A minimally disruptive method for measuring water potential in planta using hydrogel nanoreporters.

Leaf water potential is a critical indicator of plant water status, integrating soil moisture status, plant physiology, and environmental conditions. There are few tools for measuring plant water status (water potential) in situ, presenting a critical barrier for developing appropriate phenotyping (measurement) methods for crop development and modeling efforts aimed at understanding water transport in plants. Here, we present the development of an in situ, minimally disruptive hydrogel nanoreporter (AquaDust) for measuring leaf water potential. The gel matrix responds to changes in water potential in its local environment by swelling; the distance between covalently linked dyes changes with the reconfiguration of the polymer, leading to changes in the emission spectrum via Förster Resonance Energy Transfer (FRET). Upon infiltration into leaves, the nanoparticles localize within the apoplastic space in the mesophyll; they do not enter the cytoplasm or the xylem. We characterize the physical basis for AquaDust's response and demonstrate its function in intact maize (Zea mays L.) leaves as a reporter of leaf water potential. We use AquaDust to measure gradients of water potential along intact, actively transpiring leaves as a function of water status; the localized nature of the reporters allows us to define a hydraulic model that distinguishes resistances inside and outside the xylem. We also present field measurements with AquaDust through a full diurnal cycle to confirm the robustness of the technique and of our model. We conclude that AquaDust offers potential opportunities for high-throughput field measurements and spatially resolved studies of water relations within plant tissues.

Champions of winter survival: cold acclimation and molecular regulation of cold hardiness in evergreen conifers.

Evergreen conifers are champions of winter survival, based on their remarkable ability to acclimate to cold and develop cold hardiness. Counterintuitively, autumn cold acclimation is triggered not only by exposure to low temperature, but also by a combination of decreasing temperature, decreasing photoperiod and changes in light quality. These environmental cues control a network of signaling pathways that coordinate cold acclimation and cold hardiness in overwintering conifers, leading to cessation of growth, bud dormancy, freezing tolerance and changes in energy metabolism. Advances in genomic, transcriptomic and metabolomic tools for conifers have improved our understanding of how trees sense and respond to changes in temperature and light during cold acclimation and the development of cold hardiness, but there remain considerable gaps deserving further research in conifers. In the first section of this review, we focus on the physiological mechanisms used by evergreen conifers to adjust metabolism seasonally and to protect overwintering tissues against winter stresses. In the second section, we review how perception of low temperature and photoperiod regulate the induction of cold acclimation. Finally, we explore the evolutionary context of cold acclimation in conifers and evaluate challenges imposed on them by changing climate and discuss emerging areas of research in the field.

Variation in the phenology of photosynthesis among eastern white pine provenances in response to warming.

In higher-latitude trees, temperature and photoperiod control the beginning and end of the photosynthetically active season. Elevated temperature (ET) has advanced spring warming and delayed autumn cooling while photoperiod remains unchanged. We assessed the effects of warming on the length of the photosynthetically active season of three provenances of Pinus strobus L. seedlings from different latitudes, and evaluated the accuracy of the photochemical reflectance index (PRI) and the chlorophyll/carotenoid index (CCI) for tracking the predicted variation in spring and autumn phenology of photosynthesis among provenances. Seedlings from northern, local and southern P. strobus provenances were planted in a temperature-free-air-controlled enhancement (T-FACE) experiment and exposed to ET (+1.5/3°C; day/night). Over 18 months, we assessed photosynthetic phenology by measuring chlorophyll fluorescence, gas exchange, leaf spectral reflectance and pigment content. During autumn, all seedlings regardless of provenance followed the same sequence of phenological events with the initial downregulation of photosynthesis, followed by the modulation of non-photochemical quenching and associated adjustments of zeaxanthin pool sizes. However, the timing of autumn downregulation differed between provenances, with delayed onset in the southern provenance (SP) and earlier onset in the northern relative to the local provenance, indicating that photoperiod at the provenance origin is a dominant factor controlling autumn phenology. Experimental warming further delayed the downregulation of photosynthesis during autumn in the SP. A provenance effect during spring was also observed but was generally not significant. The vegetation indices PRI and CCI were both effective at tracking the seasonal variations of energy partitioning in needles and the differences of carotenoid pigments indicative of the stress status of needles. These results demonstrate that PRI and CCI can be useful tools for monitoring conifer phenology and for the remote monitoring of the length of the photosynthetically active season of conifers in a changing climate.

Photoperiod and temperature constraints on the relationship between the photochemical reflectance index and the light use efficiency of photosynthesis in Pinus strobus.

The photochemical reflectance index (PRI) is a proxy for the activity of the photoprotective xanthophyll cycle and photosynthetic light use efficiency (LUE) in plants. Evergreen conifers downregulate photosynthesis in autumn in response to low temperature and shorter photoperiod, and the dynamic xanthophyll cycle-mediated non-photochemical quenching (NPQ) is replaced by sustained NPQ. We hypothesized that this shift in xanthophyll cycle-dependent energy partitioning during the autumn is the cause for variations in the PRI-LUE relationship. In order to test our hypothesis, we characterized energy partitioning and pigment composition during a simulated summer-autumn transition in a conifer and assessed the effects of temperature and photoperiod on the PRI-LUE relationship. We measured gas exchange, chlorophyll fluorescence and leaf reflectance during the photosynthetic downregulation in Pinus strobus L. seedlings exposed to low temperature/short photoperiod or elevated temperature/short photoperiod conditions. Shifts in energy partitioning during simulated autumn were observed when the pools of chlorophylls decreased and pools of photoprotective carotenoids increased. On a seasonal timescale, PRI was controlled by carotenoid pool sizes rather than xanthophyll cycle dynamics. Photochemical reflectance index variation under cold autumn conditions mainly reflected long-term pigment pool adjustments associated with sustained NPQ, which impaired the PRI-LUE relationship. Exposure to warm autumn conditions prevented the induction of sustained NPQ but still impaired the PRI-LUE relationship. We therefore conclude that alternative zeaxanthin-independent NPQ mechanisms, which remain undetected by the PRI, are present under both cold and warm autumn conditions, contributing to the discrepancy in the PRI-LUE relationship during autumn.

Zeaxanthin-independent energy quenching and alternative electron sinks cause a decoupling of the relationship between the photochemical reflectance index (PRI) and photosynthesis in an evergreen conifer during spring.

In evergreen conifers, the winter down-regulation of photosynthesis and its recovery during spring are the result of a reorganization of the chloroplast and adjustments of energy-quenching mechanisms. These phenological changes may remain undetected by remote sensing, as conifers retain green foliage during periods of photosynthetic down-regulation. The aim was to assess if the timing of the spring recovery of photosynthesis and energy-quenching characteristics are accurately monitored by the photochemical reflectance index (PRI) in the evergreen conifer Pinus strobus. The recovery of photosynthesis was studied using chlorophyll fluorescence, leaf gas exchange, leaf spectral reflectance, and photosynthetic pigment measurements. To assess if climate change might affect the recovery of photosynthesis, seedlings were exposed to cold spring conditions or warm spring conditions with elevated temperature. An early spring decoupling of the relationship between photosynthesis and PRI in both treatments was observed. This was caused by differences between the timing of the recovery of photosynthesis and the timing of carotenoid and chlorophyll pool size adjustments which are the main factors controlling PRI during spring. It was also demonstrated that zeaxanthin-independent NPQ mechanisms undetected by PRI further contributed to the early spring decoupling of the PRI-LUE relationship. An important mechanism undetected by PRI seems to involve increased electron transport around photosystem I, which was a significant energy sink during the entire spring transition, particularly in needles exposed to a combination of high light and cold temperatures.