In vitro propagation and establishment of adventitious root cultures of Rheum emodi Wall. ex Meisn for quantification of emodin production

In Vitro Cellular & Developmental Biology - Plant - Rheum emodi Wall. ex Meisn is an important medicinal herb used in treating type II diabetes mellitus, arthritis, inflammation, and...     

大黄在体内抗柯萨奇病毒B3的实验研究

柯萨奇病毒 (Ｃｏｘｓａｃｋｉｅｖｉｒｕｓ ,ＣＶ)属小核糖核酸病毒科肠道病毒属 ,根据其对乳鼠的致病能力不同分为Ａ、Ｂ两组 ,Ａ组病毒能够引起乳鼠广泛性肌炎及坏死 ,Ｂ组病毒可致局灶性肌炎。研究表明 ,ＣＶＢ是病毒性心肌炎的主要病因[1,2 ] 。为寻找一种有效的抗ＣＶＢ3 治疗药物 ,笔者通过体外实验发现大黄注射液有抗柯萨奇病毒作用[3] ,并根据中药大黄五脏皆治的理论 ,本研究进一步利用ＣＶＢ3 病毒性心肌炎小鼠模型观察了该药的抗病毒作用。现报告如下 :1　材料与方法1.1　细胞Ｈｅｐ 2细胞由武汉大学典型培养物保藏中心提供。细…     

大黄在体内抗柯萨奇病毒B3的实验研究

In order to search a effective therapy for viral myocarditis, anti vial activity of Rheum officinale on Coxsackieivirus B3 (CVB3) was evalu ated in BALB/C mice. After being determined optium scheme with orthogonal design , mice infected with CVB3 were treated with Rheum officinale, ribavirin an d 0.9% NaCl. The survival rates of the mice were 35%,5% and 5%;Median survival time were 23,11 and 10.5 days;Infectious viral titers were 10-3.2 ,10-4.7 and 10-4.7 TCID50/0.1 mL repectively, The lesion s of cardiac tissues lightened in mice treated with Rheum offinicale. These results showed that Rheum officinale was effective in treatment of CVB3 in fected mice. The study provided scientific basis for further research and widen antiviral spectrum of Rheum officinale.     

Brown spot ofRheum officinale in indonesia with special reference to the variation in conidia of the causal fungus

Brown spot disease ofRheum officinale, a traditional medicinal plant in South East Asia, was newly recorded from the highland area of West Java Province. After confirmation of its pathogenicity towardRheum officinale and comparison of its morphology with hitherto known species, the casual fungus was identified asAscochyta rhei. It produces widely varied conidia from unicellular and bacillar conidia (Phoma-type), 1-septate oblong conidia (Ascochyta-type) to 2–3-septate cylindric conidia. These conidia were sometimes formed within the same pycnidium. They could germinate easily and produce the similar colonies to each other. Supported by “Joint Study Project for Strengthening Research on Diseases of Industrial Crops in Indonesia, Japan International Cooperation Agency (JICA)” in BALITTRO.     

Assessment of liquid culture procedures for in vitro propagation of Rheum emodi

Micropropagation of an endangered Indian medicinal plant, Rheum emodi Wall., was achieved on Murashige and Skoog's medium using different liquid culture procedures. Liquid static (submerged, semi-submerged and with filter paper bridge) and shake (80 and 120 rpm) culture procedures were assessed for their effects on growth and multiplication rates. Best results were obtained using liquid shake cultures, which resulted in 50% reduction in medium requirement, 37.5% reduction in time and 1.5–2.2 fold increase in growth and multiplication rate. Liquid culture-raised plantlets facilitated easy transplantation and 90–95% survived transfer to potting mix in glasshouse.Abbreviations BA 6-benzyladenine - IBA indole-3-butyric acid     

Preparative isolation and purification of hydroxyanthraquinones from Rheum tanguticum Maxim. on normal phase silica gel: using a Flash Master Personal system

A flash chromatography method for preparative separation and purification of five hydroxyanthraquinones from the Chinese medicinal herb Rheum tanguticum Maxim. ex Balf. was developed by using Flash Master Personal(+) systems. The purities of the products, chrysophanol, physcion, emodin, aloe-emodin, and rhein were all over 90%, determined by high performance liquid chromatography (HPLC). Their structures were ascertained by EI-MS, (1)H- and (13)C-NMR data, and by co-TLC and HPLC with the authentic samples available in our laboratory.     

Preparative Isolation and Purification of Hydroxyanthraquinones from Rheum tanguticum Maxim. on Normal Phase Silica Gel: Using a Flash Master Personal System

Abstract

A flash chromatography method for preparative separation and purification of five hydroxyanthraquinones from the Chinese medicinal herb Rheum tanguticum Maxim. ex Balf. was developed by using Flash Master Personal⁺ systems. The purities of the products, chrysophanol, physcion, emodin, aloe‐emodin, and rhein were all over 90%, determined by high performance liquid chromatography (HPLC). Their structures were ascertained by EI‐MS, ¹H‐ and ¹³C‐NMR data, and by co‐TLC and HPLC with the authentic samples available in our laboratory.     

Gene duplication and adaptive evolution of the CHS-like genes within the genus Rheum (Polygonaceae)

It is suggested that gene duplication plays an important role in adaptation and evolution of plants. In this study, we examined whether the genus Rheum, with extensive diversification in the Qinghai-Tibeten Plateau (QTP) and adjacent regions, possessed the duplication of the chalcone synthase (CHS) genes and whether it underwent positive evolution. Here we cloned CHS-like genes from 16 Rheum species. Phylogenetic analyses suggested that CHS-like genes from Rheum comprised two monophyletic lineages (subfamilies), both of which were sisters to another related genus. The results showed that a genus-specific duplication occurred after this genus originated. The comparison of non-synonymous/synonymous substitution ratios between the two lineages in Rheum further indicated that a few sites along the duplicate branch underwent positive selection. The findings indicate that the duplication of Rheum species could contribute to their adaptive ability to rapidly changing environments which was resulted from the large-scale uplifts stages of QTP.     

青藏高原丽江大黄某些生态学特性的研究

丽江大黄（Ｒｈｅｕｍ ｌｉｋｉａｎｇｅｎｓｅ）主要分布于四川青藏接壤的石灰岩山地,垂直分布范围在海拔３５００￣４４００ｍ,处在高寒灌丛草甸和山地寒温性针叶林带。其种群分布格局在大于１．５ｍ＾２尺度上为集群格局;在小于１．５ｍ＾２的尺度上,为随机分布。丽江大黄江合速率日变化在晴天呈现出典型的单峰曲线,与低海拔平原区植物明显不同。这主要缘于在植物生长季节,高原的光和温的互相补偿作用能使光反应和暗反应协     

Soluble Protein Content,Bioactive Compounds and the Antioxidant Activity in Seeds of Ten Rheum tanguticum Lines from Qinghai-Tibet Plateau

Rheum tanguticum (Rh. tanguticum) is a Chinese medicinal plant traditionally used in the treatment of constipation. As a byproduct, the seeds of this plant are rich in nutrients and phytochemicals. This study aimed to determine and assess seed germination ability, seed physical characteristics, soluble protein content, chemical constituents and antioxidant capacity from different breeding lines, to promote the development and utilization of seed resources. Significant differences were observed for the soluble protein content and antioxidant assays among the ten lines. The contents of aloe-emodin, rhein and catechins accumulated in seeds were extremely low and significantly different from those in roots. In contrast, emodin and chrysophanol were abundant in seeds, and significant differences were observed between seeds and roots. It was found that associations between gallic acid and catechins were not significant for either soluble protein or antioxidant capacity. There was a significantly positive correlation between the contents of four anthraquinones (aloe-emodin, rhein, emodin and chrysophanol) and soluble protein. Seeds have potent antioxidative capacity and relatively high levels of soluble protein content. The rich chemical composition of seeds can be widely used in the medical industry for further development.     

Molecular phylogeny, recent radiation and evolution of gross morphology of the rhubarb genus Rheum (Polygonaceae) inferred from chloroplast DNA trnL-F sequences

BACKGROUND AND AIMS: Rheum, a highly diversified genus with about 60 species, is mainly confined to the mountainous and desert regions of the Qinghai-Tibetan plateau and adjacent areas. This genus represents a good example of the extensive diversification of the temperate genera in the Qinghai-Tibetan plateau, in which the forces to drive diversification remain unknown. To date, the infrageneric classification of Rheum has been mainly based on morphological characters. However, it may have been subject to convergent evolution under habitat pressure, and the systematic position of some sections are unclear, especially Sect. Globulosa, which has globular inflorescences, and Sect. Nobilia, which has semi-translucent bracts. Recent palynological research has found substantial contradictions between exine patterns and the current classification of Rheum. Two specific objectives of this research were (1) to evaluate possible relationships of some ambiguous sections with a unique morphology, and (2) to examine possible occurrence of the radiative speciation with low genetic divergence across the total genus and the correlation between the extensive diversification time of Rheum and past geographical events, especially the recent large-scale uplifts of the Qinghai-Tibetan Plateau. METHODS: The chloroplast DNA trnL-F region of 29 individuals representing 26 species of Rheum, belonging to seven out of eight sections, was sequenced and compared. The phylogenetic relationships were further constructed based on the sequences obtained. KEY RESULTS: Despite the highly diversified morphology, the genetic variation in this DNA fragment is relatively low. The molecular phylogeny is highly inconsistent with gross morphology, pollen exine patterns and traditional classifications, except for identifying all samples of Sect. Palmata, three species of Sect. Spiciformia and a few species of Sect. Rheum as corresponding monophyletic groups. The monotypic Sect. Globulosa showed a tentative position within the clade comprising five species of Sect. Rheum. All of the analyses revealed the paraphyly of R. nobile and R. alexandrae, the only two species of Sect. Nobilia circumscribed by the possession of large bracts. The crude calibration of lineages based on trnL-F sequence differentiation implied an extensive diversification of Rheum within approx. 7 million years. CONCLUSIONS: Based on these results, it is suggested that the rich geological and ecological diversity caused by the recent large-scale uplifts of the Qinghai-Tibetan Plateau since the late Tertiary, coupled with the oscillating climate of the Quaternary stage, might have promoted rapid speciation in small and isolated populations, as well as allowing the fixation of unique or rare morphological characters in Rheum. Such a rapid radiation, combined with introgressive hybridization and reticulate evolution, may have caused the transfer of cpDNA haplotypes between morphologically dissimilar species, and might account for the inconsistency between morphological classification and molecular phylogeny reported here.     

Optical and anatomical characteristics of bracts from the Chinese "glasshouse" plant, Rheum alexandrae Batalin (Polygonaceae), in Yunnan, China

Bracts that lacked chlorophyll were compared with rosette leaves on the Chinese glasshouse plant Rheum alexandrae Batalin. The structures were analyzed anatomically and with photospectrometry. Histological features were significantly different between the bracts and the rosette leaves. Epidermal pavement cells and palisade cells were larger in the bracts than in the rosette leaves, but the ratio of the intercellular spaces to the cells in the palisade layer was the same in the two structures. Absorption spectrum analyses of the half-translucent bracts showed that, despite their thinner structure, they absorbed ultraviolet (UV) radiation more efficiently than did the green rosette leaves. The characteristics of the R. alexandrae bracts were different from those of the Himalayan glasshouse plant Rheum nobile. These results suggest that R. alexandrae evolved bracts as an adaptive strategy to protect the inflorescence from UV radiation. Received: August 22, 2001 / Accepted: October 15, 2001     

甘肃六个地区掌叶大黄的RAPD分析

目的:探讨甘肃6个地区掌叶大黄的遗传多样性.方法:采用改良CTAB法提取6个地区样品的基因组DNA,应用RAPD( Randomly amplified polymorphic DNA)技术对电泳结果进行聚类分析,得出聚合树状图.结果:10条多态性引物共扩增得到51条电泳条带,大小在250 ～1 600bp之间,其中33条为多态性条带,多态性比率为64.7％.结论:聚类分析结果表明:甘肃6个地区掌叶大黄的亲缘关系与地理距离有一定的相关性.     

Development of 10 microsatellite loci for <Emphasis Type="Italic">Rheum tanguticum</Emphasis> (Polygonaceae)

Rhubarb is an important Traditional Chinese Medicine. However, the wild resource has been declining. In order to design appropriate conservation methods for the official species across their natural distributions, it is important to characterize their genetic diversity. Here, we describe the development of 10 new microsatellite loci for AC/TG/CCA in Rheum tanguticum. The microsatellites were enriched using the combined biotin capture method. The polymorphism of each locus was further assessed in 12 individuals from four geographically distinct populations of this species. The number of alleles ranged from three to seven and the expected heterozygosity ranged from 0.53 to 0.73. All markers have been checked in the other three species in the genus and two of them together comprise the official medicinal rhubarb resource with R. tanguticum. These microsatellite markers could provide a useful tool for genetic and conservation studies of the rhubarb species.     

人工栽培唐古特大黄中蒽醌含量水平的分析

采用醋酸镁比色法测定青海省不同海拔种植基地中栽培唐古特大黄的总蒽醌含量,并和野生大黄药材进行了比较。结果表明,在海拔较高地区栽培的二、三年生唐古特大黄中,其总蒽醌含量均明显高于较低海拔地区栽培的同龄唐古特大黄,3年生人工栽培大黄总蒽醌含量只有野生大黄药材的一半左右。     

Investigation of species and environmental effects on rhubarb roots metabolome using <Superscript>1</Superscript>H NMR combined with high performance thin layer chromatography

Introduction

The pharmacological activities of medicinal plants are reported to be due to a wide range of metabolites, therein, the concentrations of which are greatly affected by many genetic and/or environmental factors. In this context, a metabolomics approach has been applied to reveal these relationships. The investigation of such complex networks that involve the correlation between multiple biotic and abiotic factors and the metabolome, requires the input of information acquired by more than one analytical platform. Thus, development of new metabolomics techniques or hyphenations is continuously needed.

Objectives

Feasibility of high performance thin-layer chromatography (HPTLC) were investigated as a supplementary tool for medicinal plants metabolomics supporting ¹H nuclear magnetic resonance (¹H NMR) spectroscopy.

Method

The overall metabolic difference of plant material collected from two species (Rheum palmatum and Rheum tanguticum) in different geographical locations and altitudes were analyzed by ¹H NMR- and HPTLC-based metabolic profiling. Both NMR and HPTLC data were submitted to multivariate data analysis including principal component analysis and orthogonal partial least square analysis.

Results

The NMR and HPTLC profiles showed that while chemical variations of rhubarb are in some degree affected by all the factors tested in this study, the most influential factor was altitude of growth. The metabolites responsible for altitude differentiation were chrysophanol, emodin and sennoside A, whereas aloe emodin, catechin, and rhein were the key species-specific markers.

Conclusion

These results demonstrated the potential of HTPLC as a supporting tool for metabolomics due to its high profiling capacity of targeted metabolic groups and preparative capability.

     

Antifungal activity of ethanolic extract of Archu (Rheum emodi) on powdery mildew (Erysiphe cichoracearum) and its role in the induction of resistance in balsam (Impatiens balsamania)

Rheum emodi, vernacularly known as Archu, is one of the important high altitude medicinal plants widely distributed in Himalayan regions. Though widely used in Ayurveda for curing various human diseases, its use in plant diseases is limited. Ethanolic extract of Rheum rhizome was assayed against spore germination of Alternaria solani, Heliminthosporium penniseti and Curvularia palliscens. The inhibition of spore germination was concentration dependent. Maximum inhibition was obtained at 4000 and 5000 ppm followed by 3000, 2000 and 1000 ppm. However, the extract was highly effective in the pre-inoculation treatment against powdery mildew (Erysiphe cichoracearum) of balsam (Impatiens balsamania) under field conditions. High performance liquid chromatographic (HPLC) analysis of balsam leaves showed increased synthesis of phenolic acids, which has been correlated with induced resistance in inhibiting the disease intensity of balsam powdery mildew.     

利用植物提取物绿色合成纳米银粒子及其对台湾乳白蚁的毒性

【目的】利用多种药用植物水提液绿色合成纳米银粒子，测定分析其对台湾乳白蚁Coptotermes formosanus的毒杀活性和作用机理，探索绿色合成的(silver nanoparticles, AgNPs)在白蚁防治方面的潜力，拓宽AgNPs在农业领域的应用前景。【方法】分别利用药用植物大黄Rheum palmatum根茎、白毛夏枯草Ajuga nipponensis全株、苦参Sophora flavescens根和鱼腥草Houttuy niacordata全株的水提取液绿色合成纳米银粒子，采用紫外 可见分光光度计(UV-vis)、扫描电子显微镜(SEM)、透射电子显微镜(TEM)、X射线能谱分析(EDS)和纳米粒度分析仪验证AgNPs的生成并表征其粒子大小、形状和聚集程度等；在实验室条件下测定AgNPs对台湾乳白蚁工蚁的毒杀效果。通过测定800 mg/L AgNPs处理7 d的台湾乳白蚁工蚁体内可溶性蛋白质含量、乙酰胆碱酯酶(AchE)活性和滤纸酶活性(filter paper activity, FPA)水平，来探析AgNPs对白蚁的毒杀作用机理。【结果】4种药用植物水提取液合成的AgNPs颗粒均呈球形，粒径在69~180 nm之间；7 d内AgNPs对台湾乳白蚁工蚁的LC₅₀值分别为150, 340, 342和309 mg/L；800 mg/L AgNPs处理7 d后台湾乳白蚁工蚁的可溶性蛋白质含量以及AchE活性和FPA与对照相比均显著降低。【结论】4种植物提取物合成的AgNPs对台湾乳白蚁工蚁均有较好的毒杀效果，它们可通过降低白蚁体内的可溶性蛋白质含量、AchE活性和FPA来影响其存活，说明绿色合成的AgNPs在防治台湾乳白蚁方面具有较大的潜力。     

Optical and anatomical characteristics of bracts from the Chinese "glasshouse" plant,Rheum alexandrae Batalin (Polygonaceae), in Yunnan,China

Bracts that lacked chlorophyll were compared with rosette leaves on the Chinese glasshouse plant Rheum alexandrae Batalin. The structures were analyzed anatomically and with photospectrometry. Histological features were significantly different between the bracts and the rosette leaves. Epidermal pavement cells and palisade cells were larger in the bracts than in the rosette leaves, but the ratio of the intercellular spaces to the cells in the palisade layer was the same in the two structures. Absorption spectrum analyses of the half-translucent bracts showed that, despite their thinner structure, they absorbed ultraviolet (UV) radiation more efficiently than did the green rosette leaves. The characteristics of the R. alexandrae bracts were different from those of the Himalayan glasshouse plant Rheum nobile. These results suggest that R. alexandrae evolved bracts as an adaptive strategy to protect the inflorescence from UV radiation.     

Proteins and peptides of the salivary gland secretion of medicinal leeches Hirudo verbana, H. medicinalis, and H. orientalis

The protein and peptide composition of medicinal leech salivary gland secretion (SGS) was analyzed in preparations obtained in July from three species--Hirudo verbana, H. medicinalis, and H. orientalis. Two-dimensional electrophoresis (molecular mass 10-150 kD and pI 3-10) revealed no distinctions in the distribution of over 100 silver-stained proteins. Differences were noted only in intensity of 10 protein spots at 30-90 kD and pI 4.7-7.5. Mass spectrometric profiling of SGS of the three leech species using the Zip-Tip/golden chip scheme and cation-exchanging chips CM-10 revealed over 50 components in SGS of each of the three leech species. It was noted that 30-40% of the individual masses of the SGS of each leech species fall within the masses present in SGS of at least one of the two other species. This rather small part of the total mass may be indicative of a high polymorphism of amino acid sequences or a high frequency of posttranslational modifications of the SGS proteins and peptides. Calculation of Jacquard's coefficient showed that H. medicinalis and H. orientalis are closest to each other in SGS composition, which is consistent with data in the literature on the phylogenetic relationship between these two species of medicinal leech. Comparison of detected molecular masses with those of six known biologically active compounds produced by medicinal leeches revealed their uneven distribution in SGS of each of the three medicinal leech species. This opens prospects for using certain species of medicinal leech for targeted therapy of various pathologies.