红麻之间仅有3个位点的变异,而加拿大麻与美国茶叶花在这3个位点却和白麻属的完全一致等。以上结果显示在ITSt、rnL内含子和trnL-F非编码区,白麻属和罗布麻属之间没有本质区别,罗布红麻与大叶白麻和白麻之间的亲缘关系可能较罗布红麻与罗布麻属内其它物种间的亲缘关系更近。遗传距离与系统树的分析结果进一步支持以上推论。建议撤销白麻属,将大叶白麻和白麻合并到罗布麻属。

微波消解-AAS法测定新疆罗布麻叶中的微量元素

微波溶样技术是近年来产生的一种崭新的极有前途的溶样技术。采用微波消解溶样,利用原子吸收法对新疆罗布麻叶中多种微量元素含量进行测定,优化了消化条件。结果表明高压微波消化法具有消解完全、结果准确的优点。

优化的HPLC-DAD法同时测定罗布麻中4种黄酮类成分的含量

目的建立优化的HPLC方法同时测定不同产地罗布麻药材中芦丁、金丝桃苷、异槲皮苷以及槲皮素含量。方法采用Shiseido C18（3.0μm,3.0 mm×100 mm）色谱柱;以乙腈：甲醇=10：1作为A相-0.1%甲酸水溶液作为B相,梯度洗脱（0~4 min,5%→16%A;4~15 min,16%→23%A;15~20 min,23%→35%A;20~22 min,35%→55%A）,流速0.6 ml.min-1;二极管阵列检测器检测波长：360 nm;柱温：30℃;进样量：5μl。结果芦丁、金丝桃苷、异槲皮苷、槲皮素在22 min内基线分离,线性范围分别为0.308 4~30.84μg.ml-1（r=0.999 9）,0.877 6~87.76μg.ml-1（r=0.999 9）,0.902 0~90.20μg.ml-1（r=0.999 9）,0.249 8~24.98μg.ml-1（r=0.999 9）。方法学考察表明,日内和日间精密度、最低检测限和定量限的范围均符合相关标准,加样回收率（n=3）分别为：芦丁102.0%、96.40%、103.8%;金丝桃苷98.50%、101.0%、102.9%;异槲皮苷98.40%、100.4%、101.4%;槲皮素104.4%、103.1%、103.5%。测定了9个产地的罗布麻中4个黄酮类成分的含量。结论本法快速,准确、简便,具有良好的重复性,可以为罗布麻药材质量控制提供依据。

三种荒漠植物幼苗对NaCl胁迫的叶绿素荧光响应

了解极干旱区沙漠过渡带植物的耐盐能力有利于揭示其耐盐机制,保护和利用这些植物。叶绿素荧光技术被使用,以研究NaCl胁迫对温室培养的三种植物骆驼刺(Alhagi sparsifolia)、罗布麻(Apocynum venetum)和盐穗木(Halostachys caspica)幼苗叶片光系统II的影响,NaCl胁迫导致骆驼刺和罗布麻叶片PSII最大量子产量(Fv/Fm)下降,但是引起下降的原因不同——骆驼刺的Fm不变而Fo增加,而罗布麻则是增加初始荧光Fo同时降低最大荧光Fm,盐胁迫对盐穗木影响不大。盐胁迫也诱导了三种植物光适应能力的降低,其中以骆驼刺和罗布麻下降明显。三种植物对NaCl胁迫的忍耐能力排序为盐穗木>骆驼刺>罗布麻。

罗布麻及其易混品的DNA分子鉴定

为从分子水平更准确地鉴别罗布麻及其易混品,本研究利用PCR产物直接测序法对罗布麻、大叶白麻和Apoacynum cannabinum的核基因组(rDNA)ITS区与叶绿体基因组(cpDNA)trnL内含子及trnL-F间隔区序列进行测序与比较。结果显示,罗布麻和大叶白麻ITS序列完全一致,与A.cannabinum在ITS1区有13个位点、在ITS2区有10个位点不同;在trnL内含子区及trnL-F间隔区,罗布麻和大叶白麻共有3个位点不同,罗布麻和A.cannabinum间共有23个位点、大叶白麻和A.cannabinum间共有20个位点不同。研究表明,依据rDNAITS区序列可鉴别A.cannabinum和国产“罗布麻”(罗布麻与大叶白麻);利用cpDNAtrnL内含子及trnL-F间隔区序列可鉴别罗布麻及其相似种。

罗布麻K^+通道编码基因AvAKT1的克隆与表达分析

罗布麻是一种典型的钾高效植物,具有很强的K^+吸收和利用效率,通过增强对K^+的选择性吸收维持体内高且稳定的K^+含量和K^+/Na^+以抵御盐和干旱胁迫。为了探究其分子机制,采用RT-PCR和RACE技术从罗布麻根中克隆得到Shaker家族成员K^+通道蛋白基因AvAKT 1,其cDNA全长序列2906 bp,编码899个氨基酸。氨基酸序列分析显示AvAKT1包含Shaker同源蛋白家族典型的6个跨膜域、1个P环以及环核苷酸结合位点(cyclic nucleotide-binding domain,cNBD)、锚蛋白区(ankyrin-related domain,ANKY)和富含疏水酸性残基区(a domain rich in hydrophobic and acidic residues,KHA)保守结构域;系统进化树将Shaker家族分为5个亚族,AvAKT1隶属第Ⅰ亚族(AKT1亚族),与烟草进化关系较近。实时荧光定量PCR分析结果表明,AvAKT 1主要在罗布麻根中表达,在茎和叶中表达量极低;AvAKT 1受5.0 mmol·L^-1 K^+的显著诱导,且在-0.2 MPa渗透胁迫和25 mmol·L^-1 NaCl处理6 h时,在根中的表达水平显著上升。AvAKT 1可能参与调控罗布麻低亲和性K^+吸收过程,并在其应答干旱和盐逆境胁迫中发挥有效作用。

Na_2CO_3、MgSO_4对罗布麻种子萌发及幼苗生长的影响

采用单盐胁迫方法,研究Na2CO3、MgSO4胁迫对罗布麻种子萌发及幼苗生长的影响。结果表明:Na2CO3对罗布麻种子的萌芽具有抑制作用,二者呈极显著的负相关关系,同时,无论Na2CO3浓度高低,已萌芽的种子均不能正常生长;MgSO4对罗布麻种子萌发具有促进作用,对地上部生长具有明显的抑制作用,二者呈极显著的负相关关系,当MgSO4浓度不高于2.0%时对罗布麻地下部生长具有促进作用,高于2.0%具有抑制作用,总的变化趋势呈负相关关系。

罗布麻总RNA提取方法比较研究

以Trizol法、改进SDS法及RNA plant plus Reagent试剂盒法提取罗布麻的总RNA,并通过紫外分光光度法和凝胶电泳检测对其提取效果进行比较分析,以筛选出罗布麻总RNA最佳提取方法。结果表明:Trizol法无法有效获得罗布麻总RNA,有较为显著的降解现象;改进SDS法提取的罗布麻RNA总量较少、浓度低,且试验重复性差;RNA plant plus Reagent试剂盒法提取的RNA质量好,纯度高,完整性强,可直接作为后续相关分子生物学试验材料。

基于ITS、psbA-trnH及matK序列的罗布麻资源分子系统学研究

目的:从分子生物学角度研究罗布麻属植物种间差异,为确定《新疆维吾尔自治区维吾尔药材标准》中的植物基原提供依据。方法:采用DNA条形码ITS、psbA-trn H及mat K序列对罗布麻、白麻及大叶白麻55份样品进行PCR扩增并双向测序,比较种内种间变异,基于K2P模型进行遗传距离分析,并构建NJ系统树。结果:罗布麻、白麻及大叶白麻ITS序列种内及种间K2P遗传距离无显著差异,NJ树聚为一支;罗布麻与白麻及大叶白麻之间psbA-trnH和matK序列种内最大K2P遗传距离明显小于种间最小K2P遗传距离,NJ树显示罗布麻与白麻及大叶白麻均单独聚为一支;白麻与大叶白麻psbA-trn H和mat K序列无差异,NJ树均为一支。结论:基于ITS、psbA-trnH及matK序列鉴定结果,《新疆维吾尔自治区维吾尔药材标准》将药材罗布麻的基原定为大叶白麻Poacynum hendersonii值得商榷。

罗布麻eIF-5A基因功能

以罗布麻为试材,采用农杆菌介导法将罗布麻eIF-5A基因导入烟草,分别对正常生长条件和NaHCO_3胁迫下转AveIF-5A基因烟草和非转基因烟草的过氧化氢酶(POD)活性、超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量进行测定,以期为罗布麻eIF-5A基因功能研究提供参考。结果表明:正常生长条件下,转AveIF-5A基因烟草和非转基因烟草POD活性和MDA含量差异不显著,转基因植株SOD活性显著高于非转基因植株;在NaHCO_3胁迫下,转AveIF-5A基因烟草POD活性和SOD活性均极显著高于非转基因植株,而MDA含量极显著低于非转基因植株。AveIF-5A基因过量表达可以提高转基因烟草的耐盐能力。

微波消解–ICP–AES法测定罗布麻叶中16种微量元素

建立微波消解–ICP–AES法测定罗布麻叶中Na,Ca,Mg,Fe,Al,P,Mn,Ni,Cu,Zn,Ti,Se,Ba,Pb,Sr,Cd 16种微量元素含量的方法。以HNO_3–H_2O_2–HF溶液(6∶4∶1)为消解体系,样品经微波消解后,用ICP–AES法对罗布麻叶中16种元素含量进行测定。各元素的质量浓度在0.01~1.00 mg/L范围内与光谱强度呈良好的线性关系,线性相关系数r>0.999,检出限在0.000 5~0.008 0 mg/kg之间。加标回收率为87.56%~101.47%,测定结果的相对标准偏差为1.2%~3.42%(n=6)。该方法操作简单,准确度好,灵敏度高,可用于罗布麻叶中微量元素分析。

罗布麻CesA基因家族的生物信息学分析

植物体中纤维素是细胞壁形成的主要成分,不仅参与细胞形态的建成,调控细胞发育,还参与细胞内多种细胞信号转导途径,进而影响植物体的生长发育。纤维素合酶是植物体合成纤维素的主要酶类。为了探究CesA基因家族对罗布麻生长发育及纤维素合成的调控机理,该文通过生物信息学分析方法,从基因家族鉴定、结构分析、蛋白理化性质与多级结构预测、亚细胞定位、信号肽、进化关系和顺式作用元件等方面,对罗布麻CesA基因家族进行系统鉴定和分子特征分析。结果表明:基于全基因组测序,罗布麻CesA基因家族鉴定含有15个成员,分布在罗布麻11条染色体中的8条上,其编码蛋白的氨基酸数量为730~1158,相对分子质量81280.81~130123.18 kDa,理论等电点6.18~8.83。除了AvCesA3、AvCesA5、AvCesA7、AvCesA10和AvCesA11蛋白为稳定蛋白,其余成员均为不稳定蛋白;除了AvCesA12蛋白为疏水性蛋白,其余成员均为亲水性蛋白。该家族成员包含3~14个外显子,8~15个保守基序。编码蛋白主要分布于质膜与高尔基体上,无信号肽,二级结构以无规则卷曲与α-螺旋为主要构成元件。AvCesA15蛋白的跨膜结构域和三级结构与其他成员存在显著不同。罗布麻CesA基因进化时主要受纯化选择作用。对上游1500 bp区域顺式作用元件分析,结果显示罗布麻CesA基因受到光、温度、水分、氧气等环境因子及生长素、赤霉素、脱落酸、乙烯、水杨酸等植物激素调控。该研究为进一步探究罗布麻CesA基因家族的生物学功能、提高纤维品质与品种改良奠定理论基础。