云南常见药用石斛DNA条形码筛选

选择云南省内常见的12种药用石斛，采用分子生物学鉴定技术筛选其适用的DNA条形码序列。以12种药用石斛共36个样品为材料，提取样品总DNA，对核基因片段ITS和ITS2、叶绿体基因片段psbA-trnH和matK序列进行扩增、测序，结合GenBank下载部分石斛序列；利用生物信息学软件进行序列分析和系统发育分析。结果表明，4条序列扩增和测序成功率均为100%；4条序列没有明显的Barcoding Gap，但ITS序列与ITS2序列的种内和种间重叠部分较少，有偏向两端的趋势；系统发育树显示，ITS和psbA-trnH序列能成功区分12种云南常见的药用石斛，ITS2序列未能区分长距石斛(Dendrobium longicornu)和矮石斛(D. bellatulum)，matK序列仅区分6种石斛。建议以ITS和psbA-trnH序列作为云南药用石斛鉴定序列，为药用石斛的种源鉴定提供理论依据。     

DNA条形码技术应用于人参鉴定

人参(Panax ginsengC.A.Meyer),被人们称为“百草之王”,是国内外常用的珍稀名贵中草药和高级滋补品,濒临灭绝,是急需进行资源保护的珍贵物种.随着中草药市场的国际化,由于利益的驱动,市场上伪混品屡见不鲜,对其正确鉴定就成为进行资源保护的首要条件.国内外学者对人参等药用植物资源的鉴定和可持续利用进行了广泛研究.DNA条形码(DNA barcoding)技术是分子鉴定的最新发展,即通过比较一段或几段通用DNA片段,对物种进行快速、准确的识别和鉴定,是近年来生物分类和鉴定的研究热点,在物种鉴定方面显示了广阔的应用前景.在分析传统的鉴定方法在适用范围和局限性的基础上,着重介绍了新兴的DNA条形码技术及其分析方法在人参鉴定上的特点及应用.     

Application of the ITS2 Region for Barcoding Medicinal Plants of Selaginellaceae in Pteridophyta

Background

Selaginellaceae is a family of nonseed plants with special evolutionary significance. Plants of the family Selaginellaceae are similarly shaped and easily confused, complicating identification via traditional methods. This study explored, for the first time, the use of the DNA barcode ITS2 to identify medicinal plants of the Selaginellaceae family.

Methodology/Principal Findings

In our study, 103 samples were collected from the main distribution areas in China; these samples represented 34 species and contained almost all of the medicinal plants of Selaginellaceae. The ITS2 region of the genome was amplified from these samples and sequenced using universal primers and reaction conditions. The success rates of the PCR amplification and sequencing were 100%. There was significant divergence between the interspecific and intraspecific genetic distances of the ITS2 regions, while the presence of a barcoding gap was obvious. Using the BLAST1 and nearest distance methods, our results proved that the ITS2 regions could successfully identify the species of all Selaginellaceae samples examined. In addition, the secondary structures of ITS2 in the helical regions displayed clear differences in stem loop number, size, position, and screw angle among the medicinal plants of Selaginellaceae. Furthermore, cluster analysis using the ITS2 barcode supported the relationship between the species of Selaginellaceae established by traditional morphological methods.

Conclusion

The ITS2 barcode can effectively identify medicinal plants of Selaginellaceae. The results provide a scientific basis for the precise identification of plants of the family Selaginellaceae and the reasonable development of these resources. This study may broaden the application of DNA barcoding in the medicinal plant field and benefit phylogenetic investigations.     

DNA Barcoding in Endangered Mesoamerican Groups of Plants

Among the applications of DNA barcoding for plant conservation is the identification of illegally traded endangered species from small samples or vegetative specimens. DNA barcoding offers an important tool for the phytosanitary authorities to identify species belonging to groups such as the bamboos and orchids, which command high prices in the horticultural trade. In this study we created a DNA barcode library for 20 endangered Orchidaceae species and 36 species of bamboo (Bambusoideae, Poaceae) distributed in Mexico. We applied several metrics to evaluate the efficiency of the barcodes matK and rbcL and, for bamboos, that of the plastid spacer psbI-K. Our results coincide with those of previous barcoding projects in which alone matK allowed for the identification of the most orchid species. For bamboos, the psbI-K spacer retrieved more polymorphic sites and in combination with matK we were able to identify bamboos to at least the generic level.     

DNA条形码技术在植物中的研究现状

DNA条形码技术（DNA barcoding）是用短的DNA片段对物种进行识别和鉴定的分子生物学技术。在动物研究中该技术已经成功应用于利用线粒体细胞色素c氧化酶亚基I（COI）进行物种鉴定和发现隐种或新物种。相对于动物, COI基因在高等植物中进化速率较慢, 因此植物条形码研究以叶绿体基因组作为重点, 但目前还处于寻找合适的基因片段阶段。许多学者对此进行了积极的探索, 报道了多种植物条形码的候选片段或组合, 但还没有获得满足所有标准的特征位点片段。该文介绍了DNA条形码的标准、优点、工作流程及数据分析方法, 总结了DNA条形码在植物中的研究现状。     

DNA条形码技术在植物中的研究现状

DNA条形码技术(DNA barcoding)是用短的DNA片段对物种进行识别和鉴定的分子生物学技术。在动物研究中该技术已经成功应用于利用线粒体细胞色素c氧化酶亚基I(COI)进行物种鉴定和发现隐种或新物种。相对于动物, COI基因在高等植物中进化速率较慢, 因此植物条形码研究以叶绿体基因组作为重点, 但目前还处于寻找合适的基因片段阶段。许多学者对此进行了积极的探索, 报道了多种植物条形码的候选片段或组合, 但还没有获得满足所有标准的特征位点片段。该文介绍了DNA条形码的标准、优点、工作流程及数据分析方法, 总结了DNA条形码在植物中的研究现状。     

DNA Barcoding of Rhodiola (Crassulaceae): A Case Study on a Group of Recently Diversified Medicinal Plants from the Qinghai-Tibetan Plateau

DNA barcoding, the identification of species using one or a few short standardized DNA sequences, is an important complement to traditional taxonomy. However, there are particular challenges for barcoding plants, especially for species with complex evolutionary histories. We herein evaluated the utility of five candidate sequences — rbcL, matK, trnH-psbA, trnL-F and the internal transcribed spacer (ITS) — for barcoding Rhodiola species, a group of high-altitude plants frequently used as adaptogens, hemostatics and tonics in traditional Tibetan medicine. Rhodiola was suggested to have diversified rapidly recently. The genus is thus a good model for testing DNA barcoding strategies for recently diversified medicinal plants. This study analyzed 189 accessions, representing 47 of the 55 recognized Rhodiola species in the Flora of China treatment. Based on intraspecific and interspecific divergence and degree of monophyly statistics, ITS was the best single-locus barcode, resolving 66% of the Rhodiola species. The core combination rbcL+matK resolved only 40.4% of them. Unsurprisingly, the combined use of all five loci provided the highest discrimination power, resolving 80.9% of the species. However, this is weaker than the discrimination power generally reported in barcoding studies of other plant taxa. The observed complications may be due to the recent diversification, incomplete lineage sorting and reticulate evolution of the genus. These processes are common features of numerous plant groups in the high-altitude regions of the Qinghai-Tibetan Plateau.     

4种中药贯众原植物提取物的体外抗菌活性研究

用水提法和醇提法制备苏铁蕨（Brainea insighis）、华南紫萁（Osmunda vachellii）、狗脊蕨（Woodwardia japonica）和乌毛蕨（Blechnum orientale）的根状茎的提取液,应用纸片扩散法和试管二倍稀释法测定了各提取液对11种常见致病菌的抑菌圈、最低抑菌浓度（MIC）和最低杀菌浓度（MBC）。结果表明,各提取液（1g生药／mL）对表皮葡萄球菌、枯草芽孢杆菌、金黄色葡萄球菌及李斯特菌4种革兰氏阳性菌有明显抑制作用,对表皮葡萄球菌的抑制作用尤为明显,且醇提液的抑菌效果优于水提液;对大肠埃希氏菌等革兰氏阴性菌则无抑制作用。在4种贯众原植物中,苏铁蕨对革兰氏阳性菌的抑制活性最强。     

Leukocyte DNA as Surrogate for the Evaluation of Imprinted Loci Methylation in Mammary Tissue DNA

There is growing interest in identifying surrogate tissues to identify epimutations in cancer patients since primary target tissues are often difficult to obtain. Methylation patterns at imprinted loci are established during gametogenesis and post fertilization and their alterations have been associated with elevated risk of cancer. Methylation at several imprinted differentially methylated regions (GRB10 ICR, H19 ICR, KvDMR, SNRPN/SNURF ICR, IGF2 DMR0, and IGF2 DMR2) were analyzed in DNA from leukocytes and mammary tissue (normal, benign diseases, or malignant tumors) from 87 women with and without breast cancer (average age of cancer patients: 53; range: 31–77). Correlations between genomic variants and DNA methylation at the studied loci could not be assessed, making it impossible to exclude such effects. Methylation levels observed in leukocyte and mammary tissue DNA were close to the 50% expected for monoallellic methylation. While no correlation was observed between leukocyte and mammary tissue DNA methylation for most of the analyzed imprinted genes, Spearman''s correlations were statistically significant for IGF2 DMR0 and IGF2 DMR2, although absolute methylation levels differed. Leukocyte DNA methylation levels of selected imprinted genes may therefore serve as surrogate markers of DNA methylation in cancer tissue.     

Cross-cultural Comparison of Medicinal Plants Used to Treat Infections in Northern Thailand

Economic Botany - Drug resistance in pathogenic microorganisms threatens both human and animal health. This has prompted the search for new antimicrobial drugs, including the ones from...     

Concentration,Source, and Potential Human Health Risk of Heavy Metals in the Commonly Consumed Medicinal Plants

Biological Trace Element Research - A trend toward the use of traditional and herbal medicines has developed nowadays, and there is a growing concern regarding them being polluted with heavy...     

Comparative Anatomical Studies on Chinese Medicinal Plants of the Genus Aconitum

This paper reports the result of comparative anatomical studies on medicinal roots of 53 species and varieties in the genus Aconitum from China. The anatomical structures of the roots can be classified into 6 types and 18 subtypes according to the level of their evolution. The anatomy of Aconitum roots is discussed in relation to phylogeny based on chemotaxonomy, cytology and other related disciplines. It was found that the six types of Aconitum roots are correlated with chemical components, chromosomes, toxicity and the gross morphology. The relatively advanced Types I and II roots always contain highly toxic alkaloids of diterpenoid diester of aconitine type, distributed mainly in Ser. 3, 5-11 of Sect. Aconitum of Subgen. Aconitum, while the relatively primitive Types III, IV and II roots with small diameters contain mainly atisine and aminal alcohol alkaloids, which have lower toxicity, present in Subgen. Gymnaconitum and Ser. 1, 2 of Sect. Aconitum of Subgen. Aconitum. The primitive Types V and VI roots of Subgen. Lycoctonum contain mainly the most low-toxic alkaloids of lycoctonine Type. The result is of significance both in theory or practice for the development of medicinal plant resourcesand the safety of clinical usage of these of plants.     

临床常用抗生素对G-致病菌的效价分析

目的:调查我院常用抗生素的种类及其对,临床常见的G~-致病菌的效价情况,为合理选用抗生素提供临床资料。方法:从2246个样本中分离到646株G~-致病茵,用全自动微生物分析仪-VITEK 32进行菌种鉴定及20种抗生素的药敏实验。结果:646株G~-致病菌经鉴定属于14大类,其中铜绿假单胞菌160株,占24.77%;大肠埃希氏菌135株,占20.90%;鲍曼不动杆菌90株,占13.93%;克雷伯菌属116株,占17.96%;其它9种G~-致病菌所占的比例均小于7%。头孢类抗生素是目前最为常用的。在检测的20种抗生素对检测的14种G~-致病菌的效价最高的前三位分别是:亚胺培南(IMP)82.66%、美洛西林(MEZ)81.15%、环丙沙星(CIP)61.06%;效价最差的三位分别是:头孢呋辛(CFO)23.76%、头孢噻吩(CEP)26.74%、替卡西林(TIC)31.58%。结论:经典的抗生素对G~-致病菌的效价不容乐观,且非科学的联合用药可能会降低药效。     

DNA Barcoding for Identification of Sugarcane Borers in China

Sugarcane borers are economically damaging insects with species that vary in distribution patterns both geographically and temporally, and vary based on ecological niche. Currently, identification of sugarcane borers is mostly based on morphological characters. However, morphological identification requires taxonomic expertise. An alternative method to identify sugarcane borers is the use of molecular data. DNA barcoding based on partial cytochrome c oxidase subunit 1 (COI) sequences has proven to be a useful tool for rapid and accurate species determination in many insect taxa. This study was conducted to test the effectiveness of DNA barcodes to discriminate among sugarcane borer species in China. Partial sequences of the COI gene (709 bp) were obtained from six species collected from different geographic areas. Results showed that the pairwise intraspecies genetic distance was < 0.02, whereas the interspecies genetic distance ranged from 0.117 to 0.182. Results from a neighbor-joining tree showed that the six sugarcane borer species were certainly separated. These results suggested that the partial COI sequences had high barcoding resolution in discriminating among sugarcane borer species. Our study emphasized the use of DNA barcodes for identification of the analyzed sugarcane borer species and represents an important step for building a comprehensive barcode library for sugarcane borers in China.     

Screening for Endophytic Fungi with Antitumour and Antifungal Activities from Chinese Medicinal Plants

Summary One hundred and thirty endophytic fungi isolated from 12 Chinese traditional medicinal plants collected at Yuanmou county and Dawei Mountain, Yunnan province, southwest China, were tested for antitumour and antifungal activities by MTT assay on human gastric tumour cell line BGC-823 and the growth inhibition test against 7 phytopathogenic fungi. The results showed that fermentation broths from 9.2% of the isolates exhibited antitumour activity and 30% exhibited antifungal activity, moreover, some of them exhibited broad-spectrum antifungal activity. The active isolates were identified to 32 taxa. The results indicate that the endophytic fungi of Chinese traditional medicinal plants are promising sources of novel bioactive compounds.     

Medicinal Plants of the Washambaa (Tanzania): Documentation and Ethnopharmacological Evaluation

Abstract: Medicinal plants are an important local resource for the Washambaa of the Western Usambara Mountains in Tanzania. In this study the Washambaa medicinal plants are inventoried. It is based on ethnobotanical fieldwork carried out over 15 months. This study presents for the first time an analysis of medicinal plants used in Africa with a ranking of these taxa by the frequency of their reported use. A total of 328 taxa were collected and yielded 2260 individual use reports. The most popular species are Myrica salicifolia and Toddalia asiatica. Subsequently, the use reports were arranged into 9 groups of medicinal uses based on the types of illness treated. The F actor of I nformant C onsensus (F_IC) is used in order to evaluate the ethnobotanical importance of the plants. The largest number of plants and of use reports are in the group of gastrointestinal disorders. For the most commonly used taxa, an ethnopharmacological evaluation was performed. Studies to evaluate the Washambaa therapeutic claims as well as toxicological data are still lacking for many of the species. This study will form the basis for pharmacological and phytochemical research on selected Washambaa medicinal plants.     

Novel Data on the Taxonomic Diversity,Distribution, and Host Plants of Leafmining Moths of the Family Gracillariidae (Lepidoptera) in Siberia,Based on DNA Barcoding

The Gracillariidae fauna of Siberia, the region that occupies almost half of the territory of the Russian Federation, remains poorly studied. During a DNA barcoding study of Gracillariidae in Siberia, based on analysis of larvae and pupae collected from their leaf mines on woody plants, we identified 41 species. Three gracillariids were identified only to genus: Caloptilia sp. (host plant Prunus padus), Parornix sp. (Malus sp.), and Phyllonorycter sp. (Crataegus sp.), representing poorly studied or undescribed species. Six species are reported here for the first time for Siberia: Callisto insperatella (from Novosibirsk and Tomsk provinces), Caloptilia alnivorella (Buryatia), Phyllonorycter ermani (Irkutsk Province), Ph. lantanella (Novosibirsk Province), Ph. pumilae (Omsk and Irkutsk provinces), and Ph. viciae (Krasnoyarsk Territory). Parornix pfaffenzelleri, found in Khakassia, is reported as a species new to Russia. Other 15 gracillariid species previously known from Siberia were recorded in new administrative regions. The invasive lime leafminer Phyllonorycter issikii was first documented in Tomsk Province and Krasnoyarsk Territory in 2017 and 2018, respectively. Seven new gracillariid-host plant associations were found: Parectopa ononidis on Lupinaster pentaphyllus, Sauterina hofmanniella on Lathyrus gmelinii, Caloptilia stigmatella on Salix kochiana, Callisto insperatella on Prunus virginiana, Parornix scoticella on Amelanchier sp., Phyllonorycter ermani on Alnus alnobetula subsp. fruticosa, and Phyllonorycter viciae on Vicia unijuga. The Gracillariidae fauna of Siberia has 51% similarity with that of European Russia and only 38% similarity with that of the Russian Far East.