紫锥菊不定根悬浮共培养中咖啡酸衍生物积累研究

为筛选出紫锥菊不定根共培养最佳组合及其接种量,以紫松果菊(Echinacea purpurea,Epu)、白色紫锥菊(Echinacea pallida,Epa)和狭叶紫锥菊(Echinacea angustifolia,Ean)3种药用紫锥菊不定根为材料,在500 m L三角瓶中加入200 m L的0.5 MS+1.0 mg/L IBA培养基进行了悬浮培养,调查了接种量5 g/L、7 g/L和10 g/L及不定根组合方式对共培养紫锥菊不定根生物量、次生代谢产物积累及抗氧化活性的影响。结果表明,接种量为7 g/L时的Epa+Epu共培养不定根干重(6.75 g/L)最高,此组合中不仅多酚(40.31 mg/g DW)和黄酮(34.29mg/g DW)含量高,其产量(多酚272.09 mg/L,黄酮231.46 mg/L)也最高。紫锥菊不定根共培养组合中,只有Epa+Epu不定根中6种咖啡酸衍生物都具有,而且此共培养不定根还刺激产生Epa和Epu单一培养时未检测到的活性物质咖啡酸,其中接种量为7 g/L时的共培养Epa+Epu不定根中总咖啡酸衍生物产量最高,达136.22mg/L,是Epu单独培养时的1.35倍,Epa单独培养时的1.73倍。还原能力和DPPH自由基清除率也是接种量为7 g/L时的Epa+Epu不定根组合较高。此结果为大规模生产具有抗肿瘤抗氧化药用价值的紫锥菊医学药材提供技术参数。     

紫锥菊的组织培养和植株再生

1植物名称紫锥菊(Echinacea purpurea),又名紫松果菊. 2材料类别无菌苗的叶柄切段. 3培养条件(1)芽诱导及增殖培养基:MS 6-BA 1.0mg·L-1(单位下同) NAA 0.5 3%蔗糖 0.7%琼脂,pH 6.0;(2)生根培养基:B5大量元素 MS微量元素、铁盐、有机物 NAA 1.0 2%蔗糖 0.9%琼脂,pH 6.0.培养温度为(25 1)℃,光照12 h·d-1,光照度1500～2000 lx.     

大孔吸附树脂对紫锥菊提取物中菊苣酸分离纯化的研究

采用大孔吸附树脂分离纯化紫锥菊提取物中免疫活性成分菊苣酸,7％(v／v)的甲醇-水溶液洗脱,HPLC法对产品中菊苣酸含量进行检测分析。该法能把紫锥菊提取物中菊苣酸含量(4％)提高9倍左右,回收率达到95％以上,且操作简单,可用于菊苣酸的分离纯化。     

紫锥菊愈伤组织诱导及生物活性成分分析

以紫锥菊(Echinacea purpurea)无菌苗为材料,研究了不同激素配比条件下不同外值体愈伤组织的诱导及所诱导的愈伤组织中的主要成分多糖和总酚,并应用高效液相色谱法对根愈伤组织提取物与紫锥菊根提取物中的酚酸类物质进行了比较分析.结果表明,紫锥菊不同部位愈伤组织诱导的最适宜激素配比不同,其中根愈伤组织诱导的最佳激素配比为0.5 mg/L 2,4-D 0.5 mg/L 6-BA.叶、茎、根诱导愈伤组织的多糖含量分别为10.15、11.84、14.49 mg/g干重;总酚含量分别为52.16、28.144、7.99 mg/g干重.根愈伤组织总酚和多糖含量均较高,其提取物的酚酸类物质的高效液相色谱与紫锥菊根提取物图谱主峰一致,且生长速度快、质地疏松,是细胞培养获取紫锥菊主要生物活性成分的适宜材料.     

紫锥菊单咖啡酰酒石酸和菊苣酸提取工艺优化

采用正交试验设计,选取乙醇体积浓度百分比、提取温度、提取时间、抗氧化剂用量等因素,优化紫锥菊Echinacea purpurea单咖啡酰酒石酸和菊苣酸的加热回流提取工艺,并考察加入抗氧化剂对提取效果的影响。结果表明,优化的提取条件是以25%乙醇,在80 ℃回流提取90 min。抗氧化剂用量对提取效果影响不显著。优化后的加热回流提取条件对紫锥菊单咖啡酰酒石酸和菊苣酸的提取均适用,提取中无需加入抗氧化剂。     

紫锥菊多糖抑制致病性大肠埃希菌细胞黏附的试验研究

目的研究紫锥菊多糖能否影响致病性大肠埃希菌对细胞的黏附。方法使用PK-15细胞进行黏附试验及黏附抑制试验。结果发现紫锥菊多糖浓度为1.6 mg/ml时,对细菌黏附细胞的抑制作用最好,黏附率由50个细菌/细胞降低到6.8个细菌/细胞。结论紫锥菊多糖对致病性大肠埃希菌的细胞黏附具有抑制作用,提示该多糖具有调节肠道微生态的潜在应用价值。     

蔗糖浓度对三裂叶野葛毛状根生长及其次生物质产生的影响

本文研究了蔗糖浓度对发根农杆菌ATCC15834诱导产生的三裂叶野葛毛状根生长及其葛根素和异黄酮类化合物产生的影响以及液体培养基中蔗糖的消耗变化.结果表明毛状根在含5%、4%、3%和2%蔗糖的MS培养基中培养16天后的干重增殖倍数分别为11.7、11.9、10.1和5.9;其中尤以3%的蔗糖浓度最有利于毛状根中异黄酮类化合物及葛根素的积累;培养12天后,毛状根的葛根素含量达到最高,约5.147mg/g DW;而其异黄酮类化合物的含量则在培养16天后达到最高,约27.76mg/g DW.在毛状根液体培养过程中培养基的蔗糖浓度随着毛状根的生长而降低,其消耗速率与毛状根的生长速度及其可溶性总糖含量成正比.毛状根的可溶性总糖含量在培养12天时达到最高,而培养16天后培养基中的蔗糖消耗完毕.     

紫锥菊的生药学研究

采用来源鉴定、性状鉴定、显微及理化鉴定的方法研究紫锥菊的生药学内容,为制定药材质量标准、研究、开发和利用紫锥菊的药用资源提供了理论依据。     

黄芩愈伤组织培养条件的研究

在暗培养条件下,黄芩愈伤组织生长和次级代谢物合成的最佳培养条件:在基本培养基MS中氮源浓度为60mmol/L(NH4 ∶NO3-为1∶1),KH2PO41.5mmol/L,附加80g/L蔗糖,0.3mg/LIAA、2mg/L6-BA和200mg/L蛋白胨,(25±1)℃。培养40d后收获愈伤组织生物量达28.7g/L,总黄酮的含量为354.6mg/g,黄芩苷的含量为167.4mg/g。并发现蔗糖作为一种最佳碳源,在黄芩次级代谢物合成过程中起着至关重要的作用。     

紫锥菊多倍体诱导与鉴定

本实验以紫锥菊愈伤组织上刚分化出的不定芽为材料,用不同浓度秋水仙素溶液对其进行诱导,确定最佳处理浓度和处理时间,并对诱导的多倍体与二倍体进行形态、显微、染色体及过氧化氢酶(CAT)活性的比较鉴定.结果表明:0.025%秋水仙素浓度处理24h的诱导效果最好,诱导率达42.85%;多倍体植株叶片肥厚、根粗壮,气孔面积极显著地大于二倍体植株,染色体数从24～28条不等,CAT平均酶活性是二倍体的2.1倍.     

Root Growth Inhibitors from Root Cap and Root Meristem of Zea mays L.

A micro-assay based on the growth inhibition of root segmentsof the seminal roots of Zea mays has been used to investigatethe root-growth-inhibiting substances in root caps and meristemsrespectively of the roots of Zea mays. This micro-assay is sensitiveto 50 pg of IAA or less. Paper chromatography of the acid fractionof methanolic extracts shows the presence of one main inhibitorin root caps and a different main inhibitor in root meristems.Neither is IAA, whose presence in meristems is sometimes indicatedby small inhibitions (or stimulations) at the characteristicRf of IAA. A Commelina leaf-epidermis assay shows the presenceof one stomata-closing ABA-like substance in root caps and onein meristems, one corresponding in Rf to the main root-growthinhibitor from the root cap. The implications of these findingsfor the geotropic responses of roots is briefly discussed.     

Dependence of Root Cell Growth and Division on Root Diameter

Primary roots of 98 species from different families of monocotyledonous and dicotyledonous plants and adventitious roots obtained from bulbs and rhizomes of 24 monocot species were studied. Root growth rate, root diameter, length of the meristem and elongation zones, number of meristematic cells in a file of cortical cells, and length of fully elongated cells were evaluated in each species after the onset of steady growth. The mitotic cycle duration and relative cell elongation rate were calculated. In all species, the meristem length was approximately equal to two root diameters. When comparing different species, the rate of root growth increased with a larger root diameter. This was due to an increase in the number of meristematic cells in a row and, to a lesser degree, to a greater length of fully elongated cells. The duration of the mitotic cycle and the relative cell elongation rate did not correlate with the root diameter. It is suggested that the meristem size depends on the level of nutrient inflow from upper tissues, and is thereby controlled during further growth.     

Shoot and Root Growth of Lettuce Seedlings Following Root Pruning

Hydroponically-grown lettuce seedlings with 13 to 18 primarylateral roots were root pruned in one of four ways; the rootapices were removed from the main root only (1) or from allthe root membranes (2), or half the total root system was removedwith the remaining apices left intact (3) or removed (4). Duringthe following 8 d the rate of lateral root production on prunedplants increased, decreased, and then increased again relativeto the unpruned control. Conversely, the rate of increase intotal root length decreased, then increased, and if all theroot apices were removed, declined again, prior to increasingon day 8. These changes in the rates of lateral root productionand growth resulted in similar, but less pronounced, patternsof change in the total root length and the total number of lateralroots with time. The changes in total lateral root productionwere related to differences in the rates of primary, secondaryand tertiary root emergence. The shoot d. wt of the most severely root pruned seedlings (treatment4) fell below that of the control 4 d after pruning and remainedlower than the control on day 14, whereas the root d. wt hadrecovered to the control level by day 6. The root: shoot d.wt ratio, which was reduced by root pruning, rose above thatof the control on days 6 and 8. Lactuca sativa L., lettuce, root pruning, root growth, lateral root, nutrient solution     

Root Density and Water Potential Gradients near the Plant Root

The models of Gardner (1960) and Cowan (1965) for water transferto the plant root are used to estimate the differences in waterpotential between the root and the bulk soil for a wide rangeof root densities and water extraction rates at a series ofmatric potentials for a Yolo light clay. For root densities and extraction rates reported both in theliterature and in this paper there is good evidence to suggestthat the large potential gradients originally predicted by Gardnerand Cowan are restricted to situations involving very low rootdensities and high extraction rates in relatively dry soil.     

The Effect of Coumarin on Root Growth and Root Histology

The effect of coumarin on the root growth was studied on roots from intact plants, isolated roots and isolated elongating zones. All material was cultivated aseptically. A new method was developed for sterile culture of intact plants in flowing nutrient medium. The effects on cell division and cell elongation were studied separately. An effect on both these processes can be established at all concentrations that affect the root growth. The concentration-growth curve has an “all-or-none” appearance. Coumarin inhibits the transverse divisions in all cell layers; the perivascular layers seem to be more sensitive. Also the mitotic activity that is involved in the initiation of laterals is inhibited. The longitudinal divisions within the stele are enhanced. Coumarin decreases the cell length in all cell layers, most likely with greater relative sensitivity in the perivascular layers. Studies on the time course of cell elongation in both attached corn roots and isolated elongating zones reveal that the decrease in cell length is caused exclusively by a decrease in the maximal rate of elongation, whereas the duration of the elongation is unchanged. With each decrease of the cell length, the cell diameter is increased. The two changes are intimately connected within the greater part of the active region of concentration. Studies on the time course of the radial expansion in isolated elongating zones show a strict connection in time between cell elongation and radial expansion. The radial expansion leads to unchanged or increased cell volume at most concentrations and for most cell types. Coumarin causes an inhibition of the longitudinally directed processes and a stimulation of the radially directed ones. This is interpreted as indicating that the formative system is disengaged or reorientated, i.e., the polarity of the cells is changed. Through experiments partly with isolated elongating zones and partly by disruption of the linear phase by means of mannitol, the inhibitory effect of coumarin could be localized to the first non-linear phase of the elongation. The results were compared with earlier findings in the literature. The microtubuli are proposed as a conceivable main Component in the formative system common to both cell division and cell elongation. These are assumed to be affected by changes in the SH/SS balance produced by coumarin.     

Root Caps and Rhizosphere

In this paper we discuss recent work on the physiological, molecular, and mechanical mechanisms that underlie the capacity of root caps to modulate the properties of the rhizosphere and thereby foster plant growth and development. The root cap initially defines the rhizosphere by its direction of growth, which in turn occurs in response to gradients in soil conditions and gravity. The ability of the root cap to modulate its environment is largely a result of the release of exudates and border cells, and so provides a potential method to engineer the rhizosphere. Factors affecting the release of border cells from the outer surface of the root cap, and function of these cells and their exudates in the rhizosphere, are considered in detail. Release of border cells into the rhizosphere depends on soil matric potential and mechanical impedance, in addition to a host of other environmental conditions. There is good evidence of unidentified feedback signals between border cells and the root cap meristem, and some potential mechanisms are discussed. Root border cells play a significant mechanical role in decreasing frictional resistance to root penetration, and a conceptual model for this function is discussed. Root and border cell exudates influence specific interactions between plant hosts and soil organisms, including pathogenic fungi. The area of exudates and border cell function in soil is an exciting and developing one that awaits the production of appropriate mutant and transgenic lines for further study in the soil environment.     

Root Cap and Georeaction

CIRCULATING lymphoid cells suffer alterations in a localized disease, chronic viral keratitis; we have established long term lymphoctye cultures from the blood of such patients¹. We have used capillary migration technique to study cellular immunity in the disease. Leucocytes from patients were challenged with corneal antigen and with the virus which had initiated the infection^2,3. Migration inhibition is specific for the antigen to which the animal is sensitive and on contact with antigen, sensitized lymphocytes produce migratory inhibitory factor (MIF).     

Root Caps and Rhizosphere

In this paper we discuss recent work on the physiological, molecular, and mechanical mechanisms that underlie the capacity of root caps to modulate the properties of the rhizosphere and thereby foster plant growth and development. The root cap initially defines the rhizosphere by its direction of growth, which in turn occurs in response to gradients in soil conditions and gravity. The ability of the root cap to modulate its environment is largely a result of the release of exudates and border cells, and so provides a potential method to engineer the rhizosphere. Factors affecting the release of border cells from the outer surface of the root cap, and function of these cells and their exudates in the rhizosphere, are considered in detail. Release of border cells into the rhizosphere depends on soil matric potential and mechanical impedance, in addition to a host of other environmental conditions. There is good evidence of unidentified feedback signals between border cells and the root cap meristem, and some potential mechanisms are discussed. Root border cells play a significant mechanical role in decreasing frictional resistance to root penetration, and a conceptual model for this function is discussed. Root and border cell exudates influence specific interactions between plant hosts and soil organisms, including pathogenic fungi. The area of exudates and border cell function in soil is an exciting and developing one that awaits the production of appropriate mutant and transgenic lines for further study in the soil environment.     

Root Growth, Secondary Root Formation and Root Gravitropism in Carotenoid-deficient Seedlings of Zea mays L.

The effect of ABA on root growth, secondary-root formation androot gravitropism in seedlings of Zea mays was investigatedby using Fluridone-treated seedlings and a viviparous mutant,both of which lack carotenoids and ABA. Primary roots of seedlingsgrown in the presence of Fluridone grew significantly slowerthan those of control (i.e. untreated) roots. Elongation ofFluridone-treated roots was inhibited significantly by the exogenousapplication of 1 mM ABA. Exogenous application of 1 µMand 1 nM ABA had either no effect or only a slight stimulatoryeffect on root elongation, depending on the method of application.The absence of ABA in Fluridone-treated plants was not an importantfactor in secondary-root formation in seedlings less than 9–10d old. However, ABA may suppress secondary-root formation inolder seedlings, since 11-d-old control seedlings had significantlyfewer secondary roots than Fluridone-treated seedlings. Rootsof Fluridone-treated and control seedlings were graviresponsive.Similar data were obtained for vp-9 mutants of Z. mays, whichare phenotypically identical to Fluridone-treated seedlings.These results indicate that ABA is necessary for neither secondary-rootformation nor for positive gravitropism by primary roots. Zea mays, gravitropism, carotenoid-deficient, Fluridone, root growth, vp-9 mutant