Optimization of Enzymatic Extraction Process of Polysaccharides from Pseudostellaria heterophylla Fibrous Roots by Response Surface Methodology and Its Pilot Application

[Objectives]To study and optimize the process conditions of enzymatic hydrolysis technology for extracting polysaccharides from Pseudostellaria heterophylla fibrous roots and its application in workshop pilot tests.[Methods]P.heterophylla fibrous roots were taken as the matrix material,and Box Behnken design was used to analyze the extraction time,composite enzyme addition amount,and liquid-solid ratio for response surface optimization experiments,and then applied to the pilot extraction of P.heterophylla fibrous roots.[Results]Response surface analysis showed that all factors had a significant impact on the experimental indicators.The optimal extraction process conditions for polysaccharides from P.heterophylla fibrous roots were extraction time of 2.7 h,compound enzyme addition of 2.5%,and liquid-solid ratio of 32.The yield of polysaccharides from P.heterophylla fibrous roots was 4.83%.The water extraction process of P.heterophylla fibrous roots extraction pilot was used as the control group for response surface optimization of the pilot experiment.The optimization results showed that the extraction time was 3 h,the amount of composite enzyme added was 2.5%,and the liquid-solid ratio was 28.The polysaccharide yield was 4.75%,an increase of 4.63%compared to the control group.[Conclusions]This paper could provide feasibility for the innovation of enzy-matic hydrolysis technology for P.heterophylla fibrous roots and its workshop pilot practice application,as well as a reference for the industrial application of its medicinal resources.

In Vitro Probiotic Effect of Pseudostellaria heterophylla Fibrous Roots on Feed Bacillus subtilis and Preliminary Study on Its Solid Fermentation

Traditional Chinese medicine Pseudostellaria heterophylla fibrous roots have been widely studied and applied in non-antibiotic clinical breeding of livestock and poultry. In vitro probiotic effect of P. heterophylla fibrous roots extract on feed probiotics Bacillus subtilis BD-K010 was studied, and the feasibility of solid fermentation of P. heterophylla fibrous roots was preliminarily evaluated. For in vitro probiotic effect, the increased concentration of P. heterophylla fibrous roots extract dependently increased the total biomass of B. subtilis BD-K010;1.0% P. heterophylla fibrous roots extract received the best effect, and the final p H of 1.0% experimental group was closer to neutral. Meantime, B. subtilis BD-K010 with optimum concentration of P. heterophylla fibrous roots showed significantly higher in vitro antibacterial effect than the control group(P<0.01),and the antibacterial effects on Escherichia coli and Staphylococcus aureus were improved by 51.99% and 63.16%, but it was ineffective to Salmonella. For solid fermentation, the profile of substrate complex and appendage flocculent structure on substrate surface at the end of fermenta-tion in experimental groups added with B. subtilis BD-K010 and cellulase plus BD-K010 were more complex;the live bacteria number, polysaccharide content and saponin content at fermentation end-point in two experimental groups were extremely higher than those in the control group(P<0.01).P. heterophylla fibrous roots extract had good in vitro probiotic effect on B. subtilis BD-K010 and promoted its antibacterial effect, and it is feasible to use probiotics for solid fermentation of P. heterophylla fibrous roots and to improve effective components. It is of great significance to further de-velop and utilize P. heterophylla fibrous roots resources in modern animal husbandry.

A Study on the Biological Effects of Laser-induced Mutation on Fibrous Roots of Yellow Skin Onion

[ Objective] This study aimed to investigate the biological effects of laser-induced mutation on fibrous roots of yellow skin onion. [ Method] Wet seeds of two yellow skin onion cultivars were irradiated by CO2 laser and He-Ne laser at three dosage levels separately. A randomized complete block design with three replications was adopted. The biological effects of laser-induced mutation on fibrous roots of Ll-generation yellow skin onion were investigated with biostatistics and physiological and biochemical methods. [Result] Significant variations in the biological effects caused by various laser treatments were observed in the length, quantity, fresh weight and activity of onion fibrous roots. Specifically, the variation in fibrous root length induced by different types of laser reached 5% significance level; significant variation was observed in fibrous roots of different onion cultivars induced by laser, while the variation among each treatment did not reach 5% sig- nifieance level ; the variation in fibrous root quantity induced by different dosage levels of laser reached 5% significance level ; laser radiation showed stimulating effect on root activity of onion. [ Conclusion] This study provided reference for laser-induced breeding of yellow skin onion.

Effects of Different Concentrations of Radix pseudostellariae Fibrous Root Extracts on Serum Cytokines in Mice and Their Correlation Analysis

To investigate the effects of Radix pseudostellariae fibrous root extracts(RPFRE) on serum cytokines in normal and immunosuppressed mice and the correlation between indexes, KM male mice were randomly divided into four groups, which were given 0(control), 0.1, 0.2, 0.4 g/kg RPFRE by gavage. The levels of IL-2, IL-4, IL-6, and IFN-γ in the serum of mice were measured 14 d after gavage. On the 15day, the remaining mice in the experimental group were intraperitoneally injected with cyclophosphamide(CY), and samples were collected on the 18day to measure serum cytokines levels. The results showed that RPFRE improved the serum cytokine level in normal mice. 0.1 g/kg RPFRE significantly increased the content of IL-2and IFN-γ(P<0.05), while 0.4 g/kg RPFRE significantly increased the content of IL-4 and IL-6(P<0.05). Correlation analysis showed that serum cytokines IL-4 and IL-6 in normal mice had an extremely significant positive correlation with RPFRE concentration(P<0.01), IL-2 had a significant or extremely significant positive correlation with IL-4, IL-6, and IFN-γ(P<0.01or P<0.05), and IL-4 had an extremely significant positive correlation with IL-6(P<0.01). After CY injection, serum cytokine levels in mice decreased and immune function was inhibited. Compared with the control group, the experimental group recovered the level of cytokines to some extent. 0.1 g/kg RPFRE significantly increased the content of IL-2 in immunosuppressed mice(P<0.05), and 0.4 g/kg RPFRE significantly increased the content of IL-4, IL-6, and IFN-γ in immunosuppressed mice(P<0.05). Correlation analysis showed that the serum cytokines in immunosuppressed mice were significantly positively correlated with RPFRE concentration(P<0.01), and there was a certain dose dependence. Moreover, the correlation coefficient between IL-2 and IL-4, IL-6, and IFN-γ, IL-4 and IL-6, and IL-6 and IFN-γ reached an extremely significant level(P<0.01). To sum up, RPFRE could improve the serum cytokines level of mice, promote the secretion of cytokines, and have a certain immune protective effect. There was a significant positive correlation between the RPFRE concentration and the serum cytokines in mice, which provided basic data support for the further study of RPFRE immune function.

Saponins Content in Fibrous Roots of Pseudostellaria heterophylla and Optimization of the SEP Extraction Technology

[Objectives]To explore the saponins resources in fibrous roots of Pseudostellaria heterophylla and optimize the SEP technology for extracting saponins.[Methods]Saponins contents in fibrous roots and root tubers of P.heterophylla were compared;with the fibrous roots as host materials,this experiment applied Box-Behnken design to analyze the influence of steam pressure,maintenance pressure time,moisture content of host materials on the content of saponins extracts.[Results]Saponins content was higher in fibrous roots than in tuberous roots of P.heterophylla;the response surface optimization showed that SEP(steam explosion pretreatment)promoted the contents of saponins and extracts in fibrous roots,and the optimal extracting conditions were stream pressure 1.50 Mpa,maintenance pressure time 54 s,and moisture content of host materials 30%.Under this extracting conditions,the verification value of saponins content experiment was 0.441%,36.53%higher than the untreated control group.[Conclusions]This research offered a new method of exploring saponins resources from fibrous roots of P.heterophylla,and provided a reference for developing new medicinal parts of P.heterophylla.

Immunoprotection Effects of Radix Pseudostellariae Fibrous Root Extraction (RPFRE) on Mice and Its Relationship with Antioxidant Function

[Objective] The paper was to explore the immunoprotection effects of Radix Pseudostellariae fibrous root extraction(RPFRE) on experi-mental mice and its relationship with antioxidant function. [Method] KM male mice were randomly divided into 4 groups based on RPFRE gradient concentration, and the immune function indexes and antioxidant indexes of mice were determined at 14 d post intragastric administration of RPFRE.The remaining mice in the experimental group were intraperitoneally injected with cyclophosphamide(CY) from the 15^(th) to 17^(th) day, and samples were collected to determine the above indexes on the 18^(th) day. [Result] Intragastric administration of RPFRE for 14 d improved the spleen index,thymus index, T/B cell stimulation index and total antioxidant capacity(T-AOC) of normal mice, and extremely increased the thymus index and TAOC(P<0.01). The T-AOC had extremely positive correlation with the spleen index, thymus index and B cell stimulation index( P<0.01), and had significantly positive correlation with the T cell stimulation index(P<0.05). From the 15^(th) to 17^(th) day of the experiment, the immune function indexes and antioxidant indexes of mice decreased and the malondialdehyde(MDA) content increased. The experimental groups of 0.1, 0.2 and 0.4 g/kg RPFRE promoted the immunoprotection and antioxidant effect of mice. RPFRE concentration had extremely positive correlation with 4 indexes of immune function and antioxidant indexes T-AOC and SOD(P<0.01), but had extremely negative correlation with MDA(P<0.01). There was a significantly positive correlation between immune function indexes and antioxidant indexes of mice after CY intervention( P<0.05 or P<0.01). [Conclusion] RPFRE can improve the immune function indexes and antioxidant indexes of normal mice and immunosuppressed mice and has immunopro-tection effect on immunosuppressed mice, and there is a significant correlation between immune function indexes and antioxidant indexes of mice.

Effect of Root Architecture on Structural Stability and Erodibility of Topsoils during Concentrated Flow in Hilly Loess Plateau

Traditional vegetation techniques for the control of concentrated flow erosion are widely recognized, whereas only a few studies have experimentally investigated the impacts of belowground roots on the erodibility of topsoils in semi-arid areas. To quantify the effects of root architectures on soil erodibility and its relevant structural properties, simulated flow experiments were conducted at six-week intervals from 18 July to 20 October in 2012 in the hilly Loess Plateau. Five treatments were: 1) bare(control), 2) purple alfalfa(Medicago sativa), representing tap roots(T), 3) switchgrass(Panicum virgatum), representing fibrous roots(F), 4) purple alfalfa and switchgrass, representing both tap and fibrous roots(T + F), and 5) natural recovery(N). For each treatment, soil structural properties and root characteristics were measured at an interval of six weeks. Soil anti-scouribility was calculated. Results showed that grass planting slightly reduced soil bulk density, but increased soil aggregate content by 19.1%, 10.6%, 28.5%, and 41.2% in the treatments T, F, T + F, and N, respectively. Soil shear strength(cohesion and angle of internal friction(φ)) significantly increased after the grass was planted. As roots grew, soil cohesion increased by 115.2%–135.5%, while soil disintegration rate decreased by 39.0%–58.1% in the 21 th week compared with the recorded value in the 9th week. Meanwhile, root density and root surface area density increased by 64.0%–104.7% and 75.9%–157.1%, respectively. No significant differences in soil anti-scouribility were observed between the treatments of T and F or of T + F and N, but the treatments of T + F and N performed more effectively than T or F treatment alone in retarding concentrated flow. Soil aggregation and root surface-area density explained the observed soil anti-scouribility during concentrated flow well for the different treatments. This result proved that the restoration of natural vegetation might be the most appropriate strategy in soil reinforcement in the hilly Loess Plateau.

Jin Xiaobin’s Unique Root Calligraphy

SEEING is believing that cal-ligraphy can be combinedwith root art.Standing be-fore the root-calligraphy works ofJin Xiaobin, a young artist fromHuangshan City,one is filled withadmiration for his unique creations,which integrate nature and mankindand art and life.Root calligraphy

Liposoluble chemical constituents from the fibrous root of Ophiopogonjaponicas

We investigated the chemical constituents of the fibrous root of Ophiopogon japonicus and their cytotoxicities against Hela and Hep2 cells.Ten compounds were purified by various chromatographic techniques.Their structures were identified as 2 - hydroxylophiopogonone A（1）,5,8-dimethoxy-6-methyl-7-hydroxy-3-（2 -hydroxy-4 -methoxybenzyl） chroman-4-one（2）,5,7- dihydroxy-6,8-dimethyl-3-（4 -hydroxybenzyl） chroman-4-one（3）,7,4 -dihydroxy-5-methoxyflavanone（4）,N-trans-coumaroyltyramine （5）,N-trans-coumaroyloctopamine（6）, N-trans-feruloyltyramine （7）, 4-hydroxycinnamic acid （8）, caffeic acid （9）, and ferulic acid （10） on the basis of spectroscopic analyses. Compounds 4, 6, 7, 9, 10 were obtained from this genus for the first time. The cytotoxic activities of 1, 2, 4, 5, 6, 7, 9 and 10 against Hela and Hep2 cells are described.

In-situ Horizontal Extrusion Test of Herbaceous Root-Soil with Different Root Types

The influence of different types of roots on the soil is complex and still remains unclear.Four in-situ extrusion tests were conducted on two types of root systems,namely fibrous and tap root system,for three plants,Eleusine indica,Potentilla anserine,and Artemisia argyi,according to the classification in Botany,and the thrust-displacement curves and failure patterns of different samples were analysed by comparison to fill the aforementioned gap.Results reveal that the roots can reduce the characteristics of soil brittleness and enhance its capability to resist large deformation,and different root types contribute different effects to the strain-hardening behavior of the root-soil mass.The contribution of the fibrous root system to strength is limited,whilst the tap root system substantially enhances strength and stiffness.Results of failure patterns show that fibrous and tap root systems affect soil solidification and surface cracking reduction.However,the effect of the tap root system depends on the composition of lateral and tap roots:long and rich lateral roots are effective for resisting the creation of cracks,but thick tap roots with few and thin lateral roots may lead to several surface cracks.

Dynamic Microwave-Assisted Extraction of Total Ginsenosides from Ginseng Fibrous Roots

A dynamic microwave-assisted extraction（DMAE） method is established for the extraction of total ginsenosides from ginseng fibrous roots. The extraction process has been simulated and its main affection factors（liquid/solid ratio K of solvent to ginseng powders（V/m）, irradiation time, irradiation temperature, extracting solution concentration, flow rate of solvent and microwave power） have been optimized by response surface methodology（RSM）. The optimum conditions of extraction are the liquid/solid ratio of 270 m L/g, the extraction temperature of 75 ℃, the extraction time of 35 min, ethanol concentration of 70%（V/V）, the solvent flow rate of 1.3 m L/min, and the microwave power of 500 W. The yield of ginsenosides obtained by the proposed DAME method is（15.0±0.7） %, which is well agreement with the yield predicted by the model. Compared with static microwave-assisted extraction, DMAE has a higher extraction yield and can avoid the degradation of ginsenoside.

知母根的组织发育及其与生长环境的关系

观察知母根的组织发育，发现具有典型的单子叶植物根的组织特征，特别是在同一条根中初生木质部脊数可以随根的发育而增加，同一株知母不同根之间初生木质部脊数也不尽相同；内皮层细胞壁增厚由凯氏带发育为马蹄形，内皮层外侧1－3层皮层细胞内侧壁可以木栓化或强烈木化增厚，这种变化与其生长环境具有密切关系，当其移植于湿润环境中后，内皮层马蹄形增厚减弱，其外侧皮层细胞内侧壁仅发生木柱化增厚，且减弱。

尪痹胶囊的质量标准研究

目的:建立尪痹胶囊的质量标准。方法:采用薄层色谱法对方中的白芍、知母进行鉴别;采用高效液相色谱法对淫羊藿苷进行定量分析。结果:薄层分离度良好,专属性强;淫羊藿苷进样量在0.030～2.0μg范围内具有良好的线性关系,回收率96.6%。结论:所建立之方法准确、灵敏度高、专属性强,可有效控制尪痹胶囊的质量。

Ideal root architecture for phosphorus acquisition of plants under water and phos-phorus coupled stresses: From simulation to application

Under water and phosphorus (P) coupled stresses, root architecture may be related to P acquisition efficiency of plants. Understanding the relationship between root architecture and P acquisition efficiency may provide basic information for improving P acquisition efficiency of plants. In the present study, we quantitatively described the effects of root architecture on P acquisition efficiency by computer simulation together with controlled biological experiments so as to determine an ideal root architecture for efficient P acquisition under water and P coupled stresses. Our results indicate that under given soil water conditions, the ideal root architecture for P acquisition efficiency of a tap root plant (as represented by common bean) is an 搖mbrella-shape?root system whose basal roots tend to be shallow in the P-rich topsoil and tap roots tend to be deep for water in the subsoil. Meanwhile, the ideal root architecture for a fibrous root plant (as represented by upland rice) is a beard-shape?root system with the moderately dispersed yet uniformly distributed adventitious and lateral roots so as to keep most roots in the topsoil for P and a few roots in the subsoil for water.

知母须根总皂苷纯化工艺及其延长秀丽隐杆线虫寿命的作用机制研究

目的考察知母须根总皂苷富集纯化的最佳工艺,并对其延长秀丽隐杆线虫(Caenorhabditis elegans,C.elegans)寿命的作用及机制进行研究。方法以知母须根总皂苷得率、总皂苷含量等为指标,考察大孔树脂型号、上样液浓度、吸附流速、上样体积、洗脱溶剂浓度及体积对富集结果的影响。以秀丽隐杆线虫为实验模型,以0.5、1.0、2.0 mg·mL^-1知母须根总皂苷进行干预,测定秀丽隐杆线虫寿命、后代数目、运动能力;采用qPCR法测定知母须根总皂苷(1.0 mg·mL^-1)干预后秀丽隐杆线虫体内daf-16、daf-2、sod-3、ctl-2等基因的表达情况。结果最佳树脂型号为AB-8,上样液浓度为0.2 g·mL^-1,吸附流速及洗脱流速均为1.0 mL·min-1,洗脱剂为70%乙醇,洗脱体积为3倍柱体积,所富集的总皂苷纯度>50%。与空白对照组比较,1.0 mg·mL^-1浓度的知母须根总皂苷可显著延长秀丽隐杆线虫的平均寿命,提高线虫的半数致死天数(P<0.05)、产卵数目及运动频率(P<0.01);显著提高线虫体内sod-3、daf-16和ctl-2基因的表达量(P<0.01,P<0.001),降低daf-2基因的表达量(P<0.05)。结论优选的工艺稳定可行,可用于知母须根总皂苷的富集纯化;知母须根总皂苷能延长秀丽隐杆线虫的寿命,其机制可能与Insulin/IGF-1信号通路有关。

响应面法优选知母须根总黄酮提取工艺及其解热作用研究

为了研究知母须根总黄酮(TF)最佳超声提取工艺及其解热作用。在单因素实验基础上,采用响应面法研究乙醇浓度、提取时间、提取次数、料液比等四个因素对总黄酮提取量的影响,根据拟合的数学模型确定最佳提取工艺。以干酵母致热大鼠为动物模型,通过测定大鼠血清中TNF-α、PGE-2、IL-6、AVP的含量研究其解热作用。知母须根总黄酮最佳提取工艺为:乙醇浓度为70%,提取时间40 min,液料比为50 m L/g,提取次数2次。解热实验表明知母须根总黄酮提取物可显著降低发热大鼠模型体温及血清、下丘脑中TNF-α、PGE-2、IL-6的含量,升高发热大鼠模型血清及下丘脑中AVP的含量。本研究表明知母须根总黄酮具有一定的解热作用,本文可为知母须根开发利用提供参考依据。

知母须根化学成分的研究

目的研究知母Anemarrhena asphodeloides Bge.须根的化学成分。方法知母须根70%乙醇提取物采用硅胶、ODS、半制备HPLC进行分离纯化,根据理化性质及波谱数据鉴定所得化合物的结构。结果从中分离得到8个化合物,分别鉴定为7-hydroxy-3-(4-methoxybenzyl)-chroman(1)、4′-hydroxyphenyl ethyl vanillate(2)、methyl 4-hydroxybenzoate(3)、1,3,6,7-tetrahydroxy-9H-xanthen-9-one(4)、6′,7′-dihydroxy-2-methoxy-3′,4′-dihydro-2′H-spiro[cyclohexane-1,1′-naphthalene]-2,5-dien-4-one(5)、michehedyoside D(6)、demethyleugenolα-D-glucopyranoside(7)、2′,3′-dihydroxyl-osthole(8)。结论化合物3~8均为首次从该植物中分离得到。

知母须根化学成分研究

目的对知母(Anemarrhena asphodeloide Bunge.)的干燥须根进行化学成分研究。方法采用硅胶柱色谱、Sephadex LH-20柱色谱及重结晶等手段进行分离纯化,运用波谱学方法鉴定化合物的结构。结果从知母须根中分离得到15个化合物,分别鉴定为胡萝卜苷(1)、菝葜皂苷元(2)、芒果苷(3)、7-hydroxy-3-(4-methoxybenzyl)-chroman(4)、3′-hydroxy-4′-methoxy-4-dehydroxynyasol(5)、5,7-dihydroxy-3-(4-methoxybenzyl)-chroman-4-one(6)、2,4′-dihydroxy-4-methoxy benzophenone(7)、1,3-Bis(4-hydroxyphenyl)pent-4-en-1-one(8)、(+)-syringaresinol(9)、ent-16,17-dihydroxy-kauran-19-oic acid(10)、(+)-pinoresinol(11)、对羟基苯甲酸(12)、新芒果苷(13)、丁香酸(14)、4′,6-dihydroxy-4-methoxy-benzophenone-2-O-(2″),3-C-(1″)-1-desoxy-α-L-fructofuranoside(15)。结论化合物5~11、14、15为首次从知母须根中分离得到。

不同产地知母须根中3个酮类成分含量测定

目的测定不同产地知母须根中新芒果苷、芒果苷和异芒果苷的含量。方法采用高效液相二极管陈列检测法,Shim-pack色谱柱(4.6mm×250mm,5μm);乙腈-0.4%冰醋酸水溶液梯度洗脱,流速1.0mL/min;柱温25℃;检测波长为258nm。结果所收集的知母须根样品中新芒果苷的含量为1.2%~1.9%,芒果苷的含量为1.0%~1.6%,异芒果苷的含量为0.4%~0.6%。结论知母须根中酮成分含量较高,在知母须根的综合开发利用中可考虑此类成分。

Evaluation of high yielding soybean germplasm under water limitation

Limited information is available for soybean root traits and their plasticity under drought stress. To date, no studies have focused on examining diverse soybean germ- plasm for regulation of shoot and root response under water limited conditions across varying soil types. In this study, 17 genetically diverse soybean germplasm lines were selected to study root response to water limited conditions in clay （trial 1） and sandy soil （trial 2） in two target environments. Physiological data on shoot traits was measured at multiple crop stages ranging from early vegetative to pod filling. The phenotypic root traits, and biomass accumulation data are collected at pod filling stage. In trial 1, the number of lateral roots and forks were positively correlated with plot yield under water limitation and in trial 2, lateral root thickness was positively correlated with the hill plot yield. Plant Introduction （PI） 578477A and 088444 were found to have higher later root number and forks in clay soil with higher yield under water limitation, in sandy soil, P1458o2o was found to have a thicker lateral root system and higher yield under water limitation. The genotypes identified in this study could be used to enhance drought tolerance of elite soybean cultivars through improved root traits specific to target environments.