不同海拔云南黄连生物量和主要有效成分变化

研究了不同海拔(2 100～2 700 m)下,野生和人工栽培云南黄连的生物量、主要有效成分含量及产量.结果表明:野生云南黄连根茎和根生物量沿海拔梯度呈上升趋势,但无显著性差异(P>0.05);人工栽培云南黄连根茎生物量平均值在海拔2 600 m和2 700 m处分别为87.5 kg·hm^-2和97.0 kg·hm^-2,显著高于海拔2 300 m处(34.8 kg·hm^-2,P<0.05),且海拔2 300、2 600和2 700 m的人工栽培云南黄连根茎和根生物量均大于野生云南黄连,但无显著性差异(P>0.05). 野生云南黄连的根茎和根生物量均与全株生物量呈显著正相关. 野生云南黄连根茎和根小檗碱含量在海拔2 700 m处最高,分别为4.60%和1.93%; 根茎巴马汀和药根碱含量、根药根碱含量在海拔2 600～2 700 m处最高;根巴马汀含量在2 300 m处最高.人工云南黄连根茎和根小檗碱含量在海拔2 600 m处最高,分别为4.41%和1.90%; 根茎巴马汀含量,根小檗碱、巴马汀和药根碱含量在海拔2 600～2 700 m处最高;根茎药根碱含量在海拔2 300 m处最高.海拔2 600～2 700 m处野生云南黄连根茎和根中各有效成分产量显著高于海拔2 100和2 300 m处(P<0.05). 野生云南黄连分株的根茎生物量、根生物量、叶生物量、总生物量、高度和冠幅沿海拔梯度呈先升后降趋势.增大种植密度和加强人工管理可以提高云南黄连生物量和主要有效成分产量.     

模拟连续逆流提取黄柏中小檗碱工艺研究

本文优化实验室模拟连续逆流提取设备工艺对黄柏中小檗碱进行提取,并用连续逆流提取设备进行中试放大。在单因素设计实验基础上采用正交设计实验优化实验室小试模拟连续逆流提取黄柏中小檗碱的工艺(提取温度、硫酸浓度和提取时间),为新型连续逆流提取设备中试条件优化提供一种实验室模拟方法,并采用中试连续逆流提取设备对黄柏中小檗碱提取工艺进行验证。正交设计实验结果和极差分析结果表明三个提取因素对提取效果的影响大小顺序是:提取温度硫酸浓度提取时间。最佳提取条件为:提取温度90℃,8倍体积的0.5%稀硫酸溶液作为提取溶剂,提取4 min,换料次数12次。在优化提取条件下小檗碱提取得率为3.6%,小檗碱提取率达到90.3%。根据实验室小试模拟所得各项实验参数,预设中试连续逆流提取器各项提取参数:提取温度90℃,转速4 rpm,总提取时长20 h,小檗碱提取率达90.5%。该工艺较小檗碱提取传统工艺能显著提高提取效率和有效降低提取溶剂用量。     

用电导率法研究低频超声提取小檗碱成分的产额

采用电导率测试法研究了超声提取对药材黄柏中小檗碱成分产额的影响,和煎煮法,浸泡法相比,低频超声辐照可增强从川黄柏中所提得的小檗碱成分的产额,证实了电导率测定法可作为瞬时,动态反馈药材中有效成分提取规律的基本方法。     

不同植被地区天然黄檗生物碱含量对季节的响应

黄檗(Phellodendron amurense Rupr.)不仅为中国特有的濒危植物,还是我国著名中药材"关黄柏"的药源植物,由于其珍贵的药用价值而被过度开采。本研究采用HPLC检测不同植被地区天然黄檗种群中药根碱、掌叶防己碱和小檗碱等生物碱的含量,分析不同部位,不同季节对生物碱分布的影响。研究表明,药根碱在黄檗各部位的分布为:根皮茎皮多年生枝一年生枝叶片。掌叶防己碱含量分布为:茎皮多年生枝皮根皮一年生枝叶片。小檗碱在不同部位的分布规律表现为:根皮茎皮多年生枝一年生枝叶片。根皮和茎皮中的药根碱含量随着春、夏、秋季节的推移呈逐渐上升的趋势,表现为秋季夏季春季,而多年生枝皮、一年生枝、叶片中的药根碱则是秋季春季夏季。各部位的掌叶防己碱均随着季节推移而升高,表现为秋季夏季春季。根皮、茎皮和多年生枝皮中小檗碱含量随着季节变化表现为夏季秋季春季。一年生枝中小檗碱在夏季降低,而到了秋季再升高。     

季节和地理差异对天然黄檗小檗碱和药根碱含量的影响

用高效液相色谱法测定了黄檗根皮、茎皮、多年生枝皮、1年生枝和叶片中小檗碱和药根碱的含量,研究黄檗不同部位两种生物碱的季节和地理差异。结果表明:不同季节,天然黄檗各部位的小檗碱、药根碱含量均表现为:根皮茎皮多年生枝皮1年生枝叶片。小檗碱含量随着季节变化在各部位呈现夏季升高,到秋季下降的趋势。而药根碱在不同部位中的含量随着季节变化的规律不同。根皮、茎皮中的药根碱含量随着季节推移逐渐增多,在夏季升高不明显,而到秋季大量积累。多年生枝皮、1年生枝、叶片中的药根碱在3个季节表现则为先降后升。从纬度水平上看,不同地域间黄檗各部位的生物碱差异显著,在调查的40°—50°N范围内,黄檗两种生物碱的含量普遍在纬度低的地区较高,随着纬度增大先逐渐降低,到N44°左右达到最低值,到了高纬度地区略有回升。     

粉叶小檗细胞的悬浮培养及某些相关的生理生化特性

研究了粉叶小檗细胞悬浮培养过程中细胞生长与小檗碱合成的时间进程,以及某些生理生化特性与上述二者之间的关系。结果表明,细胞生长速度为0．7585g（DW）·L-1（培养基）·d-1,细胞产率在第13d达最大值,为986g·L-1（培养基）。小檗碱含量在第10d最高,为2.51％;小檗碱最大产率出现在第13d,为200.2mg·L-1培养基）。细胞生长和小檗碱积累与培养液中的可溶性精、总磷含量和电导率的变化是负相关,与过氧化物酶活性变化相平行,细胞内可溶性蛋白含量的增加比细胞生长提前。     

小檗红碱的化学与药理

小檗红碱在黄连等中药材中的自然含量很低,但相关药材中的小檗碱在加热条件下可以转化成小檗红碱而使其含量增加.随着对传统中药材微量成分药理活性和药用安全性的日益关注,小檗红碱在相关中药中的治疗作用及安全性已经引起天然药物学领域专家的重视,本文在参考最新文献的基础上,对小檗红碱的化学合成方法、含量分析方法和药理作用等方面进行综述,拟为相关中药质量研究与药理研究提供参考     

不同理化因子对黄连愈伤组织生长和生物碱合成的影响

古蔺野连（又称串珠连Ｃｏｐｔｉｓｇｕｌｉｎｅｎｓｉｓ）为黄连属植物中的一个野生珍稀品种,只生长在我国西南少数山区,其根茎可作中药黄连,含有与商品黄连相同的小檗碱等化学成分^[1].作者在对黄连细胞培养研究中,通过目视法和TLC比较,从古蔺野连的愈伤组织中初步筛选到了生物量增长较快、生物碱合成能力较为稳定的无性系H292,进行了其生物量及生物碱含量的动态测定。本文报道植物激素、温度、光照等对黄连愈伤组织生长和生物碱合成能力的影响。     

蛇伤药酒的质量控制

目的　建立蛇伤药酒的质量控制方法。　方法　用薄层色谱法对蛇伤药酒制剂中黄连、黄柏、大黄进行定性鉴别 ;用紫外分光光度法测定黄连、黄柏中有效成分小檗碱的含量。测定波长为 ( 2 64± 1 ) nm。　结果　定性鉴别色谱特征明显 ,易于区别 ;盐酸小檗碱的含量测定线性范围在 1 .42～ 7.1 0 μg,r=0 .9996,平均回收率为 99.1 4% ,RSD为 1 .3 4%。　结论　该方法可有效的控制蛇伤药酒的质量。     

氮素形态对黄檗幼苗三种生物碱含量的影响

通过改变水培溶液中NH 4-N和NO3--N的比例,探讨了氮素形态对黄檗幼苗主要药用成分小檗碱、药根碱及掌叶防己碱含量的影响。结果表明,小檗碱的含量与氮素形态有明显关系,在根和茎中都表现为NO3--N比例高更有利于小檗碱的积累。单纯的NO3--N供给最有利于药根碱在黄檗幼苗根中的积累,而处理46d后,NH4 -N/NO3--N为75/25和25/75时黄檗幼苗茎中的药根碱含量最高。黄檗幼苗根和茎中的掌叶防己碱含量对营养液中NH4 -N和NO3--N的比例反应类似,在处理46d后,掌叶防己碱含量由高到低所对应的NH4 -N/NO-3-N为25/75、50/50、75/25、0/100、100/0。处理33d后,黄檗幼苗根中的小檗碱、药根碱、掌叶防己碱含量随处理天数的增加呈下降趋势,而在茎中呈上升趋势。如果同时考虑氮素形态对黄檗幼苗生物量的影响,那么NH4 -N/NO3--N为0/100时其小檗碱产量(含量与生物量乘积)最高,而NH4 -N/NO-3-N为25/75时药根碱和掌叶防已碱的产量最高。     

Selective inhibition of vascular smooth muscle cell proliferation by coptisine isolated from Coptis rhizoma, one of the crude drugs composing Kampo medicines Unsei-in.

Acceleration of vascular smooth muscle cell (VSMC) proliferation is closely linked to the pathogenesis of vascular diseases. We, therefore, focused on traditional Japanese herbal medicines (Kampo medicines) used to ameliorate the impairment of microcirculation or blood stasis and screened them for their ability to inhibit rat VSMC proliferation. Among them, Unsei-in was found to effectively suppress VSMC proliferation, and Coptis rhizome was the responsible constituent crude drug. The extract of Coptis rhizome inhibited VSMC proliferation with the GI(50) value of 4.4 microg/ml, which was much lower than those against the proliferation of 3Y1, dRLh-84, B16, and HeLa cells. The Coptis rhizome extract inhibited the progression of VSMC arrested at G(0)/G(1) phase from G(0)/G(1) to S phase, but not that of 3Y1 cells. Biological assay-guided fractionation revealed that an alkaloid of Coptis rhizome, coptisine, was the active ingredient in selectively preventing VSMC proliferation with GI(50) of 3.3 microM (1.2 microg/ml). When the structurally-related isoquinoline alkaloids of protoberberine class were studied for their inhibitory activities, berberine decreased the VSMC proliferation with GI(50) of 95.1 microM (35.4 microg/ml), about 30 times higher concentration than coptisine, while palmatine failed to show any activity. This study provides evidence that coptisine, an ingredient of Unsei-in, prevents VSMC proliferation selectively at lower concentrations compared with various cells or other structurally related alkaloids.     

Memorization of nuclear atmospheric tests by rhizomes and scales of the mediteranean seagrass Posidonia oceanica (Linnaeus) Delile

Determination of the age of rhizome sections of Posidonia oceanica (Linnaeus) Delile (Potamogetonaceae) by the examination of their dead leaf scales (lepidochronology), makes it possible to cut out the rhizomes in precisely dated 1- or 5-year sections. With increasing age of the section, the annual section length and the C:N ratio increase, while the dry weight per unit length and the ratio of dry weight:ash weight decrease. At a site where the only significant ¹³⁷Cs contamination can be ascribed to global nuclear fallout bombs, the activity of ¹³⁷Cs was measured i n 5-year age groups of rhizomes and scales separately. The maximum activity of ¹³⁷Cs in scales occurred in groups produced between 1960 and 1964, a period during which a peak of activity occurred in fallout. The distribution of ¹³⁷Cs activity in rhizome age groups indicates an apparent lag, perhaps owing to transport of material in the rhizome. In situ dead rhizome scales of P. oceanica thus constitute a memory, at least 30-years long, of an environmental chemical pulse. Posidonia oceanica could prove a valuable tool for marine-pollution surveys.     

Rhizome development ofPhragmites australis in a reed community

Variations in the height, shoot density, biomasses of above- and below-ground parts and rhizome distributions ofPhragmites australis were investigated along a line-transect in a reed community at Yufutsu Mire, Hokkaido. Relationships of performance of the reed plants to soil conditions and species compositions were also examined. Three types of rhizome development were recognized in reed plants; (1) the central part of the reed community, characterized by well developed rhizomes and dense aerial shoots, (2) the intermediate part, characterized by development of rhizomes along both the peat and surface layers and very dense aerial shoots, and (3) the marginal part, characterized by development of rhizomes only along the peat layer and sparse aerial shoots. Observation showed that rhizomes in the surface layer actively produced aerial shoots, whereas rhizomes in the peat layer contributed to the spreading of their distribution range. With the growth of rhizomes, organic debris originating from dead rhizomes and roots accumulated in the mineral soil to promote organic soil formation. In dense parts of the reed stand, species composition was poor because of the shading and litter accumulation by reed plants. The effects of microtopography and water level on the establishment of reed seedlings were also considered.     

In vitro propagation of the endangered orchid,Geodorum densiflorum (Lam.) Schltr. through rhizome section culture

Micropropagation of the endangered terrestrial orchid, Geodorum densiflorum (Lam.) Schltr. was achieved through rhizome section culture from in vitro seed- derived rhizomes. Rhizome sections were cultured on Murashige and Skoog (MS) and Knudson C(KC) media supplemented with various growth regulators and 0.1% activated charcoal. The rhizome sections responded on MS medium. Naphthaleneacetic acid (NAA) at 2.0 μM stimulated rhizome growth. However, benzyladenine (BA) at 5.0 μM induced multiple shoots within four weeks of culture and inhibited rhizome growth. The regenerated shoots rooted on MS only or with NAA at 1.0 μM. Well-developed plantlets were transferred to community pots and then to a greenhouse where the plants have been acclimatised. This revised version was published online in June 2006 with corrections to the Cover Date.     

Structural and Histochemical Features of the Slow-Growing Perennial <i>Coptis chinensis</i> Franch. (Ranunculaceae)

Huanglian (Coptis chinensis Franch.) is a slow-growing perennial medicinal herb with considerable economic value. This study aimed to determine the structural characteristics and the levels of berberine deposits in the organs and tissues of Huanglian using light and epifluorescence microscopy. The adventitious roots are composed of primary and secondary structures with endodermis, exodermis, and phellem. The rhizome structures are composed of primary and secondary structures with cuticle and phellem. The leaves are composed of sclerenchymatous rings, isolateral mesophyll, and thin cuticles. We detected berberine in the xylem walls of the roots and rhizomes as well as in the sclerenchymatous rings of the petioles. We postulate that as the exodermis is developed, the deposition of berberine in the xylem closest to the root tips may affect water and nutrient absorption and transfer. Leaf blades had a thin cuticle and isolateral mesophyll, suggesting shade tolerance. These structural and histochemical features suggest that Huanglian is adapted to the slow growing nature of a shady environment.     

Vegetative propagation and sexual reproduction in the woodland understorey pseudo-annual Circaea lutetiana L.

On the forest floor of deciduous woodlands, relative light intensity gradually declines during the early growing season. The woodland understorey pseudo-annual Circaea lutetiana L. completes its life-cycle at the end of summer. These pseudo-annuals are clonal plants which survive the winter only as seeds and as hibernacles produced by the rhizome apices. In this paper, we asked several questions related to the life-history of C. lutetiana. It was found that shoot formation in early spring did not exhaust the old hibernacle. A trade-off between rhizome + hibernacle number and weight might be expected when plants grow under resource limitation. It was hypothesised that both number and weight of rhizomes and hibernacles will be affected by light availability. Since the effect of resource supply on the size number trade-off will depend on the developmental pattern of the rhizome system, rhizome development was studied as well. Soon after the shoots emerged, 1st order rhizomes were formed in May on the nodes of the old hibernacle. First-order rhizomes branched in June and 2nd order rhizomes (side-branches) were continuously produced throughout the growing season. The phenology and developmental rate of plants growing in different light treatments were plastic. On average 30% of rhizome biomass was formed during the vegetative phase, and rhizome and fruit production were only partly separated in time. The ratio of total rhizome biomass to total fruit biomass was not affected by light. Also flower bud removal did not lead to an increase in rhizome production, which suggests that division of biomass to both reproductive modes is rather rigid. The number of 1st order rhizomes was not affected by the light treatments. Under light limitation, both rhizome number and weight of single rhizomes were reduced. In contrast, fruit number, but not weight of single fruits, was limited.     

Evolutionary and organismic constraints on the relationship between spacer length and environmental conditions in clonal plants

The rate of lateral spread of clonal plants is considered a plastic trait that responds directly to environmental conditions. However, this response is likely constrained by evolutionary history and other species attributes. Here we ask how the relationships between lateral spread and the distribution of herbaceous plant species in different environments are influenced by the type of spacer (epigeogenous versus hypogeogenous rhizome), its persistence (integrator versus splitter), and the height and phylogeny of the plant. Analysis of spacer length of 367 species from temperate to arctic Europe revealed that other plant characteristics modulate the relationship between spacer length and environmental conditions. While plants with epigeogenous rhizomes, which have usually shorter spacers than hypogeogenous rhizomes, were associated with illuminated habitats, plants with hypogeogenous rhizomes were associated with warm habitats. These relationships were also shown within the specific rhizome type, as within the group of species with epigeogenous rhizomes, those having short spacers were associated with more light. The trend toward long spacers in warm environment was detected within both epigeogenous and hypogeogenous rhizome bearing species. Splitters were found to be associated with wet, nutrient rich habitats irrespective of rhizome type. When plant height was accounted for by using the ratio of spacer length to plant height (L:H) instead of absolute spacer length, no relationship with environmental variables was found until phylogenetic relationships among the species were taken into account. This implies that overall variability in L:H ratio in the dataset masked trends common for different taxonomic groups. Future comparative studies relating particular clonal growth forms or individual traits to environmental conditions should consider the constraints arising from other growth characteristics and evolutionary history of the subject species.     

盐酸黄连素对蚜虫生物活性的研究

黄连素为黄连Coptis chinensis Franch体内的主要生物碱成份。用采自贵阳花溪附近的新鲜黄连主根提取盐酸黄连素,将粗品重结晶一次后,用 HITACHI-220紫外可见分光光度计测得一次重结晶产品含盐酸黄连素91%,黄连主根中的黄连素含量为11.56%。室内生物测定结果表明,盐酸黄连素对甘蓝蚜Brevicoryne brassicae (L.)具有很强的触杀作用和较好的拒食活性。1%盐酸黄连素的乙醇溶液点滴蚜虫,96 h死亡率高达92.5%,校正死亡率为91.4%, LC₅₀值为(2.289±0.0125)g/L,回归方程Y=7.712+1.027X,相关系数r为0.958,达显著水平;1%盐酸黄连素的乙醇溶液处理叶片,24 h处理和对照叶面的蚜虫栖息率分别为20.6%和79.4%,拒食效果为74.04%, AFC₅₀值为(4.055±0.0215) g/L ,回归方程Y＝9.115+1.719X,相关系数r为0.998,达极显著水平。     

The effects of 1-naphthaleneacetic acid and N6-benzyladenine on the growth of Cymbidium forrestii rhizomes in vitro

An Asiatic orchid, Cymbidium forrestii, was clonally propagated using seed-derived rhizomes as explants. The rhizomes were cultured and proliferated on Murashige and Skoog medium supplemented with various growth substances. Auxins stimulated rhizome growth by increasing branching and fresh weight of the explant, with 1-naphthaleneacetic acid (NAA) being the most effective auxin. All auxins tested suppressed normal shoot formation. The apical meristem of the rhizome reacted to exogenously applied auxin by reducing the cytoplasmic zone of the apical meristem and causing meristem derivatives to rapidly differentiate into vacuolated parenchyma cells. Leaf formation and development was retarded in the presence of auxin. Cytokinins generally reduced rhizome growth and the number of branches, but benzyladenine (BA) can induce shoot formation in vitro. BA induced the cytoplasmic zone of the apical meristem to enlarge and enhanced leaf development. A 5% (w/v) sucrose concentration was most effective in shoot induction when combined with 5 mg1^-1 BA. Activated charcoal promoted rhizome growth; however, shoot formation was inhibited.     

A new estimation method for plant cell viability by determining electron transport activity

A new method with which to estimate the viability of tissue-cultured plant cells was developed. In this method, electrons from the electron transport system of Coptis japonica cells were trapped by artificial electron acceptors, and the color of the reduced acceptor was monitored with a spectrophotometer through an optical fiber as surface-reflected light. Cell viability is represented by the amount of increased reflected light per unit time as electron transport activity (ETA). The electron transport activities of cultured Coptis japonica cells that had been effected in viability by the addition of different concentrations of a microbial broth, were related to the ability of the cells to proliferate. When various microbial broths were added to our Coptis japonica cultures, there was a negative correlation between electron transport activity and the amount of berberine released. During usual subculture, electron transport activity increased from the onset of culture, reached a maximum in the late log phase, then decresed rapidly.