Comparison of the Susceptibility of Quercus petraea, Q. robur and Q. rubra to Collybia fusipes

Collybia fusipes is the cause of a root rot of Quercus petraea (sessile oak), Q. robur (pedunculate oak) and Q. rubra (red oak). This parasite is often reported to cause problems in stands of red oaks and field evidence suggests that this North American oak species is more susceptible than the two European oak species. Young saplings of the three oak species and of chestnut, Castanea sativa, in the glasshouse, and also mature sessile and red oaks growing in the same stand were inoculated with C. fusipes to compare their susceptibility. Red oak, both as young seedlings and mature trees, was more susceptible to C. fusipes than sessile oak. Chestnut seedlings were as susceptible as sessile oak. Susceptibility of pedunculate oak seedlings was intermediate between red oak and sessile oak. In one experiment this species was significantly less susceptible than red oak, and in the other it was as susceptible.     

Characterizing the variation in aggressiveness and sporulation of the NA1 and EU1 lineages of Phytophthora ramorum in Oregon

Phytophthora ramorum, the cause of sudden oak death, is an invasive pathogen present in parts of coastal California and south-western Oregon forests. The majority of these forest infestations have been caused by the NA1 clonal lineage. In 2015, the EU1 lineage of P. ramorum was isolated from a tanoak (Notholithocarpus densiflorus) tree located in a mixed-conifer forest of Curry County, Oregon. In order to evaluate the threat to Oregon forests of the EU1 lineage relative to the established NA1 lineage, a series of experiments was conducted comparing aggressiveness and sporulation of NA1 and EU1 isolates on logs and seedlings in the growth chamber and forest. There was no significant difference in lesion size on logs inoculated with NA1 and EU1 isolates for any of the tree species tested. Across all seedling experiments differences among isolates within lineage, in terms of both aggressiveness and sporulation, were more commonly observed than differences among lineages. Site to site variation in tanoak sporulation, as measured by rain bucket baiting, appears to be correlated with the number of P. ramorum-positive seedlings detected at each site.     

Aerial spraying ofBacillus thuringiensis var.kurstaki for the control ofThaumetopoea processionea in Turkey oak woods

Thaumetopoea processionea L. is an important oak defoliator whose outbreaks have become increasingly frequent in Europe and thus have received a great deal of attention from foresters. Field trials were carried out in central Italy (Tuscany) to test the efficacy of aerial spraying ofBacillus thuringiensis var.kurstaki [Btk] in early spring for the control of processionary infestations inQuercus cerris L. woods. The experimental sites were located in a hilly area of 1640 ha witha maximum altitude of 593 m a.s.l. and a yearly mean temperature of 10°C. Turkey oak woods were divided into three areas to be treated with 31.75 BIU ha⁻¹ (2.5l ha⁻¹) on 578 ha, 44.45 BIU ha⁻¹ (3.5l ha⁻¹) on 306 ha and 57.15 BIU ha⁻¹ (4.5l ha⁻¹) on 756 ha, respectively. Five days after treatment, larval mortality was less than 40% in the control plot, but over 60% in the treated areas. Thirteen days after treatment, larval mortality varied from 75.05% to 96.42% in the three treated areas. Surveys conducted 2 months after the treatment showed a strong decline in the number of oak processionary nests in all the treated plots. It was possible to control the pest effectively in turkey oak woods usingBtk at 31.75 BIU ha⁻¹ distributed at ultra-low volume at the time of bud opening when nonurticating larvae were present. http://www.phytoparasitica.org posting March 12, 2008.     

Endophytic-Host Selectivity of Discula umbrinella on Quercus alba and Quercus rubra Characterized by Infection, Pathogenicity and Mycelial Compatibility

The fungal endophytic–host relationships of Discula umbrinella and two oak species, Quercus alba and Quercus rubra, were characterized on the basis of endophytic infection, pathogenicity, and mycelial compatibility. Isolates of D. umbrinella were cultured from leaves of Q. alba and Q. rubra collected from a hardwood forest located in Patuxent Wildlife Research Center in Laurel, Maryland, USA. Endophytic infection was observed on sterile leaf discs and living 2-month-old seedlings of Q. alba and Q. rubra. Fungal-host reciprocal inoculations revealed the presence of conidiomata on both hosts but conidiomata production was more abundant on Q. alba. Isolates from Q. rubra produced milder disease symptoms on both oak species. Mycelial compatibility studies identified seven different MCG groups. MCG groups 1–3 contained isolates from both oak species whereas MCG groups 4–7 contained host specific isolates. Field studies monitored the seasonal appearance of the sexual fruiting structures, perithecia, as a possible source of new genetic variation that might alter host specificity/pathogenicity of the D. umbrinella isolates on Q. alba and Q. rubra hosts. Only 1–2% of the leaves contained perithecia throughout the sampling period (April–September). Isolates collected from Q. alba differed from those collected from Q. rubra in endophytic infection, pathogenic response, and perithecia production. The results of this study suggest that the endophyte–host relationship is one of host selective preference for Q. alba, but that the endophyte has the ability to maintain the endophytic/pathogenic life cycle on the less preferred host species, Q. rubra.     

Bacterial community associated with the ambrosia beetle Platypus cylindrus on declining Quercus suber trees in the Alentejo region of Portugal

In Portugal, the oak pinhole borer Platypus cylindrus and its mycobiota have been associated with cork oak (Quercus suber) death, but no knowledge exists regarding the associated bacterial community. However, it is known that some bacteria are important for ambrosia beetle symbiosis and play a role in oak tree health. To explore the bacteria associated with this beetle and its host, with the ultimate goal of highlighting potential roles in oak decline, this study used a culture-dependent approach for strain isolation and phylogenetic identification using 16S rRNA gene sequencing and multilocus sequence analysis (MLSA). The bored galleries of different cork oak trees from a cork stand in Alentejo, together with the body and mycangia of adult beetles, were investigated. The samples revealed a diverse community comprising 500 isolates with 64 distinct types of bacterial colonies. Sixty-eight strains were selected for sequencing and used for phylogenetic analysis, 40 from wood galleries and 28 from beetles. Thirty-two genera of bacteria were identified, 18 of which were described for the first time within oak–beetle interactions. Major taxonomic groups were Actinobacteria in beetles and Enterobacterales in wood galleries. Although specific oak bacterial pathogens were not detected, a group of distinct strains detected in wood galleries, potentially belonging to a new Pectobacteriaceae species, were able to produce mild symptoms on cork oak plantlets. This study reports for the first time the biodiversity of culturable bacteria associated with the Q. suber–P. cylindrus interaction, their relevance to both organisms and the possible contribution to oak decline.     

Laboratory and field evaluation of spinosad against the gypsy moth,Lymantria dispar

The toxicity of the naturally derived insecticide spinosad was tested against the gypsy moth, Lymantria dispar. Bioassays using red oak leaf disks, treated with spinosad in a Potter spray tower, yielded an LC₅₀ value of 0.0015 µg AI cm⁻² (3‐day exposure; 13‐day evaluation; 2nd instar larvae). Applied to foliage to run‐off in the laboratory (potted red oak seedlings) and the field (4 m‐tall birch trees), spinosad effectively controlled 2nd instar larvae at concentrations ranging from 3 to 50 mg litre⁻¹. Toxicity in the laboratory, and efficacy and persistence in the field, were comparable to those achieved with the insecticide permethrin. Laboratory studies supported field observations that control was achieved in part by knockdown due to paralysis. In addition, laboratory results demonstrated that crawling contact activity may play an important role in field efficacy; 50% of treated larvae were paralyzed 16 h after a 2‐min crawling exposure to glass coated with a 4 mg litre⁻¹ spinosad solution. © 2000 Society of Chemical Industry     

Fitness characteristics of the European lineages of Phytophthora ramorum

As an introduced pathogen, Phytophthora ramorum exists as four near-clonal evolutionary lineages, of which only EU1 and EU2 are established in the UK. EU1 has become widespread since the first findings in 2002 whereas EU2, detected in 2011, has a more limited distribution. Both lineages are epidemic in plantation-grown larch, sporulating asexually on needles, but also causing heavy dieback and mortality. To understand whether EU1 and EU2 pose different threats to forest health, we compared their growth characteristics on agar, pathogenicity on several hosts, and sporulation on Japanese larch needles. When pathogenicity was evaluated by measuring colonization at 20 °C in mature bark (phloem) of Japanese and European larch (Larix kaempferi and L. decidua), English oak (Quercus robur), and beech (Fagus sylvatica), Japanese larch was the most susceptible and oak the least susceptible. On average, EU2 isolates produced significantly larger lesions than EU1 isolates in Japanese larch and oak although not in the other hosts. With tests using young saplings of Japanese and European larch, damaging bark lesions formed at both 10 °C and 20 °C, but EU2 was significantly more pathogenic at 20 °C on both hosts compared with EU1. In contrast, both lineages caused similar amounts of necrosis on inoculated leaves of rhododendron (Rhododendron ponticum). Moreover, EU2 isolates usually sporulated less abundantly on larch needles compared with EU1 isolates, suggesting a trade-off in pathogenicity and sporulation between lineages. As EU2 tends to have smaller sporangia than EU1, this could also reduce the inoculum potential of EU2.     

Calcium fertilizers induce soil suppressiveness to <Emphasis Type="BoldItalic">Phytophthora cinnamomi</Emphasis> root rot of <Emphasis Type="BoldItalic">Quercus ilex</Emphasis>

Based on the observation that the root disease caused by P. cinnamomi on Q. ilex has a low incidence and severity in soils with medium-high Ca²⁺ content, we studied the ability of Ca²⁺ fertilizers to induce soil suppressiveness to the pathogen. Studies on cultures of P. cinnamomi exposed to different Ca²⁺ fertilizers in vitro showed significant inhibition of sporangial, chlamydospore and zoospore production at millimolar concentrations while mycelial growth was mainly unaffected. Experiments performed with artificially infested soil showed that some Ca²⁺ fertilizers induce a significant decrease on chlamydospore viability. Additionally, greenhouse experiments using artificially infested soils showed a significant reduction of foliar and root symptom severities in Holm oak seedlings growing in soils amended with Ca²⁺ fertilizers. We suggest that limestone amendments in oak rangelands could enhance the suppressiveness of soils to P. cinnamomi, and it is likely that the inhibition of sporangial production was the main mechanism involved.     

Aerial Application of Bacillus thuringiensis (Dipel) against Colotois (Himera) pennaria L. in Comparison with ULV-Application of Various Insecticides 1)

In spring 1972 experiments were carried out in Lower Austria with the Bacillus thuringiensis isolate Dipel in order to assess its effectiveness in controlling Colotois pennaria L. infesting oak and other broad-leaved trees. The results obtained compared favourably with those produced by the insecticides Nexion²), malathion and Dipterex³), although both malathion and Dipterex yielded higher mortality. Similar results were obtained when B. thuringiensis was used to control Stilpnotia salicis L., feeding on poplar trees. Neodiprion sertifer Geoffr. and Pris-tiphora abietina Christ., however, were not affected.     

Sensitivity of European Plasmopara viticola populations to cymoxanil

Between 1995 and 1997, 278 grape downy mildew (Plasmopara viticola) populations originating from European vineyards were characterised for their sensitivity to cymoxanil in a leaf-disc assay. The sensitivity profile revealed a wide distribution, with minimum inhibitory concentrations (MIC) ranging from 10 to more than 800 mg litre⁻¹. EC₅₀ values ranged from 1 to more than 800 mg litre⁻¹ with an average of 125 mg litre⁻¹. The sensitivity distribution was stable between 1995 and 1997. Surprisingly, populations from Portugal appeared significantly more sensitive than those from France or Italy, which could not be linked to differential cymoxanil usage in these countries. P viticola populations collected outside Europe and never exposed to cymoxanil appeared significantly more sensitive than exposed European populations, with an average EC₅₀ value of 10 mg litre⁻¹. The level of sensitivity of European P viticola populations was relatively unaffected by the number of cymoxanil applications made during a season or by the number of years of cymoxanil use. No link was found between the level of sensitivity in the leaf-disc assay and the level of performance of the cymoxanil mixtures used in the fields where the populations originated. Specific field trials conducted in Italy and Portugal have shown that the performance of cymoxanil-based mixtures remained good even on populations of the grape downy mildew fungus characterised as less sensitive in the leaf-disc assay. While there are no baseline sensitivity data for pre-commercialisation P viticola populations, the results of our study suggest that a shift in sensitivity (12.5-fold) may have occurred in some areas since introduction of cymoxanil on grapes nearly 20 years ago. Because cymoxanil is never used alone, it is difficult to determine whether or not practical resistance is occurring in European vineyards. © 1999 Society of Chemical Industry     

Microsatellite and mating type markers reveal unexpected patterns of genetic diversity in the pine root‐infecting fungus Grosmannia alacris

Grosmannia alacris is a fungus commonly associated with root‐infesting bark beetles occurring on Pinus spp. The fungus has been recorded in South Africa, the USA, France, Portugal and Spain and importantly, has been associated with pine root diseases in South Africa and the USA. Nothing is known regarding the population genetics or origin of G. alacris, although its association with root‐infesting beetles native to Europe suggests that it is an invasive alien in South Africa. In this study, microsatellite markers together with newly developed mating type markers were used to characterize a total of 170 isolates of G. alacris from South Africa and the USA. The results showed that the genotypic diversity of the South African population of G. alacris was very high when compared to the USA populations. Two mating types were also present in South African isolates and the MAT1‐1/MAT1‐2 ratio did not differ from 1:1 (χ² = 1·39, P = 0·24). This suggests that sexual reproduction most probably occurs in the fungus in South Africa, although a sexual state has never been seen in nature. In contrast, the large collection of USA isolates harboured only a single mating type. The results suggest that multiple introductions, followed by random mating, have influenced the population structure in South Africa. In contrast, limited introductions of probably a single mating type (MAT1‐2) may best explain the clonality of USA populations.     

From horticulture and biofuel to invasion: the spread of Miscanthus taxa in the USA and Europe

Species of the genus Miscanthus (Poaceae) have recently become widely planted for ornamental and industrial uses. Yet these large perennial grasses have several traits that suggest a high invasion potential. To inform the debate on the benefits and risks of introducing these new biofuel crops, this publication summarises the invasion of Miscanthus species in Europe and the USA. Databases from North America were interrogated, and occurrence data from a variety of European sources were examined. In both continents, M. sinensis and M. sacchariflorus have recently spread rapidly, although invasion started earlier in the USA and both species are more widespread there. Both species mostly colonise grasslands and ruderal habitats. In addition, some escapes of the hybrid M. × giganteus, which has a particularly strong invasion potential because of its vigorous growth and greater capacity to establish in cool climates, have already been recorded. We conclude that the invasion potential of Miscanthus species needs to be thoroughly tested, the incipient invasions in Europe and the USA monitored and, if required, planting restrictions should be adopted.     

Sporulation potential,symptom expression and detection of Phytophthora ramorum on larch needles and other foliar hosts

Phytophthora ramorum has caused extensive dieback and mortality of commercially grown Japanese larch (Larix kaempferi) in many parts of the UK, as infected foliage generates spores that then cause bark lesions and girdling cankers on trees. Following inoculation, individual needles of Japanese, European (L. decidua) and hybrid (L. × eurolepis) larch infected with P. ramorum can produce thousands of sporangia. Mean numbers of sporangia ranged from 806 to 1778 per cm² (hybrid larch and Japanese larch, respectively), surpassing mean sporulation levels on foliar hosts previously associated with P. ramorum outbreaks in Britain, namely Rhododendron ponticum, Castanea sativa and Vaccinium myrtillus. Sporulation on larch even exceeded that of California bay laurel (Umbellularia californica), which drives the sudden oak death epidemic in California. Inoculation of foliage selected at different times of year revealed that foliage age significantly affected sporulation levels, but this varied with host species. However, symptom development and sporulation were often not correlated. Symptoms on larch were frequently insignificant or even absent at certain times of year, with sometimes the only evidence of infection being the emergence of sporangia from needles, without any sign of discolouration or necrosis. Plating infected but symptomless needles onto Phytophthora selective medium also often failed to yield the pathogen. Symptomless infection of larch needles apparently occurs, but is only detectable with microscopy. More generally, it is suggested that diagnosis of Phytophthora infection in conifers is often underestimated due to isolation difficulties and delayed symptom expression.     

Population genetic structure of Raffaelea quercus-mongolicae indicates a recent fungal introduction event to Jeju Island from inland areas of South Korea

Raffaelea quercus-mongolicae, a mycangial symbiont of a wood-boring ambrosia beetle, Platypus koryoensis (Coleoptera: Curculionidae: Platypodidae), has been associated with the mass mortality of Korean oak trees in South Korea. Systemic wilt from the inoculation of oak trees with R. quercus-mongolicae has caused significant damage to forest ecosystems in the country. Despite the continued spread of this fungus across the country since the first discovery of the disease, little is known regarding its biology. During a recent survey carried out to investigate the status of oak wilt diseases on Jeju Island, South Korea, visible signs of bark beetle attack, including bark beetle frass and galleries, were consistently observed in Quercus serrata, and R. quercus-mongolicae was successfully recovered from the trees. To determine whether the pathogen was introduced from inland areas of the country to Jeju Island, the genetic diversity and population structure of R. quercus-mongolicae isolates collected from various geographical regions and hosts in the country were investigated using a combination of eight microsatellite markers designed from the genome of the fungus in this study. The results showed lower levels of genetic diversity for the population from Jeju Island compared to those from inland areas of the country. This suggests that a limited number of individuals were recently introduced to Jeju Island, resulting in a new disease problem in the area.     

Distribution and activity of spray deposits in an oak canopy following aerial application of diluted and undiluted formulations of Bacillus thuringiensis Berliner against the gypsy moth,Lymantria dispar L. (Lepidoptera: Lymantriidae)

The distribution and biological activity of spray deposits resulting from aerial applications of diluted and undiluted Bacillus thuringiensis, ‘Dipel 64AF’ against the gypsy moth, Lymantria dispar L., were examined in oak stands in south-eastern Ontario, Canada. The sprays were applied by fixed-wing aircraft equipped with four ‘Micronair AU4000’ atomizers. Application of diluted formulation at 30 BIU ha^?1 in 6.0–6.4 litre generally resulted in a higher droplet density (10–28 cm ^?2 leaf) than application of undiluted product at the same dosage rate in 1.8 litre ha^?1 (4–10 cm ^?2). However, spray deposits of undiluted product with a volume median diameter (D_v.5) of 90–130 μm caused as much mortality of gypsy moth larvae in bioassays of sprayed foliage as deposits of diluted product with a D_v.5 of 150–350 μm despite a two- to three-fold reduction in droplet density. Our data suggest that by using fine spray atomization, undiluted application of these formulations can offer the same efficacy against gypsy moth as coarsely atomized sprays of diluted product.     

A multiplex real-time PCR assay enables simultaneous rapid detection and quantification of bacteria associated with acute oak decline

Acute oak decline (AOD) is a syndrome affecting mature oak trees and is characterized by stem bleeds from vertical fissures on trunks, and inner bark necrosis caused by a polybacterial consortium, in which Gibbsiella quercinecans and Brenneria goodwinii, and to a lesser extent Rahnella victoriana and Lonsdalea britannica, play key roles. Here we report a novel multiplex real-time PCR assay that enables simultaneous and rapid detection and quantification of these four bacterial species from stem bleed swabs. Experiments with axenic cultures were performed to determine specificity and sensitivity of the multiplex quantitative PCR (qPCR). Whilst the primer/probe set for B. goodwinii was species-specific, primer/probe sets for the other three species were able to identify other members of their respective genera. There was no cross detection of genera within the multiplex qPCR, and non-target bacteria were not detected. The multiplex AOD assay had differential sensitivity for each bacterial species. The assay was evaluated on swab samples collected from stem bleeds of declining oak trees at a site in south-east England and was able to detect all four bacterial species. Absolute quantification of the bacteria from swab samples was possible through the inclusion of a standard curve prepared from dilutions of gene copy standards. This diagnostic tool will facilitate rapid detection of AOD-associated bacteria from samples that can easily be taken by non-specialists without specific training, and will also find application in other experimental work such as pathogenicity and control trials.     

Occurrence of bacterial black shoot disease of European pear in Yamagata Prefecture

Black lesions on shoots of European pear trees observed in an orchard in Yamagata Prefecture in May 2007 were suspected to be caused by a bacterial pathogen. The surface of the colonies isolated on a high sucrose medium did not have the crater morphology that is characteristic of E. amylovora bvs. 1–3, and a specific DNA fragment was amplified from the isolates in the PCR using the EprpoD primer set. The partial sequences of the 16S rRNA gene placed the isolates in the genus Erwinia. The isolates differed serologically from E. amylovora biovars and E. pyrifoliae in an Ouchterlony double-diffusion test although their bacterial properties suggested that they are closely related to E. amylovora biovars and E. pyrifoliae. In a DNA–DNA hybridization test, the relatedness between the isolates and E. amylovora biovars or E. pyrifoliae did not exceed 70% level, indicating that they are independent species. Thus, the isolates belongs to the genus Erwnia but are not E. amylovra or E. pyrifoliae. After succulent pear shoots were injected with bacterial suspensions (10⁹, 10⁸, 10⁷ and 10⁶ cfu/ml) of the isolates, lesions formed with 10⁹ and 10⁸ cfu/ml, but the disease incidence with 10⁸ cfu/ml was much lower than with E. amylovora and E. pyrifoliae. Virulence of the present isolates is thus thought to be very weak. On the basis of these results, we consider that this is a new shoot disease of European pear. In the 2007 season, all affected trees were pulled out after harvest. No symptoms have been observed in field surveys since the fruitlet season in 2007.     

Vegetative Compatibility Groups in Verticillium dahliae Isolates from the Netherlands as Compared to VCG Diversity in Europe and in the USA

In a study of vegetative compatibility in Verticillium dahliae in the Netherlands, a collection of 45 isolates including representatives from woody hosts, several horticultural crops and from the soil of potato fields was examined. In addition an effort was made to compare vegetative compatibility groups (VCGs) from different countries. The results of this study indicate that VCG diversity in V. dahliae in the Netherlands is limited. Only two VCGs were detected: VCG NL-I and VCG NL-II. The former is the predominant VCG for isolates from tree hosts. However, Verticillium wilt in trees can be caused by isolates from both VCGs. It is suggested that the predominance of VCG NL-I in tree hosts is the result of the origin of the tree and the cropping history of its growing site, rather than trees being preferential hosts for isolates from this VCG. Comparison of VCG testers from the Netherlands, from several other European countries and from the USA show that in Europe two major VCGs are present. The first one, including NL-I, is compatible with USA VCG 3 and VCG 4, whereas the second one, including NL-II, is compatible with USA VCG 1 and VCG 2. These groups are not completely separated; in some cases, testers formed heterokaryons with VCG testers from both main groups. Because of the presence of these bridge isolates and because mutants from the same isolate differ in ability to form heterokaryons, it is emphasised that careful selection of isolate testers is an essential step to get a clear picture of VCG diversity.     

Red Oak Borer, Enaphalodes rufulus (Coleoptera: Cerambycidae), in the Ozark Mountains of Arkansas, U.S.A.: An Unexpected and Remarkable Forest Disturbance

A complex interaction of multiple factors has resulted in an oak decline event in oak-hickory forests of the Ozark Mountains of Arkansas and Missouri, U.S.A. The most striking feature of this situation is an unprecedented population explosion of red oak borer, a species of cerambycid beetle, Enaphalodes rufulus (Haldeman), which appears to be causing extensive mortality to mature red oaks (Quercus, subgenus Erythrobalanus). The insect is a native species, historically a minor pest of oaks, found throughout the eastern United States. Beetles normally reproduce in living oaks, as larvae initially feed in phloem tissue and subsequently bore into xylem where pupation occurs. The life cycle is two years in length and synchronous adult emergence occurs in odd-numbered years. Data from previous research indicate average attack densities of less than four per tree with a high of 71 on a single tree. Historical emergence densities are similarly low and the highest reported was 15 adults from one tree. Our research is concerned with understanding factors contributing to this outbreak; developing sampling methods for red oak borer; assessing oak mortality; and evaluating site and stand conditions associated with the current outbreak. Results of our initial sampling reveal dramatically higher average attack densities of 244 per m2 and emergence densities of 18 per m2 of bark surface area. We confirm a three-week period of adult emergence and activity during mid-June to early July. We also report on possible management responses by federal and state agencies to this remarkable epidemic and oak mortality crisis.