Leaf acclimation to soil flooding and light availability underlies photosynthetic capacity of Lindera melissifolia, an endangered shrub of bottomland forests in the Mississippi Alluvial Valley, USA.

Lindera melissifolia is an endangered shrub indigenous to the broadleaf forest of the Mississippi Alluvial Valley (MAV). In this region, extant colonies of the species are found in periodically ponded habitats where a diversity of broadleaf trees can form well-developed overstory and sub-canopies-these habitat characteristics suggest that soil flooding and light availability are primary drivers of L. melissifolia ecophysiology. To understand how these two factors affect its photosynthetic capacity, we quantified leaf characteristics and photosynthetic response of plants grown in a large-scaled, field setting of three distinct soil flooding levels (no flood, 0 day; short-term flood, 45 days; and extended flood, 90 days) each containing three distinct light availability levels (high light, 30% shade cloth; intermediate light, 63% shade cloth; and low light, 95% shade cloth). Lindera melissifolia leaves showed marked plasticity to interacting effects of flooding and light with lamina mass per unit area (Lm/a) varying 78% and total nitrogen content per unit area (Na) varying 63% from the maximum. Photosynthetic capacity (A1800-a) ranged 123% increasing linearly with Na from low to high light. Extended flooding decreased the slope of this relationship 99% through a reduction in N availability and metabolic depression of A1800-a relative to Na. However, neither soil flooding nor light imposed an additive limitation on photosynthetic capacity when the other factor was at its most stressful level, and the A1800-a-Na relationship for plants that experienced short-term flooding suggested post-flood acclimation in photosynthetic capacity was approaching the maximal level under respective light environments. Our findings provide evidence for wide plasticity and acclimation potential of L. melissifolia photosynthetic capacity, which supports active habitat management, such as manipulation of stand structure for improved understory light environments, to benefit long-term conservation of the species in the MAV.

Biomarker Metabolites Discriminate between Physiological States of Field, Cave and White-nose Syndrome Diseased Bats.

Analysis of volatile organic compound (VOC) emissions using electronic-nose (e-nose) devices has shown promise for early detection of white-nose syndrome (WNS) in bats. Tricolored bats, Perimyotis subflavus, from three separate sampling groups defined by environmental conditions, levels of physical activity, and WNS-disease status were captured temporarily for collection of VOC emissions to determine relationships between these combinations of factors and physiological states, Pseudogymnoascus destructans (Pd)-infection status, and metabolic conditions. Physiologically active (non-torpid) healthy individuals were captured outside of caves in Arkansas and Louisiana. In addition, healthy and WNS-diseased torpid bats were sampled within caves in Arkansas. Whole-body VOC emissions from bats were collected using portable air-collection and sampling-chamber devices in tandem. Electronic aroma-detection data using three-dimensional Principal Component Analysis provided strong evidence that the three groups of bats had significantly different e-nose aroma signatures, indicative of different VOC profiles. This was confirmed by differences in peak numbers, peak areas, and tentative chemical identities indicated by chromatograms from dual-column GC-analyses. The numbers and quantities of VOCs present in whole-body emissions from physiologically active healthy field bats were significantly greater than those of torpid healthy and diseased cave bats. Specific VOCs were identified as chemical biomarkers of healthy and diseased states, environmental conditions (outside and inside of caves), and levels of physiological activity. These results suggest that GC/E-nose dual-technologies based on VOC-detection and analyses of physiological states, provide noninvasive alternative means for early assessments of Pd-infection, WNS-disease status, and other physiological states.

Assessment of the Portable C-320 Electronic Nose for Discrimination of Nine Insectivorous Bat Species: Implications for Monitoring White-Nose Syndrome.

The development of new C-320 electronic-nose (e-nose) methods for pre-symptomatic detection of White-Nose Syndrome (WNS) in bats has required efficacy studies of instrument capabilities to discriminate between major sources of volatile organic compounds (VOCs) derived from clinical samples. In this phase-2 study, we further tested this e-nose for capabilities to distinguish between bat species based on differences in whole-body VOC emissions. Live healthy individuals of nine bat species were temporarily captured outside of caves in Arkansas and Louisiana. VOC emissions from bats were collected using newly developed portable air collection and sampling-chamber devices in tandem. Sensor-array output responses to bat VOC emissions were compared to those of 22 pure VOC analytical standards from five chemical classes. Distinct smellprint signatures were produced from e-nose analyses of VOC metabolites derived from individual bat species. Smellprint patterns were analyzed using 2-dimensional and 3-dimensional Principal Component Analysis (PCA) to produce aroma map plots showing effective discrimination between bat species with high statistical significance. These results demonstrate potential instrument efficacy for distinguishing between species-specific, bat-derived VOC metabolite emissions as major components of clinical samples collected from bats in caves for disease detection prior to symptom development. This study provided additional information required to fully test the efficacy of a portable e-nose instrument for diagnostic applications in subsequent phase-3 testing of noninvasive, early WNS disease detection in intra-cave hibernating bats.

Detection of Emerald Ash Borer Infestations in Living Green Ash by Noninvasive Electronic-Nose Analysis of Wood Volatiles.

The emerald ash borer (EAB) has been the most destructive and costly nonnative insect to threaten the health of ash (Fraxinus) species in North America for at least the past 25 years. The development of methods for detecting visually-hidden EAB galleries at early stages of infestation would provide a useful tool to more effectively facilitate the planning and implementation of targeted EAB pest-suppression and management activities. We tested the efficacy of using a dual-technology electronic-nose (e-nose)/gas chromatograph device as a means for detection of EAB infestations in green ash trees in different EAB-decline classes by analysis of VOC emissions in sapwood. We found significant differences in VOC profiles for trees from the four decline classes. The VOC composition, quantities, and types of volatile metabolites present in headspace volatiles varied considerably across sample types, and resulted in distinct e-nose smellprint patterns that were characteristic of each unique chemical composition. In addition, specific VOC metabolites were identified as potential healthy and EAB-infestation biomarkers, indicative of the health states of individual trees. Few significant differences in major bark phenolic compounds were found between ash decline classes using LC-MS. The e-nose was effective in discriminating between uninfested and EAB-infested trees based on sapwood VOC emissions.

New secondary metabolites from bioactive extracts of the fungus Armillaria tabescens.

Ethyl acetate extracts of Armillaria tabescens (strain JNB-OZ344) showed significant fungistatic and bacteristatic activities against several major human pathogens including Candida albicans, Cryptococcus neoformans, Escherichia coli and Mycobacterium intracellulare. Chemical analysis of these extracts led to the isolation and identification of four new compounds, emestrin-F (1), emestrin-G (2), 6-O-(4-O-methyl-ß-D-glucopyranosyl)-8-hydroxy-2,7-dimethyl-4H-benzopyran-4-one (3) and cephalosporolide-J (4), along with five other previously known compounds, emestrin (5), cephalosporolide-E (6), decarestrictine-C2 (7), ergosterol and brassicasterol. Structural elucidation of all compounds was carried out by NMR and MS analyses. Antimicrobial assays revealed that compounds 1 and 5 were responsible for the observed growth inhibitory activities of the fungal extracts against the human pathogens tested.