Comparison of Herbarium Label Data and Published Medicinal Use: Herbaria as an Underutilized Source of Ethnobotanical Information

The use of herbarium specimens as vouchers to support ethnobotanical surveys is well established. However, herbaria may be underutilized resources for ethnobotanical research that depends on the analysis of large datasets compiled across multiple sites. Here, we compare two medicinal use datasets, one sourced from published papers and the other from online herbaria to determine whether herbarium and published data are comparable and to what extent herbarium specimens add new data and fill gaps in our knowledge of geographical extent of plant use. Using Brazilian legumes as a case study, we compiled 1400 use reports from 105 publications and 15 Brazilian herbaria. Of the 319 species in 107 genera with cited medicinal uses, 165 (51%) were recorded only in the literature and 55 (17%) only on herbarium labels. Mode of application, plant part used, or therapeutic use was less often documented by herbarium specimen labels (17% with information) than publications (70%). However, medicinal use of 21 of the 128 species known from only one report in the literature was substantiated from independently collected herbarium specimens, and 58 new therapeutic applications, 25 new plant parts, and 16 new modes of application were added for species known from the literature. Thus, when literature reports are few or information-poor, herbarium data can both validate and augment these reports. Herbarium data can also provide insights into the history and geographical extent of use that are not captured in publications.     

Application of Glycol Methacrylate Embedding to Herbarium Specimens of Fruits

Aldehyde fixation and glycol methacrylate embedding were applied to herbarium specimens of fruits of the Compositae. Sections 1-2 μm thick were cut with glass knives. Softening was unnecessary and the hydrophilic properties of the resin permitted staining with a number of dyes. Specimens were examined with bright field and polarized light microscopy. The technique gives good structural preservation and resolution even with 81-year-old herbarium material.     

DNA damage in plant herbarium tissue

Dried plant herbarium specimens are potentially a valuable source of DNA. Efforts to obtain genetic information from this source are often hindered by an inability to obtain amplifiable DNA as herbarium DNA is typically highly degraded. DNA post-mortem damage may not only reduce the number of amplifiable template molecules, but may also lead to the generation of erroneous sequence information. A qualitative and quantitative assessment of DNA post-mortem damage is essential to determine the accuracy of molecular data from herbarium specimens. In this study we present an assessment of DNA damage as miscoding lesions in herbarium specimens using 454-sequencing of amplicons derived from plastid, mitochondrial, and nuclear DNA. In addition, we assess DNA degradation as a result of strand breaks and other types of polymerase non-bypassable damage by quantitative real-time PCR. Comparing four pairs of fresh and herbarium specimens of the same individuals we quantitatively assess post-mortem DNA damage, directly after specimen preparation, as well as after long-term herbarium storage. After specimen preparation we estimate the proportion of gene copy numbers of plastid, mitochondrial, and nuclear DNA to be 2.4-3.8% of fresh control DNA and 1.0-1.3% after long-term herbarium storage, indicating that nearly all DNA damage occurs on specimen preparation. In addition, there is no evidence of preferential degradation of organelle versus nuclear genomes. Increased levels of C→T/G→A transitions were observed in old herbarium plastid DNA, representing 21.8% of observed miscoding lesions. We interpret this type of post-mortem DNA damage-derived modification to have arisen from the hydrolytic deamination of cytosine during long-term herbarium storage. Our results suggest that reliable sequence data can be obtained from herbarium specimens.     

Metagenomic analysis of historical herbarium specimens reveals a postmortem microbial community

Advances in DNA extraction and next‐generation sequencing have made a vast number of historical herbarium specimens available for genomic investigation. These specimens contain not only genomic information from the individual plants themselves, but also from associated microorganisms such as bacteria and fungi. These microorganisms may have colonized the living plant (e.g., pathogens or host‐associated commensal taxa) or may result from postmortem colonization that may include decomposition processes or contamination during sample handling. Here we characterize the metagenomic profile from shotgun sequencing data from herbarium specimens of two widespread plant species (Ambrosia artemisiifolia and Arabidopsis thaliana) collected up to 180 years ago. We used blast searching in combination with megan and were able to infer the metagenomic community even from the oldest herbarium sample. Through comparison with contemporary plant collections, we identify three microbial species that are nearly exclusive to herbarium specimens, including the fungus Alternaria alternata, which can comprise up to 7% of the total sequencing reads. This species probably colonizes the herbarium specimens during preparation for mounting or during storage. By removing the probable contaminating taxa, we observe a temporal shift in the metagenomic composition of the invasive weed Am. artemisiifolia. Our findings demonstrate that it is generally possible to use herbarium specimens for metagenomic analyses, but that the results should be treated with caution, as some of the identified species may be herbarium contaminants rather than representing the natural metagenomic community of the host plant.     

The use of herbarium specimens in DNA phylogenetics: Evaluation and improvement

During the last few years we have been confronted with the need to use herbarium specimens in the molecular phylogeny studies, since it is generally difficult to obtain living material of some rare species. Ancient DNA has been sequenced, and there are also reports on successful DNA amplification from herbarium specimens. However, it is not easy to obtain amplified DNA from the first herbarium sample tested. In this paper, experiments are described about trials of DNA amplification from two to 151-year-old herbarium specimens of plant species we needed for our projects. Of the 17 herbarium samples tested only two allowed DNA amplification under standard DNA isolation conditions. Different types of PCR inhibiting activities were demonstrated in DNA extracts. In some of the extracts there was extremely low concentration of template with satisfactory quality. In some instances, PCR inhibiting activities were successfully removed by treating them either with insoluble polyvinylpyrrolidone or by adding bovine serum albumin (BSA) to the amplification mixture. However, some PCR-inhibiting activities were resistant to the treatments described above. When the concentration of template was very low, a second PCR amplification with internal primers was necessary to increase the amount of DNA for sequencing. Nevertheless, contamination of either DNA extract or amplification mixture were sometimes observed, and consequently precautions were taken to minimize them. Finally, successful amplification was obtained in eight samples out of the 17 examined.     

Changes in plant collection practices from the 16th to 21st centuries: implications for the use of herbarium specimens in global change research

Background and AimsHerbaria were recently advertised as reliable sources of information regarding historical changes in plant traits and biotic interactions. To justify the use of herbaria in global change research, we asked whether the characteristics of herbarium specimens have changed during the past centuries and whether these changes were due to shifts in plant collection practices.MethodsWe measured nine characteristics from 515 herbarium specimens of common European trees and large shrubs collected from 1558 to 2016. We asked botanists to rank these specimens by their scientific quality, and asked artists to rank these specimens by their beauty.Key ResultsEight of 11 assessed characteristics of herbarium specimens changed significantly during the study period. The average number of leaves in plant specimens increased 3-fold, whereas the quality of specimen preparation decreased. Leaf size negatively correlated with leaf number in specimens in both among-species and within-species analyses. The proportion of herbarium sheets containing plant reproductive structures peaked in the 1850s. The scientific value of herbarium specimens increased until the 1700s, but then did not change, whereas their aesthetic value showed no systematic trends.ConclusionsOur findings strongly support the hypothesis that many characteristics of herbarium specimens have changed systematically and substantially from the 16th to 21st centuries due to changes in plant collection and preservation practices. These changes may both create patterns which could be erroneously attributed to environmental changes and obscure historical trends in plant traits. The utmost care ought to be taken to guard against the possibility of misinterpretation of data obtained from herbarium specimens. We recommend that directional changes in characters of herbarium specimens which occurred during the past 150‒200 years, primarily in specimen size and in the presence of reproductive structures, are accounted for when searching for the effects of past environmental changes on plant traits.     

Comparative analysis of different DNA extraction protocols in fresh and herbarium specimens of the genus Dalbergia

Five published DNA extraction protocols were compared for their ability to produce good quality DNA from fresh and herbarium leaves of several species of the genus Dalbergia. The leaves of these species contain high amounts of secondary metabolites, which make it difficult to perform a clean DNA extraction and thereby interfering with subsequent PCR amplification. The protocol that produced the best DNA quality in most of the Dalbergia species analyzed, utilizes polyvinylpyrrolidone to bind the phenolic compounds, a high molar concentration of NaCl to inhibit co-precipitation of polysaccharides and DNA, and LiCl for removing RNA by selective precipitation. The DNA quality of herbarium specimens was worse than that for fresh leaves, due to collecting conditions and preservation of samples. We analyzed 54 herbarium specimens, but the recovered DNA allowed successful PCR amplification in only eight. For the genus Dalbergia, the herbarium is an important source of material for phylogenetic and evolutionary studies; due to the occurrence of the different species in various geographical regions in Brazil, it is difficult to obtain fresh material in nature. Our results demonstrated that for Dalbergia species the methods used for the collection and preservation of herbarium specimens have a mayor influence on DNA quality and in the success of phylogenetic studies of the species.     

凤仙花属两种植物花粉的扫描电镜制样研究

采用腊叶标本花粉直接制样法、烘干法、FAA固定液复原法(酒精梯度脱水、自然干燥)和FAA固定新鲜花药法(脱水、干燥处理同前)四种不同处理方法对凤仙花属(Impatiens L.)植物红纹凤仙花(I.rubro-striata Hook.f.)和长角凤仙花(I.longicornut Y.L.Chen)的花粉进行了制样研究。结果表明:经后两种方法处理的花粉形状不变,且外壁网纹清晰,可获得质量较高的照片,观察时间也不受季节和产地的影响,既可在野外固定保存新鲜花蕾备用,又可利用标本馆现有的腊叶标本花粉经固定液复原后制样观察,操作简便,是进行花粉形态研究的一种较理想的方法。     

Reconstructing the spread of invasive plants: taking into account biases associated with herbarium specimens

Aim To reconstruct the spread of invasive wetland species using herbarium specimens and to develop a method that accounts for the biases associated with this type of historical record. Location Southern Québec. Methods The temporal and spatial distribution of herbarium specimens of vascular plants was examined. Six invasive species that are mainly found in wetlands were compared with five native, non‐expanding hydrophytes. The cumulative number of locations was plotted against time to construct invasion curves. For native species, this ‘invasion curve’ indicates the spatiotemporal distribution of the sampling for herbarium specimens of wetland plants. It also represents the history of the state of knowledge of the distribution of native species. Deviation from the native species invasion curve can be used to describe the spread of invasive species. For every year on record, the cumulative number of locations of each exotic species was divided by the cumulative number of locations of all native species, and the proportions obtained were plotted against time. Periods of invasiveness were then delineated. Results During the twentieth century, two periods of intensive herbarium specimen collection in Québec can be clearly distinguished, i.e. from 1930 to 1940 and from 1950 to 1985. Several periods of invasiveness were delineated for Butomus umbellatus L. (1922–35), Hydrocharis morsus‐ranae L. (1957–96), Lycopus europaeus L. (1963–2000), Lythrum salicaria L. (1890–1905, 1923–46), Phragmites australis (Cav.) Trin. ex Steudel (1963–84) and Rorippa amphibia (L.) Bess. (1929–34, 1943–52, 1971–79). Main conclusions Accounting for sampling biases associated with herbarium specimens is important in order to objectively delineate periods of invasiveness for exotic species. The spread of wetland vascular plant species can be reconstructed using herbarium specimens, even when the phenomenon is rapid, but the reconstruction is certainly more accurate when the invasion occurred during an active sampling period. The highly variable sampling effort for herbarium specimens in Québec and in other regions strongly cautions against using herbarium data without correction procedures.     

Biological collections in an ever changing world: Herbaria as tools for biogeographical and environmental studies

Plant specimens stored in herbaria are being used as never before to document the impacts of global change on humans and nature. However, published statistics on the use of biological collections are rare, and ecologists lack quantitative data demonstrating the relevance to science of herbarium specimens. I found 382 studies with original data that used herbarium specimens to document biogeographical patterns or environmental changes. Most studies are less than 10 years old, and only 1.4% of the herbarium specimens worldwide have been used to answer biogeographical or environmental questions. The vast majority (82%) of papers dealt with vascular plants, but some studies also used bryophytes, lichens, seaweeds and fungi. The herbarium specimens were collected from all continents, but most of the studies used specimens from North America (40% of studies) or Europe (28%). Many types of researches (conservation, plant disease, plant invasion, pollution, etc.) can be conducted using herbarium specimens. Climate change, and especially phenological reconstructions, are clearly emerging research topics. By group, small herbaria (<100,000 specimens) are consulted as often as very large herbaria (>1,000,000 specimens) for biogeographical and environmental research, but in most cases, only large facilities provide specimens collected worldwide. The median number of specimens per study in papers using computerized collections (15,295) was much higher than for papers that did not include electronic data (226). The use of molecular analyses to investigate herbarium specimens is still relatively unexplored, at least from biogeographical and environmental points of view. Combined with recently developed procedures to correct biases, herbarium specimens might provide in the near future exciting additional spatio-temporal insights that are currently unimaginable.     

植物标本馆防虫方法

虫害是植物标本保藏的最大威胁。防虫一直是植物标本馆管理工作的重点。在中国科学院华南植物研究所标本馆70多年管理经验的基础上,近年来在标本保藏技术上不断改进,探索出一些实用的防虫措施和方法,如：驱虫剂、低温冷冻净化法、熏蒸法等。     

湖北虎耳草科(广义)新记录

报道湖北省虎耳草科(Saxifragaceae)植物3种新记录——芽虎耳草(Saxifraga gemmigera var. gemmigera Engl. ex Diels)、双喙虎耳草(Saxifraga davidii Franch.)和宽叶梅花草(Parnassia dilatata Hand.-Mazz.)。凭证标本存放于中国科学院武汉植物园标本馆(HIB)、吉首大学植物标本馆(JIU)和后河国家级自然保护区植物标本室(HHB)。     

Amplification of DNA of Xanthomonas axonopodis pv. citri from historic citrus canker herbarium specimens

Herbaria are important resources for the study of the origins and dispersal of plant pathogens, particularly bacterial plant pathogens that incite local lesions in which large numbers of pathogen genomes are concentrated. Xanthomonas axonopodis pv. citri (Xac), the causal agent of citrus bacterial canker disease, is a notable example of such a pathogen. The appearance of novel strains of the pathogen in Florida and elsewhere make it increasingly important to understand the relationships among strains of this pathogen. USDA-ARS at Beltsville, Maryland maintains approximately 700 herbarium specimens with citrus canker disease lesions up to 90 years old, originally collected from all over the world, and so is an important resource for phytogeographic studies of this bacterium. Unfortunately, DNA in herbarium specimens is degraded and may contain high levels of inhibitors of PCR. In this study, we compared a total of 23 DNA isolation techniques in combination with 31 novel primer pairs in order to develop an efficient protocol for the analysis of Xac DNA in herbarium specimens. We identified the most reliable extraction method, identified in terms of successful amplification by our panel of 31 primer pairs. We also identified the most robust primer pairs, identified as successful in the largest number of extracts prepared by different methods. We amplified Xac genomic sequences up to 542 bp long from herbarium samples up to 89 years old. Primers varied in effectiveness, with some primer pairs amplifying Xac DNA from a 1/10,000 dilution of extract from a single lesion from a citrus canker herbarium specimen. Our methodology will be useful to identify pathogens and perform molecular analyses of bacterial and possibly fungal genomes from herbarium specimens.     

Sampling procedures and species estimation: testing the effectiveness of herbarium data against vegetation sampling in an oceanic island

Questions: What is the relationship between species assemblages in herbarium collections and species abundances in the field, and how trustworthy are herbarium data in vegetation science? Location: Guadalupe Island, Baja California, Mexico. Methods: We compared species‐abundance distribution and evenness in 110 vegetation plots in Guadalupe Island against data from four herbaria. We tested whether the relative frequencies derived from herbarium specimens differed significantly from species frequencies in the field. We compared the rarefaction curves for both field and herbarium data sets, and tested whether taxonomic collectors accumulated new species at a higher rate than that observed in ecological plot sampling. Results: At any given sampling effort, the total number of observed species was higher in herbarium data. The relative abundance of common species in the field was higher, and the evenness of the distribution was lower, than in herbarium data. There was no significant correlation between species abundances in the field and in the herbaria. By selectively targeting rare species, collectors accumulate previously unseen species much faster than through ecological sampling. Conclusions: Because collectors aim for the rarer species and avoid the more common ones, the relative abundance of species in herbarium collections cannot be interpreted as a predictor of their true abundance in the field. Any statistical procedure that requires the sample to be representative of the true abundance distribution is likely to show errors when applied to herbarium data. However, because collectors actively search for rare species their rate of species accumulation is higher and their floristic lists are more complete than those obtained through ecological field sampling.     

高校新型植物标本馆的建立与发展——以河南农业大学植物标本馆为例

高校标本馆作为重要的科普和教学实践基地,是连接大学与社会的桥梁,在提升高校社会影响力和加快高校发展中起着重要作用。目前,高校标本馆普遍存在经费少和缺乏管理等问题。如何立足高校现状,有效解决标本采集和标本数字化过程中耗时耗力的难题,建设具有地方特色的新型植物标本馆,是高校标本馆建设关注的焦点之一。该文以河南农业大学植物标本馆新馆建设为例,介绍了河南农业大学标本馆成立以来,充分发挥高校的人力资源优势,让学生通过实验课或野外实习积极参与新型植物标本馆的建设,基本实现了标本采集、制作、鉴定和数字化的同步进行。在这一过程中,既有效锻炼了学生能力,加深了学生对植物形态的认识,发挥了植物标本馆在教学、科学研究及科学普及中的积极作用,又极大地丰富了标本馆馆藏量,有效地促进了高校新型植物标本馆的发展。     

Is Shape of a Fresh and Dried Leaf the Same?

Plants kept as dried herbarium specimens share many characteristics with their living counterparts, but there are some substantial differences between them. Due to dehydration, leaves of herbarium specimens change not only their mass and colour, but in many cases change their dimensions, too. The present study aimed to determine whether leaf shape changes during the drying process. A total of 794 pairs of fresh and dried leaves or leaflets of 22 plant taxa were studied. The shape of the blades was quantified using elliptic Fourier analysis combined with principal component analysis. In addition, area and mass of the leaves were measured. Statistical tests were applied for comparing fresh and dried leaves. The results indicate that the preservation process of pressing and drying plants for herbarium purposes causes changes in leaf shape. In general, the shape changes were directional. As the shape of fresh and dried plants is different, it is strongly recommended that shape analyses should be performed on datasets containing either of the leaf types.     

The herbarium of Trinity College, Dublin: its history and contents

The herbarium was established in 1840, with the personal herbarium of Thomas Coulter as its nucleus. Extensive purchases were made by him and by his successor, W. H. Harvey, whose personal herbarium was also incorporated. Harvey also received numerous donations, partly through his friendship with W. J. Hooker and Asa Gray, but also from his position as joint author of Flora Capensis. After Harvey's death in 1866 growth continued at a slower rate till 1950, when it once more accelerated. The herbarium contains c. 206000 specimens in all: 166000 seed plants, 8000 pteridophytes, 8000 bryophytes, 20 000 algae and 4000 lichens. An analysis of the geographical origin of the specimens is given, and a list of the chief collectors. The regions most fully represented are Europe (especially the west and south), India, South Africa, North and South America. There is relatively little from China and Tropical Africa. Among the largest collections are those of Spruce (Amazon and Andes), Harvey (world-wide, especially Australian algae), A. F. G. Kerr (Thailand), Drege (South Africa) and Kotschy (Orient).     

The Henry Parker Sartwell herbarium of Hamilton College

H. P. Sartwell's private herbarium was donated to the New York Botanical Garden in 1983 by Hamilton College of Clinton, New York. This herbarium was assembled by Dr. Sartwell (1792–1867) throughout his carcer and was eventually sold to Hamilton College. The collection is particularly rich in type material, western New York plants, and carices. Dr. Sartwell had an active exchange program with his contemporaries and, therefore, his herbarium also has collections from many parts of the world. An index to collectors is presented, as well as a list of verified and probable types.     

Historical herbarium specimens in plant molecular systematics — an example from the fern genus <Emphasis Type="Italic">Lindsaea</Emphasis> (Lindsaeaceae)

We extracted, amplified and sequenced DNA from historical herbarium specimens and silica-dried samples of the fern genus Lindsaea in order to study the sequencing success between the two kinds of samples. High quality sequences were obtained from 57% of the herbarium samples. The specimens age was found to be of little importance for sequencing success when less than 75 years, but the colour of a specimen was found more indicative of sequencing success. Shorter DNA fragments were sequenced successfully twice as often as longer fragments from the herbarium material; in relatively recently collected silica-dried material longer sequences were obtained almost as frequently as short ones. No obvious differences in sequencing success between material originating from different herbaria was observed. We conclude that by using specifically designed DNA extraction protocols and by sequencing short DNA fragments from carefully selected specimens, herbarium material and type specimens can be successfully used in molecular systematics. Typical material or specimens from the type locality (topotypes) should be preferred, when placing a species in a phylogeny.