The effects of calcium on stem lesions of silver birch seedlings

In this study, we tested the hypothesis that decreased liming of growth medium has a role in the increase of stem lesions and top dying caused by Phytophthora cactorum in containerized silver birch seedlings (Betula pendula) in Finnish forest nurseries. The effect of limestone dose rates on growth and the nutrient status was also monitored. An index based on severity of symptom expression was used to compare the effect of different liming treatments on P. cactorum infection. Limestone amended into the sphagnum peat growth medium increased the amount of calcium in the seedling stems. Liming did not significantly decrease the disease severity although index values in most cases decreased with the increased limestone dose rates. In general, the lesions were restricted after out‐planting and the mortality of seedlings was low. Only inoculated seedlings on which the lesions had spread around the stem in the nursery died. Phytophthora cactorum appears to be a nursery pathogen, as it did not survive under conditions present in the field. Four years after out‐planting, the tallest birches were those grown in sphagnum peat amended with the highest limestone dose of 8 kg m⁻³.     

Phytopbthora cactorum on walnut seedlings in Italian nurseries

Phytophthora cactorum was consistently isolated from basal stem tissues of walnut seedlings affected by collar and root rot. The isolates were identified on the basis of morphological and cultural characteristics and of electrophoretic patterns in polyacrylamide slab gels of total native mycelial proteins and isozymes (arylesterase, malate dehydrogenase, acid and alkaline phosphatase). The electrophoretic phenotypes of walnut isolates were identical or very similar to those of P. cactorum isolates from different hosts (apple, loquat, pineapple-guava and strawberry). In pathogenicity tests, P. cactorum isolates from walnut proved to be highly pathogenic for seedlings of English walnut (Juglans regia) and moderately pathogenic for seedlings of eastern black walnut (Juglans nigra); moreover, they did not rot apple seedlings, suggesting host-specificity. Poor soil drainage and excessive irrigation can be considered as predisposing factors to collar and root rot incited by P. cactorum.     

Isolation,identification and pathogenicity of Phytophthora species from declining oak stands

Within the scope of a research project on the condition of roots of declining oaks (Quercus robur, Quercus petraea), samples of fine roots and surrounding soil, specimens of stripe cankers on the stem base, and samples of stream water were examined for the presence of Phytophthora species using both baiting methods and selective agar media. At 27 sites in Germany (Bavaria, Rheinland-Pfalz, Schleswig-Holstein), Switzerland, Hungary, Italy and Slovenia the following species were isolated (mainly from soil): Phytophthora citricola, Phytophthora cactorum, Phytophthora cambivora, Phytophthora gon-apodyides, Phytophthora undulata, a species with affinity to Phytophthora drechsleri, and two additional species with close affinity to the Phytophthora cactorum group. Moreover, Pythium group P, Pythium anandrum, Pythium chamaehyphon, and many other Pythium species that have not yet been identified could be recovered. In a soil infestation test most isolates induced dieback of long root tips and necrotic lesions in the root cortex and at the root collar of Quercus robur seedlings. All Phytophthora species tested and Pythium group P caused cortical necrosis after stem inoculation of young Quercus robur trees. It could be shown in vitro that Phytophthora gonapodyides and Pythium group P were able to produce a wilting toxin. Nitrogen input and climatic changes are discussed as predisposing factors for root damage observed in the field.     

Oomycota species associated with deciduous and coniferous seedlings in forest tree nurseries of Western Turkey

Occurrence and pathogenicity of Oomycota species causing root rot were investigated in 10 forest tree nurseries in western Turkey. Soil samples (129 in total) taken from the rhizosphere of symptomatic seedlings were baited for Oomycota using young leaves of Quercus suber, Rhododendron simsii and R. ponticum. Oomycota (178 isolates) were obtained by culturing on selective media, and identified using morphological methods followed by PCR and sequencing of the ITS rDNA and cox1 regions. Phytophthora aff. cactorum, P. citricola sensu lato, P. crassamura, P. syringae, Pythium aphanidermatum, Py. intermedium, Py. irregulare, Py. ultimum and Phytopythium vexans were common amongst the isolates. The highest diversity of Oomycota was found in the forest nurseries at Adapazar?‐Hendek and ?zmir‐Torbal?. Pathogenicity tests showed that the isolates caused lesions on a range of host plants. The importance of these nursery infections in transferring potentially damaging oomycete species to Turkish forests is discussed.     

Susceptibility of common tree species in Sweden to Phytophthora cactorum,P. cambivora and P. plurivora

In Sweden, invasive Phytophthora pathogens have been recognized as a growing threat to urban and production forests, calling for an urgent update of regeneration strategies for infested areas. Stem inoculation tests were performed to test the relative susceptibility of common conifer and broadleaved tree species Pinus sylvestris, Picea abies, Larix x eurolepis, Betula pendula, Quercus robur, Fagus sylvatica, Populus trichocarpa and Tilia cordata to the root pathogens Phytophthora cactorum, P. cambivora and P. plurivora commonly isolated from Swedish soils. Results indicate that all the species tested were susceptible and formed lesions following stem inoculation with all three Phytophthora species, but to varying degrees. Of particular interest are the high levels of susceptibility in P. trichocarpa to all three Phytophthora species compared to other tested tree species.     

First report of Calonectria montana causing damping‐off disease on pine and spruce seedlings in Europe

In 2015–2016, damping‐off symptoms were observed on Pinus sylvestris and Picea abies seedlings in four forest nurseries in Poland. Some Calonectria isolates were obtained from symptomatic seedlings by culturing pieces of necrotic tissue on malt extract agar.Morphological and molecular analyses revealed that isolates represent Calonectria montana, a recently described species from natural forests in China. The isolates represent the first detection of this species outside of Asia. Pathogenicity tests demonstrated that C. montana is a severe pathogen of P. sylvestris seedlings.     

Pathogenicity of Malaysian Phytophthora palmivora on cocoa,durian, rubber and oil palm determines the threat of bud rot disease

The epidemic of bud rot disease affecting oil palm in Colombia is primarily caused by Phytophthora palmivora. The pathogen has a cosmopolitan presence that includes Southeast Asia, but to date, bud rot has not been reported in this region. This study provides an overview of the potential risk of Malaysian P. palmivora isolates cross‐infecting other host species, including cocoa, durian, rubber and Malaysian oil palm planting materials (Dura × Pisifera, D × P). On cocoa pods, the durian isolate PP7 caused dark brown necrotic lesions. Detached leaf bioassays showed that P. palmivora isolates PP3 and PP7 infected different hosts, except rubber foliage without wounding. Inoculation tests on cocoa, durian and rubber seedlings caused brown necrotic lesions when stems were wounded, with 10% mortality in cocoa and durian at 17 days post‐inoculation (dpi). However, no further infection was observed, and lesions closed within 14–28 dpi on the non‐wounded seedlings. Pathogenicity tests of oil palm seedlings inoculated with isolates PP3 and PP7 indicated that Malaysian P. palmivora isolates were not pathogenic to oil palms based on localized infection observed only through wounding. Overall, the work demonstrated that Malaysian P. palmivora isolates were able to cross‐infect multiple hosts but did not show severe infections on oil palms.     

New records of Phytophthora on trees in Britain

Phytophthora megasperma var. megasperma and unidentified isolates of Pbytophthora rescmbling P. cambivora were obtained from dead and dying roots of Horsc chestnut trccs with sparse foliage, small chlorotic leaves and branch dieback. P. citricola was isolatcd from soil around dead roots of similarly affected trccs and from around dcad feeder roots of a tree suffering from the common but unexplained Leaf scorch (marginal leaf necrosis) of Horse chestnut. Both P. cactorum and P. citricola were isolated from oozing acrial stem lesions on Horse chestnut trees. Inoculations with these species reproduced the damage.     

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.     

Diplodia shoot blight and asymptomatic persistence of Diplodia pinea on or in stems of jack pine nursery seedlings

Diplodia pinea (syn. Sphaeropsis sapinea) is known as a major cause of damage to red pine (Pinus resinosa) seedlings in nurseries. The fungus can also be a latent pathogen of red pine seedlings, persisting in the absence of gross symptoms and later proliferating under conditions that induce host stress. In the fall of 2004, three nurseries in Wisconsin were surveyed to determine the potential for the occurrence of Diplodia shoot blight on jack pine (Pinus banksiana) seedlings and the persistence of D. pinea on or in asymptomatic seedlings of this species. Incidence of shoot blight was quantified in five 1 m long segments of an interior row in each of two survey areas in each nursery. The pathogen was identified on symptomatic seedlings collected in these areas on the basis of presence of characteristic pycnidia and conidia. Five groups of 20 asymptomatic seedlings were also collected in each of the two survey areas in each nursery. A segment of the lower stem/root collar from each of these asymptomatic seedlings was surface‐disinfested and culturally assayed using tannic acid agar. The mean incidence of shoot blight (as high as 9%) and mean frequency of cultural detection from asymptomatic seedlings (as high as 20%) were greatest in proximity to red pine windbreaks which are a source of inoculum. Only D. pinea was confirmed from subsets of symptomatic and asymptomatic seedlings which were tested using mt SSU rDNA polymerase chain reaction (PCR) primers that allow differentiation of D. pinea from D. scrobiculata and other fungi in the genus Botryosphaeria and related anamorphic fungi. Jack pine seedlings inoculated with D. pinea isolates obtained from asymptomatic nursery seedlings developed shoot blight symptoms in greenhouse trials. Thus, the ability of D. pinea to damage jack pine seedlings in nurseries has been documented and the potential for virulent strains of this latent pathogen to be distributed on asymptomatic jack pine seedlings from nurseries has been confirmed.     

Charcoal root and stem rot of Eucalyptus

Macrophomina phaseolina (Tassi) Goid. caused a serious root and stem rot in juvenile seedlings of Eucalyptus in nurseries. This is a new host record. The pathogen produced systemic infection in the seedlings. The symptoms and histopathology and factors influencing the disease were studied. Of five fungicides tried in the laboratory and in the field, bavistin and dithane M-45 at 0.2% showed promising results for the control of the disease.     

Susceptibility of silver birch and black alder to several Phytophthora species isolated from soils in declining broadleaf forests in western Ukraine

In declining broadleaf forests in western Ukraine, several Phytophthora species including P. plurivora, P. bilorbang, P. polonica, P. gonapodyides and P. cactorum were recovered using soil baiting assays and identified using morphological and molecular methods. Pathogenicity tests of selected isolates were performed on black alder (Alnus glutinosa (L.) Gaerth.) and silver birch (Betula pendula Roth.) to assess susceptibility of these two tree species to the newly detected Phytophthora species. Phytophthora plurivora, P. bilorbang and P. polonica showed higher pathogenicity in both alder and birch compared to the other tested Phytophthora species.     

Pathogenicity of three Phytophthora spp. causing late seedling rot of Quercus ilex ssp. ballota

Within a research project on quality of plants for forestation of agricultural lands, we studied the aetiology of a late seedling rot affecting holm oak (Quercus ilex ssp. ballota) in two forest nurseries in southern Spain. Major disease symptoms were foliage wilting and necrosis of feeder roots. Phytophthora cinnamomi, Phytophthora cryptogea and Phytophthora drechsleri were isolated from necrotic roots of holm oaks. Selected isolates of the three Phytophthora species were pathogenic to Quercus ilex ssp. ballota and Quercus suber seedlings in artificial inoculations. Soil flooding conditions were essential for infection and root rot development. There was no host specificity among the species, the isolates of P. cinnamomi being the most virulent in all inoculated plants. In these inoculations, Q. ilex ssp. ballota plants were more susceptible than those of Q. suber. This work is the first report of P. cinnamomi, P. drechsleri and P. cryptogea affecting Q. ilex ssp. ballota in forest nurseries.     

Root diseases in bareroot and container nurseries of the Pacific Northwest: epidemiology, management, and effects on outplanting performance

In forest and conservation nurseries in the Pacific Northwest USA, seedling production can be limited by root diseases caused by fungi in the genera Fusarium Link:Fr., Cylindrocarpon Wollenw., Phytophthora de Barry, and Pythium Pringsh. Fusarium, Cylindrocarpon, and Pythium are the most ubiquitous, whereas incidence of Phytophthora is mostly associated with coastal bareroot nurseries. All of these root pathogens are encouraged by water saturated soils or media. Seedlings infected with Fusarium, Phytophthora, or Pythium often appear chlorotic or necrotic with extensive root decay. Cylindrocarpon often causes serious root decay without shoot symptoms. The best approach to mitigate losses from these diseases is to use a holistic integrated pest management program. This program should combine chemical controls with cultural practices, particularly those that increase soil permeability and drainage and reduce potential sources of inoculum, especially by disinfesting seeds and containers reused for crops. In general, we found that seedlings meeting nursery specifications for outplanting on forest soil (proper height, root collar diameter, healthy shoot color, lack of disease symptoms) but having these disease organisms on their root systems perform as well as non-infected seedlings.     

Surveys for asymptomatic persistence of Sphaeropsis sapinea on or in stems of red pine seedlings from seven Great Lakes region nurseries

The fungus Sphaeropsis sapinea persists on or in stems of asymptomatic red pine (Pinus resinosa) nursery seedlings, and proliferates to cause collar rot and mortality after planting. In the spring of 2002, seven nurseries were surveyed to determine the potential range in frequency of asymptomatic persistence: three operated by Wisconsin Department of Natural Resources (DNR), two by Minnesota DNR, one by Michigan DNR, and one by USDA Forest Service (in Michigan). At each nursery five groups of 20 asymptomatic red pine seedlings were collected near an inoculum source (red pine windbreak), if present, and five groups of 20 asymptomatic seedlings were collected away from such a source (1400 seedlings total). A segment of the lower stem/root collar from each seedling was surface disinfested and incubated on tannic acid agar. Transfers were made from resulting colonies and the pathogen identified from pycnidia and conidia produced in culture. The pathogen was identified from asymptomatic seedlings collected in all Wisconsin and Minnesota nurseries, but was never detected from seedlings from the Michigan DNR or USDA Forest Service nurseries. Frequencies of detection were greater (as high as 88%) from asymptomatic seedlings near red pine windbreaks including diseased trees than from seedlings distant from such windbreaks. A subset of isolates from asymptomatic seedlings was characterized using inter‐simple sequence repeat–polymerase chain reaction analysis. Most isolates were the A group of S. sapinea, but B group isolates (recently named Diplodia scrobiculata) were also obtained from one nursery. One Minnesota nursery was more extensively sampled in 2003, with 17–44 groups of five asymptomatic red pine seedlings collected in four separate fields (525 seedlings total). The mean frequency of detection of the pathogen in these four fields ranged from 40 to 71%. Persistence of S. sapinea on or in asymptomatic seedlings continues to be problematic, not only because of the potential for subsequent seedling mortality, but also as a means by which a pathogen may be widely distributed.     

Occurrence and pathogenicity of Phytophthora × cambivora on Prunus laurocerasus in Serbia

Cherry laurel (Prunus laurocerasus) is a native plant species in Serbian forests, but is also widely used for ornamental plantings. Following two extremely wet summers in 2014 and 2015, in spring and summer of 2016 and 2017, numerous cherry laurel plants with symptoms indicative for Phytophthora diseases, like wilting and chlorosis of leaves, dieback and bleeding bark necroses, were recorded in a park in Belgrade and in two ornamental nurseries in central Serbia. From necrotic bark samples and rhizosphere soil, self‐sterile Phytophthora isolates with woolly colonies were obtained. Due to the production of ellipsoid and elongated, non‐papillate sporangia in water and of ornamented oogonia with two‐celled antheridia in mating tests with tester strains of both Phytophthora × cambivora and P. cryptogea, these isolates were identified as P. ×cambivora which was confirmed by ITS sequence analysis. Pathogenicity of P. ×cambivora from cherry laurel (PCCL) was tested by inoculating one‐year‐old seedlings of cherry laurel under the bark. P. ×cambivora from European beech (PCB), and isolates of P. cactorum (CAC), P. cryptogea (CRY), P. plurivora (PLU) and P. ×serendipita (SER) were included as comparison. Three and a half months after inoculation, nine of the twelve plants in PCB, three in PCCL and CAC and two in PLU declined with longitudinal necroses and chlorosis, wilting and premature shedding of leaves. These results demonstrate the ability of P. ×cambivora to infect and cause decline of cherry laurel plants. The particularly high aggressiveness of the P. ×cambivora isolate from beech shows that this pathogen poses a serious risk to cherry laurel in the rare natural communities of cherry laurel and beech in Serbia.     

Pathogenicity of Phytophthora alni complex and P. plurivora in Alnus glutinosa seedlings

The ecological importance of riparian forests is well known. However, these forest habitats have been disturbed by human activities over the past century as a result of the introduction of flow regulations. Mortality of the riparian alder population caused by Phytophthora has become an important issue in Europe in the past two decades. The main objective of this study was to examine the pathogenicity of the Phytophthora alni complex (P. xalni, P. uniformis and P. xmultiformis) and P. plurivora in Alnus glutinosa seedlings. Phytophthora alni complex has traditionally been identified as the main causal agent of alder decline; however, in this study, P. plurivora was found to cause as much damage in inoculated seedlings as P. alni complex. In fact, Spanish isolates of P. xalni caused mortality rates of ca. 30%, whereas P. plurivora killed ca. 50% of seedlings. Likewise, there were no differences between P. xalni ‐ and P. plurivora‐inoculated seedlings in either the length of lesion or biomass allocation. The pathogenicity of the species within P. alni complex did not differ significantly. The findings confirm that P. plurivora, which is one of the most common species of Phytophthora found in European nurseries, is highly pathogenic to alder seedlings. Urgent implementation of measures to prevent the risk of spread of the pathogen from nurseries to natural ecosystems is required. Further studies are required to clarify the role that P. plurivora is playing in alder decline both individually and in combination with P. alni complex.     

Effects of various stress treatments on growth and ethylene evolution in seedlings and sprouts of betula pendula Roth and B. pubescens Ehrh.

Experiments were conducted to determine growth and ethylene evolution in seedlings and coppiced plants of Betula pendula Roth and B. pubescens Ehrh. when subjected to stresses relevant to the maintenance of natural forests, and especially in fuelwood plantations, i.e. cutting of the stem, thinning, bending, flooding and various combinations of these. Most of the experiments were carried out in the laboratory using 1‐year‐old seedlings or 1‐month‐old sprouts. Height and diameter growth, biomass production, morphology and ethylene evolution were studied for 1–2 months. Material for comparison was obtained from young sprouts on the stumps of fully developed trees growing under natural conditions and natural seedlings of a comparable age. Exposure of seedlings and coppiced plants to stress factors usually altered growth and increased ethylene evolution. Cutting of the stem and thinning had similar effects in that they stimulated transient ethylene evolution by both roots and stems. Bending retarded the height growth to some extent, while increasing the ethylene content especially on the lower side of the horizontal stem. Flooding arrested height growth and increased ethylene evolution in the roots. The two species reacted basically in similar ways, but B. pendula produced more ethylene. though at a slower rate, and also showed a more pronounced retardation of growth. The sprouts had a higher ethylene content than the seedlings and also increased their ethylene evolution more rapidly. The changes in ethylene evolution and growth are discussed from the point of view of the resilience of the plants and as an indication of stress factors in experimental plots.     

Pathogenicity of Heterobasidion annosum (Fr.) Bref. sensu stricto on coniferous tree species in Turkey

Two‐year‐old seedlings of Pinus brutia, P. brutia var. eldarica, Pinus pinea and 3‐year‐old seedlings of Pinus radiata, Pinus sylvestris, Pinus nigra and Cedrus libani were inoculated on the lower stem with isolates of Heterobasidion annosum s.s. collected from the Black Sea and Mediterranean regions of Turkey. In total, 315 seedlings were inoculated in April 2014 and incubated in a growth chamber for 7 weeks at 18–20°C. All isolates were pathogenic on the seven different hosts and had the ability to grow in living sapwood. The isolates had a greater growth on C. libani, P. sylvestris and P. radiata seedlings compared to plants of the other species tested. The least affected species were P. brutia and P. nigra. The isolates originating from the Black Sea region caused longer lesions on the hosts. Overall mortality during 7 weeks of incubation was 4%.     

Association of seed‐borne Fusarium species on Pinus ponderosa with germination and seedling viability in Argentina

Pinus ponderosa seedlots from eight seed orchards in Western Patagonia, Argentina, were evaluated for the presence of Fusarium species, and isolates obtained were subjected to pathogenicity tests. The following species were isolated: Fusariumoxysporum, Fusarium equiseti, Fusarium verticillioides, Fusarium incarnatum, Fusarium acuminatum, Fusarium graminearum and Fusarium proliferatum. With the exception of F. proliferatum and F. graminearum all Fusarium species were recovered from P. ponderosa seedlings with symptoms of damping‐off. Five Fusarium species were re‐isolated from necrotic roots of seedlings showing chlorosis (F. oxysporum, F. equiseti, F. incarnatum, F. acuminatum and F. verticillioides). High damping‐off losses occurred in Fusarium‐contaminated seed of P. ponderosa and low disease incidence occurred on less contaminated seed. The role of seed‐borne inoculum in disease spread within forest nurseries is discussed.