Electrical Signaling and Its Functions Under Conditions of Abiotic Stress: A Review of Methodological Approaches and Physiological Implications.

In contrast to chemical messengers, electrical signals such as action potentials and variation potentials can transmit information much faster over long distances. Electrical signals can be triggered by various abiotic stress factors and are propagated via plasmodesmata over short distances and within the phloem over long distances. Thus, in addition to assimilate transport from sources to sinks, the phloem serves as a communication highway for various types of information. Key factors for systemic signaling in the phloem are peptides, RNAs, hormones, and electrical signals. In recent years, there has been increasing evidence that rapid communication by means of electrical signals is essential for various plant physiological processes. Thus, this chapter focuses on electrical signaling and various associated physiological effects, such as regulation of leaf movements, assimilate transport, photosynthesis, and gas exchange, as well as plant water status.

Wood profiling by non-targeted liquid chromatography high-resolution mass spectrometry: Part 2, Detection of the geographical origin of spruce wood (Picea abies) by determination of metabolite pattern.

A non-targeted metabolomics-based approach using liquid chromatography high-resolution mass spectrometry was used to authenticate spruce wood (Picea abies) from two geographic source areas. The two sample sites were located in Germany and only 250 km apart. In order to achieve the highest possible metabolite coverage, the spruces samples were measured with four different methods using liquid chromatography high-resolution mass spectrometry. In this way, a total of approximately 4,100 features were detected, which included non-polar, polar, and intermediate-polar metabolites. Using supervised multivariate methods, a distinction between the two sample groups could be achieved on the basis of non-polar data sets. The major metabolites contributing to differentiation were identified by MS/MS experiments and were from the following classes of compounds: ceramides, fatty acids, glycerolipids, and phytosterols. Based on the soil descriptions of the two sites, it was concluded that there is probably a close relationship between nutrient availability and the differences in concentration of the marker compounds. The results show that a metabolomics-based approach is also suitable for differentiation of origin, even if the sample sites are close to each other.

Effect of Elevated CO2 and Drought on Biomass, Gas Exchange and Wood Structure of Eucalyptus grandis.

Juvenile Eucalyptus grandis were exposed to drought and elevated CO2 to evaluate the independent and interactive effects on growth, gas exchange and wood structure. Trees were grown in a greenhouse at ambient and elevated CO2 (aCO2, 410 ppm; eCO2, 950 ppm), in combination with daily irrigation and cyclic drought during one growing season. The results demonstrated that drought stress limited intercellular CO2 concentration, photosynthesis, stomatal conductance, and transpiration, which correlated with a lower increment in height, stem diameter and biomass. Drought also induced formation of frequent and narrow vessels accompanied by a reduction in vessel lumen area. Conversely, elevated CO2 increased intercellular CO2 concentration as well as photosynthesis, and partially closed stomata, leading to a more efficient water use, especially under drought. There was a clear trend towards greater biomass accumulation at eCO2, although the results did not show statistical significance for this parameter. We observed an increase in vessel diameter and vessel lumen area at eCO2, and, contrarily, the vessel frequency decreased. Thus, we conclude that eCO2 delayed the effects of drought and potentialized growth. However, results on vessel anatomy suggest that increasing vulnerability to cavitation due to formation of larger vessels may counteract the beneficial effects of eCO2 under severe drought.

Under salt stress guard cells rewire ion transport and abscisic acid signaling.

Soil salinity is an increasingly global problem which hampers plant growth and crop yield. Plant productivity depends on optimal water-use efficiency and photosynthetic capacity balanced by stomatal conductance. Whether and how stomatal behavior contributes to salt sensitivity or tolerance is currently unknown. This work identifies guard cell-specific signaling networks exerted by a salt-sensitive and salt-tolerant plant under ionic and osmotic stress conditions accompanied by increasing NaCl loads. We challenged soil-grown Arabidopsis thaliana and Thellungiella salsuginea plants with short- and long-term salinity stress and monitored genome-wide gene expression and signals of guard cells that determine their function. Arabidopsis plants suffered from both salt regimes and showed reduced stomatal conductance while Thellungiella displayed no obvious stress symptoms. The salt-dependent gene expression changes of guard cells supported the ability of the halophyte to maintain high potassium to sodium ratios and to attenuate the abscisic acid (ABA) signaling pathway which the glycophyte kept activated despite fading ABA concentrations. Our study shows that salinity stress and even the different tolerances are manifested on a single cell level. Halophytic guard cells are less sensitive than glycophytic guard cells, providing opportunities to manipulate stomatal behavior and improve plant productivity.

Wood profiling by non-targeted high-resolution mass spectrometry: Part 1, Metabolite profiling in Cedrela wood for the determination of the geographical origin.

The determination of the geographical origin of wood can be highly relevant for several reasons: On the one hand, it can help to prevent illegal logging and timber trade, on the other hand, it is of special interest for archaeological artefacts made of wood, as well as for a variety of biological questions. For this reason, different extraction methods were first tested for the analysis of polar and non-polar metabolites using liquid chromatography coupled electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS). A two-phase extraction with chloroform, methanol and water proved to be particularly successful. Subsequently, cedrela (Cedrela odorata) samples from South America were measured to distinguish geographic origin. Using multivariate data analysis, numerous origin-dependent differences could be extracted. The identification of the marker substances indicated that several metabolic pathways were affected by the geographical influences, some of them probably indicating pest infections.

Not stealing from the treasure chest (or just a bit): Analyses on plant derived writing supports and non-invasive DNA sampling.

Written communication plays a crucial role in the history of modern civilizations as manuscripts do not only exist contemporarily, but are passed on to subsequent generations. Besides a document's content, information is stored in the materials used for its production. Analyses of the composition allow, for example, identifying the biological origins of materials, dating, and help to understand degradation patterns. A combination of microscopic and DNA approaches was applied in order to analyze various plant derived writing sheets. Given their diversity and abundance in museum collections, plant based writing supports are yet an underexplored target for DNA studies. DNA retrieval of paper is low compared to raw paper plant material, which is likely due to the loss of organic components during paper production. Optimizing DNA extraction for each respective material drastically increased DNA recovery. Finally, we present a non-invasive DNA sampling method that utilizes nylon membranes, commonly used for bacterial DNA sampling and that is applicable to delicate material. Although bacterial infestation was visible on one sample, as indicated by scanning electron microscopy, endogenous DNA was retrieved. The results presented here are promising as they extend the scope of sources for DNA analyses by demonstrating that DNA molecules can be retrieved from a variety of plant derived writing supports. In future, such analyses can help to explore the biological diversity not only of plants and of additives utilized for producing writing supports, but also of the plenty products made from paper.

A Tandem Amino Acid Residue Motif in Guard Cell SLAC1 Anion Channel of Grasses Allows for the Control of Stomatal Aperture by Nitrate.

The latest major group of plants to evolve were the grasses. These became important in the mid-Paleogene about 40 million years ago. During evolution, leaf CO2 uptake and transpirational water loss were optimized by the acquisition of grass-specific stomatal complexes. In contrast to the kidney-shaped guard cells (GCs) typical of the dicots such as Arabidopsis, in the grasses and agronomically important cereals, the GCs are dumbbell shaped and are associated with morphologically distinct subsidiary cells (SCs). We studied the molecular basis of GC action in the major cereal crop barley. Upon feeding ABA to xylem sap of an intact barley leaf, stomata closed in a nitrate-dependent manner. This process was initiated by activation of GC SLAC-type anion channel currents. HvSLAC1 expressed in Xenopus oocytes gave rise to S-type anion currents that increased several-fold upon stimulation with >3 mM nitrate. We identified a tandem amino acid residue motif that within the SLAC1 channels differs fundamentally between monocots and dicots. When the motif of nitrate-insensitive dicot Arabidopsis SLAC1 was replaced by the monocot signature, AtSLAC1 converted into a grass-type like nitrate-sensitive channel. Our work reveals a fundamental difference between monocot and dicot GCs and prompts questions into the selective pressures during evolution that resulted in fundamental changes in the regulation of SLAC1 function.

Effect of simultaneously induced environmental stimuli on electrical signalling and gas exchange in maize plants.

Electrical signalling in response to environmental stimuli is a well-known phenomenon in higher plants. For example, in maize, different stimuli, such as wounding or re-irrigation after drought, incite characteristic electrical signals which have quite particular effects on gas exchange. What is less well understood is how plants (specifically maize) respond when two different environmental stimuli are applied simultaneously. To explore this, a three-stage experiment was designed. In the first stage, drought conditions were simulated by decreasing the soil water content to 30-40 % of field capacity. In these conditions, and in contrast to well-watered plants, the maize exhibited only 60-70% of the original level of stomatal conductance and 50-60 % of the original photosynthesis rate. In the second stage of the experiment the plants were re-irrigated and heat stimulated separately. Re-irrigation led to specific electrical signals followed by a gradual increase of gas exchange. In contrast, after heat stimulation of a leaf an electrical signal was evoked that reduced the net CO2-uptake rate as well as stomatal conductance. In the third stage, to elucidate how plants process simultaneous re-irrigation and heat stimulation, the drought-stressed maize plants were re-watered and heat-stimulated at the same time. Results showed a two phase response. In the first phase there was a rapid decrease in both the CO2 uptake rate and the stomatal conductance, while in the second phase each of these parameters increased gradually. Thus, the results strongly support the view that the responses from both stimuli were combined, indicating that maize plants can process simultaneously applied stimuli.

The desert plant Phoenix dactylifera closes stomata via nitrate-regulated SLAC1 anion channel.

Date palm Phoenix dactylifera is a desert crop well adapted to survive and produce fruits under extreme drought and heat. How are palms under such harsh environmental conditions able to limit transpirational water loss? Here, we analysed the cuticular waxes, stomata structure and function, and molecular biology of guard cells from P. dactylifera. To understand the stomatal response to the water stress phytohormone of the desert plant, we cloned the major elements necessary for guard cell fast abscisic acid (ABA) signalling and reconstituted this ABA signalosome in Xenopus oocytes. The PhoenixSLAC1-type anion channel is regulated by ABA kinase PdOST1. Energy-dispersive X-ray analysis (EDXA) demonstrated that date palm guard cells release chloride during stomatal closure. However, in Cl- medium, PdOST1 did not activate the desert plant anion channel PdSLAC1 per se. Only when nitrate was present at the extracellular face of the anion channel did the OST1-gated PdSLAC1 open, thus enabling chloride release. In the presence of nitrate, ABA enhanced and accelerated stomatal closure. Our findings indicate that, in date palm, the guard cell osmotic motor driving stomatal closure uses nitrate as the signal to open the major anion channel SLAC1. This initiates guard cell depolarization and the release of anions together with potassium.

Insect haptoelectrical stimulation of Venus flytrap triggers exocytosis in gland cells.

The Venus flytrap Dionaea muscipula captures insects and consumes their flesh. Prey contacting touch-sensitive hairs trigger traveling electrical waves. These action potentials (APs) cause rapid closure of the trap and activate secretory functions of glands, which cover its inner surface. Such prey-induced haptoelectric stimulation activates the touch hormone jasmonate (JA) signaling pathway, which initiates secretion of an acidic hydrolase mixture to decompose the victim and acquire the animal nutrients. Although postulated since Darwin's pioneering studies, these secretory events have not been recorded so far. Using advanced analytical and imaging techniques, such as vibrating ion-selective electrodes, carbon fiber amperometry, and magnetic resonance imaging, we monitored stimulus-coupled glandular secretion into the flytrap. Trigger-hair bending or direct application of JA caused a quantal release of oxidizable material from gland cells monitored as distinct amperometric spikes. Spikes reminiscent of exocytotic events in secretory animal cells progressively increased in frequency, reaching steady state 1 d after stimulation. Our data indicate that trigger-hair mechanical stimulation evokes APs. Gland cells translate APs into touch-inducible JA signaling that promotes the formation of secretory vesicles. Early vesicles loaded with H+ and Cl- fuse with the plasma membrane, hyperacidifying the "green stomach"-like digestive organ, whereas subsequent ones carry hydrolases and nutrient transporters, together with a glutathione redox moiety, which is likely to act as the major detected compound in amperometry. Hence, when glands perceive the haptoelectrical stimulation, secretory vesicles are tailored to be released in a sequence that optimizes digestion of the captured animal.

Interaction Effect between Elevated CO2 and Fertilization on Biomass, Gas Exchange and C/N Ratio of European Beech (Fagus sylvatica L.).

The effects of elevated CO2 and interaction effects between elevated CO2 and nutrient supplies on growth and the C/N ratio of European beech (Fagus sylvatica L.) saplings were studied. One-year-old beech saplings were grown in a greenhouse at ambient (385 ppm) and elevated CO2 (770 ppm/950 ppm), with or without fertilization for two growing seasons. In this study, emphasis is placed on the combined fertilization including phosphorus, potassium and nitrogen with two level of elevated CO2. The fertilized plants grown under elevated CO2 had the highest net leaf photosynthesis rate (Ac). The saplings grown under elevated CO2 had a significantly lower stomatal conductance (gs) than saplings grown under ambient air. No interaction effect was found between elevated CO2 and fertilization on Ac. A interaction effect between CO2 and fertilization, as well as between date and fertilization and between date and CO2 was detected on gs. Leaf chlorophyll content index (CCI) and leaf nitrogen content were strongly positively correlated to each other and both of them decreased under elevated CO2. At the end of both growing seasons, stem dry weight was greater under elevated CO2 and root dry weight was not affected by different treatments. No interaction effect was detected between elevated CO2 and nutrient supplies on the dry weight of different plant tissues (stems and roots). However, elevated CO2 caused a significant decrease in the nitrogen content of plant tissues. Nitrogen reduction in the leaves under elevated CO2 was about 10% and distinctly higher than in the stem and root. The interaction effect of elevated CO2 and fertilization on C/N ratio in plants tissues was significant. The results led to the conclusion that photosynthesis and the C/N ratio increased while stomatal conductance and leaf nitrogen content decreased under elevated CO2 and nutrient-limited conditions. In general, under nutrient-limited conditions, the plant responses to elevated CO2 were decreased.

SLAH3-type anion channel expressed in poplar secretory epithelia operates in calcium kinase CPK-autonomous manner.

Extrafloral nectaries secrete a sweet sugar cocktail that lures predator insects for protection from foraging herbivores. Apart from sugars and amino acids, the nectar contains the anions chloride and nitrate. Recent studies with Populus have identified a type of nectary covered by apical bipolar epidermal cells, reminiscent of the secretory brush border epithelium in animals. Border epithelia operate transepithelial anion transport, which is required for membrane potential and/or osmotic adjustment of the secretory cells. In search of anion transporters expressed in extrafloral nectaries, we identified PttSLAH3 (Populus tremula × Populus tremuloides SLAC1 Homologue3), an anion channel of the SLAC/SLAH family. When expressed in Xenopus oocytes, PttSLAH3 displayed the features of a voltage-dependent anion channel, permeable to both nitrate and chloride. In contrast to the Arabidopsis SLAC/SLAH family members, the poplar isoform PttSLAH3 is independent of phosphorylation activation by protein kinases. To understand the basis for the autonomous activity of the poplar SLAH3, we generated and expressed chimera between kinase-independent PttSLAH3 and kinase-dependent Arabidopsis AtSLAH3. We identified the N-terminal tail and, to a lesser extent, the C-terminal tail as responsible for PttSLAH3 kinase-(in)dependent action. This feature of PttSLAH3 may provide the secretory cell with a channel probably controlling long-term nectar secretion.

Phenylcoumaran benzylic ether reductase prevents accumulation of compounds formed under oxidative conditions in poplar xylem.

Phenylcoumaran benzylic ether reductase (PCBER) is one of the most abundant proteins in poplar (Populus spp) xylem, but its biological role has remained obscure. In this work, metabolite profiling of transgenic poplar trees downregulated in PCBER revealed both the in vivo substrate and product of PCBER. Based on mass spectrometry and NMR data, the substrate was identified as a hexosylated 8-5-coupling product between sinapyl alcohol and guaiacylglycerol, and the product was identified as its benzyl-reduced form. This activity was confirmed in vitro using a purified recombinant PCBER expressed in Escherichia coli. Assays performed on 20 synthetic substrate analogs revealed the enzyme specificity. In addition, the xylem of PCBER-downregulated trees accumulated over 2000-fold higher levels of cysteine adducts of monolignol dimers. These compounds could be generated in vitro by simple oxidative coupling assays involving monolignols and cysteine. Altogether, our data suggest that the function of PCBER is to reduce phenylpropanoid dimers in planta to form antioxidants that protect the plant against oxidative damage. In addition to describing the catalytic activity of one of the most abundant enzymes in wood, we provide experimental evidence for the antioxidant role of a phenylpropanoid coupling product in planta.

Involvement of respiratory processes in the transient knockout of net CO2 uptake in Mimosa pudica upon heat stimulation.

Leaf photosynthesis of the sensitive plant Mimosa pudica displays a transient knockout in response to electrical signals induced by heat stimulation. This study aims at clarifying the underlying mechanisms, in particular, the involvement of respiration. To this end, leaf gas exchange and light reactions of photosynthesis were assessed under atmospheric conditions largely eliminating photorespiration by either elevated atmospheric CO2 or lowered O2 concentration (i.e. 2000 µmol mol(-1) or 1%, respectively). In addition, leaf gas exchange was studied in the absence of light. Under darkness, heat stimulation caused a transient increase of respiratory CO2 release simultaneously with stomatal opening, hence reflecting direct involvement of respiratory stimulation in the drop of the net CO2 uptake rate. However, persistence of the transient decline in net CO2 uptake rate under illumination and elevated CO2 or 1% O2 makes it unlikely that photorespiration is the metabolic origin of the respiratory CO2 release. In conclusion, the transient knockout of net CO2 uptake is at least partially attributed to an increased CO2 release through mitochondrial respiration as stimulated by electrical signals. Putative CO2 limitation of Rubisco due to decreased activity of carbonic anhydrase was ruled out as the photosynthesis effect was not prevented by elevated CO2 .

Electrical signaling along the phloem and its physiological responses in the maize leaf.

To elucidate the role of electrical signaling in the phloem of maize the tips of attached leaves were stimulated by chilling and wounding. Two different signals were detected in the phloem at the middle of the leaf using the aphid stylet technique: (1) action potentials (AP) arose in the phloem after chilling; and (2) variation potentials (VPs) were evoked after wounding the leaf tip. Combined electric potential and gas exchange measurements showed that while the wound-induced VP moved rapidly towards the middle of the leaf to induce a reduction in both the net-CO2 uptake rate and the stomatal conductance, there was no response in the gas exchange to the cold-induced AP. To determine if electrical signaling had any impact on assimilate transport the middle of the leaf was exposed to (14)CO2. Autoradiography of labeled assimilates provided evidence that phloem and intercellular transport of assimilates from mesophyll to bundle sheath cells was strongly reduced while the cold-induced AP moved through. In contrast, wound-induced VP did not inhibit assimilate translocation but did reduce the amount of the labeled assimilate in phloem and bundle sheath cells. Biochemical analysis revealed that callose content increased significantly in chilled leaves while starch increased in chilled but decreased in wounded leaves. The results led to the conclusion that different stimulation types incite characteristic phloem-transmitted electrical signals, each with a specific influence on gas exchange and assimilate transport.

Photosynthetic responses of soybean (Glycine max L.) to heat-induced electrical signalling are predominantly governed by modifications of mesophyll conductance for CO(2).

In recent years, the effect of heat-induced electrical signalling on plant photosynthetic activity has been demonstrated for many plant species. However, the underlying triggers of the resulting transient inhibition of photosynthesis still remain unknown. To further investigate on this phenomenon, we focused in our present study on soybean (Glycine max L.) on the direct effect of signal transmission in the leaf mesophyll on conductance for CO(2) diffusion in the mesophyll (g(m) ) and detected a drastic decline in g(m) following the electrical signal, whereas the photosynthetic electron transport rate (ETR) was only marginally affected. In accordance with the drop in net photosynthesis (A(N) ), energy dispersive X-ray analysis (EDXA) revealed a shift of K, Mg, O and P on leaf chloroplasts. Control experiments under elevated CO(2) conditions proved the transient reduction of A(N) , ETR, the chloroplast CO(2) concentration (C(c) ) and g(m) to be independent of the external CO(2) regime, whereas the effect of the electrical signal on stomatal conductance for CO(2) (g(s) ) turned out much less distinctive. We therefore conclude that the effect of electrical signalling on photosynthesis in soybean is triggered by its immediate effects on g(m) .

Poplar wood rays are involved in seasonal remodeling of tree physiology.

Understanding seasonality and longevity is a major challenge in tree biology. In woody species, growth phases and dormancy follow one another consecutively. In the oldest living individuals, the annual cycle may run for more than 1,000 years. So far, however, not much is known about the processes triggering reactivation from dormancy. In this study, we focused on wood rays, which are known to play an important role in tree development. The transition phase from dormancy to flowering in early spring was compared with the phase of active growth in summer. Rays from wood samples of poplar (Populus × canescens) were enriched by laser microdissection, and transcripts were monitored by poplar whole-genome microarrays. The resulting seasonally varying complex expression and metabolite patterns were subjected to pathway analyses. In February, the metabolic pathways related to flower induction were high, indicating that reactivation from dormancy was already taking place at this time of the year. In July, the pathways related to active growth, like lignin biosynthesis, nitrogen assimilation, and defense, were enriched. Based on "marker" genes identified in our pathway analyses, we were able to validate periodical changes in wood samples by quantitative polymerase chain reaction. These studies, and the resulting ray database, provide new insights into the steps underlying the seasonality of poplar trees.

Poplar extrafloral nectaries: two types, two strategies of indirect defenses against herbivores.

Many plant species grow extrafloral nectaries and produce nectar to attract carnivore arthropods as defenders against herbivores. Two nectary types that evolved with Populus trichocarpa (Ptr) and Populus tremula × Populus tremuloides (Ptt) were studied from their ecology down to the genes and molecules. Both nectary types strongly differ in morphology, nectar composition and mode of secretion, and defense strategy. In Ptt, nectaries represent constitutive organs with continuous merocrine nectar flow, nectary appearance, nectar production, and flow. In contrast, Ptr nectaries were found to be holocrine and inducible. Neither mechanical wounding nor the application of jasmonic acid, but infestation by sucking insects, induced Ptr nectar secretion. Thus, nectaries of Ptr and Ptt seem to answer the same threat by the use of different mechanisms.

Influence of over-expression of the Flowering Promoting Factor 1 gene (FPF1) from Arabidopsis on wood formation in hybrid poplar (Populus tremula L. × P. tremuloides Michx.).

Constitutive expression of the FPF1 gene in hybrid aspen (Populus tremula L. × P. tremuloides Michx.) showed a strong effect on wood formation but no effect on flowering time. Gene expression studies showed that activity of flowering time genes PtFT1, PtCO2, and PtFUL was not increased in FPF1 transgenic plants. However, the SOC1/TM3 class gene PTM5, which has been related to wood formation and flowering time, showed a strong activity in stems of all transgenic lines studied. Wood density was lower in transgenic plants, despite significantly reduced vessel frequency which was overcompensated by thinner fibre cell walls. Chemical screening of the wood by pyrolysis GC/MS showed that FPF1 transgenics have higher fractions of cellulose and glucomannan products as well as lower lignin content. The latter observation was confirmed by UV microspectrophotometry on a cellular level. Topochemical lignin distribution revealed a slower increase of lignin incorporation in the developing xylem of the transgenics when compared with the wild-type plants. In line with the reduced wood density, micromechanical wood properties such as stiffness and ultimate stress were also significantly reduced in all transgenic lines. Thus, we provide evidence that FPF1 class genes may play a regulatory role in both wood formation and flowering in poplar.

Salt stress induces the formation of a novel type of 'pressure wood' in two Populus species.

• Salinity causes osmotic stress and limits biomass production of plants. The goal of this study was to investigate mechanisms underlying hydraulic adaptation to salinity. • Anatomical, ecophysiological and transcriptional responses to salinity were investigated in the xylem of a salt-sensitive (Populus × canescens) and a salt-tolerant species (Populus euphratica). • Moderate salt stress, which suppressed but did not abolish photosynthesis and radial growth in P. × canescens, resulted in hydraulic adaptation by increased vessel frequencies and decreased vessel lumina. Transcript abundances of a suite of genes (FLA, COB-like, BAM, XET, etc.) previously shown to be activated during tension wood formation, were collectively suppressed in developing xylem, whereas those for stress and defense-related genes increased. A subset of cell wall-related genes was also suppressed in salt-exposed P. euphratica, although this species largely excluded sodium and showed no anatomical alterations. Salt exposure influenced cell wall composition involving increases in the lignin : carbohydrate ratio in both species. • In conclusion, hydraulic stress adaptation involves cell wall modifications reciprocal to tension wood formation that result in the formation of a novel type of reaction wood in upright stems named 'pressure wood'. Our data suggest that transcriptional co-regulation of a core set of genes determines reaction wood composition.