Patterns and dynamics of canopy-root coupling in tropical tree saplings vary with light intensity but not with root depth.

Carbon (C) dynamics in canopy and roots influence whole-tree carbon fluxes, but little is known about canopy regulation of tree-root activity. Here, the patterns and dynamics of canopy-root C coupling are assessed in tropical trees. Large aeroponics facility was used to study the root systems of Ceiba pentandra and Khaya anthotheca saplings directly at different light intensities. In Ceiba, root respiration (Rr ) co-varied with photosynthesis (An ) in large saplings (3-to-7-m canopy-root axis) at high-light, but showed no consistent pattern at low-light. At medium-light and in small saplings (c. 1-m axis), Rr tended to decrease transiently towards midday. Proximal roots had higher Rr and nonstructural carbohydrate concentrations than distal roots, but canopy-root coupling was unaffected by root location. In medium-sized Khaya, no Rr pattern was observed, and in both species, Rr was unrelated to temperature. The early-afternoon increase in Rr suggests that canopy-root coupling is based on mass flow of newly fixed C in the phloem, whereas the early-morning rise in Rr with An indicates an additional coupling signal that travels faster than the phloem sap. In large saplings and potentially also in higher trees, light and possibly additional environmental factors control the diurnal patterns of canopy-root coupling, irrespective of root location.

Regulation of Sucrose Transporters and Phloem Loading in Response to Environmental Cues.

Suc transporters (SUTs) play a key role in the allocation and partitioning of photosynthetically fixed carbon in plants. While a function could be assigned to many members of the SUT family, almost no information is available on their regulation. Here, the transcriptional regulation of SUTs in response to various environmental stimuli in the leaves of five dicots (Arabidopsis [Arabidopsis thaliana], soybean [Glycine max], potato [Solanum tuberosum], tomato [Solanumlycopersicum], and poplar [Populus spp.]) and four monocots (maize [Zeamays], rice [Oryza sativa], wheat [Triticum aestivum], and barley [Hordeum vulgare]) was investigated. Extensive data on expression of SUTs in relation to changes of environmental conditions were obtained through a global analysis of 168 transcriptomics data sets. Results were validated by quantitative PCR measurements and extended by the measurement of photosynthesis rate and phloem sugar content to draw insight on the correlation of SUT expression and sugar export from leaves. For the apoplasmic phloem loaders, a clear difference in transcriptional regulation in response to different environmental stimuli was observed. The consistent patterns of SUT expression under abiotic stress indicates which types of SUTs are involved in the regulation of leaf sugar status and in stress signaling. Furthermore, it is shown that down-regulation of phloem loading is likely to be caused by transcriptional regulation of SUTs, while up-regulation depends on post-transcriptional regulation. In poplar, expression of PtaSUT4 was found to consistently respond to environmental stimuli, suggesting a significant role in the regulation of sugar export from leaves in this passive symplasmic phloem loader.

The sweet cherry (Prunus avium) FLOWERING LOCUS T gene is expressed during floral bud determination and can promote flowering in a winter-annual Arabidopsis accession.

KEY MESSAGE: FT gene is expressed in leaves and buds and is involved in floral meristem determination and bud development in sweet cherry. In woody fruit perennial trees, floral determination, dormancy and bloom, depends on perception of different environmental and endogenous cues which converge to a systemic signaling gene known as FLOWERING LOCUS T (FT). In long-day flowering plants, FT is expressed in the leaves on long days. The protein travels through the phloem to the shoot apical meristem, where it induces flower determination. In perennial plants, meristem determination and flowering are separated by a dormancy period. Meristem determination takes place in summer, but flowering occurs only after a dormancy period and cold accumulation during winter. The roles of FT are not completely clear in meristem determination, dormancy release, and flowering in perennial plants. We cloned FT from sweet cherry (Prunus avium) and analyzed its expression pattern in leaves and floral buds during spring and summer. Phylogenetic analysis shows high identity of the FT cloned sequence with orthologous genes from other Rosaceae species. Our results show that FT is expressed in both leaves and floral buds and increases when the daylight reached 12 h. The peak in FT expression was coincident with floral meristem identity genes expression and morphological changes typical of floral meristem determination. The Edi-0 Arabidopsis ecotype, which requires vernalization to flower, was transformed with a construct for overexpression of PavFT. These transgenic plants showed an early-flowering phenotype without cold treatment. Our results suggest that FT is involved in floral meristem determination and bud development in sweet cherry. Moreover, we show that FT is expressed in both leaves and floral buds in this species, in contrast to annual plants.

Functional ratios among leaf, xylem and phloem areas in branches change with shade tolerance, but not with local light conditions, across temperate tree species.

Leaf, xylem and phloem areas drive the water and carbon fluxes within branches and trees, but their mutual coordination is poorly understood. We test the hypothesis that xylem and phloem areas increase relative to leaf area when species are selected for, or branches are exposed to, higher levels of light intensity. Trees of 10 temperate, broadleaved and deciduous, tree species were selected. Fifty-centimetre-long branches were collected from shaded and exposed conditions at a height of 3-4 m. We measured the total leaf area, xylem area, phloem area and leaf traits, as well as the area of the constituent cell types, for a stem section at the branch base. Xylem area : leaf area and phloem area : leaf area ratios did not differ consistently between sun and shade branches, but, as expected, they decreased with species' shade tolerance. Similar trends were observed for conductive cell areas in xylem and phloem. Trees of light-demanding species maintain higher water loss and carbon gain rates per leaf area by producing more xylem area and phloem area than shade-tolerant species. We call for more comparative branch studies as they provide an integrated biological perspective on functional traits and their role in the ecology of tree species.

Mathematical Modeling of the Dynamics of Shoot-Root Interactions and Resource Partitioning in Plant Growth.

Plants are highly plastic in their potential to adapt to changing environmental conditions. For example, they can selectively promote the relative growth of the root and the shoot in response to limiting supply of mineral nutrients and light, respectively, a phenomenon that is referred to as balanced growth or functional equilibrium. To gain insight into the regulatory network that controls this phenomenon, we took a systems biology approach that combines experimental work with mathematical modeling. We developed a mathematical model representing the activities of the root (nutrient and water uptake) and the shoot (photosynthesis), and their interactions through the exchange of the substrates sugar and phosphate (Pi). The model has been calibrated and validated with two independent experimental data sets obtained with Petunia hybrida. It involves a realistic environment with a day-and-night cycle, which necessitated the introduction of a transitory carbohydrate storage pool and an endogenous clock for coordination of metabolism with the environment. Our main goal was to grasp the dynamic adaptation of shoot:root ratio as a result of changes in light and Pi supply. The results of our study are in agreement with balanced growth hypothesis, suggesting that plants maintain a functional equilibrium between shoot and root activity based on differential growth of these two compartments. Furthermore, our results indicate that resource partitioning can be understood as the emergent property of many local physiological processes in the shoot and the root without explicit partitioning functions. Based on its encouraging predictive power, the model will be further developed as a tool to analyze resource partitioning in shoot and root crops.

Synthesis of amphiphilic polysuccinimide star copolymers for responsive delivery in plants.

While polymeric nanocarriers are widely used in medicine for controlled release and site-specific delivery, few reports have applied such delivery methods within agriculture, despite the urgent need for specific delivery of pesticides and nutrients. We report the synthesis of stimuli-responsive and biodegradable polymeric nanocarriers designed for delivery to the phloem of plants and describe methods employed to evaluate their toxicity in plant cells.

Systemic signalling in photosynthetic induction of Rumex†K-1 (Rumex patientia × Rumex tianschaious) leaves.

The rapid induction of photosynthesis is critical for plants under light-fleck environment. Most previous studies about photosynthetic induction focused upon single leaf, but they did not consider the systemic integrity of plant. Here, we verified whether systemic signalling is involved in photosynthetic induction. Rumex K-1 (Rumex patientia × Rumex tianschaious) plants were grown under light-fleck condition. After whole night dark adaptation, different numbers of leaves (system leaf or SL) were pre-illuminated with light, and then the photosynthetic induction of other leaves (target leaf or TL) was investigated. This study showed that the pre-illumination of SL promoted photosynthetic induction in TL. This promotion was independent of the number of SL, the light intensity on SL and the distance between SL and TL, indicating that this systemic signalling is non-dose-dependent. More interestingly, the photosynthetic induction was promoted by only the pre-illumination of morphological upper leaf rather than the pre-illumination of morphological lower leaf, indicating that the transfer of this signal is directional. The results showed that the transfer of this systemic signalling depends upon the phloem. This systemic signalling helps plants to use light energy more efficiently under light flecks.

Light intensity and temperature affect systemic spread of silencing signal in transient agroinfiltration studies.

RNA silencing is a sequence-specific post-transcriptional gene inactivation mechanism that operates in diverse organisms and that can extend beyond its site of initiation, owing to the movement of the silencing signal, called non-autonomous gene silencing. Previous studies have shown that several factors manifest the movement of the silencing signal, such as the size (21 or 24 nucleotides) of the secondary small interfering RNA (siRNA) produced, the steady-state concentration of siRNAs and their cognate messenger RNA (mRNA) or a change in the sink-source status of plant parts affecting phloem translocation. Our study shows that both light intensity and temperature have a significant impact on the systemic movement of the silencing signal in transient agroinfiltration studies in Nicotiana benthamiana. At higher light intensities (≥ 450 µE/m(2)/s) and higher temperatures (≥ 30 °C), gene silencing was localized to leaf tissue that was infiltrated, without any systemic spread. Interestingly, in these light and temperature conditions (≥ 450 µE/m(2) /s and ≥ 30 °C), the N. benthamiana plants showed recovery from the viral symptoms. However, the reduced systemic silencing and reduced viral symptom severity at higher light intensities were caused by a change in the sink-source status of the plant, ultimately affecting the phloem translocation of small RNAs or the viral genome. In contrast, at lower light intensities (<300 µE/m(2)/s) with a constant temperature of 25 °C, there was strong systemic movement of the silencing signal in the N. benthamiana plants and reduced recovery from virus infections. The accumulation of gene-specific siRNAs was reduced at higher temperature as a result of a reduction in the accumulation of transcript on transient agroinfiltration of RNA interference (RNAi) constructs, mostly because of poor T-DNA transfer activity of Agrobacterium, possibly also accompanied by reduced phloem translocation.

High light decreases xylem contribution to fruit growth in tomato.

Recently, contradicting evidence has been reported on the contribution of xylem and phloem influx into tomato fruits, urging the need for a better understanding of the mechanisms involved in fruit growth. So far, little research has been performed on quantifying the effect of light intensity on the different contributors to the fruit water balance. However, as light intensity affects both transpiration and photosynthesis, it might be expected to induce important changes in the fruit water balance. In this study, tomato plants (Solanum lycopersicum L.) were grown in light and shade conditions and the fruit water balance was studied by measuring fruit growth of girdled and intact fruits with linear variable displacement transducers combined with a model-based approach. Results indicated that the relative xylem contribution significantly increased when shading lowered light intensity. This resulted from both a higher xylem influx and a lower phloem influx during the daytime. Plants from the shade treatment were able to maintain a stronger gradient in total water potential between stem and fruits during daytime, thereby promoting xylem influx. It appeared that the xylem pathway was still functional at 35 days after anthesis and that relative xylem contribution was strongly affected by environmental conditions.

Blue light-promoted rice leaf bending and unrolling are due to up-regulated brassinosteroid biosynthesis genes accompanied by accumulation of castasterone.

In this study the relationship between blue light- and brassinosteroid-enhanced leaf lamina bending and unrolling in rice was investigated. Twenty-four hours (h) irradiation with white or blue light increased endogenous brassinosteroid levels, especially those of typhasterol and castasterone, in aerial tissues of rice seedlings. There was an accompanying up-regulation of transcript levels of CYP85A1/OsDWARF, encoding an enzyme catalyzing C-6 oxidation, after 6h under either white or blue light. These effects were not observed in seedlings placed under far-red or red light regimes. It was concluded that blue light up-regulates the levels of several cytochrome P450 enzymes including CYP85A1, thereby promoting the synthesis of castasterone, a biologically active brassinosteroid in rice. Based on these findings, it is considered that blue light-mediated rice leaf bending and unrolling are consequences of the enhanced biosynthesis of endogenous castasterone. In contrast to aerial tissues, brassinosteroid synthesis in roots appeared to be negatively regulated by white, blue and red light but positively controlled by far-red light.

phytochrome B Is required for light-mediated systemic control of stomatal development.

Stomata are pores found on the surfaces of leaves, and they regulate gas exchange between the plant and the environment [1]. Stomatal development is highly plastic and is influenced by environmental signals [2]. Light stimulates stomatal development, and this response is mediated by plant photoreceptors [3-5], with the red-light photoreceptor phytochrome B (phyB) having a dominant role in white light [3]. Light also regulates stomatal development systemically, with the irradiance perceived by mature leaves modulating stomatal development in young leaves [6, 7]. Here, we show that phyB is required for this systemic response. Using a combination of tissue-specific expression and an inducible expression system in the loss-of-function phyB-9 mutant [8], we show that phyB expression in the stomatal lineage, mesophyll, and phloem is sufficient to restore wild-type stomatal development. Induction of PHYB in mature leaves also rescues stomatal development in young untreated leaves, whereas phyB mutants are defective in the systemic regulation of stomatal development. Our data show that phyB acts systemically to regulate cell fate decisions in the leaf epidermis.

Proteins from the FLOWERING LOCUS T-like subclade of the PEBP family act antagonistically to regulate floral initiation in tobacco.

Flowering is an important agronomic trait that often depends on the integration of photoperiod, vernalization, gibberellin and/or autonomous signaling pathways by regulatory proteins such as FLOWERING LOCUS T (FT), a member of the phosphatidylethanolamine-binding protein (PEBP) family. Six PEBP family proteins control flowering in the model plant Arabidopsis thaliana, and their regulatory functions are well established, but variation in the number and structural diversity of PEBPs in different species means their precise functions must be determined on a case-by-case basis. We isolated four novel FT-like genes from Nicotiana tabacum (tobacco), and determined their expression profiles in wild-type plants and their overexpression phenotypes in transgenic plants. We found that all four genes were expressed in leaves under short-day conditions, and at least NtFT3 expression was restricted to phloem companion cells. We also found that the NtFT1, NtFT2 and NtFT3 proteins are floral inhibitors (atypical for FT-like proteins), whereas only NtFT4 is a floral inducer. We were unable to detect the expression of these genes under long-day conditions, suggesting that all four tobacco FT-like proteins may control flowering in response to short days. Phylogenetic analysis of PEBP family proteins and their functions in different solanaceous species confirmed that gene duplication and divergence within the FT-like clade has led to the evolution of antagonistic regulators that may help to fine-tune floral initiation in response to environmental cues.

More than just a vulnerable pipeline: xylem physiology in the light of ion-mediated regulation of plant water transport.

Major restrictions to the hydraulic conductance of xylem (K(XYL)) in vascular plants have traditionally been attributed to anatomical constraints. More recently, changes in the cationic concentration of xylem sap have been suggested to be responsible for short-term changes in K(XYL) based on data for 35 dicot species, and very few gymnosperms and ferns, indicating that xylem water transport may no longer be considered as an entirely passive process. Recent studies have revealed that this so-called ionic effect: (i) varies from little or no increase to >30%, (ii) is species specific, (iii) changes on a seasonal basis, (iv) depends on the cationic concentration, (v) is enhanced in embolized stems, and (vi) is positively correlated with vessel grouping. Furthermore, the ionic effect has been suggested to play functional roles in planta with respect to: (i) phloem-mediated control of xylem hydraulic properties, (ii) compensation of cavitation-induced loss of hydraulic conductance, with the result of optimizing light and water utilization, and (iii) differential regulation of water delivery to branches exposed to different levels of light. Pits are likely to play a key role in the ionic effect, which has largely been explained as a consequence of the poly-electrolytic nature and hydrogel properties of the pectic matrix of interconduit pit membranes, despite little evidence that pit membrane pectins remain present after cell hydrolysis. More research is needed to address the ionic effect in more species, physico-chemical properties of pit membranes, and how the ionic effect may increase xylem hydraulic conductance 'on demand'.

Within-canopy and ozone fumigation effects on delta13C and Delta18O in adult beech (Fagus sylvatica) trees: relation to meteorological and gas exchange parameters.

In this study, the effects of different light intensities either in direct sunlight or in the shade crown of adult beech (Fagus sylvatica L.) trees on delta13C and Delta18O were determined under ambient (1 x O3) and twice-ambient (2 x O3) atmospheric ozone concentrations during two consecutive years (2003 and 2004). We analysed the isotopic composition in leaf bulk, leaf cellulose, phloem and xylem material and related the results to (a) meteorological data (air temperature, T and relative humidity, RH), (b) leaf gas exchange measurements (stomatal conductance, g(s); transpiration rate, E; and maximum photosynthetic activity, A(max)) and (c) the outcome of a steady-state evaporative enrichment model. Delta13C was significantly lower in the shade than in the sun crown in all plant materials, whilst Delta18O was increased significantly in the shade than in the sun crown in bulk material and cellulose. Elevated ozone had no effect on delta13C, although Delta18O was influenced by ozone to varied degrees during single months. We observed significant seasonal changes for both parameters, especially in 2004, and also significant differences between the study years. Relating the findings to meteorological data and gas exchange parameters, we conclude that the differences in Delta18O between the sun and the shade crown were predominantly caused by the Péclet effect. This assumption was supported by the modelled Delta18O values for leaf cellulose. It was demonstrated that independent of RH, light-dependent reduction of stomatal conductance (and thus transpiration) and of A(max) can drive the pattern of Delta18O increase with the concomitant decrease of delta13C in the shade crown. The effect of doubling ozone levels on time-integrated stomatal conductance and transpiration as indicated by the combined analysis of Delta18O and delta13C was much lower than the influence caused by the light exposure.

Morphology of nectaries and biology of nectar production in the distylous species Fagopyrum esculentum.

BACKGROUND AND AIMS: The mechanisms of floral nectar production in buckwheat (Fagopyrum esculentum, Polygonaceae), a distylous pseudo-cereal, have received relatively little attention, prompting an investigation of the factors that regulate this process. The aim was to perform a refined study of the structures that secrete nectar and of the internal and external parameters influencing nectar volumes and sugar concentrations. METHODS: In order to control environmental parameters, plants were cultivated in growth rooms under controlled conditions. The structure of nectaries was studied based on histological sections from flowers and flower buds. Nectar was extracted using glass micropipettes and the sugar concentration was measured with a hand refractometer. Sugar concentration in the phloem sap was measured using the anthrone method. To test the influence of photosynthesis on nectar production, different light and defoliation treatments were applied. KEY RESULTS: Unicellular trichomes were located in the epidermis at the ventral part of eight nectary glands situated on the flower receptacle alternately with stamens. Vascular bundles consisting of both phloem and xylem were identified at the boundary between a multilayered nectary parenchyma and a sub-nectary parenchyma with chloroplasts. A higher volume of nectar in thrum morphs was observed. No other difference was found in morphology or in sugar supply to inflorescences between morphs. Nectar secretion was strongly influenced by plant age and inflorescence position. Nectar volumes were higher in the upper inflorescences and during the flowering peak. Light had a dual role, (1) acting directly on reproductive structures to trigger flower opening, which conditions nectar secretion, and (2) stimulating photosynthetic activity, which regulates nectar accumulation in open flowers. CONCLUSIONS: In buckwheat, nectar is secreted by trichomes and probably proceeds, at least in part, from phloem sap. Nectar secretion is strongly influenced by floral morph type, plant age, inflorescence position and light.

Photosynthetic acclimation in the context of structural constraints to carbon export from leaves.

The potential role of foliar carbon export features in the acclimation of photosynthetic capacity to differences and changes in light environment was evaluated. These features included apoplastic vs. symplastic phloem loading, density of loading veins, plasmodesmatal frequency in intermediary cells, and the ratio of loading cells to sieve elements. In initial studies, three apoplastic loaders (spinach, pea, Arabidopsis thaliana) exhibited a completely flexible photosynthetic response to changing light conditions, while two symplastic loaders (pumpkin, Verbascum phoeniceum), although able to adjust to different long-term growth conditions, were more limited in their response when transferred from low (LL) to high (HL) light. This suggested that constraints imposed by the completely physical pathway of sugar export might act as a bottleneck in the export of carbon from LL-acclimated leaves of symplastic loaders. While both symplastic loaders exhibited variable loading vein densities (low in LL and high in HL), none of the three apoplastic loaders initially characterized exhibited such differences. However, an additional apoplastic species (tomato) exhibited similar differences in vein density during continuous growth in different light environments. Furthermore, in contrast to the other apoplastic loaders, photosynthetic acclimation in tomato was not complete following a transfer from LL to HL. This suggests that loading vein density and loading cells per sieve element, and thus apparent loading surface capacity, play a major role in the potential for photosynthetic acclimation to changes in light environment. Photosynthetic acclimation and vein density acclimation were also characterized in the slow-growing, sclerophytic evergreen Monstera deliciosa. This evergreen possessed a lower vein density during growth in LL compared to HL and exhibited a more severely limited potential for photosynthetic acclimation to increases in light environment than the rapidly-growing, mesophytic annuals.

Dynamics of a mobile RNA of potato involved in a long-distance signaling pathway.

BEL1-like transcription factors interact with Knotted1 types to regulate numerous developmental processes. In potato (Solanum tuberosum), the BEL1 transcription factor St BEL5 and its protein partner POTH1 regulate tuber formation by mediating hormone levels in the stolon tip. The accumulation of St BEL5 RNA increases in response to short-day photoperiods, inductive for tuber formation. RNA detection methods and heterografting experiments demonstrate that BEL5 transcripts are present in phloem cells and move across a graft union to localize in stolon tips, the site of tuber induction. This movement of RNA to stolon tips is correlated with enhanced tuber production. Overexpression of BEL5 transcripts that include the untranslated sequences of the BEL5 transcript endows transgenic lines with the capacity to overcome the inhibitory effects of long days on tuber formation. Addition of the untranslated regions leads to preferential accumulation of the BEL5 RNA in stolon tips under short-day conditions. Using a leaf-specific promoter, the movement of BEL5 RNA to stolon tips was facilitated by a short-day photoperiod, and this movement was correlated with enhanced tuber production. These results implicate the transcripts of St BEL5 in a long-distance signaling pathway that are delivered to the target organ via the phloem stream.