Physiological responses of cork oak and holm oak to infection by fungal pathogens involved in oak decline

The aim of this research was to study the changes in net photosynthesis and stomatal conductance values in 3‐year‐old cork oak and holm oak seedlings growing in natural conditions and inoculated with Apiognomonia quercina, Biscogniauxia mediterranea, Botryosphaeria corticola and Pleurophoma cava. Throughout the 4‐month experimental period, the evolution of visual external symptoms and the values of physiological variables were periodically recorded. All pathogens caused stem lesions around the infection point; however, the lesions caused by B. corticola were longer in both oak species. On cork oak seedlings, all pathogens induced a significant and gradual reduction in net photosynthesis and stomatal conductance values, whereas other physiological disturbances were induced only by B. corticola infections on holm oak seedlings.     

Seasonal and spatial mortality patterns of holm oak seedlings in a reforested soil infected with Phytophthora cinnamomi

The viability of 1-year-old holm oak (Quercus ilex) seedlings in a soil naturally infected with Phytophthora cinnamomi was studied during 2 consecutive years in a plot located in south-western Spain. In both years, total mortality during autumn and winter was not noticeable (<2.1%). In spring, mortality levels were higher (8.3–4.6%), especially the first year. A steep increase in total mortality occurred in summer, both in the first (11.4%) and second (24.2%) year, but mortality attributable to P. cinnamomi was 1.9 and 7.6%, respectively. Thus, 2 years after planting, total cumulative mortality was 43.4%, and that attributable to P. cinnamomi 9.6% (i.e. 22.1% of total mortality). Fungus-derived mortality followed a spatially aggregated pattern in the reforestation plot, suggesting a clustered distribution of the inoculum in the soil. Furthermore, mortality by P. cinnamomi was also associated with nearness of infected adult trees in the plot. Results obtained are discussed in the framework of seasonal water deficit, P. cinnamomi damage, weed competition and sanitation techniques to be used in declined holm oak stands in Spain.     

Viability of holm and cork oak seedlings from acorns sown in soils naturally infected with Phytophthora cinnamomi

The emergence and survival of pregerminated holm oak (Quercus ilex) and cork oak (Quercus suber) acorns from two ecologically different dehesas (Mediterranean open woodlands) were studied in two soils from these stands naturally infected with Phytophthora cinnamomi, and in the same soils previously sterilized in the autoclave. Phytophthora cinnamomi was consistently isolated from the radicles of all unemerged and all emerged but dead seedlings from the unsterilized substrates. Seedlings of holm oak were more susceptible to P. cinnamomi than those of cork oak. Mortality of holm oak seedlings was significantly different depending only on soil treatment (sterilized or unsterilized), and it was 100% in unsterilized soils, independent of acorn provenance and soil origin. Mortality of cork oak seedlings was significantly different depending on the acorn origin and soil treatment, and on the interactions acorn origin × soil origin and soil origin × soil treatment. The demonstrated high susceptibility of holm and cork oak young seedlings to P. cinnamomi could be a limiting factor in Mediterranean open woodlands (dehesas) not only in natural regeneration processes but also when reforestation by direct sowing is implemented.     

Susceptibility of cork oak (Quercus suber) to canker disease caused by Diplodia corticola: when time is of the essence

New Forests - Cork is a renewable resource mainly produced in Mediterranean countries thanks to sustainable management of cork oak (Quercus suber) stands. The infection caused by the ascomycete...     

Influence of water stress on susceptibility of red oak (Quercus rubra) to Phytophthora cinnamomi

Influence of water status of oak trees (Quercus rubra) on the development of Phytophthora cinnamomi lesions was studied. On agar media or excised inner bark, growth of P. cinnumomi was reduced by low water potentials. In contrast, on voung saplings or mature oak trees the development of P. cinnamomi lesions was not affected during a period of water stress. But after the end of the water stress treatment, susceptibility of the young saplings to P. cinnamoni increased.     

The oak decline: a new problem in Italy

The Authors observed, in Italy, over a number of years, isolated and grouped trees of the genus Quercus that declined over one or more vegetative seasons. The spread was very fast; in fact the trees declined in the course of 2–3 vegetative seasons. A series of symptoms was observed: production of epicormic shoots, yellowing of leaves, laceration of the bark, bleeding from the trunk. Many fungi were isolated: Armillaria spp., Cephalosporium spp., Cladosporium spp., Cylindrocarpon spp., Diplodia mutila, Hypoxylon mediterraneum, Phoma cavae, Pbomopsis quercina, Sporotrix spp. Future research will attempt to ascertain the possible causes of the decline.     

Diplodia twig canker (Diplodia gallae) of northern red oak (Quercus rubra) in the northeastern United States

The 2015–2018 outbreak of spongy moth (Lymantria dispar) in southern New England initiated a severe oak decline and mortality event. While defoliation was a primary driver, increased secondary pest and pathogen activity contributed to decline and death. Following this large defoliation event, Diplodia was frequently recovered from northern red oaks (Quercus rubra) with serious outbreaks of twig cankering. Given the many recent reports of Diplodia corticola on oak in eastern North America, it was presumed to be the causal agent. To confirm, a limited survey was conducted from five states in the region (Maine, Massachusetts, New Hampshire, New York, and Vermont). Based on ITS, tef1 and tub2 sequences generated from 28 isolates collected primarily from northern red oak, Diplodia gallae, two potentially novel Diplodia species and Diplodia sapinea were identified. Surprisingly, D. corticola was not found in this study. ITS sequences alone failed to discriminate among isolates of D. gallae and D. corticola, creating uncertainty over previous reports of D. corticola in eastern North America. Only a combined ITS + tef1 dataset successfully distinguished D. gallae and D. corticola along with two other closely related species that also occur on oak (Diplodia quercicola and Diplodia quercivora). Additional cankering and endophytic fungi (Coryneum, Dendrostoma, Gnomoniopsis, Pestalotiopsis and Tubakia) were also found on symptomatic oaks in the region. Identification of Diplodia isolates from non-Quercus hosts also detected Diplodia neojuniperi on Juniperus chinensis and Microbiota decussata, which has not been reported previously in North America.     

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.     

Influence of site factors on the impact of Phytophthora cinnamomi in cork oak stands in Portugal

Although decline of cork (Quercus suber) and holm oak trees (Quercus rotundifolia) has been described in Portugal in the late years of the 19th century, its development has become a motive of high concern during the last two decades. The presence of Phytophthora cinnamomi in cork and holm oak stands was surveyed in four different regions of the country (Trás‐os‐Montes, Alentejo, Ribatejo and Algarve) during 1995–98. Tree decline severity, sudden death and site characteristics were assessed in 56 sites representing varied conditions. The pathogen was isolated from oak roots and rhizosphere samples in 27 of those places. Other plant species from natural vegetation were sampled in three active disease centres. This survey showed that 56% of the surveyed species of shrub flora were infected with P. cinnamomi, which was detected mainly on species belonging to the families Ericaceae, Cistaceae and Leguminosae. Recovery of P. cinnamomi was more frequent in shallow soils (Leptosols and complex Leptosols and Luvisols). These soils are more common in the south (Algarve), where decline has a high impact. Soils with low fertility and low mineral nutrient levels, particularly phosphorus, seemed to favour infection. Site aspect and topographic tree situation were also evaluated. Sites facing south showed higher occurrence of P. cinnamomi, which was also more frequent in slopes and valleys than on hilltops. In Algarve, a relationship could be established between the crown status and the presence of P. cinnamomi in roots and rhizosphere. Different morphotypes of P. cinnamomi could be distinguished in vitro, and their occurrence in the field was correlated with particular site characteristics. Further research needs and management strategies to limit the extension of the disease are discussed.     

Incidence of Phytophthora species in oak forests in Austria and their possible involvement in oak decline

A survey on the occurrence of Phytophthora species in oak ecosystems in Austria was conducted from April to May 1999 and in June 2000. The investigations were carried out at 35 study sites distributed throughout the zone of oak forests in eastern Austria. Four oak species, including Quercus robur, Q. petraea, Q. cerris and Q. pubescens were considered in the survey. Rhizosphere soil samples were taken from sample trees, which consisted of healthy and declining trees as indicated by their crown transparency. Young oak leaflets were used as baits to recover Phytophthora species. The assemblage of Phytophthora spp. detected in Austrian oak forests consisted of five species, including Phytophthora quercina, P. citricola, P. gonapodyides, P. europaea and P. syringae. P. quercina and P. citricola were isolated from 11 and seven sites, respectively, and were thus the most common and most widely distributed species. The three other species were recovered only sporadically. P. citricola could be separated into two morphologically and genetically well‐characterized types (A and B). Phytophthora species, in particular the common P. quercina and P. citricola occurred on sites showing a wide variety of soil types, soil textures and moisture classes. There was mild evidence for connection between deteriorating crown status and the presence of Phytophthora spp. Furthermore, significant differences in contents of magnesium, as well as calcium, aluminium, nitrogen and carbon at different soil depths (0–10, 10–20 and 20–40 cm) were detected between Phytophthora‐infested and Phytophthora‐free sites. The results of the present study provide circumstantial evidence that Phytophthora species are involved in oak decline at certain sites in Austria.     

Evaluation of fungicides for the control of Botryosphaeria corticola on cork oak in Catalonia (NE Spain)

Botryosphaeria canker of cork oak (Quercus suber L.), caused by Botryosphaeria corticola, is a well-known factor contributing to the decline of this forest species and is a serious disease in the main cork-producing countries of the Western Mediterranean basin. In this work, fungicides with the potential for controlling the disease were selected using a combination of in vitro tests and field trials. Fourteen fungicides in different chemical families were tested in vitro to evaluate their effect on inhibition of mycelial growth. The most effective fungicides were benomyl, carbendazim, cyprodinil + fludioxonil, thiabendazol and thiophanate-methyl; these compounds were then further tested under field conditions. In the field trials, carbendazim and thiophanate-methyl were the most effective fungicides, as shown by efficacy indices, in reducing debarked trunk surface affected by cankers (76% and 50–65%, respectively). These results suggest that the occurrence of Botryosphaeria canker on cork oak can be reduced effectively with the application of selected fungicides after cork removal from the trees.     

Fine root dynamics of oak saplings in response to Phytophthora cinnamomi infection under different temperatures and durations

The belowground effects of Phytophthora cinnamomi on 1‐year‐old saplings of two common oak species in mid‐Atlantic US forests, white (Quercus alba) and red oak (Q. rubra), were examined after incubation in pathogen‐infested soilless potting mix. Fine root lengths (0–1.5 mm in diameter) of both oak species were quantified after incubation at successive 30‐day intervals up to 300 days, for a total of 10 incubation periods. In addition, colony‐forming units (CFU) of P. cinnamomi were quantified after white oak saplings were incubated in infested soilless potting mix at different temperature/duration combinations that reflect soil conditions present in the mid‐Atlantic United States. Impact of P. cinnamomi on fine root lengths of red and white oak saplings varied considerably over time. Significant periods of fine root loss occurred primarily during spring, when bud break and leaf flush began for both oak species. Red oaks had 17% fine root loss on average, while white oaks appeared more resistant to P. cinnamomi infection with a 2% decrease in fine roots over the course of the experiment. Phytophthora cinnamomi CFU declined significantly with exposure to all incubation temperatures except 8°C. This was in contrast to in vitro experiments, where the optimum temperature for mycelial growth was determined to be 21°C and above. Significant fine root loss caused by P. cinnamomi depended on plant phenology and the oak species tested. Extreme soil temperatures have a significant adverse impact on temporal changes of P. cinnamomi population.     

Susceptibility of the Quercus rubra root system to Phytophthora cinnamomi; comparison with chestnut and other oak species

The susceptibility of oak seedlings (Quercus palustris, Quercus robur, Quercus rubra) and chestnut seedlings (Castanea sativa) to Phytophthora cinnamomi was tested. The dynamics of infection was examined in plant material raised in a rhizotron. In the oak species, primary root tissues were susceptible whereas secondary cortical tissues showed some resistance to P. cinnamomi. Secondary cortical tissues of the tap root in C. sativa were susceptible. Inoculations with P. cinnamomi were performed both in situ and on excised roots of mature Q. rubra. In both cases, the resistance of Q. rubra roots and shoots was negatively correlated with diameter at the inoculation point. Small roots (l–5-cm diameter) were resistant, whereas collar and trunk were susceptible. In contrast to oak, small excised roots of mature C. sativa (0.7–2-cm diameter) were susceptible to P. cinnamomi. This may explain why P. cinnamomi does not induce a decline of the attacked oaks, but rather a trunk canker.     

Bacillus pumilus and Stenotrophomonas maltophilia as two potentially causative agents involved in Persian oak decline in Zagros forests (Iran)

The oak decline is known as one of the most destructive complex diseases causing high economic losses around the world, especially in Iran. The main objective of the present study was to investigate the possible role of bacteria as causative agents of oak decline in the Zagros forests of Iran. To do this, stem, root and leaf samples were taken from symptomatic Persian oak trees (Quercus brantii) in different zones of Zagros forests (Ilam Province, Iran). From 150 bacterial isolates, 20 showed pathogenicity against Geranium seedlings. Among 20 hypersensitivity test positive strains, four strains showed pathogenicity against oak saplings. Based on morphological and 16S rRNA gene sequence analysis, three strains were identified as Bacillus pumilus and one strain as non‐sporulating Gram‐negative Stenotrophomonas maltophilia. Pathogenicity studies of different B. pumilus and S. maltophilia strains revealed that they have potential to cause the disease in oak saplings and symptoms of disorder in Persian oak trees. To our knowledge, there are no previous records of B. pumilus and S. maltophilia causing decline on Fagaceous trees like Q. brantii. More detailed field and molecular studies are required to confirm the absolute role of such bacteria in occurrence of oak decline in Zagros forests.     

Drought tolerance and transplanting performance of holm oak (Quercus ilex) seedlings after drought hardening in the nursery

Drought stress is the main cause of mortality of holm oak (Quercus ilex L.) seedlings in forest plantations. We therefore assessed if drought hardening, applied in the nursery at the end of the growing season, enhanced the drought tolerance and transplanting performance of holm oak seedlings. Seedlings were subjected to three drought hardening intensities (low, moderate and severe) for 2.5 and 3.5 months, and compared with control seedlings. At the end of the hardening period, water relations, gas exchange and morphological attributes were determined, and survival and growth under mesic and xeric transplanting conditions were assessed. Drought hardening increased drought tolerance primarily by affecting physiological traits, with no effect on shoot/root ratio or specific leaf mass. Drought hardening reduced osmotic potential at saturation and at the turgor loss point, stomatal conductance, residual transpiration (RT) and new root growth capacity (RGC), but enhanced cell membrane stability. Among treated seedlings, the largest response occurred in seedlings subjected to moderate hardening. Severe hardening reduced shoot soluble sugar concentration and increased shoot starch concentration. Increasing the duration of hardening had no effect on water relations but reduced shoot mineral and starch concentrations. Variation in cell membrane stability, RT and RGC were negatively related to osmotic adjustment. Despite differences in drought tolerance, no differences in mortality and relative growth rate were observed between hardening treatments when the seedlings were transplanted under either mesic or xeric conditions.     

Species-specific drought effects on flower and fruit production in a Mediterranean holm oak forest

A holm oak forest was exposed to an experimental drought (reductionof 15 per cent soil moisture as predicted for this area forthe next decades by General Circulation Models and ecophysiologicalmodels) during 7 years to elucidate the reproductive responsesof the dominant species Quercus ilex L., Arbutus unedo L. andPhillyrea latifolia L. Soil moisture was partially reduced byplastic strips intercepting rainfall and by ditch exclusionof water runoff. During the period studied, meteorological conditionsand soil moisture were continuously monitored, together withflower and fruit production in the three dominant species. InQ. ilex and A. unedo, flower and specially fruit productionwere strongly correlated with annual rainfall, but not in P.latifolia. The experimental drought reduced flower and fruitproduction in Q. ilex by 30 per cent and 45 per cent, respectively.Reductions in flower and fruit production were not significantin A. unedo and were not observed in P. latifolia. A decreasein production of reproductive structures and the different responseof the species studied to a decrease in water availability couldinduce important changes in the competitive ability of the differentspecies and in the long term in the community species compositionand future distribution of these Mediterranean species.     

Wild boars (Sus scrofa) affect the recruitment rate and spatial distribution of holm oak (Quercus ilex)

The role of herbivores in controlling plant population abundance and distribution is unclear. We experimentally determine the effect of damage by wild boars (Sus scrofa) in recruitment rate and spatial pattern of a Mediterranean tree, the holm oak (Quercus ilex). We monitored oak establishment in the Sierra Nevada of southeastern Spain during 4 years (1999–2002) in four plots, two fenced and two unfenced that were used as controls. In addition, we planted 1-year-old seedlings in all microhabitats, both in fenced and control plots, to experimentally determine the effect of wild boar on their microhabitat-dependent survival. Despite seedling abundance being similar inside and outside the fences, sapling abundance diminished up to 50% in unfenced plots compared to fenced plots. Wild boars also seemed to modify the spatial distribution of seedlings and saplings. Whereas seedlings were most abundant under pines both in fenced and control plots, sapling abundance was highest under pine trees in fenced plots, but similarly abundant under pines or shrubs in control plots. This change in spatial pattern was due to the effect of wild boars killing seedlings when looking for food under pines. The effects of the different mortality factors were not additive. Where ungulates are absent, recruitment is high and occurs under oaks and especially under pines. Where wild boars are present, recruitment diminishes and occurs mostly under shrubs and pines. Wild boars arrest population growth and modify the spatial pattern of recruitment.     

Phosphite impact on the in vitro production and viability of selfed oospores by Phytophthora cinnamomi

Four isolates of Phytophthora cinnamomi were able to produce selfed oospores when cultured in vitro on modified Ribeiro’s minimal medium. Phosphite inhibited the production of selfed oospores at 100 μg phosphite/ml, lower concentrations had no effect. Selfed oospores were considered dormant as 16–47% of selfed oospores were found viable using viability staining techniques such as thiazolyl blue tetrazolium bromide and 4′,6‐diamidino‐2‐phenylindole·2HCl yet they did not germinate on a range of media. Phosphite up to 100 μg/ml did not change the proportion of viable selfed oospores. This study shows that phosphite does not affect the production of selfed oospores.     

The role of yellow lupin (Lupinus luteus) in the decline affecting oak agroforestry ecosystems

Phytophthora cinnamomi is a soilborne pathogen causing root rot in Mediterranean Quercus species growing in ‘dehesa’ rangeland ecosystems. Recently, it has been reported causing wilting and death of Lupinus luteus (yellow lupin), a spontaneous plant in southern Spain rangelands, but also frequently sowed for livestock grazing. In soils artificially infested with P. cinnamomi chlamydospores and planted with different cultivars of yellow lupin, a significant increase in the density of propagules was detected in comparison with the initial levels of inoculum and with the infested but not planted soil (control). In oak‐rangelands in which yellow lupine was planted, isolation and counting of colonies of P. cinnamomi from soil samples have shown the ability of this plant to maintain or even increase the inoculum density and thus facilitate the infection of trees. Results suggested that cultivation of yellow lupin in oak‐rangeland ecosystems should be avoided whether oak trees are affected by root disease caused by P. cinnamomi or not. This leguminous plant can act as an inoculum reservoir or even enhance inoculum soil levels available for oak root infections, exacerbating the oak decline severity in the region.