We present a 6-gene, 420-species maximum-likelihood phylogeny of Ascomycota, the largest phylum of Fungi. This analysis is the most taxonomically complete to date with species sampled from all 15 currently circumscribed classes. A number of superclass-level nodes that have previously evaded resolution and were unnamed in classifications of the Fungi are resolved for the first time. Based on the 6-gene phylogeny we conducted a phylogenetic informativeness analysis of all 6 genes and a series of ancestral character state reconstructions that focused on morphology of sporocarps, ascus dehiscence, and evolution of nutritional modes and ecologies. A gene-by-gene assessment of phylogenetic informativeness yielded higher levels of informativeness for protein genes (RPB1, RPB2, and TEF1) as compared with the ribosomal genes, which have been the standard bearer in fungal systematics. Our reconstruction of sporocarp characters is consistent with 2 origins for multicellular sexual reproductive structures in Ascomycota, once in the common ancestor of Pezizomycotina and once in the common ancestor of Neolectomycetes. This first report of dual origins of ascomycete sporocarps highlights the complicated nature of assessing homology of morphological traits across Fungi. Furthermore, ancestral reconstruction supports an open sporocarp with an exposed hymenium (apothecium) as the primitive morphology for Pezizomycotina with multiple derivations of the partially (perithecia) or completely enclosed (cleistothecia) sporocarps. Ascus dehiscence is most informative at the class level within Pezizomycotina with most superclass nodes reconstructed equivocally. Character-state reconstructions support a terrestrial, saprobic ecology as ancestral. In contrast to previous studies, these analyses support multiple origins of lichenization events with the loss of lichenization as less frequent and limited to terminal, closely related species.

Biochar cation exchange capacity (CEC) is a key property central to better retain soil nutrients and reduce fertilizer runoff. This paper reports a breakthrough progress in improving biochar CEC value by a factor of nearly 10 through biochar surface oxygenation by ozonization. The CEC value of the untreated biochar was measured to be anywhere between 14 and 17 cmol/kg. A 90-minutes dry ozonization treatment resulted in an increased biochar CEC value of 109~152 cmol/kg. Simultaneously, the biochar ozonization process resulted in a reduction of biochar pH from 9.82 to as low as 3.07, indicating the formation of oxygen-functional groups including carboxylic acids on biochar surfaces. Using the techniques of X-ray photoelectron spectroscopy (XPS), the formation of oxygen-functional groups including carboxylic acids on biochar surfaces have been observed at a nanometer molecular scale upon biochar surface oxygenation through the ozonization treatment. The molar O:C ratio (0.31:1) on ozonized biochar surface as analyzed by XPS was indeed significantly higher than that (0.16:1) of the control biochar surface. Surprisingly, the molar O:C ratio from the Elemental Analysis data also showed an increase from the non-ozonized sample (0.077:1) to the dry-ozonized sample (0.193:1). Fourier-transform infrared (FTIR) spectroscopy analysis also showed an increase in the content of oxygen-functional groups in the form of carbonyl groups on biochar surfaces upon ozonization, which can also produce certain amount of oxygenated biochar molecular fragments that may be solubilized by liquid water, potentially leading to greater effects upon application of biochar in soil.

Biochar may be a tool for mine spoil remediation; however, its mechanisms for achieving this goal remain unclear. In this study, Miscanthus (Miscanthus giganteus) biochar was evaluated for its ability to reclaim acidic mine spoils (pH < 3) through reducing metal availability, improving soil microbial enzymatic activity, and initial growth of grass seedlings. Biochar was applied at 0, 1, 2.5 and 5% (w/w) along with lime/no lime and fertilizer additions. Blue Wildrye (Elymus glaucus cv. 'Elkton') was planted and later the shoots and roots were collected and metal concentrations determined. Afterwards, each pot was leached with deionized water, and the leachate analyzed for pH, electrical conductivity (EC), dissolved organic carbon (DOC) and soluble metal concentrations. After drying, the spoil was extracted with 0.01 M CaCl2 and Mehlich 3 (M3) to determine extractable Al, Cu, and Zn concentrations. Additionally, microbial activity was measured using a fluorescent β-glucosidase and N-acetyl-β-d-glucosaminidase assay. Spoil treated with lime and biochar had significantly greater pH and EC values. Significantly greater β-glucosidase activity occurred only in the 5% biochar plus lime treatment, while N-acetyl-β-d-glucosaminidase activities were not altered. Metal concentrations in rye shoot and roots were mixed. Lime additions significantly reduced extractable metal concentrations. Increasing biochar rates alone significantly reduced leachate DOC concentrations, and subsequently reduced leachable metal concentrations. Surprisingly, miscanthus biochar, by itself, was limited at mitigation, but when combined with lime, the combination was capable of further reducing extractable metal concentrations and improving β-glucosidase enzyme activity.

Biochars can improve soil health but have been widely shown to reduce plant-available nitrogen (N) owing to their high carbon (C) content, which stimulates microbial N-immobilization. However, because biochars contain large amounts of C that are not microbially available, their total elemental C:N ratio does not correspond well with impacts on soil N. We hypothesized that impacts on soil plant-available N would relate to biochar mineralizable-C (C_min) content, and that C:N ratios of the mineralizable biochar component could provide a means for predicting conditions of net soil N-mineralization or -immobilization. We conducted two laboratory experiments, the first measuring biochar C_min from respiration of isotopically labeled barley biochars manufactured at 300, 500, and 750 °C, and the second characterizing C_min by proxy measurements for ten biochars from six feedstocks at several temperatures. For both experiments, soils were incubated with 2% biochar by mass to determine impacts to soil N-mineralization. Contrary to expectation, all the biochars increased soil N-mineralization relative to unamended soils. Also unexpected, higher temperature (500 and 700 °C) barley biochars with less C_min stimulated more soil decomposition and more soil N-mineralization than a 350 °C barley biochar. However, across diverse biochar feedstocks and production methods, none of the biochar characteristics correlated with soil N-mineralization. The finding of improved soil N-mineralization adds complexity to the range of soil N responses that can be expected in response to biochar amendment. Because of the limited ability to predict soil N responses from biochar properties, users should monitor soil N to manage soil fertility.