Pathogenicity of Phytophthora multivora to Eucalyptus gomphocephala and Eucalyptus marginata

Phytophthora multivora is associated with the rhizosphere of declining Eucalyptus gomphocephala, Eucalyptus marginata and Agonis flexuosa. Two pathogenicity experiments were conducted. The first experiment examined the pathogenicity of five P. multivora isolates and one Phytophthora cinnamomi isolate on the root systems of E. gomphocephala and one P. multivora isolate on the root system of E. marginata. In the second experiment, the pathogenicity of P. multivora to E. gomphocephala and E. marginata saplings was measured using under‐bark stem inoculation. In Experiment 1, the P. cinnamomi isolate was more aggressive than all P. multivora isolates causing significant loss of fine roots and plant death. Two P. multivora isolates and the P. cinnamomi isolate caused significant losses of E. gomphocephala fine roots 0–2 mm in diameter and significantly reduced the surface area of roots 0–1 mm in diameter. One P. multivora and the P. cinnamomi isolate significantly reduced the surface area of roots 1–2 mm in diameter. Two of the P. multivora isolates significantly reduced the number of E. gomphocephala root tips. In E. marginata, the length and surface area of roots 0–1 mm in diameter and number of root tips were significantly reduced by P. multivora infestation. Rhizosphere infestation with the P. multivora isolates and P. cinnamomi isolate on E. gomphocephala, and one P. multivora isolate on E. marginata, did not significantly influence the foliar nutrient concentrations. In Experiment 2, under‐bark inoculation with P. multivora caused significant lesion extension in E. gomphocephala and E. marginata saplings, compared to the control. We propose that P. multivora is inciting E. gomphocephala and E. marginata decline by causing fine root loss and subsequently interfering with nutrient cycling throughout the plant. The impact of fine root loss on the physiology of plants in sites infested with P. multivora requires further research.     

Susceptibility of Eucalyptus grandis and Acacia mearnsii seedlings to five Phytophthora species common in South African plantations

Eucalyptus grandis and its hybrids, as well as Acacia mearnsii, are important non‐native trees commonly propagated for forestry purposes in South Africa. In this study, we conducted pathogenicity trials to assess the relative importance of five commonly isolated Phytophthora spp. (Phytophthora alticola, P. cinnamomi, P. frigida, P. multivora and P. nicotianae) from the plantation environment on E. grandis and A. mearnsii seedlings. Overall E. grandis was more susceptible to the tested Phytophthora spp. than A. mearnsii. Phytophthora cinnamomi was the only pathogen that had a significant negative effect on both the host tree species, leading to a reduction in root and shoot weight as well as to death in the case of E. grandis. Phytophthora alticola and P. nicotianae exclusively affected E. grandis and A. mearnsii, respectively. This study updated the current knowledge on the pathogenicity of Phytophthora spp. on two important non‐native commercially propagated tree species from South Africa.     

Evaluation of Juglans species for resistance to Phytophthora cinnamomi: differences in isolate virulence and response to fosetyl‐Al

Phytophthora is considered as an important pathogen on walnut, and severe losses are reported in European as well as in American walnut stands. Though several Phytophthora spp. are known to attack walnut, P. cinnamomi is considered the most virulent and widespread in southern Europe. Up to now, no walnut species or hybrid is known to have a high resistance level towards P. cinnamomi. Efforts are addressed in finding rootstock material graft compatible with English walnut and resistant/tolerant to P. cinnamomi. The extension of P. cinnamomi lesions on five Juglans species was studied to find out sources of resistance/tolerance to this pathogen. Walnut species clustered into two main groups, J. hindsii, J. nigra, and J. mandshurica were the less susceptible to the colonization of P. cinnamomi, while J. regia and J. sieboldiana were the most susceptible. On this account, J. mandshurica represents the best alternative as rootstock because its employment overcomes the risk of the occurrence of black line disease, it has good level of resistance to Agrobacterium temefaciens and Brenneria nigrifluens, and it is tolerant to Xanthomonas arboricola pv. juglandis. J. mandshurica is also compatible in cross‐pollinations with J. regia and J. nigra. Differences in virulence of P. cinnamomi isolates was assessed and a marked interaction between species and isolate emerged. Treatment with fosetyl‐Al by dipping was mainly efficient in reducing the length P. cinnamomi lesions, and an interaction between species and treatment was evident with the highest efficacy on J. regia and J. sieboldiana.     

Influence of temperature on germination of Quercus ilex in Phytophthora cinnamomi,P. gonapodyides,P. quercina and P. psychrophila infested soils

The influence of temperature on germination of Quercus ilex acorns in Phytophthora infested soils was quantified for the first time. Radicle damage and mortality of Q. ilex seeds germinating at 17, 20, 23 and 26°C in Phytophthora cinnamomi, P. gonapodyides, P. quercina and P. psychrophila infested soils were assessed and related to in vitro mycelium growth of the same isolates of the pathogens. The optimum growth temperatures of isolates of P. cinnamomi, P. gonapodyides, P. quercina and P. psychrophila were 20–23, 23–26, 20–23 and 20°C, respectively. At 17 and 20°C, all four Phytophthora species caused 100% acorn mortality, whereas at 26°C, acorn mortality was 100, 10, 25 and 0% in P. cinnamomi, P. gonapodyides, P. quercina and P. psychrophila infested soils, respectively. At 23°C, P. cinnamomi and P. gonapodyides reduced acorn radicle length more than P. quercina and P. psychrophila, whereas at 26°C, only P. cinnamomi caused further reduction in radicle length. The higher susceptibility of germinating acorns in comparison to seedlings reported in the literature indicates age‐related susceptibility of Q. ilex to Phytophthora. The seedling/pathogen growth ratio was inversely related to the reduction in radicle length at different temperatures ( = 0.84, p < 0.0001), suggesting that rapid germination may allow seedlings to escape from infection. Increasing temperatures had different effects on damage to acorns depending on the pathogen present in the soil, indicating that Phytophthora species × temperature interactions determined Q. ilex germination. The effects of temperature on the impacts of Phytophthora species based on climate change predictions for Mediterranean countries are discussed.     

Involvement of Phytophthora species in white oak (Quercus alba) decline in southern Ohio

This study was initiated to investigate the possible role of Phytophthora species in white oak decline (Quercus alba) in southern Ohio at Scioto Trail State Forest. Surveys demonstrated the presence of four species of Phytophthora including one novel species. By far, the most common species was P. cinnamomi; P. citricola and P. cambivora were isolated infrequently. In few instances, P. cinnamomi was isolated from fine roots and necroses on larger roots. No special pattern of incidence was found, but P. cinnamomi was more commonly isolated from greater Integrated Moisture Index values suggesting moist lower bottomlands favour this Phytophthora species. When tree crown condition was examined relative to the presence of Phytophthora, no significant association was found. However, roots of declining P. cinnamomi‐infested trees had 2.5 times less fine roots than non‐infested and healthy trees, which was significantly different. The population densities of P. cinnamomi from declining trees were significantly greater than from healthy trees, suggesting increased pathogen activity that has the potential to cause dieback and decline and possibly the cause of a reduced fine root amount found on declining trees.     

Rapid diagnosis of pathogenic Phytophthora species in soil by real‐time PCR

Real‐time PCR assays based on the TaqMan system and using ITS sequences were developed for the identification of Phytophthora species, including P. cactorum, P. megasperma, P. plurivora, P. pseudosyringae and P. quercina, all of which are currently causing significant damage to roots of forest trees in both managed stands and natural ecosystems. Total genomic DNA was extracted from mycelia of aforementioned Phytophthora isolates. Species‐specific primers for P. cactorum, P. megasperma, P. plurivora, P. pseudosyringae and P. quercina were designed based on ITS sequences of rDNA. The amplification efficiency of target DNA varied from 93.1% (P. pseudosyringae) to 106.8% (P. quercina). The limit of the detection was calculated as 100 – 1,000 fg DNA, depending on the Phytophthora species. In mixed soil samples, all Phytophthora species were detected for Ct values shifted by 0.7 – 2.1 cycles. Based on these real‐time PCR assays we were able to identify the five Phytophthora species. These techniques will be of value in the identification of these pathogens, which may cause up to 80 – 90% fine root loss in oak stands.     

Pathogenicity to alder of Phytophthora species from riparian ecosystems in western Oregon

Described as one of the most destructive pathogens of agricultural crops and forest trees, Phytophthora is a genus of microorganisms containing over 100 known species. Phytophthora alni has caused collar and root disease in alders throughout Europe, and a subspecies has recently been isolated in North America. Reports of canopy dieback in red alder, Alnus rubra, prompted a survey of their overall health and to determine whether P. alni was present in western Oregon riparian ecosystems. Over 1100 Phytophthora isolates were recovered, representing 20 species and 2 taxa. Phytophthora‐type cankers were observed in many trees, and their incidence was positively correlated with canopy dieback. High levels of mortality for red alder were not observed, which suggests these Phytophthora species are not aggressive pathogens. To test this hypothesis, three stem wound inoculations and one root dip were conducted on red alder seedlings using 13 Phytophthora species recovered from the riparian survey. Ten of the 13 Phytophthora species produced significant lesions in at least one pathogenicity test. Phytophthora siskiyouensis produced the largest lesions on red alder from the two stem wound inoculation tests conducted under summer conditions, while P. taxon Pgchlamydo caused the largest lesions during the winter stem wound inoculation test. Phytophthora gonapodyides, P. taxon Pgchlamydo and P. siskiyouensis have previously been found associated with necrotic alder roots and bole cankers in the field, and with the pathogenicity results reported here, we have established these species as causes of Phytophthora root disease and Phytophthora bole canker of alder in Oregon. While none of the Phytophthora species were especially aggressive towards red alder in the pathogenicity tests, they did cause localized disease symptoms. By weakening the root systems or boles of alders, the Phytophthoras could be leaving alders more susceptible to other insects and pathogens.     

Involvement of Phytophthora spp. in chestnut decline in the Black Sea region of Turkey

Chestnut blight caused by Cryphonectria parasitica is a serious disease of Castanea sativa in the Black Sea region of Turkey. During disease surveys, dieback and decline symptoms were observed on trees without apparent blight and ink disease symptoms. Black necroses, similar to those caused by Phytophthora infections, were noted on some of the chestnut coppices and saplings in one nursery in Ordu and led to an investigation into this disease complex. Only symptomatic plants showing dieback symptoms were investigated. Soil samples together with fine roots were collected from two directions, north and north‐east, approximately 150 cm away from the main stems. Phytophthora spp. were baited with young chestnut leaves. Three Phytophthora spp., P. cambivora, P. cinnamomi and P. plurivora, were identified from 12 soil samples collected from 73 locations, while from the nurseries, only P. cinnamomi was obtained. Phytophthora cinnamomi was the most common species, obtained from seven locations in five provinces and from four nurseries having similar symptoms mentioned above in different locations. Phytophthora cambivora and P. plurivora were less frequently obtained, from three to two stands, respectively. Phytophthora cinnamomi and P. cambivora were the most aggressive species when inoculated at the stem base on 3‐year‐old chestnut saplings, killing six saplings of eight inoculated in 2 months. The three Phytophthora species were first recorded on chestnut in Black sea region of Turkey with the limited samples investigated in a large area about 150 000 ha chestnut forest.     

Occurrence of Phytophthora species in riparian stands of black alder (Alnus glutinosa) in Slovakia

During the monitoring period of alder decline phenomenon in Slovakia, indicative symptoms of Phytophthora diseases were observed in riverside stands in Slovakia. The study aimed to test the presence and diversity of Phytophthora species in declining alder stands. The samples were collected from six stands situated by rivers/streams in the central and eastern parts of Slovakia. Phytophthora plurivora and P. cactorum were detected in soil, root and water samples. The most isolated species was P. plurivora. Both Phytophthora species have been recognized in the literature as the perpetrators of black alder dieback together with other species, such as P. alni or P. polonica. To our knowledge, this is the first report of P. plurivora and P. cactorum in riverside stands with the main share of black alder in Slovakia.     

First records of soilborne Phytophthora species in Swedish oak forests

Thirty‐two oak stands in southern Sweden, 27 with predominantly declining trees and five with a higher proportion of healthy trees were investigated regarding the presence of soilborne Phytophthora species. Phytophthora quercina, an oak‐specific fine root pathogen, was isolated from rhizosphere soil samples in 10 of the 27 declining stands. Additionally, P. cactorum and P. cambivora were recovered from one stand each. No Phytophthora species were isolated from the healthy oak stands. The soil conditions at the sites from which Phytophthora spp. were recovered ranged from mesic sediments to moraines, with clayey to silty textures and with soil pH (BaCl₂) between 3.5 and 5.0. The results show that P. quercina is geographically widespread in oak stands in southern Sweden and indicate that this pathogen may be one of the factors involved in oak decline in Northern Europe as has already been shown for western, Central and parts of southern Europe.     

Diplodia corticola and Phytophthora cinnamomi: the main pathogens involved in holm oak decline on Caprera Island (Italy)

Since 2008, severe and widespread tree decline and mortality has been observed at the main growing Quercus ilex L. (holm oak) forest on Caprera Island, Italy. To clarify the symptomatology and aetiology of this phenomenon, field surveys and isolations from symptomatic trees were carried out in summer 2010. Affected trees exhibited crown thinning, branch dieback, sunken cankers, epicormic shoots, exudates on branches and trunk, root losses and sudden death symptoms. Four fungal species belonging to Botryosphaeriaceae family, namely Botryosphaeria dothidea, Diplodia corticola, D. seriata and Neofusicoccum parvum, were isolated from cankers on trunk and branches, whereas three species of Phytophthora, namely P. cinnamomi, P. cryptogea and P. gonapodyides, were isolated from fine roots and rhizosphere soil samples. Isolates were identified using both morphological analysis and DNA‐based techniques. Pathogenicity trials on holm oak seedlings showed that all the isolated species are pathogenic. D. corticola proved to be the most aggressive species. Our results provide the first evidence for a combined involvement of D. corticola and P. cinnamomi in the aetiology of holm oak decline in Italy and suggest that these pathogens are not only important contributing factors in the onset of long‐term tree decline, but also may cause the rapid devastation of extensive oak ecosystems.     

Relative susceptibility of oaks to seven species of Phytophthora isolated from oak forest soils

Isolates of Phytophthora cambivora, P. cinnamomi, P. citricola, P. europaea, P. quercetorum and two unidentified species were tested for their pathogenicity to eastern US oak species by root and stem inoculations. Experiments were conducted during two different periods and included 1‐, 2‐ and 20‐year‐old oaks grown under greenhouse and field conditions. Species of Phytophthora were pathogenic in varying degrees to the oak species tested. All species were pathogenic to fine and taproots of at least one oak species. The fine root damage caused by the species of Phytophthora ranged from 9 to 55% when compared to the controls. Roots were more susceptible during the fall inoculation period than the summer. With exception of Phytophthora sp1 and P. quercina‐like, all species of Phytophthora were pathogenic to oak stems with P. cinnamomi and P. citricola being the most aggressive. Quercus montana and Q. rubra were the most susceptible oak species to stem inoculation. Lesion sizes were considerably larger when 20‐year‐old trees were inoculated. Generally, no significant differences in lesion sizes were detected in greenhouse tests when the summer and fall inoculation periods were compared. However, on 2‐year‐old field‐grown seedlings, lesion sizes were considerably smaller or not significantly different from controls during the fall inoculation period, suggesting lower, late season temperatures may restrict lesion development.     

A severe canker disease of Corymbia ficifolia caused by Quambalaria coyrecup in native and urban forests of Western Australia

A serious canker disease is affecting the health of Corymbia ficifolia in urban areas of Perth, Western Australia. Quambalaria coyrecup was frequently associated with diseased trees and its presence confirmed by morphological characters and DNA sequencing. A number of Quambalaria species have previously been identified as causal agents of canker and shoot blight diseases on a range of Eucalyptus and Corymbia species. It was therefore the aim, using glasshouse studies, to determine whether Q. coyrecup was the primary causal agent of the cankers on C. ficifolia and whether other Quambalaria species could also be associated with the disease. All seedlings inoculated with Q. coyrecup produced canker symptoms within 1 month after inoculation. Canker lesions typical of those observed in the field also occurred in the four subsequent months, after which time the trial was terminated. Inoculation with Q. cyanescens and Q. pitereka (isolated from C. ficifolia shoots) did not result in lesion development. This study establishes that Q. coyrecup is a serious pathogen of C. ficifolia and that wounding is required for canker development to occur. The management of Quambalaria canker must therefore include the minimization of artificial wounding of trees in the nursery and field.     

Pathogenicity of Phytophthora species to Pacific Northwest Conifers

Phytophthora root rot is described for the first time killing sugar pine (Pinus lambertiana) in a seed orchard and four species of true fir (Abies spp.) in a forest nursery. P. cactorum was recovered from true firs and P. megasperma was recovered from sugar pine. P. cryptogea was recovered from sugar pine and true fir but isolates from the two locations differed from each other in pathogenicity and colony appearance. Isolates recovered from these hosts and isolates of 6 Phytophthora species previously recovered from Douglas-fir (Pseudotsuga menziesii) were then tested for pathogenicity on seedlings of 9 Northwest conifers. P. megasperma Group 1, P. cryptogea, and P. cinnamomi were pathogenic to all tree species except western redcedar (Thujaplicata). Western hemlock (Tsuga heterophylla) and true firs were susceptible to most species tested; ponderosa (P. ponderosa) and sugar pines were damaged only by P. cryptogea and P. cinnamomi; western redcedar was resistant to all isolates.     

Identification of Phytophthora species baited and isolated from forest soil and streams in northwestern Yunnan province,China

Phytophthora species were surveyed by collecting soil samples and placing bait leaves in selected streams during June–October in the years 2005, 2006 and 2010 at three sites in oak forests in Diqing Tibetan Autonomous Prefecture of NW Yunnan province, China. Seventy‐three isolates of Phytophthora spp. were recovered from 135 baited leaf samples and 81 soil samples. Eight Phytophthora species were identified by observation of morphological features and ITS1‐5.8S‐ITS2 rDNA sequence analysis. The eight taxa included two well‐known species P. gonapodyides and P. cryptogea, two recently described species P. gregata and P. plurivora, two named but as yet undescribed taxa, P. taxon PgChlamydo and P. taxon Salixsoil, and two previously unrecognized species, Phytophthora sp.1 and P. sp.2. The most numerous species, P. taxon PgChlamydo, and the second most abundant species, P. taxon Salixsoil, were recovered at all three sites. Phytophthora cryptogea was detected only once at site Nixi. Phytophthora gregata and P. sp.2 were isolated from a stream only at site Bitahai, while the other three species were each found at two sites. Phylogenetic analysis revealed that the isolates belonged to three ITS clades, one species including six isolates in clade 2, six species including 66 isolates in clade 6 and one species in clade 8. There was a relatively rich species and genetic diversity of Phytophthora detected in the investigated regions where the forest biotic and abiotic factors affecting the growth and evolution of Phytophthora populations were diverse.     

Phytophthora root and collar rot of mature Fraxinus excelsior in forest stands in Poland and Denmark

In recent years, Common ash (Fraxinus excelsior) throughout Europe has been severely impacted by a leaf and twig dieback caused by the hyphomycete Chalara fraxinea. The reasons for its current devastating outbreak, however, still remain unclear. Here, we report the presence of four Phytophthora taxa in declining ash stands in Poland and Denmark. Phytophthora cactorum, Phytophthora plurivora, Phytophthora taxon salixsoil and Phytophthora gonapodyides were isolated from rhizosphere soil samples and necrotic bark lesions on stems and roots of mature declining ash trees in four stands. The first three species proved to be aggressive to abscised roots, twigs and leaves of F. excelsior in inoculation experiments. Soil infestation tests also confirmed their pathogenicity towards fine and feeder roots of ash seedlings. Our results provide first evidence for an involvement of Phytophthora species as a contributing factor in current decline phenomena of F. excelsior across Europe. Specifically, they may act as a predisposing factor for trees subsequently infected by C. fraxinea. Phytophthora species from ash stands also proved to be aggressive towards a wide range of tree and shrub species commonly associated with F. excelsior in mixed stands. Although damage varied considerably depending on the Phytophthora species/isolate–host plant combination, these results show that many woody species may be a potential source for survival and inoculum build‐up of soilborne Phytophthora spp. in ash stands and forest ecosystems in general.     

Phytophthora species associated with dieback of sweet chestnut in Western Turkey

Sweet chestnut (Castanea sativa) is an important tree species in the Marmara and Aegean regions of Turkey as these two regions produce the great majority of edible nuts, especially those used for marron glacé production. Chestnut forests and orchards in these regions showing severe dieback symptoms not associated with chestnut blight were investigated to determine the role of Phytophthora spp. in the decline syndrome. Soil samples were collected from around 108 symptomatic chestnut trees at 29 sites and Phytophthora spp. isolated using soil baiting technique and selective medium. Species isolated were identified by cultural characteristics and ITS sequencing. Phytophthora x cambivora was the dominant species detected in 13 sites, followed by P. cinnamomi (5 sites), P. plurivora (3 sites) and P. cryptogea (1 site). Phytophthora x cambivora was present in both regions, while P. cinnamomi was found only in the Marmara region in coastal areas around Istanbul. When inoculated at the stem bases of 3‐year‐old chestnut saplings, P. cinnamomi produced significantly longer necrotic lesions (7.8–12.0 cm) than P. x cambivora (2.6–6.3 cm) by 12 days after inoculation. Phytophthora plurivora was the least aggressive species causing only small lesions. Phytophthora cryptogea, which represents the first record on chestnut in Turkey, produced intermediate sized lesions in between P. x cambivora and P. plurivora. These results indicate that P. x cambivora and in some areas P. cinnamomi play major roles in the observed dieback of sweet chestnut in western Turkey.     

Foliar susceptibility of eastern oak species to Phytophthora infection

Seven different Phytophthora species were used to test the foliar susceptibility of the common eastern US oak species and understory plants to Phytophthora infection. The Phytophthora species employed were Phytophthora cambivora, Phytophthora cinnamomi, Phytophthora citricola, Phytophthora europaea, Phytophthora quercetorum, Phytophthora quercina‐like and Phytophthora sp1. Inoculation of detached‐leaves with agar plugs containing mycelia of Phytophthora provided an estimate of their relative susceptibility. Lesions were always greater when foliage was wounded and young. On deciduous plants, lesion sizes were considerably reduced with the increasing foliar age, although with evergreen plants lesion sizes remained similar regardless of foliar age when more aggressive isolates were tested. Infections seldom resulted when foliage was not wounded. With young and mature foliage, P. citricola usually produced the largest lesions. Young foliage of Quercus rubra was the most susceptible to infection followed by Castanea dentata for both wounded and non‐wounded inoculations. Mature foliage of Hamamelis virginiana, Kalmia latifolia and Quercus alba were the most susceptible to wound and non‐wound inoculations.