Benjamin A. Babst

Sugar demand, not auxin, is the initial regulator of apical dominance

Significance It is commonly accepted that the plant hormone auxin mediates apical dominance. However, we have discovered that apical dominance strongly correlates with sugar availability and not apically supplied auxin. We have revealed that apical dominance is predominantly controlled by the shoot tip’s intense demand for sugars, which limits sugar availability to the axillary buds. These findings overturn a long-standing hypothesis on apical dominance and encourage us to reevaluate the relationship between hormones and sugars in this and other aspects of plant development. For almost a century the plant hormone auxin has been central to theories on apical dominance, whereby the growing shoot tip suppresses the growth of the axillary buds below. According to the classic model, the auxin indole-3-acetic acid is produced in the shoot tip and transported down the stem, where it inhibits bud growth. We report here that the initiation of bud growth after shoot tip loss cannot be dependent on apical auxin supply because we observe bud release up to 24 h before changes in auxin content in the adjacent stem. After the loss of the shoot tip, sugars are rapidly redistributed over large distances and accumulate in axillary buds within a timeframe that correlates with bud release. Moreover, artificially increasing sucrose levels in plants represses the expression of BRANCHED1 (BRC1), the key transcriptional regulator responsible for maintaining bud dormancy, and results in rapid bud release. An enhancement in sugar supply is both necessary and sufficient for suppressed buds to be released from apical dominance. Our data support a theory of apical dominance whereby the shoot tip’s strong demand for sugars inhibits axillary bud outgrowth by limiting the amount of sugar translocated to those buds.

A specialist root herbivore exploits defensive metabolites to locate nutritious tissues

The most valuable organs of plants are often particularly rich in essential elements, but also very well defended. This creates a dilemma for herbivores that need to maximise energy intake while minimising intoxication. We investigated how the specialist root herbivore Diabrotica virgifera solves this conundrum when feeding on wild and cultivated maize plants. We found that crown roots of maize seedlings were vital for plant development and, in accordance, were rich in nutritious primary metabolites and contained higher amounts of the insecticidal 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) and the phenolic compound chlorogenic acid. The generalist herbivores Diabrotica balteata and Spodoptera littoralis were deterred from feeding on crown roots, whereas the specialist D. virgifera preferred and grew best on these tissues. Using a 1,4-benzoxazin-3-one-deficient maize mutant, we found that D. virgifera is resistant to DIMBOA and other 1,4-benzoxazin-3-ones and that it even hijacks these compounds to optimally forage for nutritious roots.

Lymantria dispar herbivory induces rapid changes in carbon transport and partitioning in Populus nigra

We tested for rapid changes in photosynthate transport and partitioning in response to Lymantria dispar (L.) (Lepidoptera: Lymantriidae) (gypsy moth) herbivory in Populus nigra L. (Salicaceae). Transport and partitioning of [11C]‐photosynthate from young mature leaves were measured in vivo before and 18 h after leaf chewing by gypsy moth larvae, which were caged on three older leaves. Following herbivory, there was an increase in export speed of recently fixed carbon from younger mature leaves. The increased export speed was due to a quicker transit time of 11C through the leaf, rather than a change in transport speed through the phloem. Additionally, basipetal partitioning of [11C]‐photosynthate was increased following herbivory. Neither of these changes was observed in control plants. This enhancement of export occurs even though herbivores are well known to induce increases in carbon allocation to secondary metabolites within leaves. Our results demonstrate that the use of non‐destructive imaging of 11C tracer is a powerful tool for examining plant responses to herbivory. Although the mechanisms underlying the rapid increase in carbon flux to stems and roots remain to be elucidated, our results raise the possibility of a coordinated whole plant response to herbivory. Thus, even when the herbivore specializes on only one plant tissue type, a whole plant approach may be key to understanding how plants respond to herbivory.

Differential sectoriality in long-distance transport in temperate tree species: evidence from dye flow, 15N transport, and vessel element pitting

The capture of patchily distributed nutrients by tree roots has received extensive research, but the fate of those nutrients has not. We performed experiments to determine if nutrient transport within tree species is preferentially transported from specific roots to specific branches. Saplings of five species with contrasting growth requirements were examined: two Betula species (B. papyrifera and B. lenta), Populus tremuloides, and two Acer species (A. saccharum and A. rubrum). To quantify patterns of long-distance transport, we examined the accumulation of safranin-O dye and 15N in branches when these tracers were applied to isolated lateral roots (dye and 15N) and to the main root system (15N). Because transport of nutrients between sectors requires flow through intervessel pit pairs of adjacent xylem vessel elements, we quantified the area of intervessel pits, the number of pits per unit vessel wall area, and the % vessel wall area as pits in Acer and Betula. We found that the two Betula species were integrated (tracers applied to isolated roots were likely to accumulate in all branches), while P. tremuloides and the two Acer species were sectorial (tracer accumulation was more concentrated in particular branches). Betula had the largest number of intervessel pits per unit vessel wall area and the largest percentage of vessel wall area as pits. The high density of bordered pits may explain the ease of tracer movement throughout the two Betula species. Greater integration may allow certain trees (e.g., Betula) to exploit nutritionally patchy environments such as rocky soils, and may alter plant-herbivore interactions.

Induced carbon reallocation and compensatory growth as root herbivore tolerance mechanisms

Upon attack by leaf herbivores, many plants reallocate photoassimilates below ground. However, little is known about how plants respond when the roots themselves come under attack. We investigated induced resource allocation in maize plants that are infested by the larvae Western corn rootworm Diabrotica virgifera virgifera. Using radioactive (11) CO(2), we demonstrate that root-attacked maize plants allocate more new (11) C carbon from source leaves to stems, but not to roots. Reduced meristematic activity and reduced invertase activity in attacked maize root systems are identified as possible drivers of this shoot reallocation response. The increased allocation of photoassimilates to stems is shown to be associated with a marked thickening of these tissues and increased growth of stem-borne crown roots. A strong quantitative correlation between stem thickness and root regrowth across different watering levels suggests that retaining photoassimilates in the shoots may help root-attacked plants to compensate for the loss of belowground tissues. Taken together, our results indicate that induced tolerance may be an important strategy of plants to withstand belowground attack. Furthermore, root herbivore-induced carbon reallocation needs to be taken into account when studying plant-mediated interactions between herbivores.

Constitutively Elevated Salicylic Acid Levels Alter Photosynthesis and Oxidative State but Not Growth in Transgenic Populus

This study describes transcriptional and metabolic network rewiring in Populus that overaccumulate SA by two to three orders of magnitude without negative effects on growth. The work provides in planta evidence for a direct link between SA and oxidative response and contrasts sharply with Arabidopsis where growth and SA overproduction are not compatible. Salicylic acid (SA) has long been implicated in plant responses to oxidative stress. SA overproduction in Arabidopsis thaliana leads to dwarfism, making in planta assessment of SA effects difficult in this model system. We report that transgenic Populus tremula × alba expressing a bacterial SA synthase hyperaccumulated SA and SA conjugates without negative growth consequences. In the absence of stress, endogenously elevated SA elicited widespread metabolic and transcriptional changes that resembled those of wild-type plants exposed to oxidative stress-promoting heat treatments. Potential signaling and oxidative stress markers azelaic and gluconic acids as well as antioxidant chlorogenic acids were strongly coregulated with SA, while soluble sugars and other phenylpropanoids were inversely correlated. Photosynthetic responses to heat were attenuated in SA-overproducing plants. Network analysis identified potential drivers of SA-mediated transcriptome rewiring, including receptor-like kinases and WRKY transcription factors. Orthologs of Arabidopsis SA signaling components NON-EXPRESSOR OF PATHOGENESIS-RELATED GENES1 and thioredoxins were not represented. However, all members of the expanded Populus nucleoredoxin-1 family exhibited increased expression and increased network connectivity in SA-overproducing Populus, suggesting a previously undescribed role in SA-mediated redox regulation. The SA response in Populus involved a reprogramming of carbon uptake and partitioning during stress that is compatible with constitutive chemical defense and sustained growth, contrasting with the SA response in Arabidopsis, which is transient and compromises growth if sustained.

Testing the Münch hypothesis of long distance phloem transport in plants

Long distance transport in plants occurs in sieve tubes of the phloem. The pressure flow hypothesis introduced by Ernst Münch in 1930 describes a mechanism of osmotically generated pressure differentials that are supposed to drive the movement of sugars and other solutes in the phloem, but this hypothesis has long faced major challenges. The key issue is whether the conductance of sieve tubes, including sieve plate pores, is sufficient to allow pressure flow. We show that with increasing distance between source and sink, sieve tube conductivity and turgor increases dramatically in Ipomoea nil. Our results provide strong support for the Münch hypothesis, while providing new tools for the investigation of one of the least understood plant tissues. DOI: http://dx.doi.org/10.7554/eLife.15341.001

Biosynthesis of Phenolic Glycosides from Phenylpropanoid and Benzenoid Precursors in Populus

Salicylate-containing phenolic glycosides (PGs) are abundant and often play a dominant role in plant-herbivore interactions of Populus and Salix species (family Salicaceae), but the biosynthetic pathway to PGs remains unclear. Cinnamic acid (CA) is thought to be a precursor of the salicyl moiety of PGs. However, the origin of the 6-hydroxy-2-cyclohexen-on-oyl (HCH) moiety found in certain PGs, such as salicortin, is not known. HCH is of interest because it confers toxicity and antifeedant properties against herbivores. We incubated Populus nigra leaf tissue with stable isotope-labeled CA, benzoates, and salicylates, and measured isotopic incorporation levels into both salicin, the simplest PG, and salicortin. Labeling of salicortin from [13C6]-CA provided the first evidence that HCH, like the salicyl moiety, is a phenylpropanoid derivative. Benzoic acid and benzaldehyde also labeled both salicyl and HCH, while benzyl alcohol labeled only the salicyl moiety in salicortin. Co-administration of unlabeled benzoates with [13C6]-CA confirmed their contribution to the biosynthesis of the salicyl but not the HCH moiety of salicortin. These data suggest that benzoate interconversions may modulate partitioning of phenylpropanoids to salicyl and HCH moieties, and hence toxicity of PGs. Surprisingly, labeled salicyl alcohol and salicylaldehyde were readily converted to salicin, but did not result in labeled salicortin. Co-administration of unlabeled salicylates with labeled CA suggested that salicyl alcohol and salicylaldehyde may have inhibited salicortin biosynthesis. A revised metabolic grid model of PG biosynthesis in Populus is proposed, providing a guide for functional genomic analysis of the PG biosynthetic pathway.

Vascular Constraints and Long Distance Transport in Dicots

Publisher Summary Plants are often assumed to be capable of freely translocating resources from one part of the plant to all other parts, a condition that we describe as integrated. In reality, many vascular plants are sectorial, meaning that the movement of nutrients, photosynthate, and other substances is restricted to specific vascular, xylem or phloem, pathways. Sectorial plants consist of numerous integrated physiological units (IPUs) whose pathways are relatively independent of each other. Integrated and sectorial define two ends of what in reality is a continuum among species. The patterns of sectoriality across plant species, the ecological consequences of sectoriality, or the evolutionary pressures selecting for greater sectoriality or integration have been examined. This chapter reviews why sectorial transport is common and briefly discusses how it interacts with environmental heterogeneity. It ends with a description of new techniques for quantifying sectoriality and a discussion of the ecological conditions that might favor evolution of independent sectors or increased integration. The chapter aims to stimulate research on (1) the ecological consequences of sectoriality, and (2) the evolutionary forces that may determine the extent of sectoriality. The discussion is restricted to dicots; monocots are generally integrated because of the large number of interconnected vascular traces and the numerous vascular bundles that come together in complex combinations at nodes.

Local and systemic transcriptome responses to herbivory and jasmonic acid in Populus

We used DNA microarrays to examine local and systemic transcriptional responses to herbivory by gypsy moth larvae (GM) and exogenous jasmonic acid (JAtrt) in leaves of Populus nigra L. to identify candidate signaling and defense genes and also to examine primary metabolism, as might relate to tolerance of damage. GM and JAtrt altered expression of over 800 genes, most of which have putative roles in defense signaling, secondary metabolism, and primary metabolism. Additionally, numerous uncharacterized genes responded to herbivory, providing a rich resource for future studies. There was limited overlap (14%) between the responses to GM and JAtrt. GM did, however, result in strong upregulation of genes involved not only in JA biosynthesis but also abscisic acid biosynthesis and other signaling pathways. GM induced known resistance transcripts, including polyphenolic biosynthetic genes, proteinase inhibitors, and amino acid deaminases. According to GOStats pathway level analysis, GM altered primary metabolism, including aromatic amino acid biosynthesis, fatty acid β-oxidation, and carbohydrate and organic acid metabolism. These alterations may be related to increased demands for substrate for secondary metabolites or may serve a tolerance-related role. Responses were more intense locally in treated leaves than in untreated (systemic) leaves and systemic responses were mostly a subset of the genes induced locally. A stronger local response might be needed to cope with localized stresses and wound healing. Since Populus in general and this clone in particular are known for their systemic induced resistance, genes induced both locally and systemically may be the highest quality candidates for resistance.