黑水缬草和毛节缬草中缬草素的含量测定

目的:测定黑龙江省黑水缬草和毛节缬草中主要活性成分缬草素的含量.方法:RP-HPLC法.采用岛津ODSC18色谱柱(150 mm×4.6mm,5μm);流动相为甲醇-水(体积比40:60);流速为1.0 mL/min;检测波长为256nm.结果:缬草素在12.5～300 μg/mL范围内线形良好,回归方程式为y=1 006.17X-748.90(r=0.999 3),平均回收率为97.4%,RSD为2.33%(n=5).结论:该方法简便快速、结果准确,适用于黑水缬草和毛节缬草中缬草素的定量分析.     

宽叶缬草繁殖生物学特性研究

报道了药用植物宽叶缬草自然生长的历程,并对其不同生长环境及传粉环境下收获的瘦果千粒重和发芽率进行了统计分析。结果表明在不同的生长环境中,宽叶缬草瘦果千粒重有极显著差异;而在不同传粉环境中,养蜂实验区和防蜂实验区瘦果的发芽率有极显著差异,且前者高出后者近4倍。证明了通过增加传粉媒介蜜蜂的数量能够解决宽叶缬草人工栽培中的制种问题,从而使大面积栽培宽叶缬草成为可能,实现对宽叶缬草资源的可持续利用。     

黑水缬草一专多能

黑水缬草（Ｖａｌｅｒｉａｎａａｍｕｒｅｎｓｉｓ）为败酱科多年生草本植物。高达１５０厘米,茎直立中空,有纵棱,具粗毛。根状茎有强烈的特殊气味,锥形柱状,具多数细长须根。叶对生,羽状全裂,中央裂片最大。基生叶柄长达２０厘米,茎生叶柄渐短至近无柄。花序顶生...     

中国欧缬草近缘种的分类订正

作者通过查阅国内外标本及多次野外调查采集,对我国的欧缬草近缘种做了一个初步整理。经研究发现,在我国与欧缬草相近的缬草种共有6种1变种,其中包括两个新种;此外,欧缬草在我国仅有引种栽培,而与我国广泛分布且过去被误定为欧缬草的并非一种。     

基于COI基因探索黑水鸡属(Gallinula)部分鸟类的系统发育关系

秧鸡科黑水鸡属(Gallinula)鸟类的分类地位一直存在差异,除黑水鸡、特岛骨顶和暗色水鸡外,其余物种的分属情况一直没有明确定论。本文采用线粒体细胞色素c氧化酶Ⅰ亚基(mtCOI)基因作为分子标记,通过PCR产物直接测序,获取了来自贵州草海的黑水鸡的COI序列,结合GenBank中已有序列,通过遗传距离计算,NJ法、ML法及Bayes法进行系统发育树的构建,共同探讨了黑水鸡属6种鸟类的系统发育关系。结果发现:位于美洲的黑水鸡与亚欧大陆的黑水鸡在分子系统树中呈现出姐妹群关系,考虑为板块运动造成的地理隔离所致,需进一步进行种水平的分类研究;支持小黑水鸡归入Paragallinula属、斑胁水鸡归入Porphyriops属、绿水鸡归入Tribonyx属,不支持普通水鸡在种水平的分类,应为黑水鸡;支持暗色水鸡保留其在黑水鸡属中种水平的分类地位。本研究从分子水平上理清了黑水鸡属鸟类的分类地位,将为后续更深入的研究奠定一定的基础。     

缬草-4,7(11)-二烯在大肠杆菌中的生物合成

缬草-4,7(11)-二烯是一种双环倍半萜烯类化合物,具有镇静、减缓压力、抗焦虑的作用,可用来治疗注意力缺陷多动症(ADHD)等精神疾病。旨在实现缬草-4,7(11)-二烯在大肠杆菌中的从头合成,通过在大肠杆菌BL21(DE3)过表达酵母源,甲羟戊酸途径中催化乙酰辅酶A合成焦磷酸法尼基(FPP)的8个相关基因及来源于缬草属植物(Valeriana officinalis)的缬草-4,7(11)-二烯合酶(VoTPS)基因;并采用水相和有机相双相发酵的方法,通过GC-MS在有机相中检测到了缬草-4,7(11)-二烯,实现了大肠杆菌中从头合成缬草-4,7(11)-二烯。并对IPTG浓度、蛋白诱导温度和选择不同的有机相等方面进行初步优化后,确定发酵条件为:蛋白诱导温度20℃、IPTG的浓度为0.1 mmol/L,有机相为正十二烷。     

缬草与蜘蛛香挥发油的抗菌抗氧化活性研究

通过对缬草和蜘蛛香挥发油抗菌抗氧化活性的研究,为开发天然高效的抗菌抗氧化药物提供新的选择。采用纸片扩散法和试管稀释法对缬草和蜘蛛香挥发油进行抑菌圈大小及最小抑菌浓度的测定;用DPPH法对其抗氧化能力进行研究。结果表明:缬草和蜘蛛香挥发油均具有一定的抗菌作用,细菌试验中对金黄色葡萄球菌的作用最强,二者对其最小抑菌浓度分别为3.125 mg/mL和6.25 mg/mL;对真菌的抑制作用相对较弱,其中蜘蛛香挥发油作用强于缬草挥发油。两者挥发油不同浓度对DPPH自由基有较好的清除能力,实验浓度范围内,最高清除率分别达65.35%和71.75%,均强于阳性对照品BHT。     

缬草属一些存疑种的考订

本文对缬草属的3个存疑种进行了考证和确定,它们是:鞭枝缬草Valeriana flagellifera Bet.;全叶缬草V.hiemalis Graebn.;毛口缬草V.trichostoma Hand.-Mazz.     

缬草和蜘蛛香的资源分布及民族药用调查

对药用植物缬草及蜘蛛香的资源分布以及各民族的药用情况进行了调查,结果表明缬草与蜘蛛香在我国西北及西南有广泛的分布。我国有许多民族将其作为药用物质治疗不同疾病,二者在药用方面具有一定的开发潜力。     

宽叶缬草DNA的提取及正交优化RAPD反应体系

[目的]确定适合宽叶缬草RAPD分析的DNA提取方法以及建立最佳RAPD反应体系。[方法]比较宽叶缬草基因组DNA的两种提取方法(经典CTAB法、试剂盒法);采用正交设计L16(45),针对Taq DNA聚合酶浓度,d NTP浓度,Mg2+浓度,引物浓度,DNA模板浓度进行RAPD扩增,确立最佳RAPD反应体系。[结果]综合比较,试剂盒法较适合宽叶缬草基因组DNA提取;25μl最适宽叶缬草RAPD反应体系为:2.0 U Taq DNA聚合酶、0.4 mmol/L d NTP、4.0 mmol/L Mg2+、4.0μmol/L随机引物、60 ng模板DNA、2.5μl 10×buffer。[结论]试剂盒DNA提取法和正交优化的反应体系适用于宽叶缬草的RAPD分析,为进一步研究黔产宽叶缬草药材遗传多样性奠定了基础。     

Distance-dependent pollen limitation of seed set in some insect-pollinated dioecious plants

By applying hand pollination, we studied pollen limitation of seed set of female plants in four dioecious plant species with insect-pollination. The effect of hand pollination increased with distance to the nearest male plant. Distances at which seed set was 50% of its maximal value (after hand pollination) were: 2.3 m for Valeriana dioica, 5.3 m for Salix repens, 8.5 m for Asparagus officinale and 10.6 m for Bryonia dioica. We discuss to what extent the reduced seed set was caused by either fewer visits or by visits of a lower quality. We argue that quantifying distance-dependent seed set in dioecious plants may be a good way of studying effects of habitat fragmentation on the breakdown of mutualistic pollination systems.     

Dormancy and germination: making every seed count in restoration

From 50 to 90% of wild plant species worldwide produce seeds that are dormant upon maturity, with specific dormancy traits driven by species' occurrence geography, growth form, and genetic factors. While dormancy is a beneficial adaptation for intact natural systems, it can limit plant recruitment in restoration scenarios because seeds may take several seasons to lose dormancy and consequently show low or erratic germination. During this time, seed predation, weed competition, soil erosion, and seed viability loss can lead to plant re‐establishment failure. Understanding and considering seed dormancy and germination traits in restoration planning are thus critical to ensuring effective seed management and seed use efficiency. There are five known dormancy classes (physiological, physical, combinational, morphological, and morphophysiological), each requiring specific cues to alleviate dormancy and enable germination. The dormancy status of a seed can be determined through a series of simple steps that account for initial seed quality and assess germination across a range of environmental conditions. In this article, we outline the steps of the dormancy classification process and the various corresponding methodologies for ex situ dormancy alleviation. We also highlight the importance of record‐keeping and reporting of seed accession information (e.g. geographic coordinates of the seed collection location, cleaning and quality information, storage conditions, and dormancy testing data) to ensure that these factors are adequately considered in restoration planning.     

Essential oil constituents of Valeriana italica and Valeriana tuberosa. Stereochemical and conformational study of 15-acetoxyvaleranone

The chemical composition of the essential oils obtained by hydrodistillation of roots, stems with leaves and inflorescences of Valeriana italica Lam. and Valeriana tuberosa L. were studied by GC/MS. Seventy-three and forty-one constituents were identified from each plant, respectively. The major constituent of the oil obtained from the roots of V. italica was isolated and identified as 15-acetoxyvaleranone. Its stereochemistry and conformation has been studied using NMR spectroscopy and molecular modelling. The oils obtained from V. tuberosa completely lacked the characteristic valerane or kessane sesquiterpenes.     

Mucilage secretion by seeds doubles the chance to escape removal by ants

Post-dispersal seed predation is a risk for plants in semiarid environments, leading to strategies to protect their propagules from seed collection by animals. In this study, we evaluated the importance of mucilage secretion in seeds as a mechanism to reduce seed collection by ants. We selected three Mediterranean species with strong mucilage secretion on their seeds which become sticky upon wetting. Seeds of Rosmarinus officinalis, Fumana ericoides, and Fumana thymifolia were exposed to ants and survival was compared between dry loose seeds and seeds glued to the soil with previously secreted mucilage. The study site was in the Sierra Calderona, 25?km north of Valencia (Spain). The ant?Cplant interaction was analyzed by scrutinizing seed collection by ants and by analyzing the waste piles of ant nests. To test survival, groups of 10 seeds were placed on the ground. Each group consisted of five control (dry) and five?mucilaginous seeds (previously mucilage secreted) and was covered by the cover of a Petri dish modified to permit only the entry of ants. Seeds were inspected weekly for seed disappearance and the survival function (Kaplan?CMeier estimator) was calculated. Seeds of the target species were important food items for ants and were actively collected, and more than 50?% of the experimental seeds that were glued to the ground with their own mucilage survived at the end of the study period but only 0?C20?% of the control seeds survived after the same time of exposure. The implications for plant establishment of these findings are discussed.     

Harvester ant response to spatial and temporal heterogeneity in seed availability: pattern in the process of granivory

The influence of temporal and spatial heterogeneity in seed availability on the foraging behaviour of the harvester ant Messor arenarius was studied in an arid shrubland in the Negev Desert, Israel. The study investigated the implications of behavioural responses to heterogeneity in seed availability for the seed predation process and the potential for feedback effects on vegetation. Vegetation and seed rain were monitored across two landscape patch types (shrub patches and inter-shrub patches) in 1997. Shrub patches were shown to have higher plant and seed-rain density than inter-shrub patches. Patch use and seed selection by M. arenarius foragers were monitored through the spring, summer and autumn of 1997. After a pulse of seed production in the spring, the ants exhibited very narrow diet breadth, specialising on a single annual grass species, Stipa capensis. At this time, ants were foraging and collecting seeds mainly from inter-shrub patches. In the summer, diet breadth broadened and use of shrub patches increased, although the rate of seed collection per unit area was approximately equal in the two patch types. The increase in the use of shrub patches was due to colony-level selection of foraging areas with relatively high shrub cover and an increase in the use of shrub patches by individual foragers. In the autumn, a pulse of seed production by the shrub species Atractylis serratuloides and Noaea mucronata led to a reduction in diet breadth as foragers specialised on these species. During this period, foragers exhibited a large increase in the proportion of time spent in shrub patches and in the proportion of food items collected from shrub patches. The seasonal patterns in foraging behaviour showed linked changes in seed selection and patch use resulting in important differences in the seed predation process between the two landscape patch types. For much of the study period, there was higher seed predation pressure on the inter-shrub patches, which were of relatively low productivity compared with the shrub patches. This suggests that the seed predation process may help maintain the spatial heterogeneity in the density of ephemeral plants in the landscape.     

Somatic embryogenesis in Pinus halepensis Mill.: an important ecological species from the Mediterranean forest

Pinus halepensis Mill. is a common forest species in the Mediterranean area and it is important for environmental conservation. This study established a method of regenerating Pinus halepensis Mill. through somatic embryogenesis. The effect of culture medium (mineral salts, nitrogen source and plant growth regulators), collection date and seed family on embryogenic tissue initiation and proliferation in Pinus halepensis was analysed during the first steps of embryogenesis process. This study showed a marked effect of the culture medium tested as well as some significant differences among collection dates. Furthermore, the embryogenic tissue initiation was affected by the amino acid mixture in the culture medium and the proliferation stage was significantly affected by the combination of plant growth regulators. At the end of the maturation phase the presence of activated charcoal was also evaluated. Finally, maturation of embryogenic tissue was affected by the nitrogen source in the culture medium and these results were different for high and low mature embryo producing cell lines. To the best of our knowledge, this is the first report on Aleppo pine somatic embryogenesis describing a simple and efficient procedure for large-scale somatic embryo production.     

Seed weight patterns of Acacia tortilis from seven seed provenances across Botswana

Laboratory‐based seed storage systems have been developed as an alternative to in situ conservation for indigenous woody plant species. However, interactions between seed quality and environmental variables must be known for each species prior to seed collection, storage and sowing to ensure effective conservation. This study investigated Acacia tortilis seed weight/quality patterns across seven Botswana seed provenances in relation to: soil nutrient status, altitude; latitude; slope angle; % grass cover; height and density of other woody plants nearby. The higher rainfall and relatively eutrophic seed provenances of north‐western Botswana (Chobe, Okavango and Makgadikgadi) were associated with large A. tortilis species and seeds, as well as higher densities of woody plants. Spatial variation in seed weights of A. tortilis was principally a function of rainfall and soil organic carbon. Although more work is required to establish the relationship between seed weight and germination rates for A. tortilis, this research suggests that seed collection should focus on sites with high levels of rainfall and soil organic carbon.     

Frugivory and Seed Dispersal by Northern Pigtailed Macaques (Macaca leonina), in Thailand

Tropical rain forest conservation requires a good understanding of plant–animal interactions. Seed dispersal provides a means for plant seeds to escape competition and density-dependent seed predators and pathogens and to colonize new habitats. This makes the role and effectiveness of frugivorous species in the seed dispersal process an important topic. Northern pigtailed macaques (Macaca leonina) may be effective seed dispersers because they have a diverse diet and process seeds in several ways (swallowing, spitting out, or dropping them). To investigate the seed dispersal effectiveness of a habituated group of pigtailed macaques in Khao Yai National Park, Thailand, we examined seed dispersal quantity (number of fruit species eaten, proportion in the diet, number of feces containing seeds, and number of seeds processed) and quality (processing methods used, seed viability and germination success, habitat type and distance from parent tree for the deposited seeds, and dispersal patterns) via focal and scan sampling, seed collection, and germination tests. We found thousands of seeds per feces, including seeds up to 58 mm in length and from 88 fruit species. Importantly, the macaques dispersed seeds from primary to secondary forests, via swallowing, spitting, and dropping. Of 21 species, the effect of swallowing and spitting was positive for two species (i.e., processed seeds had a higher % germination and % viability than control seeds), neutral for 13 species (no difference in % germination or viability), and negative (processed seeds had lower % germination and viability) for five species. For the final species, the effect was neutral for spat-out seeds but negative for swallowed seeds. We conclude that macaques are effective seed dispersers in both quantitative and qualitative terms and that they are of potential importance for tropical rain forest regeneration.     

Production and dormancy of wild oat (Avena fatua) seed from plants grown under soil waterstress

In 1974 wild oat plants grown from very dormant seed types fA, fB, fC originating from a single field, and in 1976 plants of type fB derived from another location were waterstressed from the time when the spikelets were just beginning to emerge until the seed was fully ripe. The seed production and dormancy of the seed were determined. Waterstress reduced the number of viable seeds per plant by 42% in 1974 and 49% in 1976. The number of viable seed produced on the tillers of the plants were reduced to a greater extent by waterstress than the number produced on their main stems. Some of the differences between the number of viable seeds produced by the three wild oat types were significant. In 1974, 78% of seed from stressed plants was dormant as compared with 90% of that from unstressed plants. Under a given soil moisture, the dormancy of the three types differed little, although seed dormancy was less in tiller seed than in main stem seed. In 1976 seed from stressed plants was 80% dormant, immediately after collection, whereas that from unstressed plants was totally dormant. Storage for 6 months at 25°C decreased these percentages to 17% and 90% and at 5°C to 72% and 98% respectively. Buried in soil immediately after collection, 66% of viable seed from stressed plants gave seedlings in the first autumn after burial as compared with 4% of seed from unstressed plants. Most of the seed from waterstressed plants gave rise to seedlings in the first autumn, and seed from non-stressed plants in the second spring. The α-amylase content was four times greater in seeds from stressed compared with non-stressed plants.     

DNA-Based Identification of <Emphasis Type="Italic">Valeriana officinalis</Emphasis> s.l.: a Multiplexed Amplification Refractory Mutation System (MARMS) and High Resolution Melting Curve Analysis (HRMA)

The wide distribution of Valeriana officinalis as a herbal remedy as well as the considerably higher concentration of putative mutagenic valepotriate metabolites in other drug-delivering valerian species like Valeriana procera Kunth and Valeriana jatamansi Jones ex Roxb. illustrate the necessity of secure authentication of roots of Valeriana officinalis s.l., especially as the morphologically similar roots of the acutely toxic Veratrum album can be mistaken for those of Valeriana officinalis. We developed two DNA-based systems, a multiplex amplification refractory mutation system (MARMS), and a high-resolution melting curve analysis (HRMA) assay, both based on a sequence mutation within the atpB-rbcL region. With both methods, identification of Valeriana officinalis s.l. was possible. With the HRMA, the characteristic melting curve of 33 samples of Valeriana officinalis s.l. and of two commercial samples of Valerianae radix was distinct from the melting curves of all other Valeriana species (60 accessions), and from the closely related genera Centranthus and Valerianella. Since adulteration of Valeriana with toxic Veratrum species was reported previously, Veratrum primers were included in a multiplex PCR-HRM analysis. This system allowed the detection of a Veratrum admixture down to the level of 0.01 %. Although the advantages, in terms of sensitivity, specificity and practicality of the HRM for analysis of degraded plant material were superior to the MARMS assay, both methods are suitable for routine analysis. The results demonstrated the general ability of HRMA to detect specific (toxic) adulterations in drugs in a semiquantitative way.