Large effect quantitative trait loci for salicinoid phenolic glycosides in Populus: Implications for gene discovery.

Genomic studies have been used to identify genes underlying many important plant secondary metabolic pathways. However, genes for salicinoid phenolic glycosides (SPGs)-ecologically important compounds with significant commercial, cultural, and medicinal applications-remain largely undescribed. We used a linkage map derived from a full-sib population of hybrid cottonwoods (Populus spp.) to search for quantitative trait loci (QTL) for the SPGs salicortin and HCH-salicortin. SSR markers and primer sequences were used to anchor the map to the V3.0 P. trichocarpa genome. We discovered 21 QTL for the two traits, including a major QTL for HCH-salicortin (R2 = .52) that colocated with a QTL for salicortin on chr12. Using the V3.0 Populus genome sequence, we identified 2,983 annotated genes and 1,480 genes of unknown function within our QTL intervals. We note ten candidate genes of interest, including a BAHD-type acyltransferase that has been potentially linked to Populus SPGs. Our results complement other recent studies in Populus with implications for gene discovery and the evolution of defensive chemistry in a model genus. To our knowledge, this is the first study to use a full-sib mapping population to identify QTL intervals and gene lists associated with SPGs.

Induced plant defenses, host-pathogen interactions, and forest insect outbreaks.

Cyclic outbreaks of defoliating insects devastate forests, but their causes are poorly understood. Outbreak cycles are often assumed to be driven by density-dependent mortality due to natural enemies, because pathogens and predators cause high mortality and because natural-enemy models reproduce fluctuations in defoliation data. The role of induced defenses is in contrast often dismissed, because toxic effects of defenses are often weak and because induced-defense models explain defoliation data no better than natural-enemy models. Natural-enemy models, however, fail to explain gypsy moth outbreaks in North America, in which outbreaks in forests with a higher percentage of oaks have alternated between severe and mild, whereas outbreaks in forests with a lower percentage of oaks have been uniformly moderate. Here we show that this pattern can be explained by an interaction between induced defenses and a natural enemy. We experimentally induced hydrolyzable-tannin defenses in red oak, to show that induction reduces variability in a gypsy moth's risk of baculovirus infection. Because this effect can modulate outbreak severity and because oaks are the only genus of gypsy moth host tree that can be induced, we extended a natural-enemy model to allow for spatial variability in inducibility. Our model shows alternating outbreaks in forests with a high frequency of oaks, and uniform outbreaks in forests with a low frequency of oaks, matching the data. The complexity of this effect suggests that detecting effects of induced defenses on defoliator cycles requires a combination of experiments and models.

Hormaphis hamamelidis fundatrices benefit by manipulating phenolic metabolism of their host.

We investigated the pattern and potential adaptive value of phenolic concentrations in galls induced by the aphid Hormaphis hamamelidis on leaves of Hamamelis virginiana. By the time that founding females began reproduction, galls had higher concentrations of condensed tannins and lower concentrations of hydrolyzable tannins than leaves. Galled and ungalled leaf laminas never differed significantly in any phenolic measure. Condensed tannin concentrations also were positively related to the number of offspring per gall when gall dry weight, another important correlate of fecundity, was accounted for. This could indicate the prior sink strength of the gall. Polyphenols may act as a repository for excess carbon drawn to the gall by increased sink strength, or be an indication of the fundatrix' ability to manipulate host physiology. This study is the first to demonstrate a tangible, quantitative association between phenolic accumulation in galls and gall-former reproductive performance, and illustrates that condensed tannins may play roles other than plant defense.

Toward a mechanistic understanding of competition in vascular-feeding herbivores: an empirical test of the sink competition hypothesis.

Recent evidence suggests that competitive interactions among herbivores are mostly indirect and mediated by plant responses to herbivory. Most studies, however, emphasize chewing insects and secondary chemistry, thus ignoring the diverse group of vascular-parasites that may be more likely to compete through induced changes in phytonutrients. Using an aboveground phloem-feeding aphid (Myzus persicae) and a belowground gall-forming nematode (Meloidogyne incognita) on tobacco plants, we assessed the importance of competition via induced host-plant sinks. In a series of experimental trials, nematode root herbivory caused 55 and 72% declines in the growth and fecundity of aphids, respectively. Aphids, on the other hand, did not impact nematode performance. Therefore, we predicted that nematodes out-compete M. persicae by attenuating the magnitude of aphid-induced sinks. Through a combination of invertase enzyme measurements and stable isotope ((13)C and (15)N) enrichment, we found evidence that both herbivores act as mobilizing sinks. Aphids attracted photoassimilates to feeding aggregations on leaves and nematode galls accumulated resources in the roots. Levels of invertase enzymes, for example, were more than fourfold higher in nematode galls than in surrounding root tissue. Yet we found no evidence supporting a sink competition model for aphid-nematode interactions. The strength of aphid-induced leaf sinks was entirely unaffected by nematode presence, and vice versa. Thus, induced host-plant sinks appear to be a common strategy employed by vascular parasites to manipulate the physiology of their host, but multi-sink competition may be limited to herbivores that co-occur on the same tissue type and/or plants under growth-limited abiotic conditions.

From genes to ecosystems: a synthesis of the effects of plant genetic factors across levels of organization.

Using two genetic approaches and seven different plant systems, we present findings from a meta-analysis examining the strength of the effects of plant genetic introgression and genotypic diversity across individual, community and ecosystem levels with the goal of synthesizing the patterns to date. We found that (i) the strength of plant genetic effects can be quite high; however, the overall strength of genetic effects on most response variables declined as the levels of organization increased. (ii) Plant genetic effects varied such that introgression had a greater impact on individual phenotypes than extended effects on arthropods or microbes/fungi. By contrast, the greatest effects of genotypic diversity were on arthropods. (iii) Plant genetic effects were greater on above-ground versus below-ground processes, but there was no difference between terrestrial and aquatic environments. (iv) The strength of the effects of intraspecific genotypic diversity tended to be weaker than interspecific genetic introgression. (v) Although genetic effects generally decline across levels of organization, in some cases they do not, suggesting that specific organisms and/or processes may respond more than others to underlying genetic variation. Because patterns in the overall impacts of introgression and genotypic diversity were generally consistent across diverse study systems and consistent with theoretical expectations, these results provide generality for understanding the extended consequences of plant genetic variation across levels of organization, with evolutionary implications.

Physiological integration of roots and shoots in plant defense strategies links above- and belowground herbivory.

Roots play a critical, but largely unappreciated, role in aboveground anti-herbivore plant defense (e.g. resistance and tolerance) and root-leaf connections may therefore result in unexpected coupling between above- and belowground consumers. Using the tobacco (Nicotiana tabacum) system we highlight two examples of this phenomenon. First, the secondary metabolite nicotine is produced in roots, yet translocated aboveground for use as a foliar resistance trait. We demonstrate that nematode root herbivory interferes with foliar nicotine dynamics, resulting in positive effects on aboveground phytophagous insects. Notably, nematode-induced facilitation only occurred on nicotine-producing plants, and not on nicotine-deficient mutants. In the second case, we use stable isotope and invertase enzyme analyses to demonstrate that foliar herbivory elicits a putative tolerance response whereby aboveground nutritional reserves are allocated to roots, resulting in facilitation of phytoparasitic nematodes. Thus, plants integrate roots in resistance and tolerance mechanisms for leaf defense, and such root-leaf connections inherently link the dynamics of above- and belowground consumers.

Developmental trajectories in cottonwood phytochemistry.

We examined the hypothesis that ecologically important phytochemical traits differ predictably among various developmental zones of trees (i.e., mature and juvenile zones of individual trees and juvenile ramets that sprout from roots) and that the slope of this phytochemical gradient represents a "developmental trajectory." We focused on Populus fremontii (Fremont cottonwood), P. angustifolia (narrowleaf cottonwood), and their natural hybrids. Two major patterns emerged. First, within narrowleaf and hybrids, concentrations of important phytochemicals (condensed tannins and phenolic glycosides) differ greatly and predictably between developmental zones. Second, developmental trajectories differ greatly among these cottonwood species and their hybrids: Fremont exhibits a flat trajectory, narrowleaf a steep trajectory, and hybrids an intermediate trajectory, suggesting an additive genetic component and an ontogenetic basis to this phytochemical variation. Because diverse herbivorous species respond to the phytochemistry of their host plants, we predict that the developmental trajectories of plants play a major role in mediating ecological interactions and structuring communities, and that biodiversity in a stand of trees is determined by both interplant genetic diversity and intraplant ontogenetic diversity.

Enhanced invertase activities in the galls of Hormaphis hamamelidis.

Invertases are sucrose hydrolyzing enzymes often associated with plant tissues acting as physiological sinks, and plant galls are physiological sinks. We investigated several types of invertase and their potential benefits in galls of the aphid Hormaphis hamamelidis. Invertase activities in galls differed from those in leaves throughout the growing season. Vacuolar invertase activities (per g FW) were always greater in galls than leaves. In contrast, cell wall invertase activities per g FW started low in galls, but increased with time and were greater than those in leaves after 1 month. Gall growth was most closely related to vacuolar invertase activity, whereas leaf growth was correlated with both vacuolar and cell wall invertase activities. In separate correlational studies of aphid fecundity and invertase activities, cell wall invertase activity per gall accounted for 15-21% of the variation in offspring per gall. Gall dry weight explained more of the variation in offspring per gall (34.2%) than did gall volume (17.8%), a likely indication of the importance of sink strength over the life of the gall. Increased invertase activity probably underlies the enhanced sink strength commonly observed in galls. Hormaphis hamamelidis fundatrices appear to maximize reproductive output by eliciting multiple beneficial responses from their hosts. Initial gall growth correlated positively with high vacuolar invertase activity, while later in the season aphid fecundity correlated positively with cell wall invertase activity.