A high-quality reference genome for Fraxinus pennsylvanica for ash species restoration and research.

Green ash (Fraxinus pennsylvanica) is the most widely distributed ash tree in North America. Once common, it has experienced high mortality from the non-native invasive emerald ash borer (EAB; Agrilus planipennis). A small percentage of native green ash trees that remain healthy in long-infested areas, termed "lingering ash," display partial resistance to the insect, indicating that breeding and propagating populations with higher resistance to EAB may be possible. To assist in ash breeding, ecology and evolution studies, we report the first chromosome-level assembly from the genus Fraxinus for F. pennsylvanica with over 99% of bases anchored to 23 haploid chromosomes, spanning 757 Mb in total, composed of 49.43% repetitive DNA, and containing 35,470 high-confidence gene models assigned to 22,976 Asterid orthogroups. We also present results of range-wide genetic variation studies, the identification of candidate genes for important traits including potential EAB-resistance genes, and an investigation of comparative genome organization among Asterids based on this reference genome platform. Residual duplicated regions within the genome probably resulting from a recent whole genome duplication event in Oleaceae were visualized in relation to wild olive (Olea europaea var. sylvestris). We used our F. pennsylvanica chromosome assembly to construct reference-guided assemblies of 27 previously sequenced Fraxinus taxa, including F. excelsior. Thus, we present a significant step forward in genomic resources for research and protection of Fraxinus species.

Diacetin, a reliable cue and private communication channel in a specialized pollination system.

The interaction between floral oil secreting plants and oil-collecting bees is one of the most specialized of all pollination mutualisms. Yet, the specific stimuli used by the bees to locate their host flowers have remained elusive. This study identifies diacetin, a volatile acetylated glycerol, as a floral signal compound shared by unrelated oil plants from around the globe. Electrophysiological measurements of antennae and behavioural assays identified diacetin as the key volatile used by oil-collecting bees to locate their host flowers. Furthermore, electrophysiological measurements indicate that only oil-collecting bees are capable of detecting diacetin. The structural and obvious biosynthetic similarity between diacetin and associated floral oils make it a reliable cue for oil-collecting bees. It is easily perceived by oil bees, but can't be detected by other potential pollinators. Therefore, diacetin represents the first demonstrated private communication channel in a pollination system.

Strong phylogenetic effects on floral scent variation of oil-secreting orchids in South Africa.

PREMISE OF THE STUDY: Evolution involves the interplay between natural selection and phylogenetic constraint. This is particularly evident among the flowering plants where form and diversity of flowers attest to the importance of both pollinator-mediated selection and phylogenetic constraint. Although this has been studied mostly using visible floral characters, invisible volatile chemicals emitted by the flowers should be subject to these same evolutionary forces. Unfortunately, most analyses of floral volatiles have over-emphasized the importance of natural selection and underplayed phylogenetic constraint without quantifying their respective roles in the evolution and composition of floral scents. METHODS: We used multivariate analyses to test the relative importance of pollinators vs. phylogeny in determining the composition of floral scents among oil-secreting orchids in southern Africa. Floral scents of 42 oil-secreting taxa/ecotypes distributed among 12 subclades in the tribe Diseae were sampled using headspace adsorption and gas chromatography-mass spectroscopy. KEY RESULTS: We identified 257 scent compounds distributed over nine different compound classes, with the majority of scents dominated by aliphatic or benzenoid compounds. The only significant predictor of floral scent among these orchids above the species level was phylogeny. Nevertheless, in two of the clades there were differences in scent profiles at the species and ecotype level that corresponded to different pollinators and were thus suggestive of pollinator-mediated selection. CONCLUSIONS: Scent variation was greater than expected and phylogeny was more important than pollinator-mediated selection in predicting the composition of floral scents of oil-secreting orchids, despite the specialized nature of the pollinator reward system.

Leaf age affects the responses of foliar injury and gas exchange to tropospheric ozone in Prunus serotina seedlings.

We investigated the effect of leaf age on the response of net photosynthesis (A), stomatal conductance (g(wv)), foliar injury, and leaf nitrogen concentration (N(L)) to tropospheric ozone (O(3)) on Prunus serotina seedlings grown in open-plots (AA) and open-top chambers, supplied with either carbon-filtered or non-filtered air. We found significant variation in A, g(wv), foliar injury, and N(L) (P < 0.05) among O(3) treatments. Seedlings in AA showed the highest A and g(wv) due to relatively low vapor pressure deficit (VPD). Older leaves showed significantly lower A, g(wv), N(L), and higher foliar injury (P < 0.001) than younger leaves. Leaf age affected the response of A, g(wv), and foliar injury to O(3). Both VPD and N(L) had a strong influence on leaf gas exchange. Foliar O(3)-induced injury appeared when cumulative O(3) uptake reached 8-12 mmol m(-2), depending on soil water availability. The mechanistic assessment of O(3)-induced injury is a valuable approach for a biologically relevant O(3) risk assessment for forest trees.

Twin oil sacs facilitate the evolution of a novel type of pollination unit (meranthium) in a South African orchid.

The unique floral morphology of the South African orchid H. pulchra, with its twin meranthia, is best explained as an adaptation to pollination by oil-collecting bees. Flowers consisting of meranthia (floral parts that function as single pollination units; commonly observed in garden Iris) are extremely rare among the angiosperms and their significance poorly understood. Unlike all other known examples of meranthia, the novel type described for H. pulchra is not bilabiate. All Huttonaea species are unique in having twin petal sacs with glandular verrucae that secrete oil and are pollinated by Rediviva (Melittidae) oil-collecting bees. But only Huttonaea pulchra has long and widely divergent petal claws that place the oil sacs well beyond the reach of a centrally positioned bee. The wide separation of these sacs forces the pollinator, R. colorata, to visit each side of the flower independently and effectively divides the flower into two meranthia. Molecular data indicate that the evolution of the Huttonaea-type meranthium was dependent on the prior evolution of the oil flower/oil bee relationship. Meranthium evolution was also facilitated by the presence of oil in two separate structures (petal sacs) that were not physically constrained to remain in close proximity.

Relationships between advance oak regeneration and biotic and abiotic factors.

Relationships between advance regeneration of four tree species (red maple (Acer rubrum L.), white oak (Quercus alba L.), chestnut oak (Q. montana Willd.) and northern red oak (Q. rubra L.)) and biotic (non-tree vegetation and canopy composition) and abiotic (soil series and topographic variables) factors were investigated in 52, mature mixed-oak stands in the central Appalachians. Aggregate height was used as a composite measure of regeneration abundance. Analyses were carried out separately for two physiographic provinces. Associations with tree regeneration were found for all biotic and abiotic factors both in partial models and full models. Red maple was abundant on most of the sites, but high red maple abundance was commonly associated with wet north-facing slopes with little or no cover of mountain-laurel (Kalmia latifolia L.) and hay-scented fern (Dennstaedtia punctilobula (Michx.) Moore). Regeneration of the three oak species was greatly favored by the abundance of overstory trees of their own kind. White oak regeneration was most abundant on south-facing, gentle, lower slopes with soils in the Buchanan series. Chestnut oak regeneration was more common on south-facing, steep upper slopes with stony soils. There was a positive association between chestnut oak and huckleberry (Gaylussacia baccata (Wangh.) Koch) cover classes. Northern red oak was more abundant on north-facing wet sites with Hazleton soil, and was associated with low occurrence of mountain-laurel and hay-scented fern.

Forest regeneration composition and development in upland, mixed-oak forests.

Advance regeneration in 52 mature mixed-oak stands was analyzed and described. Red maple (Acer rubrum L.) was the most abundant species in the study area. Among oak (Quercus) species, northern red oak (Q. rubra L.) was the most abundant within the Allegheny Plateau physiographic province, whereas chestnut oak (Q. montana L.) was the most abundant within the Ridge and Valley physiographic province. Sixteen stands, for which data are available through the fourth growing season following harvest, were used to describe stand development. Cumulative height, a composite measure of size and density, was used to describe early stand development. Black gum (Nyssa sylvatica Marsh.) and black birch (Betula lenta L.) had dramatic increases in stand density and cumulative height after overstory removal. Cumulative height of northern red oak and chestnut oak showed a faster positive response to overstory removal than red maple. Oak retained its dominance in cumulative height for at least 4 years after harvest. Red maple nevertheless remained the most abundant tree species after overstory removal. Our results suggest that the principal advantage of red maple regeneration is its ability to accumulate in large numbers prior to harvest.

OIL FLOWERS AND OIL BEES: FURTHER EVIDENCE FOR POLLINATOR ADAPTATION.

We examined foreleg length and body size variation in two species of oil-collecting bees (Rediviva; Melittidae) in southern Africa. Oil-collecting bees harvest oil from host flowers by rubbing their forelegs against oil-secreting trichomes. Significant differences in foreleg length occur among populations of both species. Rediviva "pallidula" populations vary significantly in mean foreleg length (11.34 ± 0.42 mm to 12.67 ± 0.36 mm), but not in body length (10.59 ± 0.74 to 10.80 ± 0.64), and foreleg length and body size are not significantly correlated. Instead, foreleg variation appears to be a function of host plant spur length. Ninety-two percent of the variance in foreleg length of R. "pallidula" is explained by mean Diascia spur length. Rediviva rufocincta populations vary significantly in mean foreleg length (10.12 ± 0.70 mm to 12.34 ± 0.68 mm) and in body length (9.03 ± 0.26 mm to 10.56 ± 0.24 mm). Foreleg length scales allometrically with body size in this species as 90.5% of the variance in foreleg length can be explained as a function of body length. Body size appears to be constrained by the morphology of the oil-secreting host plant. Both bees collect floral oil with specially modified setae on the tarsi of their forelegs. The length of the disti- + mediotarsus (refered to here as "tarsus") in relation to the entire foreleg is shorter in R. rufocincta and does not increase as rapidly with increasing foreleg length as for R. "pallidula." These differences in variation can be attributed to differences in position of oil within the flowers of the respective host plants. Rediviva "pallidula" collects oil from Diascia species that have the oil deeply situated in narrow floral spurs of varying length, while R. rufocincta collects oil from the broadly saccate flowers of Bowkeria verticillata and B. citrina.