Nutrients are assimilated efficiently by Lymantria dispar caterpillars from the mature leaves of trees in the Salicaceae

The efficient aquisition of nutrients from leaves by insect herbivores increases their nutrient assimilation rates and overall fitness. Caterpillars of the gypsy moth (Lymantria dispar L.) have high protein assimilation efficiencies (PAE) from the immature leaves of trees such as red oak (Quercus rubra) and sugar maple (Acer saccharum) (71–81%) but significantly lower PAE from their mature leaves (45–52%). By contrast to this pattern, both PAE and carbohydrate assimilation efficiencies (CAE) remain high for L. dispar larvae on the mature leaves of poplar (Populus alba × Populus tremula) grown in greenhouse conditions. The present study tests two alternative hypotheses: (i) outdoor environmental stresses cause decreased nutrient assimilation efficiencies from mature poplar leaves and (ii) nutrients in the mature leaves of trees in the poplar family (Salicaceae) remain readily available for L. dispar larvae. When poplar trees are grown in ambient outdoor conditions, PAE and CAE remain high (approximately 75% and 78%, respectively) in L. dispar larvae, in contrast to the first hypothesis. When larvae feed on the mature leaves of species in the Salicaceae [aspen (Populus tremuloides), cottonwood (Populus deltoides), willow (Salix nigra) and poplar], PAE and CAE also remain high (68–76% and 72–92%, respectively), consistent with the second hypothesis. Larval growth rates are strongly associated with protein assimilation rates, and more strongly associated with protein assimilation rates than with carbohydrate assimilation rates. It is concluded that tree species in the Salicaceae are relatively high‐quality host plants for L. dispar larvae, in part, because nutrients in their mature leaves remain readily available.     

Olfactory response of three parasitoid species (Hymenoptera: Braconidae) to volatiles of guavas infested or not with fruit fly larvae (Diptera: Tephritidae)

The olfactory responses of the native parasitoids Doryctobracon areolatus (Szépligeti) and Asobara anastrephae (Muesebeck) and of the exotic parasitoid Diachasmimorpha longicaudata (Ashmead) to guava (Psidium guajava L.) infested or not with fruit fly larvae were evaluated. D. areolatus and D. longicaudata females responded to the odors of uninfested rotting guavas, although D. areolatus was also attracted to fruits at the initial maturation (turning) stage. The females of these species recognized the volatiles of guavas containing Ceratitis capitata (Wied.) larvae. However, in bioassays involving fruits with larvae of different instars, D. longicaudata females were not able to separate between fruits containing C. capitata larvae at the initial instars and larvae at the third instar. In the evaluations of volatiles released by guavas containing C. capitata and Anastrepha fraterculus (Wied.) larvae, the D. longicaudata females were oriented toward the volatiles of fruits containing both host species, but differed significantly from volatiles of guavas containing C. capitata larvae. The D. areolatus females also showed responses to both species, although with a preference for volatiles of fruits containing A. fraterculus larvae. The A. anastrephae females were oriented toward the odors of fruits infested with both fruit fly species. In the shade house, D. longicaudata females were oriented to volatiles of rotting fruits containing larvae or not, but could not significantly differentiate between hosts. D. areolatus females were not attracted toward fruits on the ground in the shade house, regardless of host, suggesting that this parasitoid does not forage on fallen fruits.     

光肩星天牛体内纤维素酶活性及其与寄主选择的关系（英文）

为了揭示光肩星天牛Anoplophora glabripennis Motschulsky纤维素酶与寄主选择的关系, 以4种不同寄主树种（新疆杨Populus alba var. pyramidalis Bunge、箭杆杨Populus nigra var. thevestina (Dode) Bean、合作杨Populus simonii × Populus pyramidalis cv. opera Hsu和漳河柳Salix matsudana f. lobato-glandulosa Faug et Liu来源的光肩星天牛幼虫, 和以取食5种不同树种（臭椿Ailanthus altissima、毛白杨Populus tomentosa、合作杨、旱柳Salix matsudana Koidz.和复叶槭Acer negundo Linn.）的光肩星天牛成虫为实验对象, 测定其纤维素酶活性。结果表明: 不同寄主来源的光肩星天牛幼虫内切-β-1,4-葡聚糖酶活性在1.36~2.71 μmol葡萄糖/ (g FW·h)之间, β-葡萄糖苷酶活性在2.57~4.86 μmol葡萄糖/(g FW·h)之间;取食不同树种的光肩星天牛成虫内切-β-1,4-葡聚糖酶活性在4.08~9.27 μmol葡萄糖/(g FW·h)之间, β-葡萄糖苷酶活性在2.87~6.08 μmol葡萄糖/ (g FW·h)之间。不同寄主树种来源的光肩星天牛幼虫体内纤维素酶活性无显著性差异, 取食与否以及取食树种的不同对光肩星天牛成虫纤维素酶活有较大影响。     

Compound effects of induced plant responses on insect herbivores and parasitoids: implications for tritrophic interactions

1. Induced plant responses can affect herbivores either directly, by reducing herbivore development, or indirectly, by affecting the performance of natural enemies. Both the direct and indirect impacts of induction on herbivore and parasitoid success were evaluated in a common experimental system, using clonal poplar trees Populus nigra (Salicales: Salicaceae), the gypsy moth Lymantria dispar (L.) (Lepidoptera: Lymantriidae), and the gregarious parasitoid Glyptapanteles flavicoxis (Marsh) (Hymenoptera: Braconidae). 2. Female parasitoids were attracted to leaf odours from both damaged and undamaged trees, however herbivore‐damaged leaves were three times more attractive to wasps than undamaged leaves. Parasitoids were also attracted to herbivore larvae reared on foliage and to larval frass, but they were not attracted to larvae reared on artificial diet. 3. Prior gypsy moth feeding elicited a systemic plant response that retarded the growth rate, feeding, and survival of gypsy moth larvae, however induction also reduced the developmental success of the parasitoid. 4. The mean number of parasitoid progeny emerging from hosts fed foliage from induced trees was 40% less than from uninduced trees. In addition, the proportion of parasitised larvae that survived long enough to issue any parasitoids was lower on foliage from induced trees. 5. A conceptual and analytical model is provided to describe the net impacts of induced plant responses on parasitoids, and implications for tritrophic interactions and biological control of insect pests are discussed.     

Host-associated allozyme variation in tree cambium miners,Phytobia spp. (Diptera: Agromyzidae)

The larvae of the agromyzid flies that belong to the genus Phytobia Lioy feed by mining in the differentiating xylem just below the cambium of growing forest trees. The genus, which is apparently one of the most primitive groups in the Agromyzidae, comprises over 50 currently recognized species. Most of the species are mono- or oligophagous, and the host plants belong to numerous genera in about 60 families. Thus, Phytobia is an attractive candidate for studies on the evolution of insect-plant relationships. In spite of this, the taxonomy of Phytobia is currently poorly understood, mainly because the morphological differences between species are small. We used allozyme electrophoresis to investigate whether molecular markers could be used to separate and identify species in Phytobia, and to study the patterns of host use in the group. For this, we collected Phytobia larvae from eight host tree species occurring in southern Finland. An analysis of 10 variable allozyme loci showed that there are probably five species of Phytobia that feed on the hosts included in our study: one occurs on birches (Betula pubescens Ehrh. and B. pendula Roth) and alders (Alnus incana (L.) Moench and A. glutinosa (L.) Gaertn.), one on rowan (Sorbus aucuparia L.), and three species with overlapping feeding ranges on aspen (Populus tremula L.) and two willow species (Salix phylicifolia L. and S. caprea L.). Because birches and alders belong to the plant family Betulaceae, rowan to Rosaceae, and aspen and willows to Salicaceae, the host associations of the individual fly species can be explained by the taxonomic affinities of the hosts. However, our results also show that on a larger scale the evolution of host-plant associations in Phytobia cannot be explained by strict parallel cladogenesis (cospeciation) between the flies and their hosts.     

Notes on the biology and ecology of the parasitoids of the poplar clearwing moth, Paranthrene tabaniformis (Rott.) (Lep., Sesiidae) in Bulgaria. II. Eriborus terebrans (Gravenhorst, 1826) (Hym., Ichneumonidae)

The biology and ecology of Eriborus terebrans (Grav.), a parasitoid of the poplar clearwing moth, Paranthrene tabaniformis (Rott.), were studied during the period 1987–98. One-year-old poplar ( Populus spp.) shoots infested with P. tabaniformis larvae were collected from poplar seedlings at 11 localities in Bulgaria and examined in both field and laboratory conditions. Eriborus terebrans was recorded in seven localities as a solitary internal larval parasitoid of P. tabaniformis which developed two generations in early and mid-stage host larvae. Eriborus terebrans overwintered as a larva in P. tabaniformis overwintering larvae. In the field adult parasitoids of the overwintering generation appeared between late April/early May, and June or July. The peak activity of E. terebrans adults only coincided with the beginning of host emergence, which resulted in low levels of parasitism, being no more than 6.2%. Parasitoid adults of the summer generation appeared in late June–mid August. In this period enough larvae of the host were suitable for attacking and parasitism reached 24.4–39% in some cases. The average mortality of P. tabaniformis overwintering larvae caused by this parasitoid in Bulgaria during the period of the study was 4.7%. A significant part of the parasitized P. tabaniformis larvae constructed tunnel structures of frass and silk threads over the external openings of the galleries. It is possible that these structures protect the parasitoid cocoons from natural enemies – hyperparasitoids and predators.     

Host plant choice and location by larvae of the wheat bulb fly (Delia coarctata)

Wheat bulb fly (Delia coarctata Fallen, Diptera: Anthomyiidae) is an important pest of winter wheat in the eastern half of the UK, and in northern and eastern Europe. The larvae must find a host plant and invade a tiller soon after hatching in late January. Chemical controls are costly and weather conditions may reduce their efficacy or prevent their application. Post‐emergence control relies on organophosphate insecticides, which may soon be withdrawn due to concerns about their negative health and environmental effects. Winter wheat (Triticum aestivum L.) is the preferred cereal host, but other winter cereals and related grasses may also be attacked, while oats (Avena spp.) are shunned. In choice test bioassays, neonate larvae chose couch grass (Elytrigia repens (L.) Nevski syn. Elymus repens (L.) Gould, Agropyearon repens (L.) Beauv.) seedlings and exudates over wheat seedlings and exudates, and exhibited geotaxis and negative phototaxis. Analysis of larval trails in choice test bioassays of seedling exudates showed that couch exudates are more attractive than wheat exudates, and that wheat exudates are more arrestant than couch exudates. This suggests that infochemicals isolated from couch, wheat, and oats could be used in wheat bulb fly control; possible delivery mechanisms are discussed. These findings, previous research, and a comparison of the phenologies and geographical distributions of D. coarctata and its hosts suggest that E. repens is the natural host of D. coarctata.     

Hybrid poplar clones with Populus maximowiczii parentage demonstrate postoviposition antibiosis to Cryptorhynchus lapathi (Coleoptera: Curculionidae)

Damage caused by the poplar-and-willow borer, Cryptorhynchus lapathi (L.) (Coleoptera: Curculionidae), is reported to vary among hybrid poplar clones. We evaluated oviposition preferences and larval success in four hybrid poplars on potted and field-planted trees. Oviposition occurred somewhat less frequently and abundantly on two clones with Populus maximowiczii Henry parentage in field-planted and potted trees, and significantly fewer larvae survived to adulthood on those clones. No adults emerged from field-planted NM 6 (Populus nigra L. x P. maximowiczii) and four emerged from TM 256-28 (Populus trichocarpa Torrey & Gray x P. maximowiczii) on which male-female pairs of C. lapathi had been caged. In contrast, 50 and 140 adults emerged over the same 2-yr period from two susceptible clones (n = 20), TD 52-226 (P. trichocarpa x Populus deltoides Bartram ex Marshall) and TN 302-9 (P. trichocarpa x P. nigra), respectively. Thus, resistance expressed by clones with P. maximowiczii parentage partially involves decreased levels of oviposition, but more significantly, antibiosis in resistant clones prevents the development of larvae, probably in early spring.     

Functional analysis of the Kunitz trypsin inhibitor family in poplar reveals biochemical diversity and multiplicity in defense against herbivores

We investigated the functional and biochemical variability of Kunitz trypsin inhibitor (KTI) genes of Populus trichocarpa x Populus deltoides. Phylogenetic analysis, expressed sequence tag databases, and western-blot analysis confirmed that these genes belong to a large and diverse gene family with complex expression patterns. Five wound- and herbivore-induced genes representing the diversity of the KTI gene family were selected for functional analysis and shown to produce active KTI proteins in Escherichia coli. These recombinant KTI proteins were all biochemically distinct and showed clear differences in efficacy against trypsin-, chymotrypsin-, and elastase-type proteases, suggesting functional specialization of different members of this gene family. The in vitro stability of the KTIs in the presence of reducing agents and elevated temperature also varied widely, emphasizing the biochemical differences of these proteins. Significantly, the properties of the recombinant KTI proteins were not predictable from primary amino acid sequence data. Proteases in midgut extracts of Malacosoma disstria, a lepidopteran pest of Populus, were strongly inhibited by at least two of the KTI gene products. This study suggests that the large diversity in the poplar (Populus spp.) KTI family is important for biochemical and functional specialization, which may be important in the maintenance of pest resistance in long-lived plants such as poplar.     

A field study with geometrid moths to test the coevolution hypothesis of red autumn colours in deciduous trees

Red autumn colouration of trees is the result of newly synthesized anthocyanin pigments in senescing autumn leaves. As anthocyanin accumulation is costly and the trait is not present in all species, anthocyanins must have an adaptive significance in autumn leaves. According to the coevolution hypothesis of autumn colours, red autumn leaves warn herbivorous insects – especially aphids that migrate to reproduce in trees in the autumn – that the tree will not be a suitable host for their offspring in spring due to a high level of chemical defence or lack of nutrients. The signalling allows trees to avoid herbivores and herbivores to choose better host trees. In this study the coevolution hypothesis was tested with four deciduous tree species that have red autumn leaf colouration – European aspen (Populus tremula L.) (Salicaceae), rowan (Sorbus aucuparia L.) (Rosaceae), mountain birch [Betula pubescens ssp. czerepanovii (NI Orlova) Hämet‐Ahti], and dwarf birch (Betula nana L.) (Betulaceae), and with two generalist herbivores, the autumnal moth [Epirrita autumnata (Borkhausen)] and the winter moth [Operophtera brumata (L.)] (both Lepidoptera: Geometridae). Anthocyanin concentrations of autumn leaves were determined from leaf samples and the growth performance parameters of the moth larvae on the study trees were measured in the spring. Trees with higher anthocyanin concentration in the autumn were predicted to be low‐quality food for the herbivores. Our results clearly showed that anthocyanin concentration was not correlated with the growth performance of the moths in any of the studied tree species. Thus, our study does not support the coevolution hypothesis of autumn colours.     

Regeneration of Populus nigra transgenic plants expressing a Kunitz proteinase inhibitor (KTi3) gene

Transgenic poplar (Populus nigra, cv. Jean Pourtet) plants were recovered as a result of Agrobacterium tumefaciens-mediated transformation performed with EHA105 pBI-KUN strain. Plasmid pBI-KUN contains a 650 bp insert derived from the soybean (Glycine max L.) KTi₃, gene, coding for a Kunitz trypsin proteinase inhibitor. A total of 58 independent transgenic lines were obtained from 200 co-cultivated leaf explants. Southern blot hybridization analysis demonstrated the presence of KTi₃ gene in the poplar genome. Northern blot analysis of different kanamycin-resistant plantlets confirmed the accumulation of KTi₃ mRNA and revealed different levels of expression. The trypsin inhibitory activity was determined in poplar transgenic tissues by means of specific assay. Moreover, the trypsin-like digestive proteinases of the polyphagous moth Lymantria dispar (Lepidoptera, Lymantriidae) and Clostera anastomosis (Lepidoptera, Notodontidae) were detected and inhibited in vitro by Kunitz proteinase inhibitor from selected transgenic plants. Two insect bioassays were performed on P. nigra transgenic plant lines, using larvae of the above mentioned insects. In both cases larval mortality and growth as well as pupal weight were not significantly affected when the insects were fed on transgenic leaves and control leaves, respectively.     

The influence of plant species on attraction and host acceptance inCotesia glomerata (Hymenoptera: Braconidae)

Females of the larval parasitoidCotesia glomerata (L.) use plant-associated cues to locate their lepidopteran host,Pieris rapae L. In this study we investigated the influence of four host plant species,Brassica oleracea var.acephala (‘Vates’ kale),Tropaeolum majus (nasturtium),Lunaria annua (honesty), andCleome spinosa (spider flower), on two components of the host selection process inC. glomerata, namely, attraction and host acceptance. Choice tests in a flight tunnel showed that parasitoids were attracted to some host plant species more than to others in the absence of host larvae.B. oleracea was the most attractive plant species, followed byL. annua, T. majus, andC. spinosa. In previous studies it was shown thatB. oleracea carries highly suitable hosts forC. glomerata and that, in the field, parasitization rates on this plant were the highest. When host larvae were reared on the four host plant species and then transferred to a common substrate (B. oleracea var.capitata, cabbage), plant species that had served as diet for the hosts did not have a significant effect on acceptance for parasitization. Thus, parasitoids were attracted to host plant species differentially, but they did not discriminate among host larvae based on the dietary history of their hosts. ForC. glomerata, it appears that phytochemistry mediates host selection more by influencing parasitoid attraction than it does by affecting host acceptance.     

Use of pheromone mimic to cause the disintegration and collapse of colonies of tent caterpillars (Malacosoma spp.)

The disruption of the trail-based communication system of eastern tent caterpillars (ETC) ( Malacosoma americanum ) and forest tent caterpillars (FTC) ( M. disstria ) by spraying host trees with a trail pheromone mimic caused the disintegration and collapse of young colonies. A 1-ppm formulation of 5β-cholestan-3-one or a control formulation was sprayed on trees either before or after eclosion of the caterpillars in the spring. For pre-eclosion trials, an average of 92% of the FTC colonies on control trees, but only 17% of colonies on treated trees consisted of viable, cohesive aggregates when survival was assessed 5–23 days post-spray. The remaining colonies had either completely disappeared or were reduced to small, disintegrated fragments. For the ETC, 100% of colonies on control trees and an average of 12% of colonies on treated trees consisted of cohesive aggregates inhabiting tents when survival was assessed 3–41 days post-spray. The remaining colonies were completely destroyed, reduced to scattered individuals, or to small, disintegrated, shelter-less fragments. For post-eclosion trials with the FTC in which colonies were sprayed during their first or second larval stadium, an average of 63% of individuals in control groups but only 7% in treatment groups were still on the sprayed trees 5–18 days post-spray. For the ETC, colonies were largely unaffected when trees were not sprayed until after the caterpillars had eclosed and established tents. For all trials, the loss of individuals on treated trees was largely attributable to the inability of dispersed and isolated first or second instar caterpillars to maintain secure purchase, causing them to fall from trees and perish. To our knowledge, this is the first study to investigate the potential of manipulating populations of pest species by interfering with a trail-based, chemical communication system.     

Growth of an invertebrate shredder on native (Populus) and non-native (Tamarix, Elaeagnus) leaf litter

1. Large-scale invasions of riparian trees can alter the quantity and quality of allochthonous inputs of leaf litter to streams and thus have the potential to alter stream organic matter dynamics. Non-native saltcedar ( Tamarix sp.) and Russian olive ( Elaeagnus angustifolia ) are now among the most common trees in riparian zones in western North America, yet their impacts on energy flow in streams are virtually unknown.
2. We conducted a laboratory feeding experiment to compare the growth of the aquatic crane fly Tipula (Diptera: Tipulidae) on leaf litter from native cottonwood ( Populus ) and non-native Tamarix and Elaeagnus . Tipula showed positive growth on leaf litter of all three species; however, after 7 weeks, larvae fed Tamarix leaves averaged 1.7 and 2.5 times the mass of those fed Elaeagnus and Populus , respectively. Tipula survival was highest on Populus , intermediate on Tamarix and lowest on Elaeagnus .
3. High Tipula growth on Tamarix probably reflects a combination of leaf chemistry and morphology. Conditioned Tamarix leaf litter had intermediate carbon : nitrogen values (33 : 1) compared to Populus (40 : 1) and Elaeagnus (26 : 1), and it had intermediate proportions of structural carbon (42%) compared to Elaeagnus (57%) and Populus (35%). Tamarix leaves are also relatively small and possibly more easily ingested by Tipula than either Elaeagnus or Populus .
4. Field surveys of streams in the western U.S.A. revealed that Tamarix and Elaeagnus leaf packs were rare compared to native Populus , probably due to the elongate shape and small size of the non-native leaves. Thus we conclude that, in general, the impact of non-native riparian invasion on aquatic shredders will depend not only on leaf decomposition rate and palatability but also on rates of leaf litter input to the stream coupled with streambed retention and subsequent availability to consumers.     

Host status of grapefruit and Valencia oranges for Anastrepha serpentina and Anastrepha ludens (Diptera: Tephritidae)

Anastrepha serpentina (Wiedemann) (Diptera: Tephritidae) is sporadically captured in the Rio Grande Valley of Texas. Although its preferred hosts are in the Sapotaceae family, several varieties of Citrus, including grapefruit and oranges are listed as alternate hosts. Although Mexican fruit fly, Anastrepha ludens (Loew), is known to be a major pest of Citrus, doubt exists as to the status of Citrus as a breeding host for A. serpentina. To evaluate the host status of commercial Citrus for A. serpentina we compared oviposition and development with that of A. ludens under laboratory conditions with 'Rio Red' grapefruit (Citrus paradisi MacFayden) and 'Valencia' oranges [Citrus sinensis (L.) Osbeck] in different stages of maturity. Both fly species oviposited in early season fruit in which the eggs and larvae died in the fruit albedo. Survival of either species to the adult stage occurred in later season grapefruit. In oranges, no A. serpentina larvae survived compared with 150 A. ludens surviving to adults. Survival on both Citrus species was much lower for A. serpentina, only approximately 5% of eggs eclosed into larvae in grapefruit compared with approximatley 50% for A. ludens. In oranges approximately 16% of A. serpentina eggs eclosed compared with approximately 76% for A. ludens. In grapefruit, only one fourth as many A. serpentina larvae survived to the adult stage compared with A. ludens. Additional experiments were performed in a greenhouse on small, caged trees of la coma (Sideroxylon celastrinum H.B.K.), a Texas species of Sapotaceae. The A. serpentina females readily oviposited into these berries and normal adults emerged. The present low incidence of the adults, coupled with the high mortality during development of the larvae, suggests that Texas citrus is unlikely to support a breeding population of A. serpentina.     

Effects of leaf maturity and wind stress on the nutrition of the generalist caterpillar Lymantria dispar feeding on poplar

The growth rates of insect herbivores commonly decrease when they feed on mature leaves due to the combined effects of several nutritional and physiological mechanisms. Environmental stresses during leaf development may also decrease herbivore performance. The present study tests two main hypotheses to help clarify the importance of these factors for the nutrition and growth of an insect herbivore: (i) decreases in nutrient levels, consumption rates and nutrient assimilation efficiencies impact negatively on herbivores feeding on mature leaves and (ii) wind stress has a negative impact on herbivores feeding on mature leaves. The results show that mature poplar (Populus alba × Populus tremula) leaves have decreased levels of protein and increased levels of fibre, and that growth rates of gypsy moth (Lymantria dispar L.) are decreased on mature leaves in association with decreased consumption rates. However, in contrast to the first hypothesis, protein and carbohydrate are assimilated efficiently (74–82% and 84–87%, respectively) from immature and mature poplar leaves. The larvae are able to chew mature leaves as efficiently as immature leaves, potentially maximizing nutrient extraction. By contrast to the second hypothesis, wind‐stressed leaves have no significant detrimental effects on nutrient assimilation efficiencies, and the lower growth rates of L. dispar larvae feeding on mature wind‐stressed leaves can be explained by lower consumption rates. Therefore, the availability of nutrients to herbivores feeding on mature tree leaves is not necessarily impacted by lower assimilation efficiencies, even when leaves develop under wind stress. These results help explain some of the large variation between the nutritional qualities of trees for forest Lepidoptera.     

Mycorrhizal inoculum potentials of pure reclamation materials and revegetated tailing sands from the Canadian oil sand industry

Recent improvements in the management of oil sand tailings used by the Canadian oil sand industry have resulted in the production of composite tailing sands (CT): a new challenging material for reclamation work. Jack pine (Pinus banksiana Lamb.), hybrid poplar (Populus deltoides Bartr. ex Marsh. ×Populus nigra L.) and red clover (Trifolium pratense L.) plants were used in an 8-week greenhouse bioassay to evaluate the mycorrhizal inoculum potential of CT. This inoculum potential was compared with that of three other reclamation materials [common tailing sands (TS), deep overburden (OB) and muskeg peat (MK)], and with three sites reclaimed in 1982 (R82), 1988 (R88) and 1999 (R99). CT was devoid of active mycorrhizal propagules while all other materials showed some level of inoculum potential. Arbuscular mycorrhizal fungi were observed on roots of clover or poplar grown in TS, OB, and all substrates containing peat (MK, R82, R88 and R99). Pine roots were also colonized by vesicle-forming hyphae of an unidentified fine endophyte and by dark septate fungi. Ectomycorrhizas (ECM) were observed on pine and poplar grown in OB, MK, and in soils from the two older reclaimed sites (R82 and R88). Using morpho- and molecular typing, six ECM fungi were identified to the genus or species level: Laccaria sp., Thelephora americana, Wilcoxina sp. (E-strain), Tuber sp. (I-type), a Sebacinoid, and a Pezizales species. Laccaria sp. and Wilcoxina sp. were the most frequently observed ECM species.