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Polyphenol oxidase partial gene PG-PPO was cloned and characterized from Pennisetum glaucum (pearl millet) which showed 42% identity to a PPO sequence isolated from wheat at the region of Copper B with a score of 40 and e-value of 2.8. Multiple sequence alignment results revealed similarity to polyphenol oxidase (PPO) sequences from wheat, trifolium, lettuce, apricot, tobacco, tomato, pokeweed, apple, grape and poplar especially at the Copper B region of PPO. The 395 bp pearl millet PPO sequence was AT rich (53.3%) and contained the highly conserved amino acids of histidine-rich copper binding sites similar to PPO sequences from other crops. Results also indicated that PPO in pearl millet exists in multi copy. The role of the isolated PPO gene during pearl millet-downy mildew interaction was analyzed and the results showed significantly higher and rapid accumulation of PPO mRNAs in resistant pearl millet seedlings inoculated with Sclerospora graminicola in comparison to the susceptible control, demonstrating that the PPO plays a prominent role in pearl millet defense against pathogens, particularly downy mildew pathogen.

Plant pathogen effector proteins are key to pathogen virulence. In susceptible host Brassicas, the clubroot pathogen, Plasmodiophora brassicae, induces the production of nutrient-sink root galls, at the site of infection. Among a list of 32 P. brassiae effector candidates previously reported by our group, we identified SSPbP53 as a putative apoplastic cystatin-like protein highly expressed during the secondary infection. Here we found that SSPbP53 encoding gene is conserved among several P. brassicae pathotypes and that SSPbP53 is an apoplastic protein able to directly interact with and inhibit cruciferous papain-like cysteine proteases (PLCPs), specifically Arabidopsis XYLEM CYSTEINE PEPTIDASE 1 (AtXCP1). The severity of clubroot disease is greatly reduced in the Arabidopsis xcp1 null mutant (AtΔxcp1) after infection with P. brassicae resting spores, indicating that the interaction of P. brassicae SSPbP53 with XCP1 is important to clubroot susceptibility. SSPbP53 is the first cystatin-like effector identified and characterized for a plant pathogenic protist.

Background The oomycete pathogen secretes many effectors into host cells to manipulate host defenses. For the majority of effectors, the mechanisms related to how they alter the expression of host genes and reprogram defenses are not well understood. In order to investigate the molecular mechanisms governing the influence that the Phytophthora infestans RXLR effector Pi04089 has on host immunity, a comparative transcriptome analysis was conducted on Pi04089 stable transgenic and wild-type potato plants. Results Potato plants stably expressing Pi04089 were more susceptible to P. infestans. RNA-seq analysis revealed that 658 upregulated genes and 722 downregulated genes were characterized in Pi04089 transgenic lines. A large number of genes involved in the biological process, including many defense-related genes and certain genes that respond to salicylic acid, were suppressed. Moreover, the comparative transcriptome analysis revealed that Pi04089 significantly inhibited the expression of many flg22 (a microbe-associated molecular pattern, PAMP)-inducible genes, including various Avr9/Cf-9 rapidly elicited (ACRE) genes. Four selected differentially expressed genes (StWAT1, StCEVI57, StKTI1, and StP450) were confirmed to be involved in host resistance against P. infestans when they were transiently expressed in Nicotiana benthamiana. Conclusion The P. infestans effector Pi04089 was shown to suppress the expression of many resistance-related genes in potato plants. Moreover, Pi04089 was found to significantly suppress flg22-triggered defense signaling in potato plants. This research provides new insights into how an oomycete effector perturbs host immune responses at the transcriptome level.

Known elicitors of plant defenses against eggs of herbivorous insects are low-molecular-weight organic compounds associated with the eggs. However, previous studies provided evidence that also proteinaceous compounds present in secretion associated with eggs of the herbivorous sawfly Diprion pini can elicit defensive responses in Pinus sylvestris. Pine responses induced by the proteinaceous secretion are known to result in enhanced emission of (E)-β-farnesene, which attracts egg parasitoids killing the eggs. Here, we aimed to identify the defense-eliciting protein and elucidate its function. After isolating the defense-eliciting protein from D. pini egg-associated secretion by ultrafiltration and gel electrophoresis, we identified it by MALDI-TOF mass spectrometry as an annexin-like protein, which we named 'diprionin'. Further GC-MS analyses showed that pine needles treated with heterologously expressed diprionin released enhanced quantities of (E)-β-farnesene. Our bioassays confirmed attractiveness of diprionin-treated pine to egg parasitoids. Expression of several pine candidate genes involved in terpene biosynthesis and regulation of ROS homeostasis was similarly affected by diprionin and natural sawfly egg deposition. However, the two treatments had different effects on expression of pathogenesis-related genes (PR1, PR5). Diprionin is the first egg-associated proteinaceous elicitor of indirect plant defense against insect eggs described so far.

Wild plants produce abundant seeds and seedlings, but most die before reaching maturity, and these premature deaths are often caused by pathogens. Major genes for resistance protect some seedlings or juveniles. These selected individuals can become a resistant, mature cohort. Alternatively, plants can exhibit mature, adult plant resistance. These two explanations can be indistinguishable in the field, when epidemics in natural pathosystems occur regularly resulting in annual selection for resistance. This study included multi-year observations of a biennial plant where the distinction could be made. White rust of Lunaria annua, a pathosystem native to the Mediterranean Basin, took time in its introduced range in Idaho, USA, to generate epidemics. After years of minimal white rust, an epidemic occurred in 2017 in which first-year, juvenile plants had 20 times the sorus density of second-year, adult plants. Since white rust incidence had been minimal for years prior to 2017, the greater resistance of 2017 adults over 2017 juveniles may have been due to adult-plant resistance. This could also be due to phenology: adult plants have mature leaves, and are flowering and maturing seed, by the time that white rust begins to build up on leaves of juveniles. The juvenile-adult difference was maintained in 2018. In white blister rusts, interpretation of resistance can also be complicated by the frequency of asymptomatic infections that adult plants would pass on to the next generation. However, we found no asymptomatic infection of seeds of L. annua in our sampling of the Idaho population.

Insects tend to feed on related hosts. Coevolution tends to be dominated by interactions resulting from plant chemistry in defense strategies, and evolution of secondary metabolisms being in response to insect herbivory remains a classic explanation of coevolution. The present study examines whether evolutionary constraints existing in host associations of economically important fruit flies in the species‐rich tribe Dacini (Diptera: Tephritidae) and to what extent these species have evolved specialized dietary patterns. We found a strong effect of host phylogeny on associations on the 37 fruit flies tested, although the fruit fly species feeding on ripe commercially grown fruits that lost the toxic compounds after long‐term domestication are mostly polyphagous. We assessed the phylogenetic signal of host breadth across the fruit fly species, showing that the results were substantially different depending on partition levels. Further, we mapped main host family associations onto the fruit fly phylogeny and Cucurbitaceae has been inferred as the most likely ancestral host family for Dacini based on ancestral state reconstruction., The present study examines whether evolutionary constraints existing in host associations of economically important fruit flies in the species‐rich tribe Dacini (Diptera: Tephritidae) and to what extent these species have evolved specialized dietary patterns. We found a strong effect of host phylogeny on associations on the 37 fruit flies tested, although the fruit fly species feeding on ripe commercially grown fruits that lost the toxic compounds after long domestication are mostly polyphagous. Further, we discussed the role of secondary metabolisms in feeding preference, and other suites of traits in the specialization of fruit fly dietary patterns.

Background Sucking insect pests cause severe damage to cotton crop production. The development of insect resistant cotton cultivars is one of the most effective measures in curtailing the yield losses. Considering the role of morphological and biochemical host plant resistance (HPR) traits in plant defense, 12 cotton genotypes/varieties were evaluated for leaf area, leaf glanding, total soluble sugars, total soluble proteins, total phenolics, tannin and total flavonoids against fluctuating populations of whitefly, thrips and jassid under field conditions. Results The population of these insects fluctuated during the growing season and remained above threshold level (whitefly > 5, thrips > (8–10), or jassid > 1 per leaf) during late June and early July. Strong and negative association of whitefly (r = − 0.825) and jassid (r = − 0.929) with seed cotton yield was observed. Mean population of insects were the highest in Glandless-1 followed by NIA-82 and NIA-M30. NIAB-Kiran followed by NIAB-878 and Sadori were the most resistant, with the mean population of 1.41, 1.60, 1.66 (whitefly); 2.24, 2.32, 2.53 (thrips) and 0.37, 0.31, 0.36 (jassid), respectively. The resistant variety NIAB-Kiran showed less soluble sugars (8.54 mg·g− 1), soluble proteins (27.11 mg·g− 1) and more phenolic (36.56 mg·g− 1) and flavonoids (13.10 mg·g− 1) as compared with the susceptible check Glandless-1. Moreover, all insect populations were positively correlated with total soluble sugars and proteins. Whitefly populations exhibited negative response to leaf gossypol glands, total phenolics, tannins and flavonoids. The thrips and jassid populations had a significant and negative correlation with these four biochemical HPR traits. Conclusion The identified resistant resources and HPR traits can be deployed against sucking insect pests’ complex in future breeding programs of developing insect resistant cotton varieties.

Phenotypic variability results from interactions between genotype and environment and is a major driver of ecological and evolutionary interactions. Measuring the relative contributions of genetic variation, the environment, and their interaction to phenotypic variation remains a fundamental goal of evolutionary ecology.In this study, we assess the question: How do genetic variation and local environmental conditions interact to influence phenotype within a single population? We explored this question using seed from a single population of common milkweed, Asclepias syriaca, in northern Michigan. We first measured resistance and resistance traits of 14 maternal lines in two common garden experiments (field and greenhouse) to detect genetic variation within the population. We carried out a reciprocal transplant experiment with three of these maternal lines to assess effects of local environment on phenotype. Finally, we compared the phenotypic traits measured in our experiments with the phenotypic traits of the naturally growing maternal genets to be able to compare relative effect of genetic and environmental variation on naturally occurring phenotypic variation. We measured defoliation levels, arthropod abundances, foliar cardenolide concentrations, foliar latex exudation, foliar carbon and nitrogen concentrations, and plant growth.We found a striking lack of correlation in trait expression of the maternal lines between the common gardens, or between the common gardens and the naturally growing maternal genets, suggesting that environment plays a larger role in phenotypic trait variation of this population. We found evidence of significant genotype‐by‐environment interactions for all traits except foliar concentrations of nitrogen and cardenolide. Milkweed resistance to chewing herbivores was associated more strongly with the growing environment. We observed no variation in foliar cardenolide concentrations among maternal lines but did observe variation among maternal lines in foliar latex exudation.Overall, our data reveal powerful genotype‐by‐environment interactions on the expression of most resistance traits in milkweed., This study explores how local growing environment and genetic variation dictates plant trait expression within a small population. We measured within‐population genetic variation in resistance traits of common milkweed and then compared those traits with patterns of resistance in the naturally growing milkweed population. Our findings suggest that genotype‐by‐environment interactions and environmental variation strongly influence phenotypic expression in this population of common milkweed and that standing genetic variation alone has a limited role in phenotypic variation.

Recent updates in JA biosynthesis, signaling pathways and the crosstalk between JA and others phytohormones in relation with plant responses to different stresses. In plants, the roles of phytohormone jasmonic acid (JA), amino acid conjugate (e.g., JA-Ile) and their derivative emerged in last decades as crucial signaling compounds implicated in stress defense and development in plants. JA has raised a great interest, and the number of researches on JA has increased rapidly highlighting the importance of this phytohormone in plant life. First, JA was considered as a stress hormone implicated in plant response to biotic stress (pathogens and herbivores) which confers resistance to biotrophic and hemibiotrophic pathogens contrarily to salicylic acid (SA) which is implicated in plant response to necrotrophic pathogens. JA is also implicated in plant responses to abiotic stress (such as soil salinity, wounding and UV). Moreover, some researchers have recently revealed that JA controls several physiological processes like root growth, growth of reproductive organs and, finally, plant senescence. JA is also involved in the biosynthesis of various metabolites (e.g., phytoalexins and terpenoids). In plants, JA signaling pathways are well studied in few plants essentially Arabidopsis thaliana, Nicotiana benthamiana, and Oryza sativa L. confirming the crucial role of this hormone in plants. In this review, we highlight the last foundlings about JA biosynthesis, JA signaling pathways and its implication in plant maturation and response to environmental constraints.

Aims: The conventional management adopted in many Mediterranean olive orchards makes them more vulnerable to climate change and attacks by pathogens, due to the decreased chemical plant defenses. In this scenario, a metabolomic analysis was carried out on the xylem sap (X) of olive plants (Olea europaea L.) grown in the Salento peninsula (Italy). Methods: Trials were carried out in two olive groves, one organically and one conventionally managed (controls), successively both converted to sustainable management (i.e. frequent light pruning, soil and foliar fertilization, cover crops). The X was extracted from the shoots of olive plants using a Scholander pressure chamber pressurized with N and gas chromatography-mass spectrometry metabolite profiling was performed in the X. Results: An untargeted gas chromatography mass spectrometry (GC-MS) based metabolomic analysis of primary metabolites (including underivatized volatiles) of the X revealed relative abundances of 153 identified metabolites and 336 unknown features across the 12 samples from four groups of samples. Among them, more than half were involved in the primary metabolism. Many of the compounds with increased levels under sustainable management (such as amino acids, soluble sugars, sugar alcohols) have a well-known role as osmoprotectants or are involved in plant defense, growth and development during stress or recovery stages. Conclusions: Sustainable management in olive groves can increase the ability of plants to overcome environmental stressors and enhance ecosystem balance.

Background Aphid attack induces defense responses in plants activating several signaling cascades that led to the production of toxic, repellent or antinutritive compounds and the consequent reorganization of the plant primary metabolism. Pepper (Capsicum annuum L.) leaf proteomic response against Myzus persicae (Sulzer) has been investigated and analyzed by LC-MS/MS coupled with bioinformatics tools. Results Infestation with an initially low density (20 aphids/plant) of aphids restricted to a single leaf taking advantage of clip cages resulted in 6 differentially expressed proteins relative to control leaves (3 proteins at 2 days post-infestation and 3 proteins at 4 days post-infestation). Conversely, when plants were infested with a high density of infestation (200 aphids/plant) 140 proteins resulted differentially expressed relative to control leaves (97 proteins at 2 days post-infestation, 112 proteins at 4 days post-infestation and 105 proteins at 7 days post-infestation). The majority of proteins altered by aphid attack were involved in photosynthesis and photorespiration, oxidative stress, translation, protein folding and degradation and amino acid metabolism. Other proteins identified were involved in lipid, carbohydrate and hormone metabolism, transcription, transport, energy production and cell organization. However proteins directly involved in defense were scarce and were mostly downregulated in response to aphids. Conclusions The unexpectedly very low number of regulated proteins found in the experiment with a low aphid density suggests an active mitigation of plant defensive response by aphids or alternatively an aphid strategy to remain undetected by the plant. Under a high density of aphids, pepper leaf proteome however changed significantly revealing nearly all routes of plant primary metabolism being altered. Photosynthesis was so far the process with the highest number of proteins being regulated by the presence of aphids. In general, at short times of infestation (2 days) most of the altered proteins were upregulated. However, at longer times of infestation (7 days) the protein downregulation prevailed. Proteins involved in plant defense and in hormone signaling were scarce and mostly downregulated.

Herbivorous thrips are considered as one of the most economically important pests worldwide. They cause direct damage by feeding on plants of at least 60 families including many vegetable and ornamental crops, and indirect damage by transferring devastating plant viruses. Their high and parthenogenetic reproductive capacity and hidden lifestyle (pupation in the soil, thigmotactic behavior) make thrips hard to control. Current pest management control relies mainly on the use of chemical pesticides, to which thrips can develop resistance. Exploiting the natural defense mechanisms of plants against thrips can provide the opportunity towards more sustainable resistance breeding. The major aim of this PhD research was to explore the molecular mechanisms underlying natural defenses of the model plant Arabidopsis thaliana (Arabidopsis) in response to the Western flower thrips (Frankliniella occidentalis). To achieve this, several non-destructive bioassays were designed to assess thrips performance on single Arabidopsis leaves or whole plants. Using RNA-sequencing in a high-density time series, combined with bioinformatic analyses, we were able to capture the transcriptional dynamics and chronology of genes within the gene regulatory network that were induced in single Arabidopsis leaves upon thrips infestation. We confirmed that jasmonic acid (JA) is the predominant phytohormone modulating the induced defense response against thrips as (1) most up-regulated genes showed an overrepresentation of bHLH TF binding motifs which correlated with the activation of JA-associated processes, (2) JA-insensitive mutant coi1-34 and the MYC triple mutant myc2,3,4 showed significantly more feeding damage and a higher number of oviposited eggs compared to the Col-0 wild-type and (3) the novel regulators discovered in the thrips-induced gene regulatory network (GRN), although having distinct roles in the thrips-GRN, are closely connected to the JA pathway. However, depending on the thrips developmental stage, the predominant JA-induced response is modulated differently. For example, L1 larvae induced a relatively higher expression level of the ethylene (ET)-coregulated JA-inducible ERF-branch marker gene PDF1.2, which correlated with defense repression, while the older developmental larval stages activated higher levels of the abscisic acid (ABA)-coregulated JA-inducible MYC-branch marker gene VSP2, which was associated with enhanced plant resistance. Interestingly, although herbivory by L2 stage larvae of thrips mainly activated the expression of the MYC-branch marker gene VSP2 in the locally infested leaves, the ERF-branch marker gene PDF1.2 was mainly activated in the systemic, undamaged leaves of Arabidopsis, which correlated with enhanced preference of subsequently attacking thrips for the youngest systemic leaf tissue. In contrast, the tissue-chewing caterpillar Mamestra brassicae induced relatively higher levels of VSP2 in the local as well as the systemic, undamaged leaves of Arabidopsis, which was associated with enhanced deterrence of subsequently attacking thrips. Collectively, the work described in this thesis provides a better understanding of thrips-induced plant defense responses, which can facilitate the development of more sustainable thrips-resistance breeding.

Here we report the data associated with the article: “Comparative proteome profiling of susceptible and resistant rice cultivars identified an arginase involved in rice defense against Xanthomonas oryzae pv. oryzae” [1]. Bacterial blight disease caused by Xanthomonas oryzae pv. oryzae (Xoo) is one of the most devastating diseases of rice across the globe; however, the underlying molecular mechanism of rice-Xoo interaction is currently not well understood. In this manuscript, we report the proteome profiles of rice leaves generated using a label-free quantitative proteomic analysis using QExactive™ Orbitrap High-Resolution Mass Spectrometer, MapMan, and rice interactome viewer [1].

Summary Viral infections are accompanied by a massive production of small interfering RNAs (siRNAs) of plant origin, such as virus‐activated (va)siRNAs, which drive the widespread silencing of host gene expression, and whose effects in plant pathogen interactions remain unknown.By combining phenotyping and molecular analyses, we characterized vasiRNAs that are associated with typical mosaic symptoms of cauliflower mosaic virus infection in two crops, turnip (Brassica rapa) and oilseed rape (Brassica napus), and the reference plant Arabidopsis thaliana.We identified 15 loci in the three infected plant species, whose transcripts originate vasiRNAs. These loci appear to be generally affected by virus infections in Brassicaceae and encode factors that are centrally involved in photosynthesis and stress response, such as Rubisco activase (RCA), senescence‐associated protein, heat shock protein HSP70, light harvesting complex, and membrane‐related protein CP5. During infection, the expression of these factors is significantly downregulated, suggesting that their silencing is a central component of the plant’s response to virus infections. Further findings indicate an important role for 22 nt long vasiRNAs in the plant’s endogenous RNA silencing response.Our study considerably enhances knowledge about the new class of vasiRNAs that are triggered in virus‐infected plants and will help to advance strategies for the engineering of gene clusters involved in the development of crop diseases.

Theory predicts that herbivory should primarily determine the evolution of herbivore-induced plasticity in plant defenses, but little is known about the influence of other interactions such as pollination. Pollinators may exert negative selection on the herbivore-induced plasticity of chemical defenses when floral signals and rewards are indirectly affected, provoking deterrent effects on these mutualists. We investigated the influence of constant herbivory and pollination on the evolved patterns and degree of herbivore-induced plasticity in chemical plant defenses and floral morphometry and volatiles in fast-cycling Brassica rapa plants. To do this, we used plants from an evolution experiment that had evolved under bee/hand pollination and herbivory manipulated in a 2 × 2 factorial design during six generations, producing four selection treatments. We grew sibling plant pairs from each of the four selection treatments of the last generation and infested one group with herbivores and left the other uninfested. Herbivore-induced plasticity was analyzed within- and between-selection treatments. We found support for the hypothesis that constant herbivory favors the evolution of higher constitutive yet lower herbivore-induced plasticity in defenses. However, this only occurred in plants that evolved under hand pollination and constant herbivory. Bee pollination had a strong influence on the evolution of herbivore-induced plasticity of all traits studied. Plants that evolved under bee pollination, with and without constant herbivory, showed remarkably similar patterns of herbivore-induced plasticity in their defense- and floral traits and had a higher number of plastic responses compared to plants with hand pollination. Such patterns support the hypothesis that bee pollination influenced the evolution of herbivore-induced plasticity, most likely via indirect effects, such as links between defense- and floral traits. We conclude that interactions other than herbivory, such as pollination, may impact herbivore-induced plasticity, through indirect effects and metabolic trade-offs, when it contributes to trait evolution in plants.

The biochemical activities of dirigent proteins (DPs) give rise to distinct complex classes of plant phenolics. DPs apparently began to emerge during the aquatic-to-land transition, with phylogenetic analyses revealing the presence of numerous DP subfamilies in the plant kingdom. The vast majority (>95%) of DPs in these large multigene families still await discovery of their biochemical functions. Here, we elucidated the 3D structures of two pterocarpan-forming proteins with dirigent-like domains. Both proteins stereospecifically convert distinct diastereomeric chiral isoflavonoid precursors to the chiral pterocarpans, (–)- and (+)-medicarpin, respectively. Their 3D structures enabled comparisons with stereoselective lignan– and aromatic terpenoid–forming DP orthologs. Each protein provides entry into diverse plant natural products classes, and our experiments suggest a common biochemical mechanism in binding and stabilizing distinct plant phenol–derived mono- and bis-quinone methide intermediates during different C–C and C–O bond–forming processes. These observations provide key insights into both their appearance and functional diversification of DPs during land plant evolution/adaptation. The proposed biochemical mechanisms based on our findings provide important clues to how additional physiological roles for DPs and proteins harboring dirigent-like domains can now be rationally and systematically identified.

Plant diseases caused by pathogens and pests are a constant threat to global food security. Direct crop losses and the measures used to control disease (e.g. application of pesticides) have significant agricultural, economic, and societal impacts. Therefore, it is essential that we understand the molecular mechanisms of the plant immune system, a system that allows plants to resist attack from a wide variety of organisms ranging from viruses to insects. Here, we provide a roadmap to plant immunity, with a focus on cell-surface and intracellular immune receptors. We describe how these receptors perceive signatures of pathogens and pests and initiate immune pathways. We merge existing concepts with new insights gained from recent breakthroughs on the structure and function of plant immune receptors, which have generated a shift in our understanding of cell-surface and intracellular immunity and the interplay between the two. Finally, we use our current understanding of plant immunity as context to discuss the potential of engineering the plant immune system with the aim of bolstering plant defenses against disease.

This study evaluated the effect of silicon (Si)-supplementation in controlling the fusariosis in two Piper nigrum cultivars (Bragantina and Kottanadan). We hypothesized that susceptible cultivar improves the photosynthetic performance with Si-supplementation. Under greenhouse conditions, six-month-old plants were treated with silicon 30 and 15 d before inoculation with Fusarium solani f. sp. piperis and compared with untreated plants and with healthy control plants. All samples remained photosynthetically active after infection. Chlorophyll content, chlorophyll a fluorescence, and leaf gas-exchange parameters were less affected in Si-supplemented plants as well as healthy control plants. The net carbon assimilation, stomatal conductance, associated with higher internal carbon values suggest that the lower CO2 influx in inoculated plants of Bragantina cultivar was caused by photochemical and biochemical limitations. Si-supplementation reduced the disease severity improving protection, apparently associated with the preservation of photosynthetic performance, especially for Bragantina cultivar.

The enemy release hypothesis is often cited as a potential explanation for the success of introduced plants; yet, empirical evidence for enemy release is mixed. We aimed to quantify changes in herbivory and defense in introduced plants while controlling for three factors that might have confounded past studies: using a wide native range for comparison with the introduced range, measuring defense traits without determining whether they affect herbivore preferences, and not considering the effect of time since introduction. The first hypothesis we tested was that introduced plants will have evolved lower levels of plant defense compared to their source population. We grew South African (source) and Australian (introduced) beach daisies (Arctotheca populifolia) in a common‐environment glasshouse experiment and measured seven defense traits. Introduced plants had more ash, alkaloids, and leaf hairs than source plants, but were also less tough, with a lower C:N ratio and less phenolics. Overall, we found no difference in defense between source and introduced plants. To determine whether the feeding habits of herbivores align with changes in defense traits, we conducted preference feeding trials using five different herbivore species. Herbivores showed no overall preference for leaves from either group. The second hypothesis we tested was that herbivory on introduced plant species will increase through time after introduction to a new range. We recorded leaf damage on herbarium specimens of seven species introduced to eastern Australia and three native control species. We found no change in the overall level of herbivory experienced by introduced plants since arriving in Australia. Conclusion In the field of invasion ecology, we need to rethink the paradigm that species introduced to a new range undergo simple decreases in defenses against herbivores. Instead, plants are likely to employ a range of defense traits that evolve in both coordinated and opposing ways in response to a plethora of different biotic and abiotic selective pressures., We aimed to quantify changes in herbivory and defense in introduced plants while controlling for some factors that might have confounded past studies. We compared the introduced Australian beach daisy to its South African source population to measure seven defence traits (corroborated with herbivore preference feeding trials), and we used herbarium specimens of another seven introduced species to track herbivory over time. We found many changes in defense traits but no overall difference in defense, and no changes in herbivory over time. We show that species introduced to a new range do not always undergo a simple decrease in defense against herbivores.

The rice striped stem borer (SSB, Chilo suppressalis) is one of the most destructive pests of rice plants. Si‐mediated rice defense against various pests has been widely reported, and sodium silicate (SS) has been used as an effective source of silicon for application to plants. However, there is quite limited information about the direct effects of Si application on herbivorous insects. SSB larval performance and their insecticide tolerance were examined after they had been reared either on rice plants cultivated in nutrient solution containing 0.5 and 2.0 mM SS or on artificial diets with 0.1% and 0.5% SS. SS amendment in either rice culture medium or artificial diets significantly suppressed the enzymatic activities of acetylcholinesterase, glutathione S‐transferases, and levels of cytochrome P450 protein in the midgut of C. suppressalis larvae. Larvae fed on diets containing SS showed lower insecticide tolerance. Additionally, RNA‐seq analysis showed that SS‐mediated larval insecticide tolerance was closely associated with fatty acid biosynthesis and pyruvate metabolism pathways. Our results suggest that Si not only enhances plant resistance against insect herbivore, but also impairs the insect's capacity to detoxify the insecticides. This should be considered as another important aspect in Si‐mediated plant–insect interaction and may provide a novel approach of pest management., Sodium silicate (SS) amendments in rice plants significantly suppressed the activities of insecticide detoxification‐related enzymes acetylcholinesterase, glutathione S‐transferases, and cytochrome P450 in Chilo suppressalis larvae. Therefore, larvae fed on SS‐amended plants showed lower insecticide tolerance. Double‐check the phenomena above by using artificial diets, we found sodium silicate had directly adverse effects on the performance of C. suppressalis larvae to further insecticide treatments.

The variability in the genetic variance–covariance (G‐matrix) in plant resistance and its role in the evolution of invasive plants have been long overlooked. We conducted an additional analysis of the data of a reciprocal transplant experiment with tall goldenrod, Solidago altissima, in multiple garden sites within its native range (USA) and introduced range (Japan). We explored the differences in G‐matrix of resistance to two types of foliar herbivores: (a) a lace bug that is native to the USA and recently introduced to Japan, (b) and other herbivorous insects in response to plant origins and environments. A negative genetic covariance was found between plant resistances to lace bugs and other herbivorous insects, in all combinations of garden locations and plant origins except for US plants planted in US gardens. The G‐matrix of the resistance indices did not differ between US and Japanese plants either in US or Japanese gardens, while it differed between US and Japanese gardens in both US and Japanese plants. Our results suggested that the G‐matrix of the plant resistance may have changed in response to novel environmental differences including herbivore communities and/or other biotic and abiotic factors in the introduced range. This may have revealed a hidden trade‐off between resistances, masked by the environmental factors in the origin range. These results suggest that the stability of the genetic covariance during invasion, and the environmentally triggered variability in the G‐matrices of plant resistance may help to protect the plant against multiple herbivore species without changing its genetic architecture and that this may lead to a rapid adaptation of resistance in exotic plants. Local environments of the plant also have a critical effect on plant resistance and should be considered in order to understand trait evolution in exotic plants., This article demonstrated that the novel herbivore environment in the introduced range can alter the genetic variance–covariance of resistance traits in an invasive plant can, while it remained stable during invasion. This suggests that the stability and variability of the genetic basis of resistance as an underlying mechanism for the rapid adaptation of resistance in invasive plants.

Antioxidant enzymes play an important role in plant defense against pathogenic agents. Following the identification of the pathogen, plants produce active oxygen species (ROS) as one of their first defense responses. To maintain the balance of ROS levels and prevent their harmful effects, plants produce antioxidant peroxidase (POX), catalase (CAT), ascorbate peroxidase (APX) and superoxide dismutase (SOD) enzymes. In the present study, the resistance of bean plants cultivars, namely Sadri, Paak, Darakhshan and Dorsa, to Xanthomonas axonopodis pv. phaseoli (Xap) were studied in greenhouse conditions. The catalase, peroxidase and ascorbate peroxidase enzyme activities were studied in healthy and Xap-infected bean cultivars Sadri and Derakhshan at 0, 24, 48, 72 hours and 20 days post inoculation by a completely randomized design with 5 treatments and 4 replications. The result showed that disease symptoms appeared in all tested cultivars. Derakhshan and Sadri cultivars, with 58.33 and 80.56 percentages of infected plants 20 days after inoculation, showed the least and highest infection rates, respectively. The highest catalase and peroxidase activities were recorded 24 and 48h post inoculation. These records reduced 48 and 72 hours post inoculation, respectively. The activities of these two enzymes in the susceptible cultivar were less than those in the semi-resistant one. The chlorophyll a and chlorophyll b contents of Xap-infected plants reduced significantly. The total chlorophyll content of uninfected Sadri and Darakhshan cultivars were 2.93 and 3.23 µg/g, respectively, which reduced to 1.96 and 2.14 µg/g of leaf tissue in infected plants, respectively. Based on these results, it is suggested that the Derakhshan cultivar should be planted in disease-susceptible regions as the semi-resistant cultivar.

Beneficial soil microbes can enhance plant growth and defense, but the extent to which this occurs depends on the availability of resources, such as water and nutrients. However, relatively little is known about the role of light quality, which is altered during shading, resulting a low red: far-red ratio (R:FR) of light. We examined how low R:FR light influences arbuscular mycorrhizal fungus (AMF)-mediated changes in plant growth and defense using Solanum lycopersicum (tomato) and the insect herbivore Chrysodeixis chalcites. We also examined effects on third trophic level interactions with the parasitoid Cotesia marginiventris. Under low R:FR light, non-mycorrhizal plants activated the shade avoidance syndrome (SAS), resulting in enhanced biomass production. However, mycorrhizal inoculation decreased stem elongation in shaded plants, thus counteracting the plant’s SAS response to shading. Unexpectedly, activation of SAS under low R:FR light did not increase plant susceptibility to the herbivore in either non-mycorrhizal or mycorrhizal plants. AMF did not significantly affect survival or growth of caterpillars and parasitoids but suppressed herbivore-induced expression of jasmonic acid-signaled defenses genes under low R:FR light. These results highlight the context-dependency of AMF effects on plant growth and defense and the potentially adverse effects of AMF under shading.

Brassica napus (oilseed rape) is one of the most important oil crops worldwide, but its growth is seriously threatened by Sclerotinia sclerotiorum. The mechanism of oilseed rape response to this pathogen has rarely been studied. Here, it was identified that BnaA03.MKK5 whose expression was induced by S. sclerotiorum infection was involved in plant immunity. BnaA03.MKK5 overexpression lines exhibited decreased disease symptoms compared to wild-type plants, accompanied by the increased expression of camalexin-biosynthesis-related genes, including BnPAD3 and BnCYP71A13. In addition, two copies of BnMPK3 (BnA06.MPK3 and BnC03.MPK3) were induced by Sclerotinia incubation, and BnaA03.MKK5 interacted with BnaA06.MPK3/BnaC03.MPK3 in yeast. These interactions were confirmed using in vivo co-immunoprecipitation assays. In vitro phosphorylation assays showed that BnaA06.MPK3 and BnaC03.MPK3 were the direct phosphorylation substrates of BnaA03.MKK5. The transgenic oilseed rape plants including BnaA06.MPK3 and BnaC03.MPK3 overexpression lines and BnMPK3 gene editing lines mediated by CRISPR/Cas9 were generated; the results of the genetic transformation of BnaA06.MPK3/BnaC03.MPK3 indicate that BnMPK3 also has a positive role in Sclerotinia resistance. This study provides information about the potential mechanism of B. napus defense against S. Sclerotiorum mediated by a detailed BnaA03.MKK5-BnaA06.MPK3/BnaC03.MPK3 module.

N6-methylated adenine (m6A) is the most prevalent modification of mRNA methylation and can regulate many biological processes in plants, such as mRNA processing, development, and stress response. Some studies have increased our understanding of its various roles in model plants in recent years. Nevertheless, the distribution of m6A and the impact of m6A on the regulation of plant defense responses against pathogen inoculation are virtually unknown in pear. In this study, MeRIP-seq and RNA-seq data from healthy and inoculated plants were analyzed to assess the changes in the transcript levels and posttranscriptional modification of pear in response to the fire blight pathogen Erwinia amylovora. Following the analysis of 97,261 m6A peaks, we found that m6A preferred to modify duplicate genes rather than singleton genes and that m6A-methylated genes underwent stronger purifying selection. A total of 2,935 specific m6A sites were detected at the transcriptome level after inoculation, which may increase defense-related transcript abundance to enhance pear resistance. In addition, 1,850 transcripts were detected only in the mock-inoculated groups. The hypomethylated transcripts were mainly related to transcriptional regulation and various biological processes, such as chloroplast organization and sucrose biosynthetic processes. In addition, we found that the extent of m6A methylation was significantly positively correlated with the transcript level, suggesting a regulatory role for m6A in the plant response.

Resistance gene analogs (RGAs) comprising NBS-LRR gene family members are considered prominent candidates in the development of disease-resistant genotypes. NBS-LRR gene family comprised a very large number of genes; therefore, members of one subfamily TIR-NBS-LRR (TNL) are identified in the present study from Solanum tuberosum genome, followed by their bioinformatics characterization. The study identified a total of 44 genes encoding 60 TNL transcripts with two prominent clusters at chromosome 1 and chromosome 11. Expression analysis of 14 TNL genes after Alternaria solani infection at 1, 2, 3, 5, and 7 days post inoculation in two disease-tolerant varieties, Kufri Jyoti and Kufri Pukhraj, and one relatively susceptible variety, Kufri Chandramukhi, showed differential expression of many genes including a high expression (>15-fold) of StTNLC6G2T1 and StTNLC11G9T1. Functional characterization of one such gene, StTNLC7G2, reveals involvement in the generation of reactive oxygen species under A. solani attack, implicating its putative role in plant defense via hypersensitive response.

Plant immunity against pathogens and pests is comprised of complex mechanisms orchestrated by signaling pathways regulated by plant hormones [Salicylic acid (SA) and Jasmonic acid (JA)]. Investigations of plant immune response to phytopathogens and phloem-feeders have revealed that SA plays a critical role in reprogramming of the activity and/or localization of transcriptional regulators via post-translational modifications. We explored the contributing effects of herbivory by a phytopathogen vector [Asian citrus psyllid, Diaphorina citri] and pathogen [Candidatus Liberibacter asiaticus (CLas)] infection on response of sweet orange [Citrus sinensis (L.) Osbeck] using manipulative treatments designed to mimic the types of infestations/infections that citrus growers experience when cultivating citrus in the face of Huanglongbing (HLB) disease. A one-time (7 d) inoculation access period with CLas-infected vectors caused SA-associated upregulation of PR-1, stimulating defense response after a long period of infection without herbivory (270 and 330 days). In contrast, while repeated (monthly) ‘pulses’ of 7 d psyllid feeding injury stimulated immunity in CLas-infected citrus by increasing [SA] in leaves initially (up to 120 d), long-term (270 and 330 days) repeated herbivory caused [SA] to decrease coincident with upregulation of genes associated with SA metabolism (BMST and DMR6). Similarly, transcriptional responses and metabolite (SA and its analytes) accumulation in citrus exposed to a continuously reproducing population of D. citri exhibited a transitory upregulation of genes associated with SA signaling at 120 days and a posterior downregulation after long-term psyllid (adults and nymphs) feeding (270 and 330 days). Herbivory played an important role in regulation of SA accumulation in mature leaves of C. sinensis, whether or not those trees were coincidentally infected with CLas. Our results indicate that prevention of feeding injury inflicted by D. citri from the tritrophic interaction may allow citrus plants to better cope with the consequences of CLas infection, highlighting the importance of vector suppression as a component of managing this cosmopolitan disease.Author SummaryWe explored tritrophic interactions among an insect vector (Diaphorina citri) – phytopathogen (Candidatus Liberibacter asiaticus) – and cultivated fruit crop [sweet orange, Citrus sinensis (L) Osbeck]. Transcriptional and metabolic responses of plants were analyzed over an extended time-course of disease progression after various frequencies of herbivore feeding and durations of pathogen infection using manipulative treatments designed to mimic the types of infestations/infections that citrus growers experience when cultivating citrus in the presence of the devastating citrus disease, huanglongbing. We found that in the absence of coincident psyllid feeding damage, citrus trees could mount a defense response against the pathogen by activating the salicylic acid (SA) pathway. Repeated, monthly ‘pulses’ of herbivory led to pronounced stimulation of SA transcription that was coincident with diminished pathogen titers in plants. Although insect injury initially activated SA-dependent defense responses, continuous and/or long-term (≥ 270 d) herbivory shut down PR-1-dependent defense responses against the pathogen. Our results provide a mechanism explaining how vector suppression contributes to maintaining health of cultivated citrus in areas where huanglongbing is endemic. Our results also point to specific gene targets that may yield novel genotypes expressing tolerance against CLas after appropriate manipulations.

Terpenes are secondary metabolites produced from isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP). Considering their role against biotic and abiotic stress conditions, the D-limonene, a terpene recognized for its roles in plant defense and as pigment molecules, was tested to monitor its impact in the accumulation of carotenoid terpene pigments during tomato fruit ripening. The concentrations of 0.0, 1.0, and 2.0 mL·L–1 were sprayed weekly on tomato plants ‘Micro-Tom’ until the harvest. The fruits were harvested at the mature green stage (MG), the breaker stages (BR), three days after the breaker (BR + 3), the red ripe stage (RR), and the concentration of carotenoids was evaluated. The application of D-limonene enhanced the concentration of carotenoids in the early stages of tomato fruit ripening and ripe fruits, and the accumulation of carotenoids was greater following dose increment. This result can bring important contributions to horticulture and food nutrition by linking plant defense and the accumulation of molecules with recognized antioxidant capacity in fruit.

Tomato (Solanum lycopersicum), as an important economical vegetable, is often infected with Rhizoctonia solani, which results in a substantial reduction in production. Therefore, the molecular mechanism of biocontrol microorganisms assisting tomato to resist pathogens is worth exploring. Here, we use Bacillus amyloliquefaciens SN16-1 as biocontrol bacteria, and employed RNA-Seq technology to study tomato gene and defense-signaling pathways expression. Gene Ontology (GO) analyses showed that an oxidation-reduction process, peptidase regulator activity, and oxidoreductase activity were predominant. Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that phenylpropanoid biosynthesis, biosynthesis of unsaturated fatty acids, aldosterone synthesis and secretion, and phototransduction were significantly enriched. SN16-1 activated defenses in the tomato via systemic-acquired resistance (which depends on the salicylic acid signaling pathway), rather than classic induction of systemic resistance. The genes induced by SN16-1 included transcription factors, plant hormones (ethylene, auxin, abscisic acid, and gibberellin), receptor-like kinases, heat shock proteins, and defense proteins. SN16-1 rarely activated pathogenesis-related proteins, but most pathogenesis-related proteins were induced in the presence of the pathogens. In addition, the molecular mechanisms of the response of tomatoes to SN16-1 and R. solani RS520 were significantly different.

Soybean (Glycine max) is a major plant protein source and oilseed crop. However, plant-parasitic nematodes (PPNs) affect its annual yield. In the current study, in order to better understand the regulation of defense mechanism against PPNs in soybean, we investigated the role of long non-coding RNAs (lncRNAs) in response to two nematode species, Heterodera glycines (SCN: soybean cyst nematode) and Rotylenchulus reniformis (reniform). To this end, two publicly available RNA-seq data sets (SCN data set and RAD: reniform-associated data set) were employed to discover the lncRNAome profile of soybean under SCN and reniform infection, respectively. Upon identification of unannotated transcripts in these data sets, a seven-step pipeline was utilized to sieve these transcripts, which ended up in 384 and 283 potential lncRNAs in SCN data set and RAD, respectively. These transcripts were then used to predict cis and trans nematode-related targets in soybean genome. Computational prediction of target genes function, some of which were also among differentially expressed genes, revealed the involvement of putative nematode-responsive genes as well as enrichment of multiple stress responses in both data sets. Finally, 15 and six lncRNAs were proposed to be involved in microRNA-mediated regulation of gene expression in soybean in response to SNC and reniform infection, respectively. Collectively, this study provides a novel insight into the signaling and regulatory network of soybean-pathogen interactions and opens a new window for further research.

Wheat is a widely grown cereal crop that provides >20% daily protein intake and calories worldwide. Drought stress drastically affects the crucial physiological (relative water content; RWC), biochemical (proline content), and molecular attributes in wheat plants from germination to maturity stage and ultimately reduces the yield and quality traits. Given this, the present work was conducted to know the effect of salt seed priming on wheat crop. For this, we have used two wheat variety HUW-468 (V1), and HUW-510 (V2) primed with water (T2), Mg(NO3)2 (T3), ZnSO4 (T4), and combinations of Mg(NO3)2 and ZnSO4 salt (T5) with non-primed seed (T1) under water stress (W1) and control (W0) conditions. We have observed significant differences in plant height (84 to 90 cm), leaf number, total leaf area (431.82 to 505.46 cm2 plant−1), relative water content, chlorophyll content (4.78 to 5.86 mg g−1fresh weight), membrane stability index (MSI) (56.25 to 61.42%), superoxide dismutase (SOD) activity (133.47 to 150.10), MDA content, proline content (101.64 to 100.46), nitrate reductase activity (NRA), and nutrient (N, Zn, and Fe) contents under control and stress conditions. These results concluded that seed priming with a combination of Mg(NO3)2 and ZnSO4 salt is best for improved wheat crop traits under drought stress compared to individual salt and non-priming treatments. Overall, this analysis elucidated that seed priming strengthens the antioxidant capacity under drought conditions by enhancing antioxidant enzymes and alleviating oxidative stress.

Significance Plant genomes encode hundreds of genes controlling the detection, signaling pathways, and immune responses necessary to defend against pathogens. Pathogens, in turn, continually evolve to evade these defenses. Small RNAs, such as microRNAs (miRNAs), are one mechanism used by pathogens to overcome plant defenses and facilitate plant colonization. Mounting evidence would suggest that beneficial microbes, likewise, use miRNAs to facilitate symbiosis. Here, we demonstrate that the beneficial fungus Pisolithus microcarpus encodes a miRNA that enters plant cells and stabilizes the symbiotic interaction. These results demonstrate that beneficial fungi may regulate host gene expression through the use of miRNAs and sheds light on how beneficial microbes have evolved mechanisms to colonize plant tissues., Small RNAs (sRNAs) are known to regulate pathogenic plant–microbe interactions. Emerging evidence from the study of these model systems suggests that microRNAs (miRNAs) can be translocated between microbes and plants to facilitate symbiosis. The roles of sRNAs in mutualistic mycorrhizal fungal interactions, however, are largely unknown. In this study, we characterized miRNAs encoded by the ectomycorrhizal fungus Pisolithus microcarpus and investigated their expression during mutualistic interaction with Eucalyptus grandis. Using sRNA sequencing data and in situ miRNA detection, a novel fungal miRNA, Pmic_miR-8, was found to be transported into E. grandis roots after interaction with P. microcarpus. Further characterization experiments demonstrate that inhibition of Pmic_miR-8 negatively impacts the maintenance of mycorrhizal roots in E. grandis, while supplementation of Pmic_miR-8 led to deeper integration of the fungus into plant tissues. Target prediction and experimental testing suggest that Pmic_miR-8 may target the host NB-ARC domain containing transcripts, suggesting a potential role for this miRNA in subverting host signaling to stabilize the symbiotic interaction. Altogether, we provide evidence of previously undescribed cross-kingdom sRNA transfer from ectomycorrhizal fungi to plant roots, shedding light onto the involvement of miRNAs during the developmental process of mutualistic symbioses.

Certain plant pathogens have demonstrated potential for use as bioherbicides for weed control, and numerous studies have been published on this subject for several decades. One of the early examples of an important fungal bioherbicide is Alternaria cassiae, isolated from the weed sicklepod (Senna obtusifolia). To gain further insight into biochemical interactions of this fungus and its host weed, we examined the effects of this bioherbicide on various enzymes associated with plant defense. Young sicklepod seedlings were challenged with A. cassiae spore inoculum and enzyme activities associated with plant defense (peroxidase, proteolytic, and pectinolytic) were assayed periodically over a 96-h time course on plants grown in continuous darkness or continuous light. Peroxidase activity increased with time in untreated control seedlings in both light and dark, but the effect was greater in the light. In A. cassiae-treated plants, peroxidase was elevated above that in control tissue at all sample times resulting in a 1.5 -fold increase above control in light-grown tissue and a 2- to 3-fold increase in dark-grown tissue over 48–96 h. Differences in leucine aminopeptidase activity in control versus A. cassiae-treated tissues were not significant until 48–96 h, when activity was inhibited in fungus-treated tissues by about 32% in light-grown tissue and 27% in dark-grown tissue after 96 h. Proteolytic activity on benzoyl-arginine-p-nitroanilide was not significantly different in treated versus control tissue in either light or dark over the time course. Pectinase activity increased in treated tissues at all time points as early as 16 h after spore application in light- or dark-grown plants. The greatest increases were 1.5-fold above control levels in light-grown plants (40–64 h) and 2-fold in plants grown in darkness (72–96 h). Data suggests that peroxidase may be involved as defense mechanism of sicklepod when challenged by A. cassia and that this mechanism is operative in young seedlings under both light and dark growth conditions. Differential proteolytic activity responses on these two substrates suggests the presence of two different enzymes. Increased pectinase activity during pathogenesis suggests that A. cassiae-sicklepod interaction results in an infectivity mechanism to degrade pectic polymers important to sicklepod cell wall integrity. These studies provide important information on some biochemical interactions that may be useful for improvements to biological weed control programs utilizing plant pathogens. Such information may also be useful in genetic selection and manipulation of pathogens for weed control.

β-cyclocitral (βCC), a major apocarotenoid of β-carotene, enhances plants’ defense against environmental stresses. However, the knowledge of βCC’s involvement in the complex stress-signaling network is limited. Here we demonstrate how βCC reprograms the transcriptional responses that enable Solanum lycopersicum L. (tomato) plants to endure a plethora of environmental stresses. Comparative transcriptome analysis of control and βCC-treated tomato plants was done by generating RNA sequences in the BGISEQ-500 platform. The trimmed sequences were mapped on the tomato reference genome that identifies 211 protein-coding differentially expressed genes. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analysis and their enrichment uncovered that only upregulated genes are attributed to the stress response. Moreover, 80% of the upregulated genes are functionally related to abiotic and biotic stresses. Co-functional analysis of stress-responsive genes revealed a network of 18 genes that code for heat shock proteins, transcription factors (TFs), and calcium-binding proteins. The upregulation of jasmonic acid (JA)-dependent TFs (MYC2, MYB44, ERFs) but not the JA biosynthetic genes is surprising. However, the upregulation of DREB3, an abscisic acid (ABA)-independent TF, validates the unaltered expression of ABA biosynthetic genes. We conclude that βCC treatment upregulates multiple stress-responsive genes without eliciting JA and ABA biosynthesis.

Insect vector-borne diseases are a major constraint to a wide variety of crops. Plants integrate environmental light and internal signalings to defend dual stresses both from the vector insects and vector-transmitted pathogens. In this review, we highlight a studies that demonstrate how light regulates plants deploying mechanisms against vector-borne diseases. Four major host defensive pathways involved in the host defense network against multiple biotic stresses are reviewed: innate immunity, phytohormone signaling, RNA interference, and protein degradation. The potential with light-engineering technology with light emitting diodes (LEDs) and genome engineering technology for fine-tuning crop defense and yield are also discussed.

Resistance gene analogs (RGAs) comprising NBS-LRR gene family members are considered prominent candidates in the development of disease-resistant genotypes. NBS-LRR gene family comprised a very large number of genes; therefore, members of one subfamily TIR-NBS-LRR (TNL) are identified in the present study from Solanum tuberosum genome, followed by their bioinformatics characterization. The study identified a total of 44 genes encoding 60 TNL transcripts with two prominent clusters at chromosome 1 and chromosome 11. Expression analysis of 14 TNL genes after Alternaria solani infection at 1, 2, 3, 5, and 7 days post inoculation in two disease-tolerant varieties, Kufri Jyoti and Kufri Pukhraj, and one relatively susceptible variety, Kufri Chandramukhi, showed differential expression of many genes including a high expression (>15-fold) of StTNLC6G2T1 and StTNLC11G9T1. Functional characterization of one such gene, StTNLC7G2, reveals involvement in the generation of reactive oxygen species under A. solani attack, implicating its putative role in plant defense via hypersensitive response.

Pathogenic infestations are significant threats to vegetable yield, and have become an urgent problem to be solved. Rhizoctonia solani is one of the worst fungi affecting tomato crops, reducing yield in some regions. It is a known fact that plants have their own defense against such infestations, however, it is unclear whether any exogenous material can help plants against infestation. Therefore, we performed greenhouse experiments to evaluate the impacts of R. solani on 15- and 30-day old tomato plants after fungal infestation, and estimated the antifungal activity of nanoparticles (NPs) against the pathogen. We observed severe pathogenic impacts on the above-ground tissues of tomato plants which would affect plant physiology and crop production. Pathogenic infection reduced total chlorophyll and anthocyanin contents, which subsequently disturbed plant physiology. Further, total phenolic contents (TPC), total flavonoid contents (TFC), and malondialdehyde (MDA) contents were significantly increased in pathogen treatments. Constitutively, enhanced activities were estimated for catalase (CAT), superoxide dismutase (SOD), and ascorbate peroxidase (APX) in response to reactive oxygen species (ROS)in pathogen-treated plants. Moreover, pathogenesis-related genes, namely, chitinase, plant glutathione S-transferase (GST), phenylalanine ammonia-lyase (PAL1), pathogenesis-related protein (PR12), and pathogenesis-related protein (PR1) were evaluated, with significant differences between treated and control plants. In vitro and greenhouse antifungal activity of silver nanoparticles (Ag NPs), chitosan nanoparticles, and Ag NPs/CHI NPs composites and plant health was studied using transmission electron microscopy (TEM) and Fourier transform infrared (FTIR) spectrophotometry. We found astonishing results, namely, that Ag and CHI have antifungal activities against R. solani. Overall, plant health was much improved following treatment with Ag NPs/CHI NPs composites. In order to manage R. solani pathogenicity and improve tomato health, Ag/CHI NPs composites could be used infield as well as on commercial levels based on recommendations. However, there is an urgent need to first evaluate whether these NP composites have any secondary impacts on human health or the environment.

The present data profile the large scale transcriptome changes in Arabidopsis thaliana Col-0 seedlings exposed to mono-rhamnolipids (Mono-RLs) from Pseudomonas aeruginosa secretome. Bacterial rhamnolipids (RLs) are biosurfactant known to trigger plant defense mechanisms and have a great potential for crop culture protection as environmental-friendly biocontrol solution. They are thought to interact directly with membrane lipids to induce plant defense gene expression and protection towards phytopathogens. However, to date, data on the global transcriptomic modifications induced by these natural amphiphilic glycolipids in plants are missing. Ten-day-old seedlings were treated for 1 or 3 h with 100 µM Mono-RLs in liquid growth medium for root absorption. Total RNA samples were extracted, purified, labelled and hybridized to Agilent V4 Gene Expression Microarrays 4 × 44 K (design ID 021169) carrying 43803 ssDNA probes of 60-mer covering the entire genome of A. thaliana. The dataset was validated by quality assessments including RNA sample quality, microarray quality and global gene expression profiling. The raw and normalized formats of these transcriptomic data are available via GEO repository with the accession number GSE168830. The dataset can be used to provide insights into the plant's early and later mechanisms induced or repressed by RLs. It can be compared to data obtained with other plant defense elicitors, including the well described compounds perceived by membrane protein receptors.

Grapevine (Vitis vinifera) berries are extremely sensitive to infection by the biotrophic pathogen Erysiphe necator, causing powdery mildew disease with deleterious effects on grape and wine quality. The combined analysis of the transcriptome and metabolome associated with this common fungal infection has not been previously carried out in any fruit. In order to identify the molecular, hormonal, and metabolic mechanisms associated with infection, healthy and naturally infected V. vinifera cv. Carignan berries were collected at two developmental stages: late green (EL33) and early véraison (EL35). RNA sequencing combined with GC-electron impact ionization time-of-flight MS, GC-electron impact ionization/quadrupole MS, and LC-tandem MS analyses revealed that powdery mildew-susceptible grape berries were able to activate defensive mechanisms with the involvement of salicylic acid and jasmonates and to accumulate defense-associated metabolites (e.g. phenylpropanoids, fatty acids). The defensive strategies also indicated organ-specific responses, namely the activation of fatty acid biosynthesis. However, defense responses were not enough to restrict fungal growth. The fungal metabolic program during infection involves secretion of effectors related to effector-triggered susceptibility, carbohydrate-active enzymes and activation of sugar, fatty acid, and nitrogen uptake, and could be under epigenetic regulation. This study also identified potential metabolic biomarkers such as gallic, eicosanoic, and docosanoic acids and resveratrol, which can be used to monitor early stages of infection., This work was supported by UIDB/04046/2020 and UIDP/04046/2020 Centre grants from FCT, Portugal (to BioISI). DP and FS are recipients of fellowships from BioSys PhD program PD65-2012 (Refs PD/BD/114385/2016 and PB/BD/130976/2017) from FCT (Portugal). This work was supported by Fundação para a Ciência e Tecnologia-funded research project “GrapInfectomics” (PTDC/ASP-HOR/28485/2017). Additionally, it was partially supported by the Spanish Ministry of Economy project BIO2017-86375-R. This article/publication is based upon work from COST Action CA 17111 INTEGRAPE, supported by COST (European Cooperation in Science and Technology).

Stinging trichomes are rare in plants, occurring only in angiosperms, where they are reported for a few genera belonging to six families. Although there is no report of stinging trichomes in Apocynaceae, previous fieldwork collections of Fischeria and Matelea caused us a mild allergic reaction on the skin when we contacted the dense indumentum of the plants. This fact associated with the well-known presence of glandular trichomes with acute apex in both genera raised suspicions that stinging trichomes could be present in the family. Hence, this study aimed to investigate the likely occurrence of stinging trichomes in Fischeria and Matelea. We analyzed vegetative shoots and leaves of Fischeria stellata and Matelea denticulata through the usual procedures of light and scanning electron microscopy. We also performed several histochemical tests to investigate the chemical composition of trichome secretion. We detected that glandular trichomes occur throughout the surface of the leaf and stem. They are multicellular, uniseriate with an apical secretory cell, which has a dilated base and a needle-shaped apex. The secretion is compressed into the acuminate portion of the apical cell by a large vacuole, and crystals are deposited in the cell wall in a subapical position, providing a preferential site of rupture. The secretion, composed of amino acids and/or proteins, is released under mechanical action, causing skin irritation. Based on our detailed morphological and anatomical analyses, and in the functional aspects observed, we concluded that the glandular trichomes in Fischeria and Matelea can indeed be classified as stinging. Thus, Apocynaceae is the seventh family for which this type of trichome has been reported. We also compiled information on stinging trichomes in all families of angiosperms. Their phylogenetic distribution indicates that they have evolved at least 12 times during angiosperm evolution and may represent an evolutionary convergence of plant defense against herbivory.

Simple Summary Prosystemin is a 200 amino acid precursor that releases, upon wounding and biotic attacks, an 18 amino acid peptide called Systemin. This peptide was traditionally considered as the principal actor of the resistance of tomato plants induced by triggering multiple defense pathways in response to a wide range of biotic/abiotic stress agents. Recent findings from our group discovered the disordered structure of Prosystemin that promotes the binding of different molecular partners and the possible activation of multiple stress-related pathways. All of our recent findings suggest that Prosystemin could be more than a simple precursor of Systemin peptide. Indeed, we hypothesized that it contains other sequences able to activate multiple stress-related responses. To verify this hypothesis, we produced a truncated Prosystemin protein deprived of the Systemin peptide and the relative deleted gene. Experiments with transgenic tomato plants overexpressing the truncated Prosystemin and with plants exogenously treated with the recombinant truncated protein demonstrated that both transgenic and treated plants modulated the expression of defense-related genes and were protected against a noctuid moth and a fungal pathogen. Taken together, our results demonstrated that Prosystemin is not a mere scaffold of Systemin, but itself contains other biologically active regions. Abstract Systemin (Sys) is an octadecapeptide, which upon wounding, is released from the carboxy terminus of its precursor, Prosystemin (ProSys), to promote plant defenses. Recent findings on the disordered structure of ProSys prompted us to investigate a putative biological role of the whole precursor deprived of the Sys peptide. We produced transgenic tomato plants expressing a truncated ProSys gene in which the exon coding for Sys was removed and compared their defense response with that induced by the exogenous application of the recombinant truncated ProSys (ProSys(1-178), the Prosystemin sequence devoid of Sys region). By combining protein structure analyses, transcriptomic analysis, gene expression profiling and bioassays with different pests, we demonstrate that truncated ProSys promotes defense barriers in tomato plants through a hormone-independent defense pathway, likely associated with the production of oligogalacturonides (OGs). Both transgenic and plants treated with the recombinant protein showed the modulation of the expression of genes linked with defense responses and resulted in protection against the lepidopteran pest Spodoptera littoralis and the fungus Botrytis cinerea. Our results suggest that the overall function of the wild-type ProSys is more complex than previously shown, as it might activate at least two tomato defense pathways: the well-known Sys-dependent pathway connected with the induction of jasmonic acid biosynthesis and the successive activation of a set of defense-related genes, and the ProSys(1-178)-dependent pathway associated with OGs production leading to the OGs mediate plant immunity.

Plant-insect interactions are a determining factor for sustainable crop production. Although plants can resist or tolerate herbivorous insects to varying degrees, even with the use of pesticides, insects can reduce plant net productivity by as much as 20%, so sustainable strategies for pest control with less dependence on chemicals are needed. Selecting plants with optimal resistance and photosynthetic traits can help minimize damage and maintain productivity. Here, 27 landrace accessions of lima beans, Phaseolus lunatus L., from the Yucatan Peninsula were evaluated in the field for morphological resistance traits, photosynthetic characteristics, insect damage and seed yield. Variation was found in physical leaf traits (number, area, and dry mass of leaves; trichome density, specific leaf thickness and hardness) and in physiological traits (photosynthetic rate, stomatal conductance, intercellular carbon, water-use efficiency, and transpiration). Five accessions (JMC1325, JMC1288, JMC1339, JMC1208 and JMC1264) had the lowest index for cumulative damage with the highest seed yield, although RDA analysis uncovered two accessions (JMC1339, JMC1288) with strong positive association of seed yield and the cumulative damage index with leaf production, specific leaf area (SLA) and total leaf area. Leaf traits, including SLA and total leaf area are important drivers for optimizing seed yield. This study identified 12 important morphological and physiological leaf traits for selecting landrace accessions of P. lunatus for high yields (regardless of damage level) to achieve sustainable, environmentally safe crop production.

Several experimental studies on aquatic plants have reported the prevalence of chemical defense mechanism against herbivory, as opposed to structural, life-forms or other traits. Here, our laboratory feeding experiments and integrative analysis explored the relationship among palatability (fresh or reconstituted plants used as artificial diet) and various chemical/nutritional traits (i.e., contents of dry mass, ash, nitrogen, protein, and phenols) of diverse aquatic plants and their susceptibility to consumption by the generalist gastropod Biomphalaria glabrata. Biomphalaria glabrata consumed all of the assayed aquatic plants in a hierarchical yet generalized way, with the consumption of fresh plants, their reconstituted forms and defensive properties of lipophilic extracts not being significantly correlated with plant physical or chemical traits to determine the feeding preference of the gastropod. Our results do not reveal a prevalence for a specific plant attribute contributing to herbivory. Instead, they indicate that the susceptibility of aquatic plants to generalist consumers is probably related to a combination of their chemical and physical properties, resulting in moderate grazing rates by generalist consumers.

Pathogenic infestations are significant threats to vegetable yield, and have become an urgent problem to be solved. Rhizoctonia solani is one of the worst fungi affecting tomato crops, reducing yield in some regions. It is a known fact that plants have their own defense against such infestations; however, it is unclear whether any exogenous material can help plants against infestation. Therefore, we performed greenhouse experiments to evaluate the impacts of R. solani on 15- and 30-day old tomato plants after fungal infestation, and estimated the antifungal activity of nanoparticles (NPs) against the pathogen. We observed severe pathogenic impacts on the above-ground tissues of tomato plants which would affect plant physiology and crop production. Pathogenic infection reduced total chlorophyll and anthocyanin contents, which subsequently disturbed plant physiology. Further, total phenolic contents (TPC), total flavonoid contents (TFC), and malondialdehyde (MDA) contents were significantly increased in pathogen treatments. Constitutively, enhanced activities were estimated for catalase (CAT), superoxide dismutase (SOD), and ascorbate peroxidase (APX) in response to reactive oxygen species (ROS)in pathogen-treated plants. Moreover, pathogenesis-related genes, namely, chitinase, plant glutathione S-transferase (GST), phenylalanine ammonia-lyase (PAL1), pathogenesis-related protein (PR12), and pathogenesis-related protein (PR1) were evaluated, with significant differences between treated and control plants. In vitro and greenhouse antifungal activity of silver nanoparticles (Ag NPs), chitosan nanoparticles, and Ag NPs/CHI NPs composites and plant health was studied using transmission electron microscopy (TEM) and Fourier transform infrared (FTIR) spectrophotometry. We found astonishing results, namely, that Ag and CHI have antifungal activities against R. solani. Overall, plant health was much improved following treatment with Ag NPs/CHI NPs composites. In order to manage R. solani pathogenicity and improve tomato health, Ag/CHI NPs composites could be used infield as well as on commercial levels based on recommendations. However, there is an urgent need to first evaluate whether these NP composites have any secondary impacts on human health or the environment.

Phytopathogens and pests are two major factors that limit the growth of plants. The expression of a flavonoid regulator gene, AtMYB12(AT2G47460), has been reported to increase the endogenous flavonoid content of tobacco and tomato. Previous research has only focused on the regulation mechanism of v-myb avian myeloblastosis viral oncogene homolog (MYB) transcription factors under single stress conditions. Here, research showed that AtMYB12 was involved in regulating the resistance of tobacco to multiple biological stresses such as phytopathogens and aphid. We reported that transgenic tobacco carrying AtMYB12 was more resistant to Ralstonia solanacearum when the up-regulated expression of several defense-related markers, such as NbPR1a, NbNOA1, and NbrbohB, was activated, suggesting that the priming defense of a plant may contribute to bacterial disease resistance. The improvement of the resistance of AtMYB12-expressing transgenic tobacco is achieved by promoting the production of ROS, H2O2, and NO. AtMYB12-expressing transgenic tobacco also has resistance to fungal pathogens, such as Colletotrichum nicotianae Averna and Alternaria alternate. The enrichment of flavonols components, such as rutin, which directly inhibit the growth of C. nicotianae and A. alternate, may also contribute to the defense mediated by AtMYB12 over-expression. At the same time, the results also confirm that AtMYB12-expressing transgenic tobacco enhanced plant resistance to aphid-infested (Aphidoidea) pests. These results suggest that the AtMYB12 gene is a good candidate for pest and disease control, with limited resistance costs and enrichment in flavonols, and that AtMYB12 has a potential in the breeding of disease-resistant tobacco crops.

Pectin is a major cell wall component that plays important roles in plant development and response to environmental stresses. Arabidopsis thaliana plants expressing a fungal polygalacturonase (PG plants) that degrades homogalacturonan (HG), a major pectin component, as well as loss-of-function mutants for QUASIMODO2 (QUA2), encoding a putative pectin methyltransferase important for HG biosynthesis, show accumulation of reactive oxygen species (ROS), reduced growth and almost complete resistance to the fungal pathogen Botrytis cinerea. Both PG and qua2 plants show increased expression of the class III peroxidase AtPRX71 that contributes to their elevated ROS levels and reduced growth. In this work, we show that leaves of PG and qua2 plants display greatly increased cuticle permeability. Both increased cuticle permeability and resistance to B. cinerea in qua2 are suppressed by loss of AtPRX71. Increased cuticle permeability in qua2, rather than on defects in cuticle ultrastructure or cutin composition, appears to be dependent on reduced epidermal cell adhesion, which is exacerbated by AtPRX71, and is suppressed by the esmeralda1 mutation, which also reverts the adhesion defect and the resistant phenotype. Increased cuticle permeability, accumulation of ROS, and resistance to B. cinerea are also observed in mutants lacking a functional FERONIA, a receptor-like kinase thought to monitor pectin integrity. In contrast, mutants with defects in other structural components of primary cell wall do not have a defective cuticle and are normally susceptible to the fungus. Our results suggest that disrupted cuticle integrity, mediated by peroxidase-dependent ROS accumulation, plays a major role in the robust resistance to B. cinerea of plants with altered HG integrity.

Late blight caused by Phytophthora infestans is one of the most devastating diseases in potatoes and tomatoes. At present, several late blight resistance genes have been mapped and cloned. To better understand the transcriptome changes during the incompatible interaction process between R3a and Avr3a, in this study, after spraying DEX, the leaves of MM-R3a-Avr3a and MM-Avr3a transgenic plants at different time points were used for comparative transcriptome analysis. A total of 7,324 repeated DEGs were detected in MM-R3a-Avr3a plants at 2-h and 6-h, and 729 genes were differentially expressed at 6-h compared with 2-h. Only 1,319 repeated DEGs were found in MM-Avr3a at 2-h and 6-h, of which 330 genes have the same expression pattern. Based on GO, KEGG and WCGNA analysis of DEGs, the phenylpropanoid biosynthesis, plant-pathogen interaction, and plant hormone signal transduction pathways were significantly up-regulated. Parts of the down-regulated DEGs were enriched in carbon metabolism and the photosynthesis process. Among these DEGs, most of the transcription factors, such as WRKY, MYB, and NAC, related to disease resistance or endogenous hormones SA and ET pathways, as well as PR, CML, SGT1 gene were also significantly induced. Our results provide transcriptome-wide insights into R3a and Avr3a-mediated incompatibility interaction.

Harnessing plant-microbe interactions to advance crop resistance to pathogens could be a keystone in sustainable agriculture. The breeding of crops to maximize yield in intensive agriculture might have led to the loss of traits that are necessary for beneficial plant-soil feedback. In this study, we tested whether the soil microbiome can induce a stronger plant defense against root-lesion nematodes in ancestral genotypes of barley than in elite cultivars. Plants were grown in a sterile substrate with or without the inoculation of rhizosphere microbiomes, and Pratylenchus neglectus was inoculated to the roots. Unexpectedly, elite cultivars profited significantly more from the microbiome than ancestral genotypes, by the reduction of nematodes in roots and the increased shoot weight relative to control plants. The elite cultivars had higher microbial densities in the rhizosphere, which were correlated with root weight. The structure of the bacterial and fungal community of elite and ancestral genotypes differed, as compared by 16S rDNA or internal transcribed spacer amplicon profiles in denaturing gradient gel electrophoresis. The elite cultivars differed in responsiveness to the microbiome. For the most responsive cultivars Beysehir and Jolgeh, the strong microbe-induced suppression of nematodes coincided with the strongest microbe-dependent increase in transcripts of salicylic acid-regulated defense genes after nematode invasion, while the jasmonate-regulated genes LOX2 and AOS were downregulated in roots with the inoculated microbiome. The microbe-triggered modulation of defense gene expression differed significantly between elite and ancestral genotypes of barley. Soil microbiomes conditioned by maize roots suppressed the nematodes in elite cultivars, while the corresponding bulk soil microbiome did not. In conclusion, cultivars Beysehir and Jolgeh harbor the genetic background for a positive plant-microbiome feedback. Exploiting these traits in breeding for responsiveness to beneficial soil microbiomes, accompanied by soil biome management for compatible plant-microbe interactions, will support low-input agriculture and sustainability.

There have been efforts to develop citrus cultivars that are tolerant of Huanglongbing (HLB), a catastrophic phloem-limited disease. Previous studies demonstrated that continuous plant growth with phloem regeneration is one of the major characteristics of HLB tolerance. In this study, the metabolic mechanisms of HLB tolerance in citrus were elucidated using a multiple pathway-targeted metabolomic approach. Comparative analysis of healthy and infected HLB-tolerant and HLB-sensitive mandarin cultivars (Citrus reticulata) revealed differentially expressed metabolic responses among different groups. Pathway enrichment analysis indicated aspartate and glutamate metabolism, purine metabolism, and biosynthesis of plant hormones were upregulated in the tolerant group, except salicylic acid signaling. Catabolic pathways linked to energy-yielding metabolism were also upregulated in the tolerant group. These metabolisms and pathways were interconnected with each other, unveiling a pivotal metabolic network associated with HLB tolerance. In the network, auxins and cytokinins, the plant hormones responsible for plant growth and phloem regeneration, were accumulated. In addition, purine metabolites serving as energy carriers and nitrogen sources of plants were increased. Only salicylic acid-related metabolites for plant defense responses were decreased in the tolerant group. Our findings may evidence the strategy of HLB-tolerant cultivars that sustain plant growth and phloem formation rather than displaying direct plant defense to overcome the disease.

Plants are constantly confronted by a multitude of biotic stresses involving a myriad of pathogens. In crops, pathogen infections result in significant agronomical losses worldwide posing a threat to food security. In order to enter plant tissues and establish a successful infection, phytopathogens have to surpass several physical, and chemical defense barriers. In recent years, post-translational modification (PTM) mechanisms have emerged as key players in plant defense against pathogens. PTMs allow a highly dynamic and rapid response in front of external challenges, increasing the complexity and precision of cellular responses. In this review, we focus on the role of SUMO conjugation (SUMOylation) in plant immunity against fungi, bacteria, and viruses. In plants, SUMO regulates multiple biological processes, ranging from development to responses arising from environmental challenges. During pathogen attack, SUMO not only modulates the activity of plant defense components, but also serves as a target of pathogen effectors, highlighting its broad role in plant immunity. Here, we summarize known pathogenic strategies targeting plant SUMOylation and, the plant SUMO conjugates involved in host-pathogen interactions. We also provide a catalog of candidate SUMO conjugates according to their role in defense responses. Finally, we discuss the complex role of SUMO in plant defense, focusing on key biological and experimental aspects that contribute to some controversial conclusions, and the opportunities for improving agricultural productivity by engineering SUMOylation in crop species., This work was supported by the FEDER/Spanish Ministry of Economy, Industry and Competitiveness (MINECO; BIO2017-89874-R) and “Severo Ochoa Programme for Centres of Excellence in R&D” 2016-2019 (SEV-2015-0533), and from the Generalitat de Catalunya (2017SGR1211) to LL’s laboratory. MS was supported by “Severo Ochoa” post-doctoral fellowship program. JP-G was supported by research contracts through the CRAG. DF was supported by the pre-doctoral fellowship BES-2017-079690 from Spanish Ministry of Education, Culture and Sport.