Prevalence and incidence of the root-inhabiting fungi, Fusarium, Cylindrocarpon and Pythium, on container-grown Douglas-fir and spruce seedlings in British Columbia

Surveys were made at the end of the 1990 and 1991 growing seasons for root-inhabiting fungi in the genera Fusarium, Cylindrocarpon and Pythium from the roots of one year-old container-grown Douglas-fir and spruce seedlings grown under greenhouse conditions. In the 1990 survey of four nurseries, it was found that 61–97% of both Douglas-fir and spruce roots were colonized with Fusarium, Cylindrocarpon or Pythium. There were significantly (p0.05) more Douglas-fir roots than spruce roots colonized by Fusarium at all nurseries, however, there were significantly (p0.05) more spruce roots than Douglas-fir roots colonized by Cylindrocarpon and Pythium. Root colonization of Douglas-fir and spruce by the three fungal genera during 1991 varied from 0–82% at three nurseries, however, only at a south coastal nursery was there significantly (p0.05) more spruce than Douglas-fir roots colonized by Cylindrocarpon. Significantly more seedlings were infected in 1990 than in 1991. In 1991, there were few significant differences between Douglas-fir and spruce, in the percentage of seedlings with colonized roots and in the percentage of growth medium colonized by the fungi. However, there were significant differences between nurseries.     

Pathogenic fungi associated with pre‐ and post‐emergence seedling blight of pine and cypress in Fars Province,Iran

Seed and root rot of pine and cypress seedlings cause heavy annual losses to forest nurseries in Fars Province. Root and crown samples of various species of conifers, such as Tehran pine (Pinus eldarica), brutia pine (Pinus brutia), Arizona cypress (Cupressus arizonica), Shirazian cypress (Cupressus sempervirens var. fastigiata), common cypress (C. sempervirens var. horizontalis) and oriental arborvitae (Thuja orientalis), showing chlorosis, necrosis, stunted growth, defoliation and root and crown rot symptoms were collected from pine and cypress nurseries across Fars province at various time intervals. Infected tissues were washed and cultured on acidified potato dextrose agar (PDA) and corn meal agar (CMA) amended with Delvocide and ampicillin, with and without surface sterilization, respectively, and incubated at 25°C for 3–5 days. Decaying seeds of all plant species were also collected from nursery seed stocks and cultured on PDA. Hyphal tip isolates were used for further studies. Species of Phytophthora, Pythium, Rhizoctonia and Fusarium were isolated from symptomatic seedlings of pine and cypress at different times during the growing season. Pathogenicity of isolates of Phytophthora, Pythium and Rhizoctonia was confirmed on seedlings of all plant species, whereas Fusarium sp. isolated from the seeds of Tehran pine was only pathogenic on seedlings of Tehran pine and Arizona cypress. Virulent isolates identified as Phytophthora nicotiana var. parasitica, Pythium ultimum, Pythium paroecandrum, Rhizoctonia solani and binucleate Rhizoctonia‐like fungus caused root and crown rot of pine and cypress seedlings. Pathogenic isolates of Fusarium identified as Fusarium proliferatum also caused pre‐emergence seed decay of pine and cypress. Isolates of Phytophthora and Pythium were the most virulent of these fungal isolates.     

Effects of oak root pruning in forest nurseries on potential pathogen infections

Despite the general practice of root pruning, little is known about the potential impact of reducing shoot/root systems of oak seedlings in this way on their future susceptibility to pathogens, for example Cylindrocarpon spp., Fusarium spp., Ilyonectria spp., Pythium spp. Phytophthora spp. or Rhizoctonia spp. In this study, root‐pruned and non‐pruned seedlings of Quercus robur grown under controlled conditions were inoculated with aggressive and less‐aggressive pathogens. Results indicated, in contrast to our initial assumption, that pathogens significantly reduced lateral root biomass more in non‐pruned seedlings, the extent of the response depending on the pathogens species. In response to pathogen pressure, pruned seedlings tended to attain a higher dry stem mass fraction, but lower dry leaf mass fraction. Pathogens also suppressed leaf mass in total root dry mass fraction (dry leaf mass/total root dry mass ratio, in g × g^?1) more in pruned than in non‐pruned seedlings. These results suggest differences in growth between non‐pruned and pruned seedlings in response to pathogen stress. In nurseries, root pruning of oak trees may enhance the reduction in leaf mass in lateral roots mass fraction resulting from pathogen infections, which may decrease seedling quality. It is therefore important to ensure a low level of inoculum of soil‐borne pathogens to minimize the risk of seedling infection.     

Effect of some nursery practices on incidence and severity of diseases,and growth of Eucalyptus grandis seedlings*

Nursery practices influenced incidence and severity of diseases of Eucalyptus grandis viz. damping-off caused by Pythium sp., Rhizoctonia solani and Cylindrocladium quinqueseptatum, web might by R. solani, seedling blight by C. quinqueseptatum and shoot wilt by Sclerotium rolfsii. Growth of seedlings of E. grandis and microclimatic conditions were also affected by nursery practices. Shading with coconut leaf thatch led to low light intensity (av. 1463 lux) with high soil water potential, low soil and ambient temperatures, and high severity of damping-off and web blight diseases and poor shoot:root ratio of seedlings. Seed beds under coirmat had dispersed light (av. 22 299 lux), apparently high severity of seedling blight and shoot wilt, and good growth (snoot:root ratio) of seedlings. In both the types of shading high soil moisture regime and high seed rate contributed to high disease severity as well as low shoot:root ratio of seedlings.     

Conifer seedling root fungi and root dieback in Finnish nurseries

Fungi were isolated from the roots and growth substrate of bare‐rooted and containerized Pinus sylvestris and Picea abies nursery seedlings displaying a root dieback. Isolations were also made from visually healthy seedlings. The potential pathogenicity of all isolated species was determined in laboratory trials.

Cylindrocarpon spp., Fusarium spp. and Trichoderma viride were frequently isolated. The isolation frequency of a uninucleate Rhizoctonia‐like fungus, Pythium spp. and Phytophthora imdulatum from diseased containerized seedlings and their pathogenicity in tests suggest that these fungi are likely involved in the root dieback disease in containers. The pathogenic Rhizoctonia‐like fungus in addition to Pythium spp. was also isolated from bare‐rooted seedlings. In greenhouse tests Pythium spp. were more pathogenic to 4‐week‐old Scots pine seedlings grown before transplantation in unsterile substrate than to those seedlings grown axenically in agar. External factors are considered to have some role in the expression of disease.     

Fusarium root infection of container-grown Douglas-fir: effect on survival and growth of outplanted seedlings and persistence of the pathogen

Greenhouse-grown Douglas-fir (Pseudotsuga menziesii var.glauca [Beissn.] Franco) seedlings with roots infected with eitherFusarium oxysporum Schlect. orFusarium proliferatum (Matsushima) Nirenberg were outplanted on a forest site in northern Idaho, U.S.A. No residentFusarium populations were detected in the forest soil.Fusarium persisted the first four years on roots initiated during the greenhouse phase, but occurred sparingly or was absent on roots that grew after outplanting. Height growth was unaffected, and mortality was not often associated withFusarium. A seasonal pattern ofFusarium activity was observed. Low levels (10–40%) of initial root infection apparently have little adverse effect on outplanting performance of Douglas-fir seedlings.University of Idaho, Idaho Forest, Wildlife and Range Experiment Station Contribution No. 640.     

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.     

Fusarium spp. and Pinus strobus seedlings: root disease pathogens and taxa associated with seed

Eastern white pine (Pinus strobus L.) seeds weresown in soil infested with Fusarium proliferatum, rootnecrosis developed on seedling roots, and F. proliferatumwas reisolated from symptomatic roots; thus, demonstrating thatF. proliferatum is pathogenic toeastern white pine seedlings. Soils infested with F.acuminatum or F. sporotrichioides resulted infew diseased seedlings. Seedlings with root rot generally showed reductions inseedling height. All Fusarium species tested wererecoveredfrom rhizosphere soil samples. Three seedlots of Pinusstrobus were examined for Fusarium infestation.Fusarium species were recovered from most seeds in twoseedlots. Fusarium proliferatum and F.sporotrichioides were the predominant species isolated. Additionalspecies not previously reported from P. strobus included:F. acuminatum, F. chlamydosporum,F. equiseti, F. graminearum,F. moniliforme, F. poae, F.polyphialidicum, F. heterosporum, F.sambucinum, and F. semitectum.     

Beech decline in Central Europe driven by the interaction between Phytophthora infections and climatic extremes

During the past decade, and in particular after the wet year 2002 and the dry year 2003, an increasing number of trees and stands of European beech (Fagus sylvatica L.) in Bavaria were showing symptoms typical for Phytophthora diseases: increased transparency and crown dieback, small‐sized and often yellowish foliage, root and collar rot and aerial bleeding cankers up to stem heights of >20 m. Between 2003 and 2007 134 mature beech stands on a broad range of geological substrates were surveyed, and collar rot and aerial bleeding cankers were found in 116 (86.6%) stands. In most stands the majority of beech trees were declining and scattered or clustered mortality occurred. Bark and soil samples were taken from 314 trees in 112 stands, and 11 Phytophthora species were recovered from 253 trees (80.6%) in 104 stands (92.9%). The most frequent species were P. citricola, P. cambivora and P. cactorum. Primary Phytophthora lesions were soon infected by a series of secondary bark pathogens, including Nectria coccinea, and wood decay fungi. In addition, infected trees were often attacked by several bark and wood boring insects leading to rapid mortality. Bark necroses were examined for their probable age in order to determine whether the onset of the current Phytophthora epidemic was correlated to rainfall rates recorded at 22 Bavarian forest ecosystem monitoring stations. A small‐scale survey in nine Bavarian nurseries demonstrated regular infestations of all beech fields with the same range of Phytophthora species. The results indicate that (1) Phytophthora species are regularly associated with beech decline and may also be involved in the complex of ‘Beech Bark Disease’, (2) excessive rainfalls and droughts are triggering the disease, and (3) widespread Phytophthora infestations of nursery stock might endanger current and future silvicultural projects aiming on the replacement of non‐natural conifer stands by beech dominated mixed stands.     

Nursery soil cover influences germination of Thujopsis dolabrata var. hondai seeds

We examined the influence of nursery soil cover on germination of Thujopsis dolabrata var. hondai. Seeds were sown under seeding bed conditions: (1) nursery soil (andosol) without soil cover, (2) Kanuma pumice without soil cover, (3) nursery soil with nursery soil cover, and (4) nursery soil with sterilized sand cover. Germination percentages were compared between these conditions. The germination percentage (8.8%) at 10 weeks after sowing under condition 3 was significantly lower than under conditions 1, 2, and 4 (56.8%, 52.0%, and 47.2%, respectively). Of the nongerminated seeds under condition 3, 75.0% were decayed and several fungi were isolated from decayed seeds. An inoculation test of isolated fungi Cylindrocarpon tenue, Cylindrocarpon sp., Fusarium oxysporum, and Botrytis sp. to seeds showed that these fungi caused seed decay. Our results indicated that nursery soil cover may not be suitable for T. dolabrata var. hondai seeds sown in nursery soil because of seed decay by pathogens.     

Growth and Mycorrhizal Dependency of Acacia mangium Willd. Inoculated with Three Vesicular Arbuscular Mycorrhizal Fungi in Lateritic Soil

Three VA-mycorrhizal fungi; Glomus occultum, Glomus aggregatum (local isolates) and G. mosseae (strain from Bangalore, India) were inoculated to assess their effect on growth of Acacia mangium in lateritic soil. All inoculations enhanced growth with respect to shoot height, root diameter, leaf area, chlorophyll content and biomass of A. mangium significantly compared to uninoculated control seedlings. G. occultum proved most efficient among the three. The mycorrhizal dependency factor indicated that the growth of A. mangium was 57% dependent on G. occultum, 47% on G. mosseae and 46% on Glomus aggregatum.     

Populations of adult fungus gnats and shore flies in British Columbia container nurseries as related to nursery environment,and incidence of fungi on the insects

Adult fungus gnat and shore fly populations were monitored in four container nurseries near Victoria, British Columbia. In general, populations of both insects were larger in summer than in winter. Also, both insects were more abundant in the nursery with a plastic-covered, soil floor, which favored moisture accumulation and build up of algae, liverworts and mosses, than in nurseries with other types of floors such as cement. Results of fungus isolations revealed that gnats and flies both harbor the seedling pathogens Botrytis cinerea and species of Fusarium and Phoma, plus several non-pathogenic fungi. The results are discussed in relation to nursery sanitation practices, their effects on gnat and fly populations, and some possible roles of these insects in seedling disease epidemiology.     

Microbial Inoculants for Sustainable Forests

Abstract

Fungal root pathogens are widespread and may cause substantial seedling losses in conifer nurseries. Furthermore, poor seedling survival and growth on reforestation sites results in reduced forest regeneration. Use of microbial inoculants for disease control and plant growth promotion has become an important endeavour. A microbial culture collection of 500 strains was assessed for biological control of fungal root pathogens and/or plant growth promotion of conifer seedlings. Seven of these strains showed significant suppres-sive effects on various soil-bome fungal pathogens. On Douglas fir, two strains, RAL3 and 64-3, reduced disease caused by Fusariumby. 7-42% in repeated growth room assays. The same strains significantly increased healthy stand of white spruce seedlings inoculated with Fusariumand Pythiumin a conifer nursery, and increased the survival of bare-root white spruce seedlings planted on a reforestation site by 19-23%. Both strains also significantly increased new root and total plant dry weights. Strain RAL3 in commercial formulation maintained a viable population of about log 8-9 cfu/ml for over a year when stored at 5°C. Strain survival on seed varied with conifer species. No decreases in bacterial populations were observed on seeds of jack pine or Douglas fir after 37 to 44 days storage at 5°C, but decreases were observed on seeds of white spruce and Scots pine. This study has provided candidate beneficial microbial inocu-lants which offer promise for development of commercial inoculants for the forestry industry.     

林地土壤及其真菌对毛红椿种子发芽及幼苗保存的影响

[目的]通过不同群落毛红椿天然林林地土壤及其真菌对种子萌发和幼苗存活影响的研究,探讨影响毛红椿天然更新的障碍因素。[方法]在江西九连山国家级自然保护区内的3个毛红椿天然优势群落中,分别取距离毛红椿母树3个不同距离(2.5、5.0、7.5 m)处的根区土壤和远离毛红椿母树(25 m以外)的非根区土壤,以非林地土为对照开展室内模拟播种和以根区土壤真菌的水或根系分泌物悬液对毛红椿幼苗进行灌根接种,观察种子萌发、幼苗存活和幼苗感病情况。[结果]各群落土壤中种子发芽均呈先上升后下降规律,且基本在第8 10天达到高峰;林地土与非林地土种子发芽率差异不显著,但林地根区土幼苗的存活率极显著低于非林地土和林地非根区土;距母树3个不同距离根区土的幼苗存活率差异不显著。灌根接种不同处理间幼苗存活率差异极显著,具体表现为接种RS2、RS5根区真菌的幼苗感病率显著高于根系分泌物及无菌水空白对照,且RS2真菌的根系分泌物悬浮液幼苗感病率显著高于其无菌水悬浮液。[结论]毛红椿种子萌发不受土壤环境的影响,但幼苗建成受根区范围内土壤的影响,根系分泌物和致病菌的互作显著降低幼苗保存率。由此推断,毛红椿根区土壤内存在幼苗的潜在致病菌及根系分泌物可增强其致病性。     

Use of soil solarization to improve growth of eucalyptus forest nursery seedlings in Argentina

Damping-off and root rot are major diseases affectingseedlings of Eucalyptus species in forestnurseries in temperate regions in Argentina. The mostcommon fungi associated with these diseases andaffecting the vigor of the root system are Fusarium and Pythium species. Two forestnursery experiments were conducted in the province ofBuenos Aires, Argentina, to determine the effect ofsoil solarization on growth of Eucalyptusviminalis seedlings and relate this effect to thepresence of pathogenic and native ectomycorrhizaepopulations in roots and nutrient availability insoil. Changes in populations of soilborne pathogenswere determined by a bioassay that relates theirpotential to induce disease. Changes in nativeectomycorrhizae were assesed by measuring colonizationlevels in roots. Nutrient availability was determinedby the amount of nitrates released by solarization.Solar heating decreased pathogenic and ectomycorrhizalinoculum potential and increased soil nitrates.Seedling growth in solarized seedbeds may be relatedto a low initial pathogenic population and/or toincreases in nitrate availability. Solarization mayinduce soil suppressiveness against re-establishmentof major seedling pathogens in treated soils.     

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.     

Assessing the Quality of Seedlings in Small-scale Nurseries in the Highlands of Cameroon: The Use of Growth Characteristics and Quality Thresholds as Indicators

In developing countries seedlings are often produced in small-scale nurseries as a means to raise tree planting materials and, provided they are of high quality, these can form the basis for successful forest plantation initiatives. This study uses morphological characteristics to assess the quality of seedlings of three tree species (Persea americana, Cola acuminata and Dacryodes edulis) in the Western Highlands of Cameroon. The growth characteristics and quality indicators of seedlings produced in two nursery categories were compared—three nurseries collaborating with the tree domestication program implemented by the World Agroforestry Centre (Category 1) and three nurseries collaborating with government and non-governmental projects (Category 2). Seedlings produced in nurseries in the first nursery category were found to be uniform in terms of growth characteristics and quality indicators. In the second category, there were relatively higher seedling proportions having measured parameters out of threshold standards for root to shoot ratio and for sturdiness quotient. Significant differences between nursery categories (at α = 0.05) were detected in root collar diameter of C. acuminata and D. edulis and in shoot dry weight of D. edulis. These differences could be attributed to better nursery management as a results of training and technical backstopping received by nursery operators in Category 1. The level of experience of nursery operators in both categories was found to be insufficient and greater exposure to skills and knowledge could enhance the production of high quality seedlings.     

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.     

In vitro effects of four ectomycorrhizal fungi, Boletus edulis, Rhizopogon roseolus, Laccaria laccata and Lactarius deliciosus on Fusarium damping off in Pinus nigra seedlings

Species of Fusarium cause damping off of forest seedlings resulting in considerable losses and oftentimes fungicides are used to control the disease. Such chemicals may be detrimental to the environment. Protection of forest seedlings against this disease must be focused on integrated management for in which biological control is one of the most important tools. Mycorrhizal fungi may provide protection against pathogen invasion of seedlings. In this study, the interactions between the mycorrhizal fungi Boletus edulis, Rhizopogon roseolus, Laccaria laccata and Lactarius deliciosus and damping off pathogens (Fusarium oxysporum and F. moniliforme) were investigated in vitro and mechanisms involved in the protection of damping off in P. nigra seedlings are also discussed. The effect of filtrates from mycorrhizal species on spore germination of F. oxysporum varied throughout the tests. At the end of the assay, the inhibitory effect only could be observed in the L.␣deliciosus treatment. None of the filtrates reduced spore germination in F. moniliforme. Finally, three of the four mycorrhizal species grown for 2 months in the substrate, L. laccata, L.␣deliciosus and B. edulis, increased the survival of Fusarium inoculated P.␣nigra seedlings.     

Comparative studies on the fine root status of healthy and declining spruce and beech trees in the Bavarian Alps and occurrence of Phytophthora and Pythium species

Investigations on root and crown status of spruce and beech were carried out on selected trees in the ‘Werdenfelser Land’ area (Bavarian Alps, Southern Germany). In addition, the association of fine root pathogens of the genera Phytophthora and Pythium with the trees’ rhizosphere was studied. In a variety of stands representing various site conditions, soil and root samples were taken from 12 spruce (Picea abies) pairs and eight beech (Fagus sylvatica) pairs. Each pair consisted of a healthy and a declining tree as indicated by crown transparency. The root status was characterized using a set of parameters, and correlations between crown and fine root status were observed. In spruce, most parameters decreased significantly with increasing crown transparency, whereas in beech, correlations were less pronounced. The total number of lateral roots per cm small root (diameter 2–5 mm) was significantly lower in both species for declining trees compared to healthy trees. Pythium spp. were isolated from 15 of 24 soil samples taken from under spruce, and from eight of 16 samples from under beech. Phytophthora citricola was found in two beech stands only. Among the isolated species, Pythium anandrum, Pythium inflatum and Pythium acrogynum were identified according to morphological features. After polymerase chain reaction‐restriction fragment length polymorphism analysis, residual Pythium isolates were assigned to four different groups. No crown transparency‐dependent differences in isolation frequency were found. In soil infestation tests, all species tested caused root damage on both young spruce and beech plants, with P. citricola being the most aggressive pathogen. Additionally, Pythium‐infected beeches showed severe leaf chloroses and necroses. Due to their low isolation frequency, Phytophthora spp. are not considered to play a major role in the decline of spruce and beech in the investigated area. Pythium spp., however, were isolated frequently, showed pathogenicity towards the fine roots of spruce and beech, and are therefore considered to be at least contributing factors in the decline of Bavarian mountain forests.