Root-to-shoot signaling positively mediates source-sink relation in late growth stages in diploid and tetraploid wheat.

Non-hydraulic root source signaling (nHRS) is a unique positive response to soil drying in the regulation of plant growth and development. However, it is unclear how the nHRS mediates the tradeoff between source and sink at the late growth stages and its adaptive mechanisms in primitive wheat. To address this issue, a root-splitting design was made by inserting solid partition in the middle of the pot culture to induce the occurrence of nHRS using four wheat cultivars (MO1 and MO4, diploid; DM22 and DM31, tetraploid) as materials. Three water treatments were designed as 1) both halves watered (CK), 2) holistic root system watered then droughted (FS), 3) one-half of the root system watered and half droughted (PS). FS and PS were designed to compare the role of the full root system and split root system to induce nHRS. Leaves samples were collected during booting and anthesis to compare the role of nHRS at both growth stages. The data indicated that under PS treatment, ABA concentration was significantly higher than FS and CK, demonstrating the induction of nHRS in split root design and nHRS decreased cytokinin (ZR) levels, particularly in the PS treatment. Soluble sugar and proline accumulation were higher in the anthesis stage as compared to the booting stage. POD activity was higher at anthesis, while CAT was higher at the booting stage. Increased ABA (nHRS) correlated with source-sink relationships and metabolic rate (i.e., leaf) connecting other stress signals. Biomass density showed superior resource acquisition and utilization capabilities in both FS and PS treatment as compared to CK in all plants. Our findings indicate that nHRS-induced alterations in phytohormones and their effect on source-sink relations were allied with the growth stages in primitive wheat.

Sink strength, nutrient allocation, cannabinoid yield and associated transcript profiles vary in two drug-type Cannabis chemovars.

Cannabis sativa L. is one of the oldest domesticated crops. Hemp-type cultivars, which predominantly produce non-intoxicating cannabidiol (CBD), have been selected for their fast growth, seed, and fibre production, while drug-type chemovars were bred for high accumulation of tetrahydrocannabinol (THC). We investigated how the generation of CBD-dominant chemovars by introgression of hemp- into drug-type Cannabis impacted plant performance. The THC-dominant chemovar showed superior sink strength, higher flower biomass and demand-driven control of nutrient uptake. By contrast, the CBD-dominant chemovar hyperaccumulated phosphate in sink organs leading to reduced carbon and nitrogen assimilation in leaves, which limited flower biomass and cannabinoid yield. RNA-seq analyses determined organ- and chemovar-specific differences in expression of genes associated with nitrate and phosphate homeostasis as well as growth-regulating transcription factors that were correlated with measured traits. Among these were genes positively selected for during Cannabis domestication encoding an inhibitor of the phosphate starvation response SPX DOMAIN GENE3, nitrate reductase and two nitrate transporters. Altered nutrient sensing, acquisition or distribution are likely a consequence of adaption to growth on marginal, low-nutrient input lands in hemp. Our data provide evidence that such ancestral traits may become detrimental for female flower development and consequently overall CBD yield in protected cropping environments.

The Alteration of Tomato Chloroplast Vesiculation Positively Affects Whole-Plant Source-Sink Relations and Fruit Metabolism under Stress Conditions.

Changes in climate conditions can negatively affect the productivity of crop plants. They can induce chloroplast degradation (senescence), which leads to decreased source capacity, as well as decreased whole-plant carbon/nitrogen assimilation and allocation. The importance, contribution and mechanisms of action regulating source-tissue capacity under stress conditions in tomato (Solanum lycopersicum) are not well understood. We hypothesized that delaying chloroplast degradation by altering the activity of the tomato chloroplast vesiculation (CV) under stress would lead to more efficient use of carbon and nitrogen and to higher yields. Tomato CV is upregulated under stress conditions. Specific induction of CV in leaves at the fruit development stage resulted in stress-induced senescence and negatively affected fruit yield, without any positive effects on fruit quality. Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9 (CRISPR/CAS9) knockout CV plants, generated using a near-isogenic tomato line with enhanced sink capacity, exhibited stress tolerance at both the vegetative and the reproductive stages, leading to enhanced fruit quantity, quality and harvest index. Detailed metabolic and transcriptomic network analysis of sink tissue revealed that the l-glutamine and l-arginine biosynthesis pathways are associated with stress-response conditions and also identified putative novel genes involved in tomato fruit quality under stress. Our results are the first to demonstrate the feasibility of delayed stress-induced senescence as a stress-tolerance trait in a fleshy fruit crop, to highlight the involvement of the CV pathway in the regulation of source strength under stress and to identify genes and metabolic pathways involved in increased tomato sink capacity under stress conditions.

Sorghum in dryland: morphological, physiological, and molecular responses of sorghum under drought stress.

MAIN CONCLUSION: Droughts negatively affect sorghum's productivity and nutritional quality. Across its diversity centers, however, there exist resilient genotypes that function differently under drought stress at various levels, including molecular and physiological. Sorghum is an economically important and a staple food crop for over half a billion people in developing countries, mostly in arid and semi-arid regions where drought stress is a major limiting factor. Although sorghum is generally considered tolerant, drought stress still significantly hampers its productivity and nutritional quality across its major cultivation areas. Hence, understanding both the effects of the stress and plant response is indispensable for improving drought tolerance of the crop. This review aimed at enhancing our understanding and provide more insights on drought tolerance in sorghum as a contribution to the development of climate resilient sorghum cultivars. We summarized findings on the effects of drought on the growth and development of sorghum including osmotic potential that impedes germination process and embryonic structures, photosynthetic rates, and imbalance in source-sink relations that in turn affect seed filling often manifested in the form of substantial reduction in grain yield and quality. Mechanisms of sorghum response to drought-stress involving morphological, physiological, and molecular alterations are presented. We highlighted the current understanding about the genetic basis of drought tolerance in sorghum, which is important for maximizing utilization of its germplasm for development of improved cultivars. Furthermore, we discussed interactions of drought with other abiotic stresses and biotic factors, which may increase the vulnerability of the crop or enhance its tolerance to drought stress. Based on the research reviewed in this article, it appears possible to develop locally adapted cultivars of sorghum that are drought tolerant and nutrient rich using modern plant breeding techniques.

A whole canopy gas exchange system for the targeted manipulation of grapevine source-sink relations using sub-ambient CO2.

BACKGROUND: Elucidating the effect of source-sink relations on berry composition is of interest for wine grape production as it represents a mechanistic link between yield, photosynthetic capacity and wine quality. However, the specific effects of carbohydrate supply on berry composition are difficult to study in isolation as leaf area or crop adjustments can also change fruit exposure, or lead to compensatory growth or photosynthetic responses. A new experimental system was therefore devised to slow berry sugar accumulation without changing canopy structure or yield. This consisted of six transparent 1.2 m3 chambers to enclose large pot-grown grapevines, and large soda-lime filled scrubbers that reduced carbon dioxide (CO2) concentration of day-time supply air by approximately 200 ppm below ambient. RESULTS: In the first full scale test of the system, the chambers were installed on mature Shiraz grapevines for 14 days from the onset of berry sugar accumulation. Three chambers were run at sub-ambient CO2 for 10 days before returning to ambient. Canopy gas exchange, and juice hexose concentrations were determined. Net CO2 exchange was reduced from 65.2 to 30 g vine- 1 day- 1, or 54%, by the sub-ambient treatment. At the end of the 10 day period, total sugar concentration was reduced from 95 to 77 g L- 1 from an average starting value of 23 g L- 1, representing a 25% reduction. Scaling to a per vine basis, it was estimated that 223 g of berry sugars accumulated under ambient supply compared to 166 g under sub-ambient, an amount equivalent to 50 and 72% of total C assimilated. CONCLUSIONS: Through supply of sub-ambient CO2 using whole canopy gas exchange chambers system, an effective method was developed for reducing photosynthesis and slowing the rate of berry sugar accumulation without modifying yield or leaf area. While in this case developed for further investigations of grape and wine composition, the system has broader applications for the manipulation and of study of grapevine source-sink relations.

Direct evidence for modulation of photosynthesis by an arbuscular mycorrhiza-induced carbon sink strength.

It has been suggested that plant carbon (C) use by symbiotic arbuscular mycorrhizal fungi (AMF) may be compensated by higher photosynthetic rates because fungal metabolism creates a strong C sink that prevents photosynthate accumulation and downregulation of photosynthesis. This mechanism remains largely unexplored and lacks experimental evidence. We report here two experiments showing that the experimental manipulation of the mycorrhizal C sink significantly affected the photosynthetic rates of cucumber host plants. We expected that a sudden reduction in sink strength would cause a significant reduction in photosynthetic rates, at least temporarily. Excision of part of the extraradical mycorrhizal mycelium from roots, and causing no disturbance to the plant, induced a sustained (10-40%) decline in photosynthetic rates that lasted from 30 min to several hours in plants that were well-nourished and hydrated, and in the absence of growth or photosynthesis promotion by mycorrhizal inoculation. This effect was though minor in plants growing at high (700 ppm) atmospheric CO2 . This is the first direct experimental evidence for the C sink strength effects exerted by arbuscular mycorrhizal symbionts on plant photosynthesis. It encourages further experimentation on mycorrhizal source-sink relations, and may have strong implications in large-scale assessments and modelling of plant photosynthesis.

High carbon storage in carbon-limited trees.

The concentrations of nonstructural carbohydrates (NSCs) in plant tissues are commonly used as an indicator of total plant carbon (C) supply; but some evidence suggests the possibility for high NSC concentrations during periods of C limitation. Despite this uncertainty, NSC dynamics have not been investigated experimentally under long-term C limitation. We exposed saplings of 10 temperate tree species differing in shade tolerance to 6% of ambient sunlight for 3 yr to induce C limitation, and also defoliated one species, Carpinus betulus, in the third season. Growth and NSC concentrations were monitored to determine C allocation. Shade strongly reduced growth, but after an initial two-fold decrease, NSC concentrations of shaded saplings recovered to the level of unshaded saplings by the third season. NSC concentrations were generally more depleted under shade after leaf flush, and following herbivore attacks. Only under shade did artificial defoliation lead to mortality and depleted NSC concentrations in surviving individuals. We conclude that, irrespective of shade tolerance, C storage is maintained under prolonged shading, and thus high NSC concentrations can occur during C limitation. Yet, our results also suggest that decreased NSC concentrations are indicative of C limitation, and that additional leaf loss can lead to lethal C shortage in deep shade.

Asymmetric pruning reveals how organ connectivity alters the functional balance between leaves and roots of Chinese fir.

The functional balance between leaves and roots is believed to be mediated by the specific location of shoots and roots, i.e. differences in transport distances and degrees of organ connectivity. However, it remains unknown whether the adaptive responses of trees to biomass removal depend on the relative orientation of leaf and root pruning. Here, we applied five pruning treatments to saplings of Cunninghamia lanceolata (Chinese fir) under field and glasshouse conditions, namely no pruning (control), half of lateral branches pruned, half of lateral roots pruned, half of the branches and roots pruned on the same side of the plant, and half of the branches and roots pruned on opposite sides of the plant. The effects of pruning on the growth, carbon storage and allocation, and physiology of leaves and fine roots on the same and opposite sides of the plant were investigated. Compared with the effect of root-pruning on leaves, fine roots were more limited by carbon availability and their physiological activity was more strongly reduced by shoot pruning, especially when branches on the same side of the plant were removed. Pruning of branches and roots on the opposite side of the plant resulted in the lowest carbon assimilation rates and growth among all treatments. The results of a stable-isotope labeling indicated that less C was distributed to fine roots from the leaves on the opposite side of the plant compared to those on the same side, but N allocation from roots to leaves depended less on the relative root and leaf orientation. The results collectively indicate that the functional responses of C. lanceolata to pruning are not only determined by the source-sink balance model but are also related to interactions between leaves and fine roots. We argue that the connectivity among lateral branches and roots depends on their relative orientation, which is therefore critical for the functional balance between leaves and fine roots.

Carbon partitioning in Arabidopsis thaliana is a dynamic process controlled by the plants metabolic status and its circadian clock.

Plant growth involves the coordinated distribution of carbon resources both towards structural components and towards storage compounds that assure a steady carbon supply over the complete diurnal cycle. We used (14) CO2 labelling to track assimilated carbon in both source and sink tissues. Source tissues exhibit large variations in carbon allocation throughout the light period. The most prominent change was detected in partitioning towards starch, being low in the morning and more than double later in the day. Export into sink tissues showed reciprocal changes. Fewer and smaller changes in carbon allocation occurred in sink tissues where, in most respects, carbon was partitioned similarly, whether the sink leaf assimilated it through photosynthesis or imported it from source leaves. Mutants deficient in the production or remobilization of leaf starch exhibited major alterations in carbon allocation. Low-starch mutants that suffer from carbon starvation at night allocated much more carbon into neutral sugars and had higher rates of export than the wild type, partly because of the reduced allocation into starch, but also because of reduced allocation into structural components. Moreover, mutants deficient in the plant's circadian system showed considerable changes in their carbon partitioning pattern suggesting control by the circadian clock.

Linking microbial community coalescence to ecological diversity, community assembly and species coexistence in a typical subhumid river catchment in northern China.

Community coalescence denotes the amalgamation of biotic and abiotic factors across multiple intact ecological communities. Despite the growing attention given to the phenomenon of coalescence, there remains limited investigation into community coalescence in single and multiple source habitats and its impact on microbial community assemblages in sinks. This study focused on a major river catchment in northern China. We investigated microbial community coalescence across different habitats (i.e., water, sediment, biofilm, and riparian soil) and seasons (i.e., summer and winter). Using 16S rRNA gene amplicon sequence variants, we examined the relationship between community coalescence and microbial diversity, assembly processes, and species coexistence. The results showed that the intensity of microbial community coalescence was higher in the same habitat pairs compared to disparate habitat pairs in both summer and winter. During the occurrence of microbial community coalescence, the assembly processes regulated the intensity of coalescence. When the microbial community exhibited strong heterogeneous selection (heterogeneous environmental conditions leading to more dissimilar community structures), the intensity of community coalescence was low. With the assembly process shifted towards stochasticity, coalescence intensity increased gradually. However, when homogeneous selection (homogeneous environmental conditions leading to more similar community structures) predominantly shaped microbial communities, coalescence intensity exceeded the threshold of 0.25-0.30. Moreover, the enhanced intensity of community coalescence could increase the complexity of microbial networks, thereby enhancing species coexistence. Furthermore, the assembly processes mediated the relationship between community coalescence and species coexistence, underscoring the pivotal role of intermediate intensity of community coalescence in maintaining efficient species coexistence. In conclusion, this study highlights the crucial role of community coalescence originating from single and multiple source habitats in shaping microbial communities in sinks, thus emphasizing its central importance in watershed ecosystems.

Positive impacts of livestock and wild ungulate routes on functioning of dryland ecosystems.

Livestock grazing is often perceived as being detrimental to the quality and functioning of dryland ecosystems. For example, a study in a semiarid Kenyan savanna proposed that cattle form bare spaces throughout the landscape, which indicate ecosystem degradation. Other studies, conducted in north-eastern Spain, where climatic conditions range between semiarid and Mediterranean subhumid, reported that sheep and goat trails have increased the emergence of rill erosion processes. Sometimes, this negative perception is extended to include wild, large ungulate herbivores as well. Here, we challenge this perception by highlighting the generally nonadverse and even ameliorative impacts of moderate animal rate on geoecosystem functioning of hilly drylands. Specifically, trampling routes (also known as treading paths, livestock terracettes, cattle trails, migration tracks, cowtours, etc.) formed across hillslopes by grazing animals-being either domesticated livestock or native large herbivores-transform the original two-phase vegetation mosaic of shrubby patches and interpatch spaces into a three-phase mosaic. The animal routes increase the complexity of ecosystem, by strengthening the spatial redistribution of water and soil resources at the patch scale and decreasing hydrological connectivity at the hillslope scale. As a consequence, the animal routes improve functioning of hilly drylands and increase their resilience to long-term droughts and climatic change. Therefore, instead of viewing the animal routes as degraded spots, they should be perceived at a wider perspective that allows to properly understand their overall role in sustaining dryland geoecosystems.

Cycas micronesica Stem Carbohydrates Decline Following Leaf and Male Cone Growth Events.

The growth of synchronized leaf flushes or male cones on Cycas trees is an ephemeral event, and non-structural carbohydrates (NSCs) are likely deployed from stem and root storage tissues to support their construction. The relationships among various stem NSCs and these rapid growth events have not been studied to date. Monosaccharides, disaccharides, and starch were quantified in Cycas micronesica stem tissue prior to and immediately after the growth of leaf flushes or male cones to determine the influences on the concentration of these carbohydrates. The pre-existing leaves were removed from half of the plants to determine if the elimination of this carbon source would influence the NSC behaviors. Starch and sucrose dominated the NSC profiles, and these two NSCs declined following cone or new leaf growth. Removal of pre-existing leaves generated a greater decline in starch and sucrose for cone growth, and a greater decline in sucrose, but not starch following new leaf growth than in control trees with no leaf removal. The initial differences in starch and sucrose among cortex, vascular, and pith tissues disappeared as the concentrations declined in all three tissue categories to reach similar post-growth concentrations among the stem tissue categories. The fructose, glucose, and maltose behaviors were not consistent, and their concentrations were low such that their influence on the total NSC behaviors was minimal. These results provided indirect evidence that stem NSCs were mobilized to support ephemeral male cone and new leaf growth for this arborescent cycad. Growth of female strobili is slow and lengthy, so we did not include female trees in this study. The contributions of stem NSCs to female strobili growth remain to be studied with alternative methods.

Dynamic changes in the starch-sugar interconversion within plant source and sink tissues promote a better abiotic stress response.

Starch is a significant store of sugars, and the starch-sugar interconversion in source and sink tissues plays a profound physiological role in all plants. In this review, we discuss how changes in starch metabolism can facilitate adaptive changes in source-sink carbon allocation for protection against environmental stresses. The stress-related roles of starch are described, and published mechanisms by which starch metabolism responds to short- or long-term water deficit, salinity, or extreme temperatures are discussed. Numerous examples of starch metabolism as a stress response are also provided, focusing on studies where carbohydrates and cognate enzymes were assayed in source, sink, or both. We develop a model that integrates these findings with the theoretical and known roles of sugars and starch in various species, tissues, and developmental stages. In this model, localized starch degradation into sugars is vital to the plant cold stress response, with the sugars produced providing osmoprotection. In contrast, high starch accumulation is prominent under salinity stress, and is associated with higher assimilate allocation from source to sink. Our model explains how starch-sugar interconversion can be a convergent point for regulating carbon use in stress tolerance at the whole-plant level.

Assessing Long-Distance Carbon Partitioning from Photosynthetic Source Leaves to Heterotrophic Sink Organs with Photoassimilated [14C]CO2.

Phloem loading and long-distance transport of photoassimilate from source leaves to sink organs are essential physiological processes that contribute to plant growth and yield. At a minimum, three steps are involved: phloem loading in source organs, transport along the phloem path, and phloem unloading in sink organs. Each of these can have variable rates contingent on the physiological state of the plant, and thereby influence the overall transport rate. In addition to these phloem transport steps, rates of photosynthesis and photosynthate movement in the pre-phloem path, as well as photosynthate utilization in post phloem tissues of sink organs also contribute to phloem transport. The protocol described here estimates carbon allocation along the entire path from initial carbon fixation to delivery to sink organs after a labeling pulse: [14C]CO2 is photoassimilated in source leaves and loading and transport of the 14C label to heterotrophic sink organs (roots) is quantified by scintillation counting. This method is flexible and can be adapted to quantify long-distance transport in many plant species.

A Mechanistic Model to Predict Distribution of Carbon Among Multiple Sinks.

Modeling is a fundamental part of quantitative science used to bring together several quantitative components, often developed though detailed reductionist approach on component parts, e.g., sucrose transport through a membrane osmotic relation. It is now generally accepted that phloem transport is the result of bulk solution flow generated by the difference in osmotic pressure between source and sink tissues. However, there is still little agreement on how different sink tissues compete for available carbohydrate. Furthermore, the impact of phloem pathway leakage (unloading) and reloading on source-to-sink carbon transport remains unclear. Moreover, it is debated to what degree the interactions between phloem and xylem flows influence carbohydrate source-sink relations. These aspects are extremely difficult to research by a reductionist approach, with modeling being an important tool to examine the consequences of proposed mechanisms, which can then be tested on whole plants.Phloem/xylem modeling has been at the limits of quantitative modeling, especially when dynamic models are needed to explain tracer studies. Advances in computing now enable more realistic modeling, which are utilized by the PiafMunch approach described here. This model enables a high level of mechanistic detail to be incorporated and the observable effect of it to be tested. In the most recent version of the software with the introduction of tracer dynamics, it can now predict the effects of specific phloem mechanisms upon the shape of evolving tracer profiles.

Drought induced changes of leaf-to-root relationships in two tomato genotypes.

Water deficit triggers a dynamic and integrated cross-talk between leaves and roots. Tolerant plants have developed several physiological and molecular mechanisms to establish new cell metabolism homeostasis, avoiding and/or escaping from permanent impairments triggered by drought. Two tomato genotypes (a Southern Italy landrace called Ciettaicale and the well-known commercial cultivar Moneymaker) were investigated at vegetative stage to assess leaf and root metabolic strategies under 20 days of water deficit. Physiological and metabolic changes, in terms of ABA, IAA, proline, soluble sugars and phenols contents, occurred in both tomato genotypes under water stress. Overall, our results pointed out the higher plasticity of Ciettaicale to manage plant water status under drought in order to preserve the source-sink relationships. This aim was achieved by maintaining a more efficient leaf photosystem II (PSII) photochemistry, as suggested by chlorophyll fluorescence parameters, associated with a major investment towards root growth and activity to improve water uptake. On the contrary, the higher accumulation of carbon compounds, resulting from reduced PSII photochemistry and enhanced starch reserve mobilization, in leaves and roots of Moneymaker under drought could play a key role in the osmotic adjustment, although causing a feedback disruption of the source-sink relations. This hypothesis was also supported by the different drought-induced redox unbalance, as suggested by H2O2 and MDA contents. This could affect both PSII photochemistry and root activity, leading to a major involvement of NPQ and antioxidant system in response to drought in Moneymaker than Ciettaicale.

Assimilates mobilization, stable canopy temperature and expression of expansin stabilizes grain weight in wheat cultivar LOK-1 under different soil moisture conditions.

BACKGROUND: Grain yield of wheat is primarily determined by both grain number and grain weight, which often influence each other in response to environmental stimuli. Some of the genotypes are capable of maintaining high single grain weight (SGW) across the environments. Understanding mechanisms and factors associated with the superiority of such genotypes over others is necessary to enhance productivity of wheat. RESULTS: Experiments were conducted to elucidate the physiological basis of high SGW of LOK-1, a wheat cultivar grown in dry and hot environments in the central and peninsular zones of India. SGW of LOK-1 was least affected by removal of spikelets indicating little competition between the grains within the spike for assimilates. Reduction in SGW due to defoliation was less and the contribution of stem reserves to the grain development was high in LOK-1 relative to other cultivars. It seems that high level of expression of genes such as expansin (TaExpA6) contributes to the high SGW of LOK-1. CONCLUSIONS: Source was not a limiting factor for grain growth of LOK-1 in contrast to other cultivars, whereas sink appeared to be a limiting factor in recently released/identified cultivars. Differences in the amounts of water soluble stem carbohydrate reserves translocated to grain could be one of the factors contributing to higher grain weight in LOK-1. High level expression of TaExpA6, one of the genes contributing to the elongation of endosperm, seems to be crucial for grain growth in wheat.

How rice Glycogen Synthase Kinase-like 5 (OsGSK5) integrates salinity stress response to source-sink adaptation: A proposed model.

We have previously shown that overexpression of GSK3-like kinase 5 in rice (OsGSK5) was associated with higher starch accumulation and better growth under severe salinity stress. Short-term 14CO2 feeding experiments suggested that OsGSK5 promoted higher flux to starch accumulation in the roots under this condition and that this mechanism may help to underscore the better growth characteristics observed. Here, we expand upon this hypothesis and consider (1) how OsGSK5 action could fit into a signaling model that integrates salinity stress to changes in starch metabolism, and (2) how this would facilitate whole plant physiological adaptations in source-to-sink partitioning. We also discuss additional functions of OsGSK5, necessary to support this adaptive mechanism.

Regulation of reserve mobilisation in sunflower during late seedling establishment in continuous darkness.

Reserve mobilisation, metabolite partitioning and reserve-degrading enzyme activity were studied in sunflower seedlings cultivated in vitro under a 12-h photoperiod or in the dark to investigate the involvement of source-sink relation and carbon starvation in the regulation of reserve mobilisation under continuous darkness. Reserves, metabolites and enzyme activity were determined with standard spectrophotometric methods. At the first 24 h of treatment (acclimation phase), darkness did not affect growth, but restricted carbon and nitrogen use, as indicated by sugar and amino acid accumulation in the different seedling parts. After 5 days of treatment (survival phase), extended darkness limited growth and retarded storage lipid mobilisation due to carbon starvation, as evidenced by the depletion of carbohydrates in cotyledons and hypocotyl, as well as the consumption of amino acids in hypocotyls and roots. Alterations in the source-sink relationship might have been a response to prolonged darkness, instead of a mechanism used to regulate reserve mobilisation, as these alterations cannot be associated with negative feedback mediated by metabolite accumulation. Storage lipid degradation depends, at least in part, on mechanisms that co-ordinately regulate the activities of lipases and isocitrate lyase. Taking these results together, it is possible that reserve mobilisation in sunflower seedlings cultivated in the dark might be regulated by mechanisms that perceive the absence of light and predict carbon starvation, adjusting reserve use according to future energy demands to allow, at least in the short term, seedling survival.