Singlet oxygen signalling and its potential roles in plant biotic interactions.

Singlet oxygen (SO) is among the most potent reactive oxygen species, and readily oxidizes proteins, lipids and DNA. It can be generated at the plant surface by phototoxins in the epidermis, acting as a direct defense against pathogens and herbivores (including humans). SO can also accumulate within mitochondria, peroxisomes, cytosol and the nucleus through multiple enzymatic and nonenzymatic processes. However, the majority of research on intracellular SO generation in plants has focused on transfer of light energy to triplet oxygen by photopigments from the chloroplast. SO accumulates in response to diverse stresses that perturb chloroplast metabolism, and while its high reactivity limits diffusion distances, it participates in retrograde signalling through the EXECUTER1 sensor, generation of carotenoid metabolites and possibly other unknown pathways. SO thereby reprogrammes nuclear gene expression and modulates hormone signalling and programmed cell death. While SO signalling has long been known to regulate plant responses to high-light stress, recent literature also suggests a role in plant interactions with insects, bacteria and fungi. The goals of this review are to provide a brief overview of SO, summarize evidence for its involvement in biotic stress responses and discuss future directions for the study of SO in defense signalling.

Cardiac glycosides protect wormseed wallflower (Erysimum cheiranthoides) against some, but not all, glucosinolate-adapted herbivores.

The chemical arms race between plants and insects is foundational to the generation and maintenance of biological diversity. We asked how the evolution of a novel defensive compound in an already well-defended plant lineage impacts interactions with diverse herbivores. Erysimum cheiranthoides (Brassicaceae), which produces both ancestral glucosinolates and novel cardiac glycosides, served as a model. We analyzed gene expression to identify cardiac glycoside biosynthetic enzymes in E. cheiranthoides and characterized these enzymes via heterologous expression and CRISPR/Cas9 knockout. Using E. cheiranthoides cardiac glycoside-deficient lines, we conducted insect experiments in both the laboratory and field. EcCYP87A126 initiates cardiac glycoside biosynthesis via sterol side-chain cleavage, and EcCYP716A418 has a role in cardiac glycoside hydroxylation. In EcCYP87A126 knockout lines, cardiac glycoside production was eliminated. Laboratory experiments with these lines revealed that cardiac glycosides were highly effective defenses against two species of glucosinolate-tolerant specialist herbivores, but did not protect against all crucifer-feeding specialist herbivores in the field. Cardiac glycosides had lesser to no effect on two broad generalist herbivores. These results begin elucidation of the E. cheiranthoides cardiac glycoside biosynthetic pathway and demonstrate in vivo that cardiac glycoside production allows Erysimum to escape from some, but not all, specialist herbivores.

Cardiac glycosides protect wormseed wallflower (Erysimum cheiranthoides) against some, but not all, glucosinolate-adapted herbivores.

The chemical arms race between plants and insects is foundational to the generation and maintenance of biological diversity. We asked how the evolution of a novel defensive compound in an already well-defended plant lineage impacts interactions with diverse herbivores. Erysimum cheiranthoides (Brassicaceae), which produces both ancestral glucosinolates and novel cardiac glycosides, served as a model.We analyzed gene expression to identify cardiac glycoside biosynthetic enzymes in E. cheiranthoides and characterized these enzymes via heterologous expression and CRISPR/Cas9 knockout. Using E. cheiranthoides cardiac glycoside-deficient lines, we conducted insect experiments in both the laboratory and field.EcCYP87A126 initiates cardiac glycoside biosynthesis via sterol side chain cleavage, and EcCYP716A418 has a role in cardiac glycoside hydroxylation. In EcCYP87A126 knockout lines, cardiac glycoside production was eliminated. Laboratory experiments with these lines revealed that cardiac glycosides were highly effective defenses against two species of glucosinolate-tolerant specialist herbivores but did not protect against all crucifer-feeding specialist herbivores in the field. Cardiac glycosides had lesser to no effect on two broad generalist herbivores.These results begin elucidation of the E. cheiranthoides cardiac glycoside biosynthetic pathway and demonstrate in vivo that cardiac glycoside production allows Erysimum to escape from some, but not all, specialist herbivores.

Redox responses of Arabidopsis thaliana to the green peach aphid, Myzus persicae.

The green peach aphid (Myzus persicae) is a phloem-feeding insect that causes economic damage on a wide array of crops. Using a luminol-based assay, a superoxide-responsive reporter gene (Zat12::luciferase), and a probe specific to hydrogen peroxide (HyPer), we demonstrated that this aphid induces accumulation of reactive oxygen species (ROS) in Arabidopsis thaliana. Similar to the apoplastic oxidative burst induced by pathogens, this response to aphids was rapid and transient, with two peaks occurring within 1 and 4 hr after infestation. Aphid infestation also induced an oxidative response in the cytosol and peroxisomes, as measured using a redox-sensitive variant of green fluorescent protein (roGFP2). This intracellular response began within minutes of infestation but persisted 20 hr or more after inoculation, and the response of the peroxisomes appeared stronger than the response in the cytosol. Our results suggest that the oxidative response to aphids involves both apoplastic and intracellular sources of ROS, including ROS generation in the peroxisomes, and these different sources of ROS may potentially differ in their impacts on host suitability for aphids.

Reactive Oxygen Species in Plant Interactions With Aphids.

Reactive oxygen species (ROS) such as hydrogen peroxide and superoxide are produced in plants in response to many biotic and abiotic stressors, and they can enhance stress adaptation in certain circumstances or mediate symptom development in others. The roles of ROS in plant-pathogen interactions have been extensively studied, but far less is known about their involvement in plant-insect interactions. A growing body of evidence, however, indicates that ROS accumulate in response to aphids, an economically damaging group of phloem-feeding insects. This review will cover the current state of knowledge about when, where, and how ROS accumulate in response to aphids, which salivary effectors modify ROS levels in plants, and how microbial associates influence ROS induction by aphids. We will also explore the potential adaptive significance of intra- and extracellular oxidative responses to aphid infestation in compatible and incompatible interactions and highlight knowledge gaps that deserve further exploration.

Economic Injury Levels for Aphis glycines Matsumura (Hemiptera: Aphididae) on the Soybean Aphid Tolerant KS4202 Soybean.

The soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), is an invasive species from Asia that has been the major economic insect pest of soybeans, Glycine max (L.) Merrill, since 2000. While use of soybeans expressing antibiosis and antixenosis is a well-studied strategy to manage this pest, aphid-tolerant soybeans remain underexplored. This study examined the relationship between cumulative aphid-days (CAD) and yield loss in the tolerant soybean KS4202 during two growing seasons to determine the economic injury levels (EILs) for soybean aphids on KS4202. Soybean aphid infestations were initiated during the soybean reproductive stages. A range of CAD treatments (3,000-45,000 CADs) were applied during the growing seasons. Aphid populations reached 45,000 CAD in 2011 and 38,000 CAD in 2013 in plots that were not treated with insecticides. It was estimated that the population doubling time was 9.4 d. In infested plots, soybean yield was reduced by 1.4-13.3%, equivalent to a 3.1% yield loss for every 10,000 CAD. Overall, most CAD treatments did not affect yield parameters, although CAD > 39,000 caused a significant reduction in most yield parameters. The EILs calculated for KS4202 ranged from 526 to 2,050 aphids/plant, which were approximately 2.5-fold higher when compared to EILs previously calculated for susceptible soybean. The adoption of soybean aphid tolerant soybean with higher EILs may help mitigate treatment delay problems by lengthening the treatment lead-time and possibly reduce the number of insecticide applications.