Elevated atmospheric carbon dioxide and plant immunity to fungal pathogens: do the risks outweigh the benefits?

Anthropogenic emissions have caused atmospheric carbon dioxide (CO2) concentrations to double since the industrial revolution. Although this could benefit plant growth from the 'CO2 fertilisation' effect, recent studies report conflicting impacts of elevated CO2 (eCO2) on plant-pathogen interactions. Fungal pathogens are the leading cause of plant disease. Since climate change has been shown to affect the distribution and virulence of these pathogens, it is important to understand how their plant hosts may also respond. This review assesses existing reports of positive, negative, and neutral effects of eCO2 on plant immune responses to fungal pathogen infection. The interaction between eCO2 and immunity appears specific to individual pathosystems, dependent on environmental context and driven by the interactions between plant defence mechanisms, suggesting no universal effect can be predicted for the future. This research is vital for assessing how plants may become more at risk under climate change and could help to guide biotechnological efforts to enhance resistance in vulnerable species. Despite the importance of understanding the effects of eCO2 on plant immunity for protecting global food security, biodiversity, and forests in a changing climate, many plant-pathogen interactions are yet to be investigated. In addition, further research into the effects of eCO2 in combination with other environmental factors associated with climate change is needed. In this review, we highlight the risks of eCO2 to plants and point to the research required to address current unknowns.

Elevated CO2 alters photosynthesis, growth and susceptibility to powdery mildew of oak seedlings.

Elevated CO2 (eCO2) is a determinant factor of climate change and is known to alter plant processes such as physiology, growth and resistance to pathogens. Quercus robur, a tree species integrated in most forest regeneration strategies, shows high vulnerability to powdery mildew (PM) disease at the seedling stage. PM is present in most oak forests and it is considered a bottleneck for oak woodland regeneration. Our study aims to decipher the effect of eCO2 on plant responses to PM. Oak seedlings were grown in controlled environment at ambient (aCO2, ∼400 ppm) and eCO2 (∼1000 ppm), and infected with Erysiphe alphitoides, the causal agent of oak PM. Plant growth, physiological parameters and disease progression were monitored. In addition, to evaluate the effect of eCO2 on induced resistance (IR), these parameters were assessed after treatments with IR elicitor ß-aminobutyric acid (BABA). Our results show that eCO2 increases photosynthetic rates and aerial growth but in contrast, reduces root length. Importantly, under eCO2 seedlings were more susceptible to PM. Treatments with BABA protected seedlings against PM and this protection was maintained under eCO2. Moreover, irrespectively of the concentration of CO2, BABA did not significantly change aerial growth but resulted in longer radicular systems, thus mitigating the effect of eCO2 in root shortening. Our results demonstrate the impact of eCO2 in plant physiology, growth and defence, and warrant further biomolecular studies to unravel the mechanisms by which eCO2 increases oak seedling susceptibility to PM.

Data science approaches provide a roadmap to understanding the role of abscisic acid in defence.

Abscisic acid (ABA) is a plant hormone well known to regulate abiotic stress responses. ABA is also recognised for its role in biotic defence, but there is currently a lack of consensus on whether it plays a positive or negative role. Here, we used supervised machine learning to analyse experimental observations on the defensive role of ABA to identify the most influential factors determining disease phenotypes. ABA concentration, plant age and pathogen lifestyle were identified as important modulators of defence behaviour in our computational predictions. We explored these predictions with new experiments in tomato, demonstrating that phenotypes after ABA treatment were indeed highly dependent on plant age and pathogen lifestyle. Integration of these new results into the statistical analysis refined the quantitative model of ABA influence, suggesting a framework for proposing and exploiting further research to make more progress on this complex question. Our approach provides a unifying road map to guide future studies involving the role of ABA in defence.

Scaling-up to understand tree-pathogen interactions: A steep, tough climb or a walk in the park?

Plants have proficient tools that allow them to survive interactions with pathogens. Upon attack, they respond with specific countermeasures, which are controlled by the immune system. However, defences can fail and this failure exposes plants to fast-spreading devastation. Trees face similar challenges to other plants and their immune system allows them to mount defences against pathogens. However, their slow growth, longevity, woodiness, and size can make trees a challenging system to study. Here, we review scientific successes in plant systems, highlight the key challenges and describe the enormous opportunities for pathology research in trees. We discuss the benefits that scaling-up our understanding on tree-pathogen interactions can provide in the fight against plant pathogenic threats.

Long-Lasting Defence Priming by ß-Aminobutyric Acid in Tomato Is Marked by Genome-Wide Changes in DNA Methylation.

Exposure of plants to stress conditions or to certain chemical elicitors can establish a primed state, whereby responses to future stress encounters are enhanced. Stress priming can be long-lasting and likely involves epigenetic regulation of stress-responsive gene expression. However, the molecular events underlying priming are not well understood. Here, we characterise epigenetic changes in tomato plants primed for pathogen resistance by treatment with ß-aminobutyric acid (BABA). We used whole genome bisulphite sequencing to construct tomato methylomes from control plants and plants treated with BABA at the seedling stage, and a parallel transcriptome analysis to identify genes primed for the response to inoculation by the fungal pathogen, Botrytis cinerea. Genomes of plants treated with BABA showed a significant reduction in global cytosine methylation, especially in CHH sequence contexts. Analysis of differentially methylated regions (DMRs) revealed that CHH DMRs were almost exclusively hypomethylated and were enriched in gene promoters and in DNA transposons located in the chromosome arms. Genes overlapping CHH DMRs were enriched for a small number of stress response-related gene ontology terms. In addition, there was significant enrichment of DMRs in the promoters of genes that are differentially expressed in response to infection with B. cinerea. However, the majority of genes that demonstrated priming did not contain DMRs, and nor was the overall distribution of methylated cytosines in primed genes altered by BABA treatment. Hence, we conclude that whilst BABA treatment of tomato seedlings results in characteristic changes in genome-wide DNA methylation, CHH hypomethylation appears only to target a minority of genes showing primed responses to pathogen infection. Instead, methylation may confer priming via in-trans regulation, acting at a distance from defence genes, and/or by targeting a smaller group of regulatory genes controlling stress responses.

The Induced Resistance Lexicon: Do's and Don'ts.

To be protected from biological threats, plants have evolved an immune system comprising constitutive and inducible defenses. For example, upon perception of certain stimuli, plants can develop a conditioned state of enhanced defensive capacity against upcoming pathogens and pests, resulting in a phenotype called 'induced resistance' (IR). To tackle the confusing lexicon currently used in the IR field, we propose a widely applicable code of practice concerning the terminology and description of IR phenotypes using two main phenotypical aspects: local versus systemic resistance, and direct versus primed defense responses. Our general framework aims to improve uniformity and consistency in future scientific communication, which should help to avoid further misinterpretations and facilitate the accessibility and impact of this research field.

Chitosan primes plant defence mechanisms against Botrytis cinerea, including expression of Avr9/Cf-9 rapidly elicited genes.

Current crop protection strategies against the fungal pathogen Botrytis cinerea rely on a combination of conventional fungicides and host genetic resistance. However, due to pathogen evolution and legislation in the use of fungicides, these strategies are not sufficient to protect plants against this pathogen. Defence elicitors can stimulate plant defence mechanisms through a phenomenon known as defence priming. Priming results in a faster and/or stronger expression of resistance upon pathogen recognition by the host. This work aims to study defence priming by a commercial formulation of the elicitor chitosan. Treatments with chitosan result in induced resistance (IR) in solanaceous and brassicaceous plants. In tomato plants, enhanced resistance has been linked with priming of callose deposition and accumulation of the plant hormone jasmonic acid (JA). Large-scale transcriptomic analysis revealed that chitosan primes gene expression at early time-points after infection. In addition, two novel tomato genes with a characteristic priming profile were identified, Avr9/Cf-9 rapidly elicited protein 75 (ACRE75) and 180 (ACRE180). Transient and stable over-expression of ACRE75, ACRE180 and their Nicotiana benthamiana homologs, revealed that they are positive regulators of plant resistance against B. cinerea. This provides valuable information in the search for strategies to protect Solanaceae plants against B. cinerea.

The IBI1 Receptor of ß-Aminobutyric Acid Interacts with VOZ Transcription Factors to Regulate Abscisic Acid Signaling and Callose-Associated Defense.

External and internal signals can prime the plant immune system for a faster and/or stronger response to pathogen attack. ß-aminobutyric acid (BABA) is an endogenous stress metabolite that induces broad-spectrum disease resistance in plants. BABA perception in Arabidopsis is mediated by the aspartyl tRNA synthetase IBI1, which activates priming of multiple immune responses, including callose-associated cell wall defenses that are under control by abscisic acid (ABA). However, the immediate signaling components after BABA perception by IBI1, as well as the regulatory role of ABA therein, remain unknown. Here, we have studied the early signaling events controlling IBI1-dependent BABA-induced resistance (BABA-IR), using untargeted transcriptome and protein interaction analyses. Transcriptome analysis revealed that IBI1-dependent expression of BABA-IR against the biotrophic oomycete Hyaloperonospora arabidopsidis is associated with suppression of ABA-inducible abiotic stress genes. Protein interaction studies identified the VOZ1 and VOZ2 transcription factors (TFs) as IBI1-interacting partners, which are transcriptionally induced by ABA but suppress pathogen-induced expression of ABA-dependent genes. Furthermore, we show that VOZ TFs require nuclear localization for their contribution to BABA-IR by mediating augmented expression of callose-associated defense. Collectively, our study indicates that the IBI1-VOZ signaling module channels pathogen-induced ABA signaling toward cell wall defense while simultaneously suppressing abiotic stress-responsive genes.

Metabolomics to Exploit the Primed Immune System of Tomato Fruit.

Tomato is a major crop suffering substantial yield losses from diseases, as fruit decay at a postharvest level can claim up to 50% of the total production worldwide. Due to the environmental risks of fungicides, there is an increasing interest in exploiting plant immunity through priming, which is an adaptive strategy that improves plant defensive capacity by stimulating induced mechanisms. Broad-spectrum defence priming can be triggered by the compound ß-aminobutyric acid (BABA). In tomato plants, BABA induces resistance against various fungal and bacterial pathogens and different methods of application result in durable protection. Here, we demonstrate that the treatment of tomato plants with BABA resulted in a durable induced resistance in tomato fruit against Botrytis cinerea, Phytophthora infestans and Pseudomonas syringae. Targeted and untargeted metabolomics were used to investigate the metabolic regulations that underpin the priming of tomato fruit against pathogenic microbes that present different infection strategies. Metabolomic analyses revealed major changes after BABA treatment and after inoculation. Remarkably, primed responses seemed specific to the type of infection, rather than showing a common fingerprint of BABA-induced priming. Furthermore, top-down modelling from the detected metabolic markers allowed for the accurate prediction of the measured resistance to fruit pathogens and demonstrated that soluble sugars are essential to predict resistance to fruit pathogens. Altogether, our results demonstrate that metabolomics is particularly insightful for a better understanding of defence priming in fruit. Further experiments are underway in order to identify key metabolites that mediate broad-spectrum BABA-induced priming in tomato fruit.

Food security in 2044: How do we control the fungal threat?

Plant fungal pathogens place considerable strain on agricultural productivity and threaten global food security. In recent decades, advances in crop breeding, farming practice and the agrochemical industry have allowed crop yields to keep pace with food demand. In this opinion article, we speculate on which recent technological advances will allow us to maintain this situation into the future. We take inspiration that it is 25 y since the first plant disease resistance genes were cloned, and imagine if and how agricultural control of pathogens will be achieved by the year 2044. We examine which technologies are best poised to make the jump from lab bench to field application, and propose that future control measures will likely depend on effective integrated disease management.

The relationship between transgenerational acquired resistance and global DNA methylation in Arabidopsis.

Progeny of heavily diseased plants develop transgenerational acquired resistance (TAR). In Arabidopsis, TAR can be transmitted over one stress-free generation. Although DNA methylation has been implicated in the regulation of TAR, the relationship between TAR and global DNA methylation remains unknown. Here, we characterised the methylome of TAR-expressing Arabidopsis at different generations after disease exposure. Global clustering of cytosine methylation revealed TAR-related patterns in the F3 generation, but not in the F1 generation. The majority of differentially methylated positions (DMPs) occurred at CG context in gene bodies. TAR in F3 progeny after one initial generation of disease, followed by two stress-free generations, was lower than TAR in F3 progeny after three successive generations of disease. This difference in TAR effectiveness was proportional to the intensity of differential methylation at a sub-set of cytosine positions. Comparison of TAR-related DMPs with previously characterised cytosine methylation in mutation accumulation lines revealed that ancestral disease stress preferentially acts on methylation-labile cytosine positions, but also extends to methylation-stable positions. Thus, the TAR-related impact of ancestral disease extends beyond stochastic variation in DNA methylation. Our study has shown that the Arabidopsis epigenome responds globally to disease in previous generations and we discuss its contribution to TAR.

Fruit Decay to Diseases: Can Induced Resistance and Priming Help?

Humanity faces the challenge of having to increase food production to feed an exponentially growing world population, while crop diseases reduce yields to levels that we can no longer afford. Besides, a significant amount of waste is produced after fruit harvest. Fruit decay due to diseases at a post-harvest level can claim up to 50% of the total production worldwide. Currently, the most effective means of disease control is the use of pesticides. However, their use post-harvest is extremely limited due to toxicity. The last few decades have witnessed the development of safer methods of disease control post-harvest. They have all been included in programs with the aim of achieving integrated pest (and disease) management (IPM) to reduce pesticide use to a minimum. Unfortunately, these approaches have failed to provide robust solutions. Therefore, it is necessary to develop alternative strategies that would result in effective control. Exploiting the immune capacity of plants has been described as a plausible route to prevent diseases post-harvest. Post-harvest-induced resistance (IR) through the use of safer chemicals from biological origin, biocontrol, and physical means has also been reported. In this review, we summarize the successful activity of these different strategies and explore the mechanisms behind. We further explore the concept of priming, and how its long-lasting and broad-spectrum nature could contribute to fruit resistance.

Chemical priming of immunity without costs to plant growth.

ß-Aminobutyric acid (BABA) induces broad-spectrum disease resistance, but also represses plant growth, which has limited its exploitation in crop protection. BABA perception relies on binding to the aspartyl-tRNA synthetase (AspRS) IBI1, which primes the enzyme for secondary defense activity. This study aimed to identify structural BABA analogues that induce resistance without stunting plant growth. Using site-directed mutagenesis, we demonstrate that the (l)-aspartic acid-binding domain of IBI1 is critical for BABA perception. Based on interaction models of this domain, we screened a small library of structural BABA analogues for growth repression and induced resistance against biotrophic Hyaloperonospora arabidopsidis (Hpa). A range of resistance-inducing compounds were identified, of which (R)-ß-homoserine (RBH) was the most effective. Surprisingly, RBH acted through different pathways than BABA. RBH-induced resistance (RBH-IR) against Hpa functioned independently of salicylic acid, partially relied on camalexin, and was associated with augmented cell wall defense. RBH-IR against necrotrophic Plectosphaerella cucumerina acted via priming of ethylene and jasmonic acid defenses. RBH-IR was also effective in tomato against Botrytis cinerea. Metabolic profiling revealed that RBH, unlike BABA, does not majorly affect plant metabolism. RBH primes distinct defense pathways against biotrophic and necrotrophic pathogens without stunting plant growth, signifying strong potential for exploitation in crop protection.

Defense Priming: An Adaptive Part of Induced Resistance.

Priming is an adaptive strategy that improves the defensive capacity of plants. This phenomenon is marked by an enhanced activation of induced defense mechanisms. Stimuli from pathogens, beneficial microbes, or arthropods, as well as chemicals and abiotic cues, can trigger the establishment of priming by acting as warning signals. Upon stimulus perception, changes may occur in the plant at the physiological, transcriptional, metabolic, and epigenetic levels. This phase is called the priming phase. Upon subsequent challenge, the plant effectively mounts a faster and/or stronger defense response that defines the postchallenge primed state and results in increased resistance and/or stress tolerance. Priming can be durable and maintained throughout the plant's life cycle and can even be transmitted to subsequent generations, therefore representing a type of plant immunological memory.

Estudio clínico genético y psicosocial de familias con ceguera del municipio Kalandula, provincia Malange, Angola, 2014 / Clinical - genetic and psychosocial study of families with blindness of the Kalandula municipality, Malange province, Angola, 2014

Las alteraciones congénitas funcionales y estructurales de los ojos aparecen durante el desarrollo embrionario y fetal evidenciándose al nacimiento, los defectos de la visión producidas por estas anomalías pueden llevar a la ceguera total o parcial. Se conoció de la existencia de varias personas con ceguera pertenecientes a una misma familia en la Comuna de Santo António, municipio Kalandula, provincia de Malange, Angola. Objetivos: Contribuir a la prevención de la discapacidad mediante el Asesoramiento Genético en las familias afectadas. Método: Se realizó un estudio tipo serie de casos para el diagnóstico clínico, genético y psicosocial de familias con ceguera, que incluyó el diseño de un modelo de historia clínica y la creación de un grupo multidisciplinario, se tomó como referencia la estrategia desarrollada para el estudio de personas con discapacidades en Cuba y otros países. Resultados: Se realizó el estudio psicosocial, pedagógico y clínico genético de la familia conocida (Familia 1) y de otra familia encontrada en el curso de la investigación, ésta de la comuna de Ginga Luamba (Familia 2), ambas con varios miembros afectados de ceguera. Se llegó al diagnóstico clínico genético de los casos y las enfermedades encontradas en ambas familias fueron clasificadas como cegueras de causa genética monogénica. Conclusiones: Mediante la aplicación del método clínico, se consiguió establecer un diagnóstico inicial para ambas familias, ofrecerles asesoramiento genético y apoyo psicosocial…(AU)

Conocimientos sobre factores de riesgo genético en estudiantes de medicina de la Facultad de Medicina de Malanje, Angola, 2014 / Knowledge about genetic risk factors in students of the Faculty of Medicine of Malanje, Angola, 2014

Los servicios de Genética son más eficaces si se ofrecen en el contexto de un público educado que es capaz y está dispuesto a actuar voluntariamente, por lo tanto, la información y el conocimiento son esenciales para promover la educación pública sobre los factores de riesgo genético. Con el objetivo de evaluar el nivel de conocimiento de los estudiantes de la Facultad de Medicina de Malanje de la Universidad Lueji ANkonde, Angola, sobre los principales factores de riesgo genético y su prevención comunitaria; se realizó en 2013 - 2014 un estudio descriptivo para el cual se diseñó un cuestionario y una clave de calificación, basados en las experiencias de las autoras y en el criterio de expertos. La muestra quedó formada por un total de 152 estudiantes que voluntariamente accedieron a participar una vez informados. Como resultados se obtuvo que los estudiantes de Medicina de la Facultad de Medicina de Malanje presentaban un insuficiente y deficiente nivel de conocimientos en cuanto a factores de riesgo genético y las acciones de prevención primaria para los defectos congénitos y las enfermedades genéticas, de lo que se deduce que la situación se comporta de forma similar entre los actuales trabajadores de la salud y la población. Se evidenció en los estudiantes un interés creciente por la adquisición de estos conocimientos y su divulgación en la comunidad…(AU)

Optimizing Chemically Induced Resistance in Tomato Against Botrytis cinerea.

Resistance-inducing chemicals can offer broad-spectrum disease protection in crops, but can also affect plant growth and interactions with plant-beneficial microbes. We have evaluated different application methods of ß-aminobutyric acid (BABA) and jasmonic acid (JA) for long-lasting induced resistance in tomato against Botrytis cinerea. In addition, we have studied nontarget effects on plant growth and root colonization by arbuscular mycorrhizal fungi (AMF). Germinating seeds for 1 week in BABA- or JA-containing solutions promoted seed germination efficiency, did not affect plant growth, and induced resistance in 4-week-old plants. When formulating BABA and JA in carboxymethyl cellulose seed coating, only BABA was able to induce resistance in 4-week-old plants. Root treatment of 1-week-old seedlings with BABA or JA also induced resistance in 4-week-old plants. However, this seedling treatment repressed plant growth at higher concentrations of the chemicals, which was particularly pronounced in hydroponically grown plants after BABA treatment. Both seed coating with BABA, and seedling treatments with BABA or JA, did not affect AMF root colonization in soil-grown tomato. Our study has identified commercially feasible application methods of BABA and JA, which induce durable disease resistance in tomato without concurrent impacts on plant growth or colonization by plant-beneficial AMF.

Role of NPR1 and KYP in long-lasting induced resistance by ß-aminobutyric acid.

Priming of defense increases the responsiveness of the plant immune system and can provide broad-spectrum protection against disease. Recent evidence suggests that priming of defense can be inherited epigenetically to following generations. However, the mechanisms of long-lasting defense priming within one generation remains poorly understood. Here, we have investigated the mechanistic basis of long-lasting induced resistance after treatment with ß -aminobutyric acid (BABA), an agent that mimics biologically induced resistance phenomena. BABA-induced resistance (BABA-IR) is based on priming of salicylic acid (SA)-dependent and SA-independent defenses. BABA-IR could be detected up to 28 days after treatment of wild-type Arabidopsis. This long-lasting component of the induced resistance response requires the regulatory protein NPR1 and is associated with priming of SA-inducible genes. In contrast, NPR1-independent resistance by BABA was transient and had disappeared by 14 days after treatment. Chromatin immunoprecipitation (ChIP) assays revealed no increased acetylation of histone H3K9 at promoters regions of priming-responsive genes, indicating that this post-translational histone modification is not critical for long-term transcriptional priming. Interestingly, the kyp-6 mutant, which is affected in methyltransferase activity of H3K9, was blocked in long-lasting BABA-IR, indicating a critical requirement of this post-translational histone modification in long-lasting BABA-IR. Considering that KYP suppresses gene transcription through methylation of H3K9 and CpHpG DNA methylation, we propose that KYP enables long-term defense gene priming by silencing suppressor genes of SA/NPR1-dependent genes.