Mark P. Widrlechner

Development of a New USDA Plant Hardiness Zone Map for the United States

AbstractIn many regions of the world, the extremes of winter cold are a major determinant of the geographic distribution of perennial plant species and of their successful cultivation. In the United States, the U.S. Department of Agriculture (USDA) Plant Hardiness Zone Map (PHZM) is the primary reference for defining geospatial patterns of extreme winter cold for the horticulture and nursery industries, home gardeners, agrometeorologists, and plant scientists. This paper describes the approaches followed for updating the USDA PHZM, the last version of which was published in 1990. The new PHZM depicts 1976–2005 mean annual extreme minimum temperature, in 2.8°C (5°F) half zones, for the conterminous United States, Alaska, Hawaii, and Puerto Rico. Station data were interpolated to a grid with the Parameter-Elevation Regressions on Independent Slopes Model (PRISM) climate-mapping system. PRISM accounts for the effects of elevation, terrain-induced airmass blockage, coastal effects, temperature inversions, and...

Hypericum in infection: Identification of anti-viral and anti-inflammatory constituents.

The Iowa Center for Research on Botanical Dietary Supplements seeks to optimize Echinacea, Hypericum, and Prunella botanical supplements for human-health benefit, emphasizing anti-viral, anti-inflammatory, and anti-pain activities. This mini-review reports on ongoing studies on Hypericum. The Center uses the genetically diverse, well-documented Hypericum populations collected and maintained at the USDA-ARS North Central Regional Plant Introduction Station (NCRPIS), and the strength of research in synthetic chemistry at Iowa State University to tap natural diversity, to help discover key constituents and interactions among constituents that impact bioactivity and toxicity. The NCRPIS has acquired more than 180 distinct populations of Hypericum, with a focus on Hypericum perforatum L. (Hypericaceae), representing about 13% of currently recognized taxa. Center chemists have developed novel synthetic pathways for key flavones, acyl phloroglucinols, hyperolactones, and a tetralin that have been found in Hypericum, and these compounds are used as standards and for bioactivity studies. Both light-dependent and light-independent anti-viral activities have been identified by using bioactivity-guided fractionation of H. perforatum and a HIV-1 infection test system. Our Center has focused on light-independent activity, potentially due to novel chemicals, and polar fractions are undergoing further fractionation. Anti-inflammatory activity has been found to be light-independent, and fractionation of a flavonoid-rich extract revealed four compounds (amentoflavone, chlorogenic acid, pseudohypericin, and quercetin) that interacted in the light to inhibit lipopolysaccharide-induced prostaglandin E2 activity. The Center continues to explore novel populations of H. perforatum and related species to identify constituents and interactions of constituents that contribute to potential health benefits related to infection.

Phylogenetic, morphological, and chemotaxonomic incongruence in the North American endemic genus Echinacea

The study of recently formed species is important because it can help us to better understand organismal divergence and the speciation process. However, these species often present difficult challenges in the field of molecular phylogenetics because the processes that drive molecular divergence can lag behind phenotypic divergence. In the current study we show that species of the recently diverged North American endemic genus of purple coneflower, Echinacea, have low levels of molecular divergence. Data from three nuclear loci and two plastid loci provide neither resolved topologies nor congruent hypotheses about species-level relationships. This lack of phylogenetic resolution is likely due to the combined effects of incomplete lineage sorting, hybridization, and backcrossing following secondary contact. The poor resolution provided by molecular markers contrasts previous studies that found well-resolved and taxonomically supported relationships from metabolic and morphological data. These results suggest that phenotypic canalization, resulting in identifiable morphological species, has occurred rapidly within Echinacea. Conversely, molecular signals have been distorted by gene flow and incomplete lineage sorting. Here we explore the impact of natural history on the genetic organization and phylogenetic relationships of Echinacea.

Permeability of rosmarinic acid in Prunella vulgaris and ursolic acid in Salvia officinalis extracts across Caco-2 cell monolayers

ETHNOPHARMACOLOGICAL RELEVANCE Rosmarinic acid (RA), a caffeic acid-related compound found in high concentrations in Prunella vulgaris (self-heal), and ursolic acid (UA), a pentacyclic triterpene acid concentrated in Salvia officinalis (sage), have been traditionally used to treat inflammation in the mouth, and may also be beneficial for gastrointestinal health in general. AIM OF THE STUDY To investigate the permeabilities of RA and UA as pure compounds and in Prunella vulgaris and Salvia officinalis ethanol extracts across human intestinal epithelial Caco-2 cell monolayers. MATERIALS AND METHODS The permeabilities and phase II biotransformation of RA and UA as pure compounds and in herbal extracts were compared using Caco-2 cells with HPLC detection. RESULTS The apparent permeability coefficient (P(app)) for RA and RA in Prunella vulgaris extracts was 0.2 ± 0.05 × 10(-6)cm/s, significantly increased to 0.9 ± 0.2 × 10(-6)cm/s after β-glucuronidase/sulfatase treatment. P(app) for UA and UA in Salvia officinalis extract was 2.7 ± 0.3 × 10(-6)cm/s and 2.3 ± 0.5 × 10(-6)cm/s before and after β-glucuronidase/sulfatase treatment, respectively. Neither compound was affected in permeability by the herbal extract matrix. CONCLUSION RA and UA in herbal extracts had similar uptake as that found using the pure compounds, which may simplify the prediction of compound efficacy, but the apparent lack of intestinal glucuronidation/sulfation of UA is likely to further enhance the bioavailability of that compound compared with RA.

Assessing phenotypic, biochemical, and molecular diversity in coriander (Coriandrum sativum L.) germplasm

Our goals for this research were to elucidate phenotypic and biochemical diversity in coriander (Coriandrum sativum L.) populations maintained at the North Central Regional Plant Introduction Station in Ames, IA, and examine relationships between amplified fragment length polymorphism (AFLP) markers and patterns of phenotypic and biochemical diversity. Phenotypic and biochemical traits were evaluated, and analyses of variance and mean comparisons were performed on the resulting data sets. Euclidean distances from phenotypic (PD) and biochemical (BD) data were estimated, and modified Rogers’ distances (RD) were estimated for 80 polymorphic AFLP markers. These data were subjected to cluster analyses (CA) and principal components analyses (PCA), to reveal patterns among populations, and to analyses of molecular variance (AMOVA) for grouping patterns from PD and BD by using the 80 polymorphic AFLP markers. Resulting phenotypic, biochemical, and molecular distance matrices were also compared by applying Mantel tests. Our results describe significant differences among populations for all the phenotypic traits, and dendrograms obtained from PD and BD revealed complex phenetic patterns, as did groups from PCA. The primary seed essential oils and nearly all fatty-acid components were identified and their abundance measured; the primary chemical constituents of corresponding PCA groups are described herein. Molecular evidence supported phenotypic and biochemical subgroups. However, variation attributed among subgroups and groups was very low (∼4–6%), while variation among populations within groups was intermediate (∼24–26%), and that within populations was large (∼69–70%), reflecting weak differentiation among subgroups and groups, which was confirmed by values for fixation indices. Phenotypic subgroups described in this study differed somewhat from previous infraspecific classifications. Weak correlations were found between the phenotypic and biochemical matrices and between the biochemical and AFLP matrices. No correlation was found between the phenotypic and AFLP matrices. These results may be related to coriander’s phenotypic plasticity, its wide range in lifecycle duration, its predominantly allogamous reproductive biology, a human-selection process focused on special traits that may be controlled by few genes, and the widespread trade of coriander seeds as a spice, which may result in dynamic, poorly differentiated molecular variation, even when phenotypic and biochemical differentiation is easily documented.

Inhibition of HIV-1 infection by aqueous extracts of Prunella vulgaris L.

BackgroundThe mint family (Lamiaceae) produces a wide variety of constituents with medicinal properties. Several family members have been reported to have antiviral activity, including lemon balm (Melissa officinalis L.), sage (Salvia spp.), peppermint (Mentha × piperita L.), hyssop (Hyssopus officinalis L.), basil (Ocimum spp.) and self-heal (Prunell a vulgaris L.). To further characterize the anti-lentiviral activities of Prunella vulgaris, water and ethanol extracts were tested for their ability to inhibit HIV-1 infection.ResultsAqueous extracts contained more anti-viral activity than did ethanol extracts, displaying potent antiviral activity against HIV-1 at sub μg/mL concentrations with little to no cellular cytotoxicity at concentrations more than 100-fold higher. Time-of-addition studies demonstrated that aqueous extracts were effective when added during the first five hours following initiation of infection, suggesting that the botanical constituents were targeting entry events. Further analysis revealed that extracts inhibited both virus/cell interactions and post-binding events. While only 40% inhibition was maximally achieved in our virus/cell interaction studies, extract effectively blocked post-binding events at concentrations similar to those that blocked infection, suggesting that it was targeting of these latter steps that was most important for mediating inhibition of virus infectivity.ConclusionsWe demonstrate that aqueous P. vulgaris extracts inhibited HIV-1 infectivity. Our studies suggest that inhibition occurs primarily by interference of early, post-virion binding events. The ability of aqueous extracts to inhibit early events within the HIV life cycle suggests that these extracts, or purified constituents responsible for the antiviral activity, are promising microbicides and/or antivirals against HIV-1.

Metabolic profiling of Echinacea genotypes and a test of alternative taxonomic treatments.

The genus Echinacea is used as an herbal medicine to treat a variety of ailments. To better understand its potential chemical variation, 40 Echinacea accessions encompassing broad geographical and morphological diversity were evaluated under controlled conditions. Metabolites of roots from these accessions were analyzed by HPLC-photo diode array (HPLC-PDA), GC-MS, and multivariate statistical methods. In total, 43 lipophilic metabolites, including 24 unknown compounds, were detected. Weighted principal component analysis (WPCA) and clustering analysis of the levels of these metabolites across Echinacea accessions, based on Canberra distances, allowed us to test two alternative taxonomic treatments of the genus, with the further goal of facilitating accession identification. A widely used system developed by McGregor based primarily on morphological features was more congruent with the dendrogram generated from the lipophilic metabolite data than the system more recently developed by Binns et al. Our data support the hypothesis that Echinacea pallida is a diverse allopolyploid, incorporating the genomes of Echinacea simulata and another taxon, possibly Echinacea sanguinea. Finally, most recognized taxa of Echinacea can be identified by their distinct lipophilic metabolite fingerprints.

Medicinal Plants: A Public Resource for Metabolomics and Hypothesis Development

Specialized compounds from photosynthetic organisms serve as rich resources for drug development. From aspirin to atropine, plant-derived natural products have had a profound impact on human health. Technological advances provide new opportunities to access these natural products in a metabolic context. Here, we describe a database and platform for storing, visualizing and statistically analyzing metabolomics data from fourteen medicinal plant species. The metabolomes and associated transcriptomes (RNAseq) for each plant species, gathered from up to twenty tissue/organ samples that have experienced varied growth conditions and developmental histories, were analyzed in parallel. Three case studies illustrate different ways that the data can be integrally used to generate testable hypotheses concerning the biochemistry, phylogeny and natural product diversity of medicinal plants. Deep metabolomics analysis of Camptotheca acuminata exemplifies how such data can be used to inform metabolic understanding of natural product chemical diversity and begin to formulate hypotheses about their biogenesis. Metabolomics data from Prunella vulgaris, a species that contains a wide range ofantioxidant, antiviral, tumoricidal and anti-inflammatory constituents, provide a case study of obtaining biosystematic and developmental fingerprint information from metabolite accumulation data in a little studied species. Digitalis purpurea, well known as a source of cardiac glycosides, is used to illustrate how integrating metabolomics and transcriptomics data can lead to identification of candidate genes encoding biosynthetic enzymes in the cardiac glycoside pathway. Medicinal Plant Metabolomics Resource (MPM) [1] provides a framework for generating experimentally testable hypotheses about the metabolic networks that lead to the generation of specialized compounds, identifying genes that control their biosynthesis and establishing a basis for modeling metabolism in less studied species. The database is publicly available and can be used by researchers in medicine and plant biology.

Response of Wild and Weedy Cucurbita L. to Pathotypes of Pseudoperonospora Cubensis (Berk. & Curt.) Rostov. (Cucurbit Downy Mildew)

The genus Cucurbita includes ca 14 species native to the New World from the United States south to Argentina. It includes at least five different species domesticated before European contact (Sanjur et al., 2002). In many parts of the world, these domesticated species are widely cultivated as vegetables and, to a lesser extent, as oilseeds, animal forages, and ornamentals. Wild populations of Cucurbita often possess disease-resistance genes that are unknown or extremely rare in domesticated populations. This has been demonstrated for resistance to many viruses and fungal pathogens that infect Cucurbita (Rhodes, 1964; Provvidenti, 1990; McCreight and Kishaba, 1991; Munger, 1993; Provvidenti, 1993). Extensive research has also been conducted to transfer disease-resistance genes from wild species into modern Cucurbita cultivars (Contin and Munger, 1977; de Vaulx and Pitrat, 1979; Washek and Munger, 1983; Whitaker and Robinson, 1986; Herrington et al., 1988a, 1988b, 1989; Robinson et al., 1988; Tasaki and Dusi, 1990).

Inhibition of lentivirus replication by aqueous extracts of Prunella vulgaris

BackgroundVarious members of the mint family have been used historically in Chinese and Native American medicine. Many of these same family members, including Prunella vulgaris, have been reported to have anti-viral activities. To further characterize the anti-lentiviral activities of P. vulgaris, water and ethanol extractions were tested for their ability to inhibit equine infectious anemia virus (EIAV) replication.ResultsAqueous extracts contained more anti-viral activity than did ethanol extracts, displaying potent anti-lentiviral activity against virus in cell lines as well as in primary cell cultures with little to no cellular cytotoxicity. Time-of-addition studies demonstrated that the extracts were effective when added during the first four h of the viral life cycle, suggesting that the botanical constituents were targeting the virion itself or early entry events. Further analysis revealed that the extracts did not destroy EIAV virion integrity, but prevented viral particles from binding to the surface of permissive cells. Modest levels of anti-EIAV activity were also detected when the cells were treated with the extracts prior to infection, indicating that anti-EIAV botanical constituents could interact with both viral particles and permissive cells to interfere with infectivity. Size fractionation of the extract demonstrated that eight of the nine fractions generated from aqueous extracts displayed anti-viral activity. Separation of ethanol soluble and insoluble compounds in the eight active fractions revealed that ethanol-soluble constituents were responsible for the anti-viral activity in one fraction whereas ethanol-insoluble constituents were important for the anti-viral activity in two of the other fractions. In three of the five fractions that lost activity upon sub-fractionation, anti-viral activity was restored upon reconstitution of the fractions, indicating that synergistic anti-viral activity is present in several of the fractions.ConclusionOur findings indicate that multiple Prunella constituents have profound anti-viral activity against EIAV, providing additional evidence of the broad anti-viral abilities of these extracts. The ability of the aqueous extracts to prevent entry of viral particles into permissive cells suggests that these extracts may function as promising microbicides against lentiviruses.