Study on Extraction of Polysaccharides and Antioxidant Activity of Sagittaria sagittifolia L.Polysaccharide

[Objective] This experiment was conducted to investigate the optimal ex- traction process and antioxidant activity of polysaccharide in Sagittaria sagittifo/ia L. [Method] The effects of extraction temperature, extraction time, extraction times and material-to-liquid ratio on content of polysaccharides in S. sagittifo/ia were investi- gated. On the basis of single-factor tests, an orthogonal test was carried out to de- termine the optimal process parameters. In addition, the in-vitro antioxidant activity of S. sagittifo/ia polysaccharides was evaluated on the basis of determination of their scavenging activities against DPPH and hydroxyl free radicals and reducing power. [Result] The results showed that the optimized process parameters were as follows： material-to-liquid ratio at 1：40 （g/ml）, extraction temperature at 90 ℃, extrac- tion time of 4 h, and extraction times of 3 times. Under this condition, the content of S. sagittifo/ia polysaccharides was 29.32%. For 1 mg/ml S. sagittifo/ia polysaccha- rides, the scavenging ratios of DPPH and hydroxyl free radicals were 70.62% and 35.82%, respectively, and in the determination of reducing power, the absorbance at 700 nm was 0.453 1. [Conclusion] It could be concluded that S. sagittifolia polysac- charides have strong antioxidant capacity to have free radicals scavenged in vitro.

Cd^(2+)和Cr^(6+)对慈姑(Sagittaria sagittifolia L．)的毒害效应

以水生经济植物慈姑(Sagittaria sagittifolia L．)为实验材料,通过模拟水体Cd2+和Cr6+污染环境,研究了两种单一离子处理对慈姑的毒害影响．结果表明:随着水体Cd2+、Cr6+浓度增大,慈姑叶绿素含量与根细胞质膜透性先升高后降低;Cd2+浓度增大,根和叶的POD活性,叶的SOD活性、丙二醛(MDA)含量及根的超氧阴离子(O2+)产生速率均是先升后降;根和叶的CAT活性、根的SOD活性、MDA含量和叶的O2-产生速率则逐渐升高;而Cr6+处理时,MDA含量、根的 SOD、CAT、POD活性均随浓度增大而升高,O2-产生速率和叶的SOD、CAT、POD活性则升高后降低;Cd2+和Cr6+污染都对慈姑产生毒害,其毒害的效应既有相似性又有差异,同浓度条件下根的受害程度比叶严重．

慈姑(Sagittaria sagittifolia L．)对环境镉胁迫因子的应答

主要研究了水环境中Cd2+污染对慈姑(Sagittaria sagittifolia L.)根的毒害影响。结果表明:在Cd2+胁迫下,慈姑根出现不同程度的褐变、发黑,生长受到限制;叶和根尖细胞超微结构受损,叶绿体膨大、类囊体排列紊乱、线粒体嵴减少、空泡化、染色质凝集等;根尖细胞在扫描电镜下呈细胞壁增厚、细胞形状扭曲和排列不规则现象。能谱分析结果表明随外界Cd2+浓度增大,单个慈姑根尖中的Cd2+含量升高,其他营养元素的吸收受到不同程度的影响。Cd2+对慈姑的毒害有明显的剂量效应和时间效应。

Effects of “3414” Fertilization Effect Test on the Yield and Economic Benefit of Sagittaria sagittifolia

[Objective] To screen the optimal fertilizer formula, and to lay foundation for the research on special fertilizer for Sagittaria sagittifolia production. [Method] Based on the ＂3414＂ fertilization design, the yield of S. sagittifolia in each treatment was determined, and the effects of N, P and K treatments on the yield and eco- nomic benefit of S. sagittifolia were investigated, obtaining the optimal fertilizer appli- cation ratio. [Result] N, P and K had relatively good effects on the yield of S. sagittifolia. Treatment of N2P2K3 showed the optimal fertilizer application ratio, with the yield of 18 397.80 kg/hm2, yield increasing rate of 104.77%, and net income in- crease of 80 317.35 Yuan/hm2. N fertilizer had the greatest impacts on the yield of S. sagittifolia, followed with K fertilizer; while P fertilizer showed the poorest impacts. [Conclusion] To achieve the highest yield of S. sagittifolia, the fertilizer application was as follows： N of 446.70 kg/hm2, P20 of 568.10 kg/hm2, K2O of 534.90 kg/hm2, when the highest yield was 18 612.75 kg/hm2. Fertilizer application for the optimal economic benefit was N of 420.60 kg/hm2, P2O5 of 70.50 kg/hm2, K2O of 492.15 kg/hm2, when the optimal yield was 18 579.00 kg/hm2.

Extraction and Anti-inflammatory Effects of Colchicine from Sagittaria sagittifolia

[Objective] This study was to study the optimal extraction technology and anti-inflammatory effects of colchicine from Sagittaria sagittifolia. [Method] The ef- fects of ethanol concentration, extraction time, extraction temperature and solid-liquid ratio on the extraction rate of colchicine from S. sagittifolia were investigated. On the basis of single-factor experiments, an L9 （34） orthogonal test was carried out to optimize the extraction process. According to the optimal extraction process, the content of colchicine in S. sagittifolia was determined by high performance liquid chromatography. The anti-inflammatory ability of colchicine was evaluated through an anti-inflammatory test in vitro. [Result] The optimal extraction process of colchicine from S. sagittifolia was as follows： ethanol concentration of 60%, extraction temper- ature of 50℃, extraction time of 30 min, and solid-liquid ratio of 1：25 （g/ml）. The content of colchicine in S. sagittifolia was determined as 40.58 μg/100 mg. Com- pared with the control, the colchicine from S. sagittifolia （9.0 and 4.5 g/kg） inhibited the increase in PGE2, TNF-α and IL-1β contents in pleural fluid （P〈0.05）. High-dose colchicine inhibited the increase in TNF-α, IL-1β and MDA contents in lung tissue （P〈0.01）, and middle-dose colchicine inhibited the increase in IL-1β content in lung tissue （P〈0.01）. [Conclusion] The colchicine in S. sagittifolia has a good anti-inflam- matory effect, which may be achieved through hindering the production of inflam- matory mediators and antioxidation.

High Efficiency of Propagation for Sagittaria sagittifolia Using a Temporary Immersion Bioreactor System

Although several studies have reported on the propagation of the Sagittaria sagittifolia to date, none of these methods have efficiently achieved the mass production of these plants. The present study aimed to investigate the propagation and growth of S. sagittifolia using a temporary immersion bioreactor system （TIBS） compared with conventional semi-solid and liquid culture. The effect of different immersion frequencies and immersion times together with supplementation of various plant growth regulators to the Murashige and Skoog （MS） medium was evaluated on shoot proliferation and plant growth. The results showed that the higher immersion frequency （every 6 h） and shorter immersion time （3 min and 10 rain） in medium containing 4 mg/L BA and 0.1 mg/L NAA produced the highest multiplication rate （23）, which are significantly higher than conventional semi-solid （3.6） and liquid （4.5） method, and the best plant growth parameter. While, the lower immersion frequency and longer immersion time （30 rain every 12 h and 60 min every 24 h） induced vitrification and pollution rate in shoot tips 16.6% and 19%, 42% and 37%, respectively. There is distinct decrease in pollution rate （8.3%） in TIBS （10 min every 6 h） compared with the conventional semi-solid and liquid cultures. Medium containing 4 mg/L BA and 0.5 mg/L NAA using 10 min immersion every 6 h showed satisfaction at the rooting stage, with high shoot proliferation rate （21.6）, 100% rooting and 94% plant survival. Therefore, applying TIBS in S. sagittifolia is an efficient method for scaling up the production of plantlets with high quality seedlings.

Changes of Essential Mineral Elements Contents in Response to Cu²⁺Treatment in <i>Sagittaria sagittifolia</i>

Changes of various mineral elements (P, K, Ca, Mg, Fe, Mn, Zn and Na) contents in roots and leaves of S. sagittifolia were studied with treatment of different Cu2+ concentrations (5 μM, 10 μM, 20 μM and 40 μM) after 15 days. The results showed that: 1) Cu accumulated in roots of S. sagittifolia in large quantities, while Cu content in leaves showed no significant change;2) It can be seen from the changes of macroelements that Cu2+ treatments had inhibited the absorption of P, K, Ca, Mg in roots of S. sagittifolia, but the contents of P, K and Mg in leaves were higher than those in the roots in all Cu2+ treatment groups;3) It can be seen from the changes of microelements that Cu2+ treatment promoted the absorption of Fe, inhibited absorption of Mn, Zn and Na in roots of S. sagittifolia, and hindered the transport of various micro-elements from roots to leaves. In all the Cu2+ treatment groups, contents of Fe, Mn, Zn and Na in leaves were lower than those in the roots;4) The critical concentration of Cu2+ to S. sagittifolia was 5 μM. It could be seen from the above results that exogenous added Cu2+ of different concentrations broke the balance of various mineral elements in S. sagittifolia, which would exert a significant impact on numerous metabolic pathways and physiological processes.

慈姑多糖调控TLR4/MyD88/NF-κB抑制细胞炎症和氧化损伤对多种重金属诱导L02肝细胞凋亡的影响

目的探索慈姑多糖(Sagittaria Sagittifolia polysaccharide,SSP)是否通过介导Toll样受体4(Toll-like receptors 4,TLR4)/髓样分化因子88(myeloid differentiation factor 88,MyD88)/核转录因子-κB(nuclear transcription factor-kappa B,NF-κB)信号通路对6种重金属联合所致L02肝细胞损伤发挥保护作用。方法首先根据北京市售大米6种重金属膳食摄入比例,构建6种重金属混合染毒液所致L02细胞损伤体外模型。应用MTT法检测不同质量浓度慈姑多糖(0.25、0.5、1、2 mg/mL)提前作用24 h对L02细胞存活率的影响;Annexin V-FITC/碘化丙啶(PI)双染流式结合Hoechst 33342荧光检测细胞凋亡;DCFH-DA荧光探针检测细胞中活性氧(reactive oxygen species,ROS)含量变化;Western blot法检测细胞中多聚ADP核糖聚合酶1(poly-ADP ribose polymerase1,PARP1)、动力相关蛋白1(dynamin-related protein 1,Drp1)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素(interleukin,IL)-1β、IL-17、TLR4、MyD88、NF-κB p65蛋白表达情况。结果MTT结果显示,SSP在1 mg/mL作用24 h保护效果最佳;与模型组相比,SSP治疗组中ROS含量及细胞凋亡率均明显下降(P<0.05),Drp1、TNF-α、IL-1β、IL-17和TLR4、MyD88、NF-κB p65蛋白表达均显著降低(P<0.05),PARP1蛋白表达显著升高(P<0.01)。结论SSP可有效改善6种重金属联用诱导的L02细胞凋亡,可能与调控TLR4/MyD88/NF-κB信号通路抑制氧化损伤和炎症反应相关。

Production of Sagittaria sagittifolia Beverage and Technological Condition Optimization

[Objectives] To develop a kind of antioxidant health drink which could also regulate blood sugar.[Methods]With Sagittaria sagittifolia,hawthorn and liquorice as raw materials,the orthogonal test was done to determine the optimum preparation ratio of each raw material,optimum flavor modulation ratio and optimum stabilizer type and amount.[Results]Under the conditions of 0. 4% crude polysaccharide powder amount of Sagittaria sagittifolia,10. 0 m L hawthorn extract,2. 5 m L liquorice extract,7. 0% white sugar amount,0. 5% citric acid amount,0. 2% cyclodextrine amount,0. 05 g/L sodium alginate amount,0. 5 g/L xanthan gum amount,2. 0 g/L agar amount and 20 MPa homogenization,the beverage had optimum flavor and stability.[Conclusions]The health drink obtained under these technological conditions had delicate sweet taste and rich nutrition.

A Preliminary Study on the Tissue Culture of Sagittaria trifolia L.

[Objective] This study was to optimize the experimental conditions for large scale propagation of Sagittaria trifolia L via tissue culture.[Method] The dominant S.trifolia cultivar Baoyingziyuan introduced from Jiangsu Province was used as experimental material to study the impacts of various culture conditions on tissue culture of its stem tips and induction of stolons.[Result] Hormone combination 0.10 mg/L 6-BA ＋0.05 mg/L NAA performed best in plantlet regeneration and 2.0 mg/L 6-BA ＋0.5 mg/L NAA best in induction of stolon.Various sucrose concentrations did not show significant difference in the impact on sprouted stolons.[Conclusion] Various culture conditions could to some extent impact plantlet regeneration and stolon induction,and our results reveal the optimal hormone combinations for regeneration and stolon induction of S.trifolia.

慈菇对CCl_4致大鼠肝纤维化血清cyfra21-1和CEA影响的探讨

目的 探讨慈菇对CCl4 致大鼠肝纤维化的干预作用。方法 对 5月龄Wistar大鼠 ,用CCl4 染毒致大鼠形成肝组织纤维化 ,以ELISA法和放射免疫分析法检测大鼠血清cyfra2 1 1和CEA的浓度的变化 ,并观察慈菇的干预效果。结果 染毒组大鼠cyfra2 1 1和CEA明显高于对照组 ,慈菇可减轻CCl4致大鼠肝纤维化程度并减少cyfra2 1 1和CEA的产生。 结论 大鼠肝纤维化后血清cyfra2 1 1和CEA升高 。

慈菇下胚轴毛的形态发育研究

慈菇种子萌发前,下胚轴基部的表皮细胞分化成生毛细胞。当下胚轴穿出种皮约1—2毫米时,生毛细胞的外壁向外突出,形成下胚轴毛。开始时,其顶端膨大,呈分泌毛状,后呈根毛状。下胚轴毛的主要功能是起固着作用。下胚轴毛发育后期,在其保留细胞核的膨大的基部和突起的毛状体之间形成一细胞壁,此时毛状体便开始萎缩脱落。下胚轴毛的基部重新形成完整的表皮细胞。

慈姑活性成分对体外肝细胞损伤的保护作用

目的研究慈姑活性成分的保肝作用及其机理。方法采用系统化学溶剂萃取法制备慈姑不同活性成分,培养Hep G2型肝细胞,建立异烟肼和利福平合用致体外肝细胞损伤模型,给予慈姑不同活性成分处理后,利用MTT法检测肝细胞的存活率,并检测细胞培养上清液中丙氨酸氨基转移酶、天门冬氨酸氨基转移酶、乳酸脱氢酶、丙二醛的浓度。结果与模型组比较,慈姑乙酸乙酯组、慈姑石油醚组、慈姑总提取物组和慈姑水提组的细胞存活率均增加(P<0.01);慈姑石油醚组、慈姑总提取物组和慈姑水提组均可降低肝细胞培养上清液中的丙氨酸氨基转移酶、天门冬氨酸氨基转移酶、乳酸脱氢酶、丙二醛的水平(P<0.05),其中慈姑水提组效果最为明显(P<0.01)。结论提示慈姑水提部分对体外异烟肼和利福平合用致肝细胞损伤有较强保护作用,其作用可能与抗脂质过氧化有关。

慈姑匍匐茎的起源和球茎的膨大研究

匍匐茎的发生一般见于主茎倒二或倒三叶原基的叶腋部位。在匍匐茎发生区域的主茎一侧,匍匐茎原始细胞的基部形成壳状区;壳状区的形成对匍匐茎原基的外凸起一定作用。匍匐茎无居间分生组织;它的伸长依靠顶端分生组织细胞的横向分裂,使轴向细胞数目增多,并使细胞的轴向延伸。球茎的膨大是通过匍匐茎第8—10节基本分生组织的细胞有丝分裂,增加细胞数目,然后细胞体积的扩大来实现的。球茎中的淀粉一般为单粒淀粉;匍匐茎中的淀粉由单粒和复合两种淀粉粒组成。

蒽酮-硫酸法测定慈姑中多糖的含量

从慈姑球茎中分离提取出多糖,并用蒽酮-硫酸法对多糖含量进行测定。测定波长620nm,多糖换算因子f=1.82;在范围20μg～100μg内呈良好线性关系,r=0.9992。结果表明,供试溶液在3h内显色稳定,重现性好,平均回收率为98.04%,RSD=2.55(n=5)。

城市河道黑臭底泥对挺水植物光合荧光特性的影响

城市河道黑臭现象日益严重,探讨常见湿地植物在黑臭底泥中的生长,有利于为河道修复物种选择提供科学依据.研究黑臭底泥对3种常见湿地植物——菖蒲（Acorus calamus）、美人蕉（Canna indica）及慈姑（Sagittaria sagittifolia）生长状况和叶片叶绿素荧光参数的影响.结果表明：菖蒲和美人蕉在第30~40 d时生物量增量明显降低,慈姑的生物量增量在整个实验中持续增加;通过比较3种湿地植物的荧光参数可知,菖蒲和美人蕉的光化学淬灭系数（q P）、相对光合电子传递速率（r ETR）值在第10 d时达到最大,分别为41.33μmol/（m^2·s）和68.60μmol/（m^2·s）,后30 d一直下降;慈姑q P、r ETR值在第30 d时增加;在第40 d时,美人蕉q P值下降,非光化学淬灭系数（q N）值上升,叶片有较强的热耗散能力,而菖蒲的q P、q N值同时下降,黑臭底泥对菖蒲叶片的光合系统PSⅡ造成伤害.这说明黑臭底泥对菖蒲和美人蕉的光合能力产生较长期抑制,而慈姑可较快适应黑臭底泥的胁迫.因此利用湿地植物修复黑臭河道时,可优先选择慈姑,其次是美人蕉,最后是菖蒲.

铅对慈姑生理指标及其在球茎残留量分布的影响

以慈姑(Sagittaria sagittif olia L.)品种苏州黄和紫圆为试验材料,研究其在不同浓度Pb2+胁迫下部分生理生化指标以及产品器官中残留量变化。结果表明:叶片和根系可溶性蛋白质含量、根系活力总体上随Pb2+浓度的增加而下降,细胞膜透性和丙二醛含量则呈上升趋势。叶片SOD、POD活性在低浓度Pb2+处理下均有所上升,随着Pb2+浓度的增加,活性开始下降;但CAT活性则相反,当Pb2+浓度达到100 mg.L-1后,CAT活性开始增加。产品器官球茎中表皮残留量最高,去皮球茎最低。品种间紫圆球茎中Pb2+残留量高于苏州黄。

水稻与慈姑间作栽培对水稻病虫害和产量的影响

合理的间套作种植能够显著降低病虫害的发生,但已有相关研究主要集中在旱地作物种类或旱稻上,而水田环境下水稻间作体系的作物搭配及种植模式研究目前还十分缺乏。本文通过田间对比试验研究了水稻与慈姑间作栽培对水稻病虫害、微环境以及水稻产量的影响效应。结果表明,间作栽培模式对稻飞虱和稻纵卷叶螟无显著的防控效果。但在拔节期和抽穗期期间,间作栽培模式下水稻纹枯病病丛率分别比单作处理低64.3%和88.2%,稻瘟病病叶率在灌浆期和乳熟期显著低于单作,表明间作栽培模式显著降低了水稻稻瘟病和纹枯病的发生。间作栽培模式下水稻叶片POD、SOD和CAT酶活性与单作模式相比出现了不同程度降低,单作模式下水稻较高的病害发生程度可能是导致叶片保护酶活性增高的原因;间作栽培在早午时段降低了水稻叶片表面的空气湿度,且水稻叶片光合有效辐射强度在多数时段均高于单作栽培,说明间作栽培模式改善了稻田小气候环境,有利于降低病菌的滋生和传播;光合气体交换日进程测定结果表明,灌浆期间水稻/慈姑间作栽培模式下水稻叶片净光合速率、蒸腾速率以及气孔导度在日间大部分时段均高于单作水稻,说明间作栽培模式提高了水稻的群体光能利用率以及群体光合效率;间作栽培模式下的土地当量比为1.17,表明水稻与慈姑间作栽培模式不仅能有效控制水稻病虫害的发生,同时能起到良好的增产效果。

慈姑多糖部位颗粒剂制备工艺研究

目的:优选慈姑多糖部位的提取方法及其颗粒剂的制备工艺,为慈姑多糖部位的开发利用提供实验依据。方法:首先采用正交设计L4(23),以浸膏得率为指标,对影响慈姑提取工艺的因素进行考察并制备慈姑多糖部位;利用苯酚-硫酸法对慈姑多糖部位中的慈姑多糖含量进行测定;再以颗粒的吸湿性、成型性、溶化性、休止角及制粒的难易程度为评价指标,采用L9(33)正交试验设计对慈姑多糖部位颗粒剂的工艺条件进行优选;并采用固定漏斗法测定休止角,考察所添加辅料种类、慈姑多糖部位与辅料比例、乙醇浓度对慈姑多糖部位颗粒剂成型工艺的影响,借以优选成型工艺。结果:慈姑的最佳提取工艺为6倍量70%乙醇,冷浸提取48 h;慈姑多糖部位中慈姑多糖的含量为20.22%;优选的慈姑多糖部位颗粒剂成型工艺的条件为以乳糖作为辅料,慈姑多糖部位与辅料比例为1∶2,并使用80%乙醇作为润湿剂;在优选的条件下,颗粒的成型性好、流动性好且不易粘连,溶化速度快且静置后无沉淀。结论:优选的慈姑多糖部位的提取方法及其颗粒剂的制备工艺合理且简单可行,为慈姑多糖部位的开发利用提供了实验依据。