不同诱导子诱导虎杖愈伤组织白藜芦醇的生物合成

目的:研究不同胁迫条件对虎杖叶愈伤组织白藜芦醇含量的影响。方法:分别采用低温、热休克、紫外辐射、高盐、真菌诱激子、水杨酸(SA)、茉莉酸甲酯(MJ)处理虎杖叶愈伤组织,HPLC测定其中白藜芦醇的含量。结果:经过低温、紫外辐射、真菌诱激子、SA处理,白藜芦醇含量分别提高38%、20%、24%和18%(P<0.01),而热休克、NaCl高盐和MJ处理后,白藜芦醇含量与对照无显著差异(P>0.05)。结论:适当的胁迫条件能促进虎杖叶愈伤组织中白藜芦醇的合成。     

酶解法提高虎杖中白藜芦醇含量的研究

目的:提高虎杖中白藜芦醇的含量. 方法:采用酶解法使虎杖中白藜芦醇甙转化为白藜芦醇.对影响酶解的虎杖颗粒度、浸润用水量、酶解温度、时间4个因素进行考察,得出了最佳的酶解工艺. 结论:最佳酶解工艺是,虎杖粗碎至3 mm～10目、用3倍量的水浸润后、在(45±0.5) ℃下酶解2天.酶解后虎杖中白藜芦醇的含量提高了2.72倍.     

RP-HPLC法同时测定虎杖中白藜芦醇和白藜芦醇苷的含量

建立了国内首次用RP-HPLC同时测定虎杖中白藜芦醇和白藜芦醇苷含量的方法,并详细考察了2个指标成分的稳定性。对不同产地的虎杖药材、不同采收期的虎杖根及根茎以及虎杖的不同部位共25个样品中的白藜芦醇和白藜芦醇苷进行了含量测定,考察了两种活性成分的含量与产地、采收期及部位的关系,为虎杖资源的合理采收利用及进一步的研究提供了理论依据。     

酶解法提取纯化虎杖提取物中自藜芦醇的工艺研究

本文对虎杖提取物中虎杖苷的酶解条件及苷元白藜芦醇的提取纯化工艺进行研究,以样品中自藜芦醇的含量为指标,对纤维素酶、β-葡萄糖苷酶、复合酶进行筛选,结果表明以复合酶的水解效率最高:采用正交实验对影响复合酶酶解的因素:加酶量、温度、酶解时间进行考察;并对酶解后提取物中自藜芦醇的提取纯化工艺进行研究.得出如下较理想的酶解条件和提取纯化工艺:虎杖提取物,加水(pH 5)10倍,加20%的复合酶,于50℃保温24 h;酶解后的提取物经水、乙醇-水、碱溶液分步溶解沉淀,得白藜芦醇粗品,含量可达65%,工艺稳定可行.     

新型β-葡萄糖苷酶BglD2异源表达及水解虎杖苷能力

从海洋细菌Bacillus sp.D1中克隆、重组表达β-葡萄糖苷酶BglD2,研究其酶学性质,并对其水解虎杖苷制备白藜芦醇的能力进行分析。BglD2的最适催化温度和pH分别为45℃和6.5,在30℃和pH 6.5条件下的半衰期约为20 h。BglD2能够水解含β(1→3)、β(1→4)、β(1→6)等键型的多种底物。BglD2具有良好的糖促活特性,100 mmol/L葡萄糖和150 mmol/L木糖分别将酶活力提升2.0倍和2.3倍。BglD2具有较好的乙醇促活及耐受特性,30℃时,10%乙醇使酶活力提升1.2倍,25%乙醇存在时其仍保留60%的酶活力。BglD2具有水解虎杖苷制备白藜芦醇的能力,35℃条件下反应2 h水解率为86%。具有乙醇耐受及抗产物抑制等特性的β-葡萄糖苷酶BglD2在酶法水解虎杖苷制备白藜芦醇方面有应用潜力。     

凝胶柱层析分离虎杖中白藜芦醇的研究

本论文初步探讨了虎杖中白藜芦醇分离纯化的工艺条件,比较了不同溶剂、不同浓度、不同流速洗脱条件下白藜芦醇样品在LH-20凝胶层析柱上的分离效果,以及虎杖的乙醇提取液通过氯仿、乙酸乙酯萃取,凝胶柱层析分离后的成分变化,实验表明,当以甲醇为洗脱液,浓度为60%,流速为0.5 mL/min时,白藜芦醇样品的分离效果最好.采用中压液相色谱检测收集到的白藜芦醇纯度(以白藜芦醇峰面积占总峰面积计算)可达80.34%,回收率达86.16%.     

MPLC测定虎杖中白藜芦醇含量

建立测定虎杖中白藜芦醇含量的反相中压高效液相色谱分析方法。实验采用SOURCE 5RPC ST 4.6/150预装拄,以乙腈:水(体积比30:70 V/V)溶液为流动相、流速l mL/min、检测波长308 nm。结果表明白藜芦醇对照品浓度在0.6~1.6 mg/mL时,浓度与峰面积呈良好的线形关系(R=0.9919);加样回收率为103.0~90.4%,RSD=7.21%。采用中压高效液相色谱法测定虎杖中白藜芦醇含量,操作方便,准确性和实用性好。     

气象因子和矿质元素对虎杖根茎白藜芦醇含量的影响

对全年各月虎杖材料中白藜芦醇含量,及其Ca、Cu、Fe、K、Mg、Na、Zn 8种矿质元素积累进行检测,结合气象因子动态变化,采用主成分分析法,研究了不同季节虎杖白藜芦醇含量动态变化与其矿质元素、气象因子动态变化的关系．结果表明,Cu与Fe,Mg与K、Zn,Ca与月日照时数,K与Zn相关系数分别为0．812、0．871、0．793、0．602、0．729,呈极显著正相关;Cu、Mn与月平均气温,Fe与K相关系数分别为-0．738、-0．712、-0．766,呈极显著负相关;Cu与白藜芦醇含量呈显著负相关．找出了影响虎杖白藜芦醇含量动态变化的5个主成分,其中“酶促反应促进性矿质营养因子”,“高温、强照射气象因子”的贡献率分别为35．539％、33．358％,合计为68．897％,二者对不同季节虎杖白藜芦醇的变异贡献率最大．同时进行白藜芦醇含量的动态变化与主成分的多元相关、回归分析,建立了回归方程。     

虎杖中白藜芦醇的酶法制备

以白藜芦醇得率为指标,通过对酶制剂的筛选和酶解条件的考察,分别选出酶法提取和酶法转化制备虎杖中白藜芦醇的最佳条件,并对两种酶法进行比较.结果显示:(1)酶法提取最佳酶解条件为:酶解温度45℃,酶解时间60 min,最适pH 5.0,底物浓度17%;酶法转化最佳酶解条件为:酶解温度40℃,酶解时间48 h,最适pH 5.0,底物浓度15%.(2)与空白样品(不加酶)相比,酶法提取白藜芦醇得率增加了3.12 mg/g;酶法转化白藜芦醇得率增加了12.72 mg/g.研究表明,两种方法与不加酶提取相比均提高了白藜芦醇的得率,但酶法转化效果更显著,可用于白藜芦醇的制备.     

虎杖中白藜芦醇、白藜芦醇甙及大黄素的综合提取工艺研究

采用HPLC法,以白藜芦醇、白藜芦醇甙及大黄素的含量为指标,对虎杖中的有效成分进行了综合提取,通过正交实验,确定出乙醇溶剂提取的最佳工艺为：以80％的乙醇水溶液为提取剂,液固比为12：1,在70℃下提取2次,每次提取2h,HPLC法测定结果表明,从虎杖中提取的有效成分的含量分别可达：白藜芦醇为2．40mg／g,白藜芦醇甙为18．12mg／g,大黄素为16．82m/g。     

Comparative studies of polydatin and resveratrol on mutual transformation and antioxidative effect in vivo

BackgroundPolydatin and resveratrol are extractives of radix or rhizoma of Polygonum cuspidatum, and as the glycoside forms, it is a natural precursor of resveratrol.PurposeIn this study, we aimed to explore the mutual transformation between polydatin and resveratrol in rats, and to compare the antioxidative effect of them in vivo.Study designIn this study, we analyzed the serum molar concentration of polydatin and resveratrol after oral administration in rats and evaluated the anti-oxidative stress effects of them using a mouse model.MethodsRats were orally administered polydatin or resveratrol and the concentration of them in serum were analyzed by high performance liquid chromatography (HPLC). Their antioxidative effect was compared in mice with oxidative stress cardiomyopathy induced by doxorubicin (DOX).ResultsThe results showed that polydatin and resveratrol could mutually transform in vivo, the molar concentration of polydatin in serum was always averagely 3.35 and 4.28 times as much as resveratrol after oral administration of polydatin and resveratrol at 200 mg/kg, respectively. Both polydatin and resveratrol could significantly decrease the content of malonydialdehyde (MDA), promote the activities of total superoxide dismutase (T-SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in plasma, and increase the content of glutathione (GSH) in myocardial tissue. The effect of polydatin surpassed resveratrol, particularly embodied in increasing the activities of T-SOD and CAT, and the content of GSH.ConclusionIt illustrates that polydatin is the main substance in serum after intragastric administration with polydatin or resveratrol, and the mutual transformation between polydatin and resveratrol keeps balance; they both have the ability of antioxidative stress in vivo, and polydatin has a better effect than resveratrol, which hints that polydatin may be a substitute for resveratrol in antioxidant for clinical use.     

光强对毛脉酸模根中白藜芦醇和白藜芦醇苷的影响

在人工控制条件下,观察了光强对毛脉酸模根中白藜芦醇和白藜芦醇苷的影响.结果表明,较短时期的遮荫使毛脉酸模根中白藜芦醇和白藜芦醇苷含量随着遮荫程度的增加而增加,较长时期的遮荫则使白藜芦醇和白藜芦醇苷含量降低;随遮荫程度和遮荫时间的增加,根中白藜芦醇和白藜芦醇苷含量显著降低.根中白藜芦醇产量在8月份达最大值、白藜芦醇苷产量在9月份达最大值,遮荫降低根中白藜芦醇和白藜芦醇苷的产量.在毛脉酸模生长后期给予短期深度遮荫可以提高白藜芦醇和白藜芦醇苷产量.     

2种植物生长调节剂诱导烟草抗链格孢菌研究

以链格孢菌Alternaria alternata和烟草Nicotiana tabacum品种云烟87为试材,研究不同浓度茉莉酸甲酯(MeJA)和水杨酸(SA)处理下,链格孢菌菌丝生长情况以及成熟期烟叶防御酶活性与多酚含量变化,探讨植物生长调节剂对烟草抗链格孢菌的影响。结果表明,1 mmol·L^–1 MeJA对链格孢菌的抑制效果最好,抑菌率高达59%以上,其次是3.5 mmol·L^–1SA;MeJA和SA对链格孢菌的抑制效果随药剂浓度升高而增强;0.1 mmol·L^–1 MeJA和2.5 mmol·L^–1 SA能诱导烟草叶片SOD、POD、CAT等防御酶活性,并能降低H₂O₂活性氧含量,尤其以MeJA诱导效果较好。2种植物生长调节剂处理并接种链格孢菌能诱导提高烟叶多酚代谢相关酶PPO及PAL活性,但对烟草多酚类物质影响较小;成熟烟叶中含量较高的前3种多酚物质是绿原酸、芸香苷、隐绿原酸。     

Effects of varying air and soil moisture on the water relations and growth of sugar beet

Sugar beet were grown for short periods with different amounts of moisture in the soil and air. Growing plants in wet soil (23 % moisture on dry weight) compared with dry soil (15% moisture) increased growth of the shoots and roots and plant dry weights by 15% in young plants and 10% in mature plants. Growing plants in wet air containing 10.9 g m^-3 of water (equivalent to a saturation deficit of 2.5 mb) compared with dry air containing 6.4 g m^-3 of water (saturation deficit = 8.5 mb) increased the dry weights of both young and mature plants by 8%, mostly by increasing the sizes of their storage roots. Wet air and wet soil increased the net assimilation rates of both young and mature plants. Wet soil, but not wet air, increased leaf areas of young plants by accelerating leaf expansion, and both increased the leaf area of mature plants by slowing senescence of the older leaves. Wet soil increased the water potential of the leaves of both young and mature plants and, by doing so, increased their stomatal conductances and rates of photosynthesis. Wet air also increased stomatal conductances and rates of photosynthesis of leaves of plants of both ages, but without changing their water potentials. Stomatal conductances and photosynthetic rates were greater for young leaves than mature on the same plant and at the same water potential. It is suggested that at certain stages in the crops growth photosynthetic efficiency could be increased by applying additional water as a mist to increase the moisture content of the air around the crop.     

丛枝菌根真菌对番茄信号物质的诱导效应

盆栽番茄Lycopersicon esculentum幼苗分别接种丛枝菌根(AM)真菌摩西球囊霉Glomus mosseae、地表球囊霉G.versiforme、根内球囊霉G.intraradices、幼套球囊霉G.etunicatum及珠状巨孢囊霉Gigaspora margarita 35d后,开始测定番茄植株内源信号物质水杨酸(SA)、茉莉酸(JA)、一氧化氮(NO)和过氧化氢(H2O2)含量变化,抗性相关酶活性,丙二醛(MDA)含量以及生长量等指标。结果表明,接种AM真菌增加了番茄植株鲜重、株高、地上部和地下部干重、叶片和根系NO、JA、H2O2含量和结合态SA含量,其中,以摩西球囊霉G.mosseae诱导作用最大,叶片和根系内NO、JA、H2O2和结合态SA含量分别比对照增加了3.3和1.9倍、6.8和8.0倍、0.9和1.2倍、1.9和2.6倍,而根系中游离态SA含量一直处于较低水平,只有摩西球囊霉G.mosseae处理在诱导高峰时根系游离态SA含量比对照略有增加。接种AM真菌处理的番茄叶片和根系超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、苯丙氨酸解氨酶(PAL)活性显著增加,其中以摩西球囊霉G.mosseae的诱导效应最大,与未接种对照相比分别增加了0.6和0.3倍、7.9和3.1倍、0.4和1.2倍、2.3和1.9倍;幼套球囊霉G.etunicatum的诱导效应最小:与未接种对照相比分别增加了0.26和0.14倍、2.3和1.0倍、0.1和0.28倍、0.55和0.31倍;而MDA含量下降,分别降低了66%和68%、34%和41%、51%和50%、12%和26%、18%和29%。表明AM真菌能诱导植物同时产生多种信号物质,而且这些信号参与了AM真菌-番茄共生体系统抗性的表达。     

Effect of UV-C irradiation on stilbene synthase localization in young grape plants

Stilbene synthase (STS) is a pivotal enzyme that catalyzes the biosynthesis of resveratrol and could be induced by UV-C irradiation. However, at present the effect of UV-C irradiation on tissue and subcellular localization of STS is not studied. In this work, subcellular localization of STS in young grape (Vitis vinifera L. cv. Cabernet Sauvignon) plants exposed to UV-C was examined immunohistochemically using a polyclonal antibody raised against grape berry STS. The immunohistochemical analysis showed that the UV-induced STS occurred in palisade tissues of grape leaves and phloem tissues of grape leaf veins, stems, and roots. At the subcellular level, the enhanced STS stimulated by UV-C light was visualized in the cell walls, chloroplasts (plastids), cytoplasm, and nucleus of the phloem (stems and roots), while only in chloroplasts in mesophyll cells. This distribution pattern of STS arising in response to UV-C irradiation may be closely involved in its defense function, which needs much more in-depth evidence to confirm.     

三株长蠕孢属真菌对毛脉酸模幼苗中白藜芦醇、白藜芦醇苷含量和产量的影响

以幼苗期毛脉酸模（Rumex gmelinii Turcz．）为实验材料,研究经长蠕孢属真菌（Helminthosporium carbonum）侵染后毛脉酸模中白藜芦醇和白藜芦醇苷的含量和产量的变化。实验结果表明：幼苗期毛脉酸模中的白藜芦醇在受到外界真菌诱导时对真菌具有一定的选择性。而且同一真菌的不同菌株诱导强度也有一定的差异,主要表现在起效时间上。但白藜芦醇苷含量受外界真菌诱导的专属性不强,且影响强度与诱导浓度有关。对幼苗期毛脉酸模各种诱导处理都没有使其生物量受到明显的抑制。通过试验筛选出大斑病长蠕孢001使幼苗期毛脉酸模中白藜芦醇苷和白藜芦醇产量影响较大,与对照相比分别使其提高1．54和3．99倍。     

The influence of organophosphoric insecticides on total nitrogen and free amino acid levels in the leaves of apple seedlings

The effect of 0.1% Metation E 50 (fenitrothion) and Fosfotion 50 (malathion) on the level of total nitrogen and that of free amino acids was investigated in juvenile and mature leaves of apple seedlings in the course of six days following treatment. Fosfotion affected both nitrogen and amino acid content to a higher degree than Metation. Fosfotion increased the total nitrogen content in all leaves, but Metation induced the change only in the leaves which were at the end of the logarithmic growth phase at the time of treatment. The level of amino acids increased on the whole, with the exception of a temporary decline in the youngest leaves as well. The treatment with both insecticides affected most the content of amides: the greatest decrease of glutamine occurred in young leaves, the highest increase in asparagine content was recorded in both juvenile and mature leaves.     

增强UV-C辐射作用下小麦和豌豆幼苗光合响应及其相对抗性

以小麦和豌豆为材料,研究了UV-C辐射(波长<280 nm)对叶片光合特性及抗氧化酶活性的影响.结果表明: UV-C辐射增强,可使豌豆叶片光合速率减弱,气孔导度、胞间CO₂浓度、蒸腾速率和羧化效率明显降低,而对小麦叶片上述各项指标的影响则是先增加、后降低;在UV-C辐射下,豌豆的CO₂补偿点逐渐升高,而小麦的CO₂补偿点先降低、后升高.UV-C辐射除了使豌豆的POD活性和小麦的SOD活性逐渐降低外,其他抗氧化酶活性则呈先升高、后降低的变化趋势.小麦对短时间UV-C辐射的抗性比豌豆强,但随着UV-C辐射时间的延长,小麦和豌豆的抗氧化酶活性均降低,光合作用减弱.