M. Brian Traw

Natural allelic variation underlying a major fitness trade-off in Arabidopsis thaliana.

Plants can defend themselves against a wide array of enemies, from microbes to large animals, yet there is great variability in the effectiveness of such defences, both within and between species. Some of this variation can be explained by conflicting pressures from pathogens with different modes of attack. A second explanation comes from an evolutionary ‘tug of war’, in which pathogens adapt to evade detection, until the plant has evolved new recognition capabilities for pathogen invasion. If selection is, however, sufficiently strong, susceptible hosts should remain rare. That this is not the case is best explained by costs incurred from constitutive defences in a pest-free environment. Using a combination of forward genetics and genome-wide association analyses, we demonstrate that allelic diversity at a single locus, ACCELERATED CELL DEATH 6 (ACD6), underpins marked pleiotropic differences in both vegetative growth and resistance to microbial infection and herbivory among natural Arabidopsis thaliana strains. A hyperactive ACD6 allele, compared to the reference allele, strongly enhances resistance to a broad range of pathogens from different phyla, but at the same time slows the production of new leaves and greatly reduces the biomass of mature leaves. This allele segregates at intermediate frequency both throughout the worldwide range of A. thaliana and within local populations, consistent with this allele providing substantial fitness benefits despite its marked impact on growth.

Reduced Performance of Two Specialist Herbivores (Lepidoptera: Pieridae, Coleoptera: Chrysomelidae) on New Leaves of Damaged Black Mustard Plants

Abstract This study addressed whether prior damage to black mustard, Brassica nigra (L.) Koch, would reduce growth, herbivory, or mortality of two specialist herbivores on new leaves. Plants received either no initial damage or 12 h of feeding by two third instars of Pieris rapae (L.) or 50 adult Phyllotreta cruciferae (Goeze) when plants had four leaves. Later, the seventh leaf of plants was either harvested for measurement of trichome density and glucosinolate concentration or enclosed in a mesh cage containing two neonate P. rapae or 10 adult P. cruciferae. Caged herbivores were measured for mass gain, leaf consumption, and mortality after 1 wk. Damage by P. rapae caused substantial increases in trichome density and sinigrin concentration, whereas damage by P. cruciferae had no effect. Larvae of P. rapae grew 30% more slowly on plants initially damaged by conspecifics than on control plants. Percent herbivory by P. rapae was 33% lower on plants initially damaged by either P. rapae or P. cruciferae than on control plants. Growth rate and percent herbivory by P. cruciferae were not generally affected by prior plant damage. However, mortality of P. cruciferae was 84% higher on plants previously damaged by conspecifics than on control plants. Together, the data demonstrate that induction responses negatively affected both Pieris and Phyllotreta and suggest that trichomes may be relatively important in the increased resistance. Test herbivores generally performed similarly on plants damaged by either herbivore, suggesting a low specificity of effect for the induction response.

Salicylic Acid and Jasmonic Acid Signaling Defense Pathways Reduce Natural Bacterial Diversity on Arabidopsis thaliana

Terrestrial plants serve as large and diverse habitats for a wide range of pathogenic and nonpathogenic microbes, yet these communities are not well described and little is known about the effects of plant defense on microbial communities in nature. We designed a field experiment to determine how variation in two plant defense signaling pathways affects the size, diversity, and composition of the natural endophytic and epiphytic bacterial communities of Arabidopsis thaliana. To do this, we provide an initial characterization of these bacterial communities in one population in southwestern Michigan, United States, and we compare these two communities among A. thaliana mutants deficient in salicylic acid (SA) and jasmonic acid (JA) signaling defense pathways, controls, and plants with artificially elevated levels of defense. We identified 30 distinct bacterial groups on A. thaliana that differ in colony morphology and 16S rRNA sequence. We show that induction of SA-mediated defenses reduced endophytic bacterial community diversity, whereas plants deficient in JA-mediated defenses experienced greater epiphytic bacterial diversity. Furthermore, there was a positive relationship between total community size and diversity, indicating that relatively susceptible plants should, in general, harbor higher bacterial diversity. This experiment provides novel information about the ecology of bacteria on A. thaliana and demonstrates that variation in two specific plant-signaling defense pathways can influence bacterial diversity on plants.

Is induction response negatively correlated with constitutive resistance in black mustard

Abstract.— Black mustard, Brassica nigra, is highly variable in both constitutive resistance and induction response following damage by herbivores. A focal population from Ithaca, New York, was used to test the following two predictions of optimal defense theory: (1) that allocation to resistance will reduce plant performance in the absence of herbivores; and (2) that induction response will be negatively correlated with constitutive resistance. The experiment consisted of a half‐sib mating design with 47 paternal families and four dams per sire, fully crossed with a damage treatment consisting of 25% leaf removal by the cabbage white butterfly, Pieris rapae, when plants had four leaves. Leaf trichome density, sinigrin concentration, and glucobrassicin concentration were 38%, 19%, and 16% higher, respectively, for the seventh leaf of damaged plants. Paternal families did not vary significantly in their induction response. Narrow‐sense heritabilities were h2s= 0.51, 0.76, and 0.50 for constitutive leaf trichome density, sinigrin concentration, and glucobrassicin concentration, respectively. Positive genetic correlations were found between glucobrassicin concentration and days to first flower, suggesting a genetic cost of resistance. Induction responses were negatively correlated with constitutive allocation for leaf trichome density and sinigrin concentration. The results were therefore consistent with optimal defense theory, offering modest evidence for both predictions.

Ascorbic Acid Deficiency in Arabidopsis Induces Constitutive Priming That is Dependent on Hydrogen Peroxide, Salicylic Acid, and the NPR1 Gene

The ascorbic acid (AA)-deficient Arabidopsis thaliana vtc1-1 mutant exhibits increased resistance to the virulent bacterial pathogen Pseudomonas syringae. This response correlates with heightened levels of salicylic acid (SA), which induces antimicrobial pathogenesis-related (PR) proteins. To determine if SA-mediated, enhanced disease resistance is a general phenomenon of AA deficiency, to elucidate the signal that stimulates SA synthesis, and to identify the biosynthetic pathway through which SA accumulates, we studied the four AA-deficient vtc1-1, vtc2-1, vtc3-1, and vtc4-1 mutants. We also studied double mutants defective in the AA-biosynthetic gene VTC1 and the SA signaling pathway genes PAD4, EDS5, and NPR1, respectively. All vtc mutants were more resistant to P. syringae than the wild type. With the exception of vtc4-1, this correlated with constitutively upregulated H(2)O(2), SA, and messenger RNA levels of PR genes. Double mutants exhibited decreased SA levels and enhanced susceptibility to P. syringae compared with the wild type, suggesting that vtc1-1 requires functional PAD4, EDS5, and NPR1 for SA biosynthesis and pathogen resistance. We suggest that AA deficiency causes constitutive priming through a buildup of H(2)O(2) that stimulates SA accumulation, conferring enhanced disease resistance in vtc1-1, vtc2-1, and vtc3-1, whereas vtc4-1 might be sensitized to H(2)O(2) and SA production after infection.

Low Levels of Polymorphism in Genes That Control the Activation of Defense Response in Arabidopsis thaliana

Plants use signaling pathways involving salicylic acid, jasmonic acid, and ethylene to defend against pathogen and herbivore attack. Many defense response genes involved in these signaling pathways have been characterized, but little is known about the selective pressures they experience. A representative set of 27 defense response genes were resequenced in a worldwide set of 96 Arabidopsis thaliana accessions, and patterns of single nucleotide polymorphisms (SNPs) were evaluated in relation to an empirical distribution of SNPs generated from either 876 fragments or 236 fragments with >400 bp coding sequence (this latter set was selected for comparisons with coding sequences) distributed across the genomes of the same set of accessions. Defense response genes have significantly fewer protein variants, display lower levels of nonsynonymous nucleotide diversity, and have fewer nonsynonymous segregating sites. The majority of defense response genes appear to be experiencing purifying selection, given the dearth of protein variation in this set of genes. Eight genes exhibit some evidence of partial selective sweeps or transient balancing selection. These results therefore provide a strong contrast to the high levels of balancing selection exhibited by genes at the upstream positions in these signaling pathways.

GLUCOSINOLATES AND TRICHOMES TRACK TISSUE VALUE IN TWO SYMPATRIC MUSTARDS

Glucosinolates, trichomes, nitrogen, and carbon are not distributed uniformly through the canopies of mustards. In this study, we asked whether glucosinolate concentrations and trichome densities in two sympatric mustards, Brassica kaber and B. nigra, are highest in tissues of greatest value to the plant. We also asked whether nitrogen or carbon content is the stronger predictor of tissue value, and what fraction of each resource is incorporated in glucosinolates. To quantify tissue values, we removed three equal-area fractions (lower, middle, and upper) from the canopies of B. kaber and B. nigra in the greenhouse, as well as whole canopies of naturally growing B. nigra in the field, at two times during growth and measured reductions in their performance relative to controls. We also measured trichome density in both experiments, as well as glucosinolate, nitrogen, and carbon concentrations for the equal-area fractions in the greenhouse. We found that upper leaves had the highest glucosinolate concentrations, trichome densities, and tissue values. Furthermore, young plants in the field had higher trichome densities and tissue values than did older plants. Collectively, these data provide strong support for optimal defense theory and are among the first such evidence for glucosinolates and for physical defenses. The positive relationship between trichome density and tissue value was strong even after we accounted for the effects of leaf expansion. While nitrogen and carbon have both received attention as currencies for trade-offs, our data suggest that nitrogen concentration is a significantly better predictor of tissue value in these two mustard species. Interestingly, <1% of the nitrogen or carbon in leaves was incorporated in glucosinolates, which may explain why glucosinolates lack a consistent response to nitrogen fertilization.

ATP-Dependent Binding Cassette Transporter G Family Member 16 Increases Plant Tolerance to Abscisic Acid and Assists in Basal Resistance against Pseudomonas syringae DC3000

An ATP-dependent binding cassette transporter contributes to plant resistance to infection by Pseudomonas syringae and tolerance of abscisic acid. Plants have been shown previously to perceive bacteria on the leaf surface and respond by closing their stomata. The virulent bacterial pathogen Pseudomonas syringae pv tomato DC3000 (PstDC3000) responds by secreting a virulence factor, coronatine, which blocks the functioning of guard cells and forces stomata to reopen. After it is inside the leaf, PstDC3000 has been shown to up-regulate abscisic acid (ABA) signaling and thereby suppress salicylic acid-dependent resistance. Some wild plants exhibit resistance to PstDC3000, but the mechanisms by which they achieve this resistance remain unknown. Here, we used genome-wide association mapping to identify an ATP-dependent binding cassette transporter gene (ATP-dependent binding cassette transporter G family member16) in Arabidopsis (Arabidopsis thaliana) that contributes to wild plant resistance to PstDC3000. Through microarray analysis and β-glucuronidase reporter lines, we showed that the gene is up-regulated by ABA, bacterial infection, and coronatine. We also used a green fluorescent protein fusion protein and found that transporter is more likely to localize on plasma membranes than in cell walls. Transferred DNA insertion lines exhibited consistent defective tolerance of exogenous ABA and reduced resistance to infection by PstDC3000. Our conclusion is that ATP-dependent binding cassette transporter G family member16 is involved in ABA tolerance and contributes to plant resistance against PstDC3000. This is one of the first examples, to our knowledge, of ATP-dependent binding cassette transporter involvement in plant resistance to infection by a bacterial pathogen. It also suggests a possible mechanism by which plants reduce the deleterious effects of ABA hijacking during pathogen attack. Collectively, these results improve our understanding of basal resistance in Arabidopsis and offer unique ABA-related targets for improving the innate resistance of plants to bacterial infection.

Constitutive camalexin production and environmental stress response variation in Arabidopsis populations from the Iberian Peninsula.

Optimal defense theory predicts that induction of defensive secondary metabolites in plants will be inversely correlated with constitutive expression of those compounds. Here, we asked whether camalexin, an important defense against fungal and bacterial pathogens, support this prediction in structured natural populations of Arabidopsis thaliana from the Iberian Peninsula. In common garden experiments, we found that genotypes from the VIE population constitutively hyper-accumulated camalexin. Camalexin concentrations were not induced significantly when plants were exposed to a temperature of 10°C for 48h. However, they were induced when plants were exposed to 48h of infection by the virulent bacterial pathogen, Pseudomonas syringae pv. tomato DC3000. Genotypes from the VIE population with the hyper-accumulation of camalexin were significantly more resistant to bacterial growth. Induction of camalexin was negatively correlated with constitutive camalexin concentrations following log transformation and two different corrections for autocorrelation, thus supporting the tradeoff predicted by optimal defense theory. Constitutive overexpression of camalexin was not explained by the only known natural genetic polymorphism at the Accelerated Cell Death 6, ACD6, locus. Collectively, the results support an important role of camalexin in defense against P. syringae as well as significant structured variation in defense levels within wild populations.

Theories, Mechanisms and Patterns of Microbiome Species Coexistence in an Era of Climate Change

Understanding the mechanisms of microbiome species coexistence is one of the major challenges confronted by microbiome ecologists today. Recent research largely advocates that the niche, biogeographic, and neutral/stochastic processes interactively determine the microbiome community composition across spatio-temporal scales in the ecosystem. However, current research is mainly exploratory and descriptive, and it is still difficult to link the observed microbiome species or functional trait diversity patterns to the ecosystem functioning on local, regional, and global scales. Similarly, whether and how climate and land-use changes impact the patterns of microbiome functional ecology remains to be studied. Moreover, little is known about how climate and land-use changes are linked to evolution or loss of microbial functional traits. Future research should nevertheless investigate the patterns of microbiome functional diversity across the velocity of climate and land-use changes, and attempt to integrate the patterns with functions in broader contexts of functional traits and community ecology.