Three new hosts for Phytophthora ramorum confirmed in Washington State: Salal, Oregon grape, and red huckleberry

Salal (Gaultheria shallon), Oregon grape (Berberis aquifolium), and red huckleberry (Vaccinium parvifolium) are common shrubs in the understory of northwestern forests and are ecologically, culturally and economically important to the region. They are also sold in nurseries for use in ornamental landscapes and ecological restoration. These plant species were symptomatic for Phytophthora in nursery surveys in Washington State between 2011-2015. Symptoms observed on these three hosts resembled those of foliar Phytophthora infection on other woody broadleaf plants. The nursery plants were positive for P. ramorum and Koch’s postulates were completed on potted plants of the three host species, illustrating their potential as a pathway of spread from nurseries to wildlands. We recommend these three hosts be added to the USDA-APHIS regulated host list, which will aid in the effort to prevent movement of P. ramorum into new areas.

Phytochemical and microbial analyses of Berberis sp. extracts

In the near past the use of herbs for health enhanced and scientists are studying new anti-microbial phytochemicals. Although plants have a wide variety of secondary metabolites, very few are still used as antimicrobial. This study performes phytochemical and antibacterial analysis of ethanolic extracts from Berberis vulgaris and Berberis aquifolium. Extracts were prepared from stem and root bark of Berberis sp. with 70% ethanol. After obtaining the plant extracts qualitative and quantitative phytochemical analysis were performed through spectrophotometry, thin layer chromatography, reversed phase HPLC and UV-VIS spectra. The results showed that B. aquifolium extract has a bigger concentration of alkaloids (5.555%) than B. vulgaris extract (4.161%). The analysis from reversed phase HPLC showed that berberine concentration in B. aquifolium is 0.515 mg/ml and in B. vulgaris extract is 1.369 mg/ml, so in oregon grape is found a smaller concentration of berberine than in common barberry. The plant extracts were tested on Escherichia coli (Gram negative) and Staphylococcus aureus (Gram positive) bacteria. We found inhibition between 10-12 mm on S. aureus and on E. coli between 8-10 mm. The extracts exhibited a stronger activity versus S. aureus, which demonstrates that berberine extracts are usefull in treatment of infections.

Notes from the H. J. Andrews Experimental Forest

This chapter details the author's field notes from the H. J. Andrews Experimental Forest. The author was awarded a two-week residency in the forest through the Spring Creek Project, which is administered through Oregon State University. The project began in 2003 and will continue until 2203, fully funded. The mission is to keep a “Forest Log” of ecological reflections for two centuries. The chapter then recounts the author's identification of the trees and plants. The trees include Douglas-firs, western hemlock, western red cedars, and Pacific silver firs. Meanwhile, the plants include the Low Oregon grape, trillium, and Linnaea borealis, better known as “twinflower.” However, there were no blooms to speak of at the end of October.

First Report of Oregon Grape (Mahonia aquifolium) as an Alternate Host for the Wheat Stripe Rust Pathogen (Puccinia striiformis f. sp. tritici) Under Artificial Inoculation

As the primary host of the stripe rust pathogen, Puccinia striiformis f. sp. tritici (Pst), wheat can be infected by both aeciospores and urediniospores, and later is the host that gives rise to urediniospores and teliospores. Barberry species (e.g., Berberis vulgaris) can be infected by basidiospores, produced from the teliospores of wheat plants, and later gives rise to pycniospores and aeciospores, which has been demonstrated through artificial inoculation (3). Oregon grape (Mahonia aquifolium), closely related to Berberis, is a native evergreen shrub that is also grown as an ornamental plant in the Pacific Northwest. To determine if M. aquifolium can also serve as an alternate host for Pst, we conducted artificial inoculations under controlled conditions. Seeds of M. aquifolium collected from Pullman, WA, were sown in pots filled with soil mixture, and plants were grown in a greenhouse under wheat-growing conditions (1). In the first experiment, conducted in May to June 2011, the inoculum was telia collected from artificially inoculated wheat cv. Avocet S with urediniospores of isolate 09-134 (race PST-127) from the greenhouse. In the second experiment, conducted in July to August 2011, the inoculum was telia collected from naturally infected wheat cv. Nugaines with urediniospores from isolate 11-292 (race PST-127) from an experimental field near Pullman. For each experiment, mature teliospores of 60 telia from a single wheat plant were suspended in 1.0 ml of distilled water and inoculated with a fine paint brush onto the leaves of seven or eight 10- to 15-day-old plants of M. aquifolium. Plants were incubated initially in a dew chamber at 10°C for 72 h in darkness, then transferred to a growth chamber with a diurnal temperature cycle of 10 to 24°C and a 16 h light/8 h dark cycle (1). Reddish pycnia with nectar appeared on adaxial surfaces of inoculated leaves at 12 days post-inoculation (DPI), and reddish aecia were produced on the baxial surface at 16 DPI. All 15 M. aquifolium leaves of the 15 plants inoculated with teliospores produced pycnia and aecia. Seedlings of Nugaines and Avocet S, wheat cultivars that are susceptible to all Pst races (1), were then inoculated with a water suspension of aeciospores of 30 aecia collected from the M. aquifolium plants. Wheat plants were incubated as described above for M. aquifolium. Uredinia appeared at 15 DPI, and telia were produced after an additional 15 days. From these uredinia that formed on inoculated wheat, a total of 30 single-uredinium isolates were obtained using the standard procedure (1). Virulence tests were carried out on 20 wheat differentials for 10 randomly selected urediniospore isolates, revealing six virulence patterns. When tested with four selected Pst SSR markers (PstP001, PstP003, PstP005, PstP029) (2) and compared to other race PST-127 isolates, all 10 progeny isolates were homozygous, as were the parental isolates (09-134, 11-292). The virulence tests and marker genotypes verified that the urediniospore isolates resulted from infection by aecia, produced by parental isolate 09-134 through its sexual cycle on M. aquifolium. The study exhibited the completed sexual lifecycle of Pst through the five spore stages on wheat and M. aquifolium in a controlled setting, and suggests that under appropriate weather conditions, M. aquifolium may serve as an alternate host for Pst. Due to the wide distribution of M. aquifolium, further studies are needed to determine if the species can be infected by Pst under natural conditions. References: (1) X. M. Chen et al. Can. J. Plant Pathol. 32:315, 2010. (2) P. Cheng et al. Mol. Ecol. Resour. 12:779, 2012. (3) Y. Jin et al. Phytopathology 100:432, 2010.

First Report of Powdery Mildew of Mahonia aquifolium (Pursh.) Nutt. Caused by Erysiphe (Microsphaera) berberidis (DC.) Lév in Canada

Mahonia aquifolium (Tall Oregon Grape) is a plant native to British Columbia and the coastal Pacific Northwest of the USA. The first discovery of powdery mildew of Mahonia aquifolium in the USA was reported in Washington State in 2003. In Canada, powdery mildew of Oregon grape was discovered in the early summer of 2004 in Victoria, BC. To our knowledge, this is the first record of powdery mildew caused by E. berberidis on Oregon grape in Canada. Accepted for publication 15 May 2005. Published 21 June 2005.

Method Validation for Determination of Alkaloid Content in Goldenseal Root Powder

A fast, practical ambient extraction methodology followed by isocratic liquid chromatography (LC) analysis with UV detection was validated for the determination of berberine, hydrastine, and canadine in goldenseal (Hydrastis canadensis L.) root powder. The method was also validated for palmatine, a major alkaloid present in the possible bioadulterants Coptis, Oregon grape root, and bar-berry bark. Alkaloid standard solutions were linear over the evaluated concentration ranges. The analytical method was linear for alkaloid extraction using 0.3–2 g goldenseal root powder/100 mL extraction solvent. Precision of the method was demonstrated using 10 replicate extractions of 0.5 g goldenseal root powder, with percent relative standard deviation for all 4 alkaloids ≤1.6. Alkaloid recovery was determined by spiking each alkaloid into triplicate aliquots of neat goldenseal root powder. Recoveries ranged from 92.3% for palmatine to 101.9% for hydrastine. Ruggedness of the method was evaluated by performing multiple analyses of goldenseal root powder from 3 suppliers over a 2-year period. The method was also used to analyze Coptis root, Oregon grape root, barberry bark, and celandine herb, which are possible goldenseal bioadulterants. The resulting chromato-graphic profiles of the bioadulterants were significantly different from that of goldenseal. The method was directly transferred to LC with mass spectrometry, which was used to confirm the presence of goldenseal alkaloids tetrahydro-berberastine, berberastine, canadaline, berberine, hydrastine, and canadine, as well as alkaloids from the bioadulterants, including palmatine, jatrorrhizine, and coptisine.