Characterisation of European and North American Phytophthora ramorum isolates due to their morphology and mating behaviour in vitro with heterothallic Phytophthora species

2005 ◽  
Vol 109 (8) ◽  
pp. 860-871 ◽  
Author(s):  
Sabine Werres ◽  
Katrin Kaminski
2018 ◽  
Vol 108 (7) ◽  
pp. 829-836 ◽  
Author(s):  
Fumiaki Funahashi ◽  
Jennifer L. Parke

Research on solarization efficacy has examined the critical temperature and minimum exposure time to inactivate soilborne pathogens. Most mathematical models focus on survival of inoculum subjected to a constant heat regime rather than an intermittent heat regime that better simulates field conditions. To develop a more accurate predictive model, we conducted controlled lab experiments with rhododendron leaf disks infested with Phytophthora ramorum and P. pini. Focused in vitro experiments with P. ramorum showed significantly longer survival of inoculum exposed to intermittent versus constant heat, indicating that intermittent heat is less damaging. A similar trend was observed in soil. Damage was evaluated by comparing the reduction in subsequent survival time of inoculum subjected to different intensities of sublethal heat treatments. Inoculum exposure to continuous heat reflected an increasing rate of damage accumulation. Multiple sublethal heat events resulted in a constant rate of damage accumulation which allowed us to calculate total damage as the sum of damage from each heat event. A model including a correction for an intermittent heat regime significantly improved the prediction of thermal inactivation under a temperature regime that simulated field conditions.


Plant Disease ◽  
2003 ◽  
Vol 87 (10) ◽  
pp. 1266-1266 ◽  
Author(s):  
Sabine Werres ◽  
Daphné De Merlier

Since its original isolation in 1993, Phytophthora ramorum has become an important pathogen. Initially, it was determined to be the causal agent of a twig blight of Rhododendron spp. in Germany and the Netherlands (3). Around the same period, symptoms and mortality on oak (Quercus spp.) and tanoak (Lithocarpus densiflorus) were associated with P. ramorum in California (2), where the disease was named sudden oak death. Subsequently, P. ramorum has been detected on a wide range of forest trees and shrub species in the United States. In Europe, the pathogen has spread to many countries, primarily on nursery plants of Rhododendron and Viburnum spp., and recently, on Camellia japonica, Kalmia latifolia, Pieris formosa var. forrestii, P. japonica, Leucothoe sp., Syringa vulgaris, and Taxus baccata. P. ramorum has not been observed in European forests. P. ramorum is heterothallic, and initial in vitro mating studies on agar media suggested that only the A1 mating type occurred in Europe, while only the A2 mating type was present in the United States (4). However, an isolate collected in 2002 in Belgium (1) appears to be the A2 mating type. This isolate (CBS 110901, Centraal Bureau voor Schimmelcultures, Baarn, the Netherlands) originated from an imported V. bodnantense plant at an ornamental nursery. A hyphal tip culture (BBA 26/02) of this isolate produced no oogonia on carrot piece agar after 6 weeks in pairing tests with other Phytophthora species of mating type A2. When paired with mating type A1 of P. cambivora, P. cinnamomi, P. cryptogea, and P. drechsleri, however, oogonia were observed in all pairings within 6 weeks. The number of oogonia was low in all pairings but was highest in pairings with P. cryptogea. No oospores were produced after 6 weeks between P. ramorum isolates BBA 26/02 and BBA 9/95 (from the holotype, mating type A1), but gametangia were observed when these isolates were paired on Rhododendron sp. twigs. Normal oogonia were produced on the outgrowing mycelium when pieces from these twigs were placed on carrot piece agar. The shape and size of the oogonia produced on carrot piece agar after pairing with P. cryptogea and on Rhododendron sp. twigs after pairing with P. ramorum BBA 9/95 were similar (24 to 34 μm, mean 29.6 μm and 25 to 33 μm, mean 30.6 μm, respectively). To our knowledge, this is the first observation of P. ramorum mating type A2 in Europe. References: (1) D. De Merlier et al. Plant Dis. 87:203, 2003. (2) D. M. Rizzo et al. Plant Dis. 86:205, 2002. (3) S. Werres et al. Mycol. Res. 105:1166, 2001. (4) S. Werres and B. Zielke. J. Plant Dis. Prot. 110:129, 2003.


1983 ◽  
Vol 36 (2) ◽  
pp. 191 ◽  
Author(s):  
D Keast ◽  
C Tonkin

Soil pH, soil moisture content and soil organic matter content did not appear to influence significantly the total numbers of actinomycetes isolated from sample sites in Western Australia. However, seasonal influences exist with summer conditions leading to higher spore isolation. Substantial but non-specific antifungal activity against Phytophthora cinnamomi, P. cryptogea, P. nicotiana, Pythium proli/erum and L. laccata was detected in vitro from many of the 2367 actinomycetes isolated. Antifungal activity mayor may not occur in members of the same actinomycete group, suggesting segregation of antifungal capacity within all groups. A limited number of actinomycete groups was isolated from the rhizosphere of plants and these exhibited similar properties to their counterparts in soil or litter. Actinomycetes isolated from the rhizosphere of Pinus radiata produced a high degree of in vitro antifungal activity against the Phytophthora species but, in general, actinomycetes isolated from root surfaces exhibited antibiosis against all the fungi tested. More actinomycetes showed antifungal activity from soils where P. cinnamomi was causing dieback of jarrah and other understorey species.


2014 ◽  
Vol 67 ◽  
pp. 54-59 ◽  
Author(s):  
I.J. Horner ◽  
E.G. Hough

In kauri forest soils surveys Phytophthora taxon Agathis (PTA) P cinnamomi P multivora and P cryptogea were detected frequently In vitro and glasshouse studies determined that all four Phytophthora species produced lesions on excised kauri leaves and stems Lesion advance was significantly slower with P cinnamomi P multivora and P cryptogea than with PTA When 2yearold kauri seedlings were trunkinoculated lesion spread was rapid with PTA trunks were girdled and all trees died within 46 weeks Phytophthora cinnamomi P multivora and P cryptogea produced substantially smaller lesions than PTA no trees died and plant growth was only slightly suppressed Following soil inoculation with PTA all kauri seedlings died within 10 weeks There were no deaths following soil inoculation with P cinnamomi P multivora or P cryptogea although feeder root damage was observed and the respective pathogens were reisolated Results suggest that PTA is an aggressive pathogen and the other three species are weaker pathogens of kauri


Plant Disease ◽  
2003 ◽  
Vol 87 (2) ◽  
pp. 203-203 ◽  
Author(s):  
D. De Merlier ◽  
A. Chandelier ◽  
M. Cavelier

In the past decade, a new Phytophthora species inducing shoot canker on Rhododendron and dieback of Viburnum has been observed in Europe, mainly in Germany and the Netherlands, and California. This new pathogen has been named Phytophthora ramorum (3). In May 2002, a diseased Viburnum plant (Viburnum bodnantense) from the Plant Protection Service (Ministry of Agriculture, Belgium) was submitted to our laboratory for diagnosis. Symptoms included wilting, leaves turning from green to brown, discolored vascular tissues, and root necrosis. The plant came from a Belgian ornamental nursery that obtained supplies of stock plants from the Netherlands. Pieces of necrotic root tissue were excised, surface-disinfected, and transferred aseptically to a Phytophthora selective medium. P. ramorum was identified based on morphological characteristics, including the production of numerous, thin-walled chlamydospores (25 to 70 µm in diameter, average 43 µm) and deciduous, semi-papillate sporangia arranged in clusters. Radial growth after 6 days at 20°C on V8 juice agar was 2.8 mm per day. Random amplified microsatellite markers (RAMS) (2) from the total genomic DNA of the Belgian strain (CBS 110901) were similar to those of P. ramorum reference strains (CBS 101330, CBS 101332, and CBS 101554). Using PCR primers specific for P. ramorum, the identification was confirmed by W. A. Man in't Veld (Plantenziektenkundige Dienst, Wageningen, the Netherlands) (1). A pathogenicity test was carried out on three sterile cuttings of Rhododendron catawbiense (3). Brown lesions were observed on the inoculated cuttings after 6 to 7 days. None of the three uninoculated cuttings showed symptoms of infection. P. ramorum was reisolated from lesion margins on the inoculated cuttings. To our knowledge, this is the first report of the fungus from Belgium. Since our initial observation, we have found P. ramorum in other Belgian nurseries on R. yakusimanum. References: (1) M. Garbelotto et al. US For. Ser. Gen. Tech. Rep. PSW-GRT. 184:765, 2002. (2) J. Hantula et al. Mycol. Res. 101:565, 1997. (3) S. Werres et al. Mycol. Res. 105:1155, 2001.


Plant Disease ◽  
2014 ◽  
Vol 98 (11) ◽  
pp. 1582-1582 ◽  
Author(s):  
S. Vitale ◽  
L. Luongo ◽  
M. Galli ◽  
A. Belisario

The genus Viburnum comprises over 150 species of shrubs and small trees such as Laurustinus (Viburnum tinus L.), which is one of the most widely used ornamental plants in private and public gardens. Furthermore, it commonly forms stands of natural woodland in the Mediterranean area. In autumn 2012, a survey was conducted to determine the presence of Phytophthora ramorum on Viburnum in commercial nurseries in the Latium region where wilting, dieback, and death of twigs were observed on 30% of the Laurustinus plants. A Phytophthora species was consistently recovered from soil rich in feeder roots from potted Laurustinus plants showing symptoms. Soil samples were baited with rhododendron leaves. Small pieces of leaf tissue cut from the margin of lesions were plated on P5ARPH selective medium (4). Pure cultures, obtained by single-hypha transfers on potato dextrose agar (PDA), were petaloid. Sporangia formation was induced on pepper seeds (3). Sporangia were almost spherical, ovoid or obpyriform, non-papillate and non-caducous, measuring 36.6 to 71.4 × 33.4 to 48.3 μm (average 53.3 × 37.4 μm) with a length/width ratio of 1.4. Chlamydospores were terminal and 25.2 to 37.9 μm in diameter. Isolates were considered heterothallic because they did not produce gametangia in culture or on the host. All isolates examined had 30 to 35°C as optimum temperatures. Based on these morphological characteristics, the isolates were identified as Phytophthora hydropathica (2). Morphological identification was confirmed by internal transcribed spacer (ITS), and mitochondrial partial cytochrome oxidase subunit 2 (CoxII) with BLAST analysis in the NCBI database revealing 99% identity with ITS and 100% identity with CoxII. The sequences of the three isolates AB234, AB235, and AB236 were deposited in European Nucleotide Archive (ENA) with the accession nos. HG934148, HG934149, and HG934150 for ITS and HG934151, HG934152, and HG934153 for CoxII, respectively. Pathogenicity tests were conducted in the greenhouse on a total of six 1-year-old shoots cut from V. tinus plants with two inoculation points each. Mycelial plugs cut from the margins of actively growing 8-day-old cultures on PDA were inserted through the epidermis into the phloem. Controls were treated as described above except that sterile PDA plugs replaced the inoculum. Shoots were incubated in test tubes with sterile water in the dark at 24 ± 2°C. After 2 weeks, lesions were evident at the inoculation points and symptoms were similar to those caused by natural infection. P. hydropathica was consistently re-isolated from the margin of lesions, while controls remained symptomless. In the United States in 2008, P. hydropathica was described as spreading from irrigation water to Rhododendron catawbiense and Kalmia latifolia (2). This pathogen can also attack several other horticultural crops (1), but to our knowledge, this is the first report of P. hydropathica causing wilting and shoot dieback on V. tinus. References: (1) C. X. Hong et al. Plant Dis. 92:1201, 2008. (2) C. X. Hong et al. Plant Pathol. 59:913, 2010. (3) E. Ilieva et al. Eur. J. Plant Path. 101:623, 1995. (4) S. N. Jeffers and S. B. Martin. Plant Dis. 70:1038, 1986.


2011 ◽  
Vol 89 (6) ◽  
pp. 419-427 ◽  
Author(s):  
Misagh Alipour ◽  
Abdelwahab Omri ◽  
Zacharias E. Suntres

This study was carried out to examine the antimicrobial activity of the aqueous extract of Panax quinquefolius from North American ginseng (NAGE) root against Pseudomonas aeruginosa . The minimum inhibitory concentrations of reference and clinical isolates of Pseudomonas aeruginosa were measured by a standard agar-dilution method. At subinhibitory NAGE concentrations, the secretion of virulence factors, motility on agar, and adhesion to 96-well microplates were studied on the nonmucoid Pseudomonas aeruginosa O1 strain. At suprainhibitory concentrations, the activity of NAGE against mature biofilm complexes formed in the Calgary Biofilm Device and the Stovall flow cell were assessed. NAGE possessed an antibacterial activity against all the Pseudomonas aeruginosa strains at 1.25%–2.5% w/v. NAGE also significantly attenuated pyocyanin, pyoverdine, and lipase concentrations, stimulated twitching, and attenuated swarming and swimming motility. At 1.25% w/v, NAGE augmented adhesion, and at 5% w/v detached 1-day-old biofilms in microplates. The extract also eradicated 6-day-old mature biofilms (5% w/v), and fluorescence microscopy displayed a reduction of live cells and biofilm complexes compared with nontreated biofilms. These data suggest that the aqueous extract from North American ginseng possesses antimicrobial activities in vitro.


2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Inga Kwiecień ◽  
Beata Kawka ◽  
Halina Ekiert

American skullcap (Scutellaria lateriflora L.) has long been used as a medicine by North American Indian tribes. A monograph of Scutellariae lateriflorae herba was included in the American Herbal Pharmacopoeia in 2009. This raw material is very popular in North America because of its sedative action. It is also highly valued in Australia, while in Europe, it has been widely used until now in the United Kingdom. The main components of the herb of this plant species include flavonoids specific for the Scutellaria genus, mostly baicalein, baicalin, wogonin, scutellarin. They are accompanied by other groups of compounds: diterpenoids – neoclerodan derivatives, γ-aminobutyric acid, melatonin, serotonin, catalpol, phenolic acids, phenylpropanoid glycosides (mostly verbascoside) and essential oil. Studies of chemical composition and investigation of medicinal activity of this raw material confirmed its uses known from North American folk medicine. Study results proved most of all a significance of flavonoid compounds present in the raw material for its documented biological activity. The aim of the paper is to review the present knowledge on chemical composition and medicinal significance of this very valuable plant. The research team from the Department of Pharmaceutical Botany JU MC initiated biotechnological studies of this species. Its microshoots successfully grown in vitro produce considerable amounts of specific flavonoids (mostly baicalin and wogonoside) and verbascoside, and in future they can be an alternative rich source of these compounds for pharmaceutical and cosmetic applications.


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