Chantal D. Reid

Long-term kinetics of the light-dependent regulation of ribulose-1,5-bisphosphate carboxylase/oxygenase activity in plants with and without 2-carboxyarabinitol 1-phosphate

The light-dependent modulation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity was studied in two species: Phaseolus vulgaris L., which has high levels of the inhibitor of Rubisco activity, carboxyarabinitol 1-phosphate (CA1P), in the dark, and Chenopodium album L., which has little CA1P. In both species, the ratio of initial to fully-activated Rubisco activity declined by 40–50% within 60 min of a reduction in light from high a photosynthetic photon flux density (PPFD; >700 μmol · m−2 · s−1) to a low PPFD (65 ± 15 μmol · m−2 · s−1) or to darkness, indicating that decarbamylation of Rubisco is substantially involved in the initial regulatory response of Rubisco to a reduction in PPFD, even in species with potentially extensive CA1P inhibition. Total Rubisco activity was unaffected by PPFD in C. album, and prolonged exposure (2–6 h) to low light or darkness was accompanied by a slow decline in the activity ratio of this species. This indicates that the carbamylation state of Rubisco from C. album gradually declines for hours after the large initial drop in the first 60 min following light reduction. In P. vulgaris, the total activity of Rubisco declined by 10–30% within 1 h after a reduction in PPFD to below 100 μmol · m−2 · s−1, indicating CA1P-binding contributes significantly to the reduction of Rubisco capacity during this period, but to a lesser extent than decarbamylation. With continued exposure of P. vulgaris leaves to very low PPFDs (< 30 μmol · m−2 · s−1), the total activity of Rubisco declined steadily so that after 6–6.5 h of exposure to very low light or darkness, it was only 10–20% of the high-light value. These results indicate that while decarbamylation is more prominent in the initial regulatory response of Rubisco to a reduction in PPFD in P. vulgaris, binding of CA1P increases over time and after a few hours dominates the regulation of Rubisco activity in darkness and at very low PPFDs.

Exploring the transport of plant metabolites using positron emitting radiotracers.

Short‐lived positron‐emitting radiotracer techniques provide time‐dependent data that are critical for developing models of metabolite transport and resource distribution in plants and their microenvironments. Until recently these techniques were applied to measure radiotracer accumulation in coarse regions along transport pathways. The recent application of positron emission tomography (PET) techniques to plant research allows for detailed quantification of real‐time metabolite dynamics on previously unexplored spatial scales. PET provides dynamic information with millimeter‐scale resolution on labeled carbon, nitrogen, and water transport over a small plant‐size field of view. Because details at the millimeter scale may not be required for all regions of interest, hybrid detection systems that combine high‐resolution imaging with other radiotracer counting technologies offer the versatility needed to pursue wide‐ranging plant physiological and ecological research. In this perspective we describe a recently developed hybrid detection system at Duke University that provides researchers with the flexibility required to carry out measurements of the dynamic responses of whole plants to environmental change using short‐lived radiotracers. Following a brief historical development of radiotracer applications to plant research, the role of radiotracers is presented in the context of various applications at the leaf to the whole‐plant level that integrates cellular and subcellular signals and/or controls.

Effects of CO2 enrichment on whole-plant carbon budget of seedlings of Fagus grandifolia and Acer saccharum in low irradiance

Carbon exchange rates (CER) and whole-plant carbon balances of beech (Fagus grandifolia) and sugar maple (Acer saccharum) were compared for seedlings grown under low irradiance to determine the effects of atmospheric CO2 enrichment on shade-tolerant seedlings of co-dominant species. Under contemporary atmospheric CO2, photosynthetic rate per unit mass of beech was lower than for sugar maple, and atmospheric CO2 enrich ment enhanced photosynthesis for beech only. Aboveground respiration per unit mass decreased with CO2 enrichment for both species while root respiration per unitmass decreased for sugar maple only. Under contemporary atmoapheric CO2, beech had lower C uptake per plant than sugar maple, while C losses per plant to nocturnal aboveground and root respiration were similar for both species. Under elevated CO2, C uptake per plant was similar for both species, indicating a significant relative increase in whole-seedling CER with CO2 enrich ment for beech but not for sugar maple. Total C loss per plant to aboveground respiration was decreased for beech only because increase in sugar maple leaf mass counterbalanced a reduction in respiration rates. Carbon loss to root respiration per plant was not changed by CO2 enrichment for either species. However, changes in maintenance respiration cost and nitrogen level suggest changes in tissue composition with elevated CO2. Beech had a greater net daily C gain with CO2 enrichment than did sugar maple in contrast to a lower one under contemporary CO2. Elevated CO2 preferentially enhances the net C balance of beech by increasing photosynthesis and reducing respiration cost. In all cases, the greatest C lost was by roots, indicating the importance of belowground biomass in net C gain. Relative growth rate estimated from biomass accumulation was not affected by CO2 enrichment for either species possibly because of slow growth under low light. This study indicates the importance of direct effects of CO2 enrichment when predicting potential change in species distribution with global climate change.

Positron emission tomography detector development for plant biology

There are opportunities for the development of new tools to advance plant biology research through the use of radionuclides. Thomas Jefferson National Accelerator Facility, Duke University, West Virginia University and the University of Maryland are collaborating on the development of radionuclide imaging technologies to facilitate plant biology research. Biological research into optimizing plant productivity under various environmental constraints, biofuel and carbon sequestration research are areas that could potentially benefit from new imaging technologies. Using 11CO2 tracers, the investigators at Triangle University Nuclear Laboratory / Duke University Phytotron are currently researching the dynamical responses of plants to environmental changes forecasted from increasing greenhouse trace gases involved in global change. The biological research primary focus is to investigate the impact of elevated atmospheric CO2 and nutrients limitation on carbon and nitrogen dynamics in plants. We report here on preliminary results of 11CO2 plant imaging experiments involving barley plants using Jefferson Lab dual planar positron emission tomography detectors to image 11CO2 in live barley plants. New detector designs will be developed based on the preliminary studies reported here and further planned.

Atractiella rhizophila, sp. nov., an endorrhizal fungus isolated from the Populus root microbiome

ABSTRACT Among fungi isolated from healthy root mycobiomes of Populus, we discovered a new endorrhizal fungal species belonging to the rust lineage Pucciniomycotina, described here as Atractiella rhizophila. We characterized this species by transmission electron microscopy (TEM), phylogenetic analysis, and plant bioassay experiments. Phylogenetic sequence analysis of isolates and available environmental and reference sequences indicates that this new species, A. rhizophila, has a broad geographic and host range. Atractiella rhizophila appears to be present in North America, Australia, Asia, and Africa and is associated with trees, orchids, and other agriculturally important species, including soybean, corn, and rice. Despite the large geographic and host range of this species sampling, A. rhizophila appears to have exceptionally low sequence variation within nuclear rDNA markers examined. With inoculation studies, we demonstrate that A. rhizophila is nonpathogenic, asymptomatically colonizes plant roots, and appears to foster plant growth and elevated photosynthesis rates.

Imaging corn plants with PhytoPET, a modular PET system for plant biology

PhytoPET is a modular positron emission tomography (PET) system designed specifically for plant imaging. The PhytoPET design allows flexible arrangements of PET detectors based on individual standalone detector modules built from single Hamamatsu H8500 position sensitive photomultiplier tubes and pixelated LYSO arrays. We have used the PhytoPET system to perform preliminary corn plant imaging studies at the Duke University Biology Department Phytotron. Initial evaluation of the PhytoPET system to image the biodistribution of the positron emitting tracer 11C in corn plants is presented. 11CO2 is loaded into corn seedlings by a leaf-labeling cuvette and translocation of 11C-sugars is imaged by a flexible arrangement of PhytoPET modules on each side. The PhytoPET system successfully images 11C within corn plants and allows for the dynamic measurement of 11C-sugar translocation from the leaf to the roots.

PhytoPET: Design and initial results of modular PET for plant biology

We have developed a positron emission tomography (PET) system designed specifically for plant imaging in Phytotron at Duke University. Initial evaluation of a PhytoPET system to image the biodistribution of the positron emitting tracer 11C in live plants is presented. The mechanical arrangement of the detectors is designed to accommodate the unpredictable and random distribution in space of the plant parts such as stems, leaves, and roots. Prototyping such a system requires a completely new PET system design strategy different from preclinical and clinical applications. This PhytoPET design allows flexible arrangements of PET detectors based on individual standalone detector modules built from single 5 cm × 5 cm Hamamatsu H8500 position sensitive photomultiplier tubes. Each H8500 is coupled to a L YSO:Ce scintillator array composed of 48 × 48 elements that are 10 mm thick with a 1 mm pitch. Initial results provide planar PET images from two different arrangements (1 × 4 or 2 × 2) for flexible imaging capability.

Ethernet-based flash ADC for a plant PET detector system

We have developed a flash analog to digital (ADC) based read out system to be used for a Positron Emission Tomography (PET) system. The custom designed 16 channel 12-bit Ethernet-based flash ADC (EFADC-16) unit operates at 250 MHzls/channel utilizing a gigabit Ethernet interface to parse time-stamped event signals. Each unit allows the user to define a custom coincidence table for triggering. Each EFADC-16 unit can digitize four H8500 position sensitive photomultiplier tubes (PSPMT) equipped with a Jefferson Lab designed 4 channel resistive readout (a total of 16 channels). We present initial performance results of the EFADC-16 with four PET detector modules in a plant biology application to acquire tomographic images of the translocation of 11C within an oak seedling.

Development of PhytoPET: A plant imaging PET system

The development and initial evaluation of a high-resolution positron emission tomography (PET) system to image the biodistribution of positron emitting tracers in live plants is underway. The positron emitting 11CO2 tracer is used in plant biology research investigating carbon sequestration in biomass, optimization of plant productivity and biofuel development. This PhytoPET design allows flexible arrangements of PET detectors based on individual standalone detector modules built from single 5 cm × 5 cm Hamamatsu H8500 position sensitive photomultiplier tubes. Each H8500 is coupled to a LYSO:Ce scintillator array composed of 48×48 elements that are 10 mm thick arranged with a 1.0 mm pitch. An Ethernet based 12-bit flash analog to digital data acquisition system with onboard coincident matrix definition is under development to digitize the signals. The detector modules of the PhytoPET system can be arranged and stacked to accommodate various sized plants and plant structures.

Compact beta particle/positron imager for plant biology

The 11 CO2 tracer is used to facilitate plant biology research towards optimization of plant productivity, biofuel development and carbon sequestration in biomass. Positron emission tomography (PET) imaging has been used to study carbon transport in live plants using 11 CO2. Plants typically have very thin leaves resulting in little medium for the emitted positrons to undergo an annihilation event. For the emitted positron from 11C decay approximately 1mm of water equivalent material is needed for positron annihilation. Thus most of the positrons do not annihilate inside the leaf, resulting in limited sensitivity for PET imaging. To address this problem we have developed a compact beta-positive beta-minus particle (BPBM) imager for 11 CO2 leaf imaging. The detector is based on a Hamamatsu H8500 position sensitive photomultiplier tube optically coupled via optical grease and a 3mm thick glass plate to a 0.5mm thick Eljin EJ-212 plastic scintillator. The detector is equipped with a flexible arm to allow its placement and orientation on the leaf of the plant of interest while maintaining the leafs original orientation. We are planning to utilize the imaging device at the Duke University Phytotron to investigate dynamic carbon transport differences between invasive and native species.