John F. Walker

Hyperdiversity of ectomycorrhizal fungus assemblages on oak seedlings in mixed forests in the southern Appalachian Mountains

Diversity of ectotrophic mycobionts on outplanted seedlings of two oak species (Quercus rubra and Quercus prinus) was estimated at two sites in mature mixed forests in the southern Appalachian Mountains by sequencing nuclear 5.8S rRNA genes and the flanking internal transcribed spacer regions I and II (ITS). The seedlings captured a high diversity of mycorrhizal ITS‐types and late‐stage fungi were well represented. Total richness was 75 types, with 42 types having a frequency of only one. The first and second order jackknife estimates were 116 and 143 types, respectively. Among Basidiomycetes, tomentelloid/thelephoroid, russuloid, and cortinarioid groups were the richest. The ascomycete Cenococcum geophilum was ubiquitously present. Dominant fungi included a putative Tuber sp. (Ascomycetes), and Basidiomycetes including a putative Craterellus sp., and Laccaria cf. laccata. Diversity was lower at a drier high elevation oak forest site compared to a low elevation mesic cove — hardwood forest site. Fungal specificity for red oak vs. white oak seedlings was unresolved. The high degree of rarity in this system imposes limitations on the power of community analyses at finer scales. The high mycobiont diversity highlights the potential for seedlings to acquire carbon from mycelial networks and confirms the utility of using outplanted seedlings to estimate ectomycorrhizal diversity.

INHIBITION OF SEEDLING SURVIVAL UNDER RHODODENDRON MAXIMUM (ERICACEAE) : COULD ALLELOPATHY BE A CAUSE?

In the southern Appalachian mountains a subcanopy species, Rhododendron maximum, inhibits the establishment and survival of canopy tree seedlings. One of the mechanisms by which seedlings could be inhibited is an allelopathic effect of decomposing litter or leachate from the canopy of R. maximum (R.m.) on seed germination, root elongation, or mycorrhizal colonization. The potential for allelopathy by R.m. was tested with two bioassay species (lettuce and cress), with seeds from four native tree species, and with three ectomycorrhizal fungi. Inhibitory influences of throughfall, fresh litter, and decomposed litter (organic layer) from forest with R.m. (+R.m. sites) were compared to similar extractions made from forest without R.m. (-R.m. sites). Throughfall and leachates of the organic layer from both +R.m. and -R.m. sites stimulated germination of the bioassay species above that of the distilled water control, to a similar extent. There was an inhibitory effect of leachates of litter from +R.m. sites on seed germination and root elongation rate of both bioassay species compared with that of litter from -R.m. sites. Native tree seed stratified in forest floor material from both forest types had a slightly higher seed germination rate compared with the control. A 2-yr study of seed germination and seedling mortality of two tree species, Quercus rubra and Prunus serotina, in field plots showed no significant influence of litter or organic layer from either forest type. Incorporating R.m. leaf material into the growth medium in vitro depressed growth of one ectomycorrhizal species but did not affect two other species. Leaf material from other deciduous tree species depressed ectomycorrhizal growth to a similar or greater extent as leaf material from R.m. In conclusion, R.m. litter can have an allelopathic effect on seed germination and root elongation of bioassay species as well as some ectomycorrhizal species. However, this allelopathic affect is not manifest in field sites and is not likely to be an important cause for the inhibition of seedling survival within thickets of R.m.

Effects of Rhododendron maximum Thickets on Tree Seed Dispersal, Seedling Morphology, and Survivorship

In the southern Appalachian forests, the regeneration of canopy trees is severely inhibited by Rhododendron maximum L., an evergreen understory shrub producing dense thickets. While light availability is a major cause, other factors may also contribute to the absence of tree seedlings under R. maximum. We examined the effects of R. maximum on several life history stages of tree species, including seed dispersal, seed bank germination, seedling growth, and survivorship. We found no significant effect of R. maximum on seed reaching the forest floor for Acer rubrum, Liriodendron tulipifera, Quercus rubra, Quercus prinus, Carya spp., and Nyssa sylvatica. This indicates that either seed output of maternal trees rooted within the thicket were unaffected by R. maximum or seed dispersal from surrounding areas into thickets compensated for a lower seed production of canopy trees rooted in the thickets. Germination of tree seeds (A. rubrum, L. tulipifera, Q. rubra, and Betula lenta) from the seed bank also was not reduced by leaves and substrates within the thickets. Seedling mortality of all species (Q. rubra, Prunus serotina, and Tsuga canadensis) planted in our experimental plots was up to fivefold higher in thickets of R. maximum compared with those outside the thickets. The order of mortality under the R. maximum thickets, \documentclass{aastex} \usepackage{amsbsy} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{bm} \usepackage{mathrsfs} \usepackage{pifont} \usepackage{stmaryrd} \usepackage{textcomp} \usepackage{portland,xspace} \usepackage{amsmath,amsxtra} \usepackage[OT2,OT1]{fontenc} \newcommand\cyr{ \renewcommand\rmdefault{wncyr} \renewcommand\sfdefault{wncyss} \renewcommand\encodingdefault{OT2} \normalfont \selectfont} \DeclareTextFontCommand{\textcyr}{\cyr} \pagestyle{empty} \DeclareMathSizes{10}{9}{7}{6} \begin{document} \landscape

Suppression of ectomycorrhizae on canopy tree seedlings in Rhododendron maximum L. (Ericaceae) thickets in the southern Appalachians

Prunus> Quercus> Tsuga

Variation in Soil and Forest Floor Characteristics Along Gradients of Ericaceous, Evergreen Shrub Cover in the Southern Appalachians

\end{document} , was consistent with the shade tolerance ranking of these species. Loss of Tsuga seedlings was attributed to burial by litter rather than shade. Surviving seedlings of Quercus and Prunus in R. maximum thickets were taller than those outside the thickets, but the seedlings in R. maximum thickets produced significantly fewer leaves, smaller total leaf area, leaf mass, and stem mass. Leaf N (%) was significantly higher in Quercus seedlings in R. maximum thickets compared with seedlings outside the thickets. Moreover, no difference was found in leaf N (%) between forest types for Prunus and Tsuga, indicating that seedlings in R. maximum thickets were not N limited. Rather, light limitation, herbivory, and litter fall contributed to the lack of tree regeneration under R. maximum thickets.

Ectomycorrhizal sporophore distributions in a southeastern Appalachian mixed hardwood/conifer forest with thickets of Rhododendron maximum

The potential for seasonal dynamics in ectomycorrhizal (EM) fungal assemblages has important implications for the ecology of both the host trees and the fungal associates. We compared EM fungus distributions on root systems of out-planted oak seedlings at two sites in mixed southeastern Appalachian Mountain forests at the Coweeta Hydrologic Laboratory in North Carolina, from samples taken in mid-July and early September. Species level EM fungus type specificity, and identification in some cases, was enabled by direct sequencing of the mycobionts from the seedling roots. Seventy-four EM fungal ITS types were documented, most of which occurred only in the midsummer or early-fall samples, respectively. Cenococcum geophilum (morphotyped) was ubiquitously present and accounted for the majority of root tips sampled. Abundance and relative frequency of types other than C. geophilum were significantly higher in the July samples, while C. geophilum was significantly more frequent and abundant in September. Several generalistic dominants were found fairly equally at both sites and on both sample dates. Other taxa with relatively high frequency were recovered from both sites and tree seedling species, but were reliable indicators occurring primarily in the July sample (e.g., Laccaria cf laccata). Notable shifts in mycobiont dominance were apparent in relation to sample date, including increases in Cortinarius spp. richness, decreases in Thelephoraceae richness, and the disappearance of Amanita spp. types in the early fall compared to midsummer samples. However, diversity and rarity were high and differences in overall community composition (other than C. geophilum) by season were not significant based on multi-response permutation procedures. Although these results based on a single growing season are preliminary, changes in abundance and frequency, detection of significant indicator species, and the apparent systematic affinities of shifting EM types support the potential for seasonal variability in EM associations in this system.

Application of fungistatics in soil reduces N uptake by an arctic ericoid shrub (Vaccinium vitis-idaea)

Abstract Thickets of Rhododendronmaximum (Ericaceae) (Rm) in the southern Appalachians severely limit regeneration of hardwood and coniferous seedlings. Experimental blocks were established in and out of Rm thickets in a mature, mixed hardwood/conifer forest in Macon County, N.C. Litter and organic layer substrates were removed, composited and redistributed among plots within the blocks (except for control plots). Seedlings of northern red oak (Quercusrubra) and eastern hemlock (Tsugacanadensis) were planted in the plots and harvested at the end of the first and second growing seasons. Litter manipulation had no effect on total mycorrhizal colonization, but the distribution of Cenococcumgeophilum mycorrhizae was altered. After the first year, percent mycorrhizal colonization of hemlocks not in Rm thickets (62%) was at least three times higher than in Rm thickets (19%), and the ramification index (no. of mycorrhizae cm–1) had increased by more than a factor of four (2.83 versus 0.61). In addition, colonization of 1-year-old hemlocks by C. geophilum was significantly higher within blocks with (10.4%) than without (4.6%) Rm. Differences in mycorrhizal colonization, ramification indices and colonization by C.geophilum were absent or less pronounced on 2-year-old hemlocks and 1- and 2-year-old oak seedlings. The biomasses of first year oak roots and shoots and second year shoots were 50% less in Rm thickets. Biomasses of first year hemlock roots and second year shoots were also reduced. Mycorrhizal parameters were correlated with some growth parameters only for hemlock seedlings, but did not explain most of the variation observed.

A molecular phylogeny of the Russulales including agaricoid, gasteroid and pleurotoid taxa

Inhibition of canopy tree recruitment beneath thickets of the evergreen shrubs Rhododendron maximum L. and Kalmia latifolia L. has long been observed in Southern Appalachian forests, yet the mechanisms of this process remain unresolved. We present a first-year account of suppression of oak seedlings in relation to Rhododendron and Kalmia basal area, light and resource availability, seedling performance and the rates of seedling damage (i.e., herbivory). We found no evidence of first-year seedling suppression or significant resource deficiencies beneath thickets of K. latifolia in mature mixed hardwood stands. Suppression beneath R. maximum was apparent during the first growing season. We found that seedling biomass, light availability prior to canopy closure, and seedling tissue C:N ratios were negatively correlated with R. maximum basal area. Basal area of R. maximum was positively correlated with seedling mortality rates, soil [Al], and early-growing season leaf herbivory rates. Seedling growth was positively correlated with light and tissue C:N, while negatively correlated with soil [Al]. Overall, our results support the inhibition model of shade-mediated carbon limitation beneath dense understory shrubs and indicate the potential importance of herbivory and aluminum toxicity as components of a suppression mechanism beneath R. maximum thickets. We present a causal model of first year inhibition beneath R. maximum in the context of our findings and the results of prior studies.