Plant Defense Pathways Research Articles

Overexpression of BnNAC19 in Brassica napus enhances resistance to Leptosphaeria maculans, the blackleg pathogen of canola

AbstractLeptosphaeria maculans is a fungal pathogen that causes blackleg disease in canola (Brassica napus), resulting in significant yield and economic losses in Canada and many parts of the world. Plant NAC transcription factors play critical roles in plant development and response to biotic or abiotic stress. In this study, we identified and characterized a BnNAC19 gene from Brassica napus. The overexpression of BnNAC19 in transgenic canola plants contributed to the improvement of seedling resistance against L. maculans. The mycelial growth of a green fluorescent protein‐tagged strain of L. maculans and production of pycnidiospores were shown to be inhibited in the transgenic canola plants overexpressing BnNAC19. In addition, the canola transgenic line overexpressing BnNAC19 showed increased disease resistance in the adult plant, which was determined by quantitative resistance. Both increased seedling and adult plant resistance in transgenic canola plants overexpressing BnNAC19 indicate that the BnNAC19 gene plays a positive role against L. maculans. The expression pattern of genes BnNAC19 upstream and downstream of BnNAC19 that participate in plant defence pathways were investigated to elucidate the B. napus resistance mechanisms to L. maculans infection, and hence to aid the long‐term blackleg disease‐resistant breeding programmes.

A snapshot of the transcriptome of Medicago truncatula (Fabales: Fabaceae) shoots and roots in response to an arbuscular mycorrhizal fungus and the pea aphid (Acyrthosiphon pisum) (Hemiptera: Aphididae).

Plants simultaneously interact with belowground symbionts such as arbuscular mycorrhizal (AM) fungi and aboveground antagonists such as aphids. Generally, plants gain access to valuable resources including nutrients and water through the AM symbiosis and are more resistant to pests. Nevertheless, aphids' performance improves on mycorrhizal plants, and it remains unclear whether a more nutritious food source and/or attenuated defenses are the contributing factors. This study examined the shoot and root transcriptome of barrel medic (Medicago truncatula Gaertn.) plants highly colonized by the AM fungus Rhizophagus irregularis (Blaszk., Wubet, Renker, and Buscot) C. Walker and A. Schüßler (Glomerales: Glomeraceae) and exposed to 7 days of mixed age pea aphid (Acyrthosiphon pisum (Harris)) herbivory. The RNA-seq samples chosen for this study showed that aphids were heavier when fed mycorrhizal plants compared to nonmycorrhizal plants. We hypothesized that (i) insect-related plant defense pathways will be downregulated in shoots of mycorrhizal plants with aphids compared to nonmycorrhizal plants with aphids; (ii) pathways involved in nutrient acquisition, carbohydrate-related and amino acid transport will be upregulated in shoots of mycorrhizal plants with aphids compared to nonmycorrhizal plants with aphids; and (iii) roots of mycorrhizal plants with aphids will exhibit mycorrhiza-induced resistance. The transcriptome data revealed that the gene repertoire related to defenses, nutrient transport, and carbohydrates differs between nonmycorrhizal and mycorrhizal plants with aphids, which could explain the weight gain in aphids. We also identified novel candidate genes that are differentially expressed in nonmycorrhizal plants with aphids, thus setting the stage for future functional studies.

Dicer-like 3a mediates intergenerational resistance against root-knot nematodes in rice via hormone responses.

In a continuously changing and challenging environment, passing down the memory of encountered stress factors to offspring could provide an evolutionary advantage. In this study, we demonstrate the existence of 'intergenerational acquired resistance' in the progeny of rice (Oryza sativa) plants attacked by the belowground parasitic nematode Meloidogyne graminicola. Transcriptome analyses revealed that genes involved in defense pathways are generally downregulated in progeny of nematode-infected plants under uninfected conditions but show a stronger induction upon nematode infection. This phenomenon was termed "spring loading" and depends on initial downregulation by the 24nt siRNA biogenesis gene dicer-like 3a (dcl3a) involved in the RNA-directed DNA methylation pathway. Knock-down of dcl3a led to increased nematode susceptibility and abolished intergenerational acquired resistance, as well as jasmonic acid/ethylene spring loading in the offspring of infected plants. The importance of ethylene signaling in intergenerational resistance was confirmed by experiments on a knock-down line of ethylene insensitive 2 (ein2b), which lacks intergenerational acquired resistance. Taken together, these data indicate a role for DCL3a in regulating plant defense pathways during both within-generation and intergenerational resistance against nematodes in rice.

Plant Microbiome: An Ocean of Possibilities for Improving Disease Resistance in Plants.

Plant diseases pose a serious threat to crop production and the agricultural economy across the globe. Currently, chemical pesticides are frequently employed to combat these infections, which cause environmental toxicity and the emergence of resistant pathogens. Moreover, the genetic manipulation of plant defense pathways and the breeding of resistant genes has attained limited success due to the rapid evolution of pathogen virulence and resistance, together with host range expansion. Additionally, due to climate change and global warming, the occurrence of multiple stresses during disease outbreak has further impacted overall crop growth and productivity, posing a serious threat to food security. In this regard, harnessing the plant beneficial microbiome and its products can provide novel avenues for disease resistance in addition to boosting agricultural output, soil fertility and environmental sustainability. In plant-beneficial microbiome interactions, induced systemic resistance (ISR) has emerged as a key mechanism by which a beneficial microbiome primes the entire plant system for better defense against a wide range of phytopathogens and pests. In this review, we provide the recent developments on the role of plant beneficial microbiomes in disease resistance. We also highlight knowledge gaps and discuss how the plant immune system distinguishes pathogens and beneficial microbiota. Furthermore, we provide an overview on how immune signature hormones, such as salicylic acid (SA), jasmonic acid (JA) and ethylene (ET), shape plant beneficial microbiome. We also discuss the importance of various high-throughput tools and their integration with synthetic biology to design tailored microbial communities for disease resistance. Finally, we conclude by highlighting important themes that need future attention in order to fill the knowledge gaps regarding the plant immune system and plant-beneficial-microbiome-mediated disease resistance.

Genetic basis of resistance to southern corn leaf blight in the maize multi-parent population and diversity panel.

Southern corn leaf blight (SLB), caused by the necrotrophic pathogen Cochliobolus heterostrophus, is one of the maize foliar diseases and poses a great threat to corn production around the world. Identification of genetic variations underlying resistance to SLB is of paramount importance to maize yield and quality. Here, we used a random-open-parent association mapping population containing eight recombinant inbred line populations and one association mapping panel consisting of 513 diversity maize inbred lines with high-density genetic markers to dissect the genetic basis of SLB resistance. Overall, 109 quantitative trait loci (QTLs) with predominantly small or moderate additive effects, and little epistatic effects were identified. We found 35 (32.1%) novel loci in comparison with the reported QTLs. We revealed that resistant alleles were significantly enriched in tropical accessions and the frequency of about half of resistant alleles decreased during the adaptation process owing to the selection of agronomic traits. A large number of annotated genes located in the SLB-resistant QTLs were shown to be involved in plant defence pathways. Integrating genome-wide association study, transcriptomic profiling, resequencing and gene editing, we identified ZmFUT1 and MYBR92 as the putative genes responsible for the major QTLs for resistance to C.heterostrophus. Our results present a comprehensive insight into the genetic basis of SLB resistance and provide resistant loci or genes as direct targets for crop genetic improvement.

Aged sweet corn (Zea mays L. saccharata Sturt) seeds trigger hormone and defense signaling during germination

Abstract: Seed ageing during storage or shipping can affect the yield and cost of agricultural production. Sweet corn is a naturally mutated corn cultivar with weak seed vitality during storage. In this study, we have shown that endogenous hormone level dynamics of fresh and aged sweet corn seeds were different during early geminating process. Transcriptomic assembling uncovered several novel transcripts during early stage of sweet corn germination in signal transduction and RNA processing pathways. Comparative transcriptomic profiling revealed that expression of many genes was activated earlier in aged seeds than fresh ones. Functional analyses showed that, as two major networks, plant hormone signaling and defense pathways were different between fresh and aged sweet corn seeds. Auxin, cytokinin, gibberellin, salicylic acid and jasmonic acid signaling pathways were activated earlier in aged seeds than fresh ones on the first day after imbibition. These results provided knowledge for understanding the ageing process of sweet corn seeds and several candidate genes for genetic study of seed ageing mechanism.

An unexpected role for tomato threonine deaminase 2 in host defense against bacterial infection.

The hormones salicylic acid (SA) and jasmonic acid (JA) often act antagonistically in controlling plant defense pathways in response to hemibiotrophs/biotrophs (hemi/biotroph) and herbivores/necrotrophs, respectively. Threonine deaminase (TD) converts threonine to α-ketobutyrate and ammonia as the committed step in isoleucine (Ile) biosynthesis and contributes to JA responses by producing the Ile needed to make the bioactive JA-Ile conjugate. Tomato (Solanum lycopersicum) plants have two TD genes: TD1 and TD2. A defensive role for TD2 against herbivores has been characterized in relation to JA-Ile production. However, it remains unknown whether TD2 is also involved in host defense against bacterial hemi/biotrophic and necrotrophic pathogens. Here, we show that in response to the bacterial pathogen-associated molecular pattern (PAMP) flagellin flg22 peptide, an activator of SA-based defense responses, TD2 activity is compromised, possibly through carboxy-terminal cleavage. TD2 knockdown (KD) plants showed increased resistance to the hemibiotrophic bacterial pathogen Pseudomonas syringae but were more susceptible to the necrotrophic fungal pathogen Botrytis cinerea, suggesting TD2 plays opposite roles in response to hemibiotrophic and necrotrophic pathogens. This TD2 KD plant differential response to different pathogens is consistent with SA- and JA-regulated defense gene expression. flg22-treated TD2 KD plants showed high expression levels of SA-responsive genes, whereas TD2 KD plants treated with the fungal PAMP chitin showed low expression levels of JA-responsive genes. This study indicates TD2 acts negatively in defense against hemibiotrophs and positively against necrotrophs and provides insight into a new TD2 function in the elaborate crosstalk between SA and JA signaling induced by pathogen infection.

A potential role of salicylic acid in the evolutionary behavior of Trichoderma as a plant pathogen: from Marchantia polymorpha to Arabidopsis thaliana

Main conclusionRecognition of the interaction of Trichoderma during the evolution of land plants plays a potential key role in the development of the salicylic acid defense pathway and the establishment of a mutualistic relationship.Marchantia polymorpha is a common liverwort considered in recent years as a model plant for evolutionary studies on plant–microorganism interactions. Despite the lack of research, remarkable results have been reported regarding the understanding of metabolic and evolutionary processes of beneficial and/or harmful interactions, owing to a better understanding of the origin and evolution of different plant defense pathways. In this study, we have carried out work on the direct and indirect interactions (exudates and volatiles) of M. polymorpha with different species of the fungal genus Trichoderma. These interactions showed different outcomes, including resistance or even growth promotion and disease. We have analyzed the level of tissue colonization and defense-related gene expression. Furthermore, we have used the pteridophyte Dryopteris affinis and the angiosperm Arabidopsis thaliana, as subsequent steps in plant evolution, together with the plant pathogen Rhizoctonia solani as a control of plant pathogenicity. Trichoderma virens, T. brevicompactum and T. hamatum are pathogens of M. polymorpha, while exudates of T. asperellum are harmful to the plant. The analysis of the expression of several defense genes in M. polymorpha and A. thaliana showed that there is a correlation of the transcriptional activation of SA-related genes with resistance or susceptibility of M. polymorpha to Trichoderma. Moreover, exogenous SA provides resistance to the virulent Trichoderma species. This beneficial fungus may have had an evolutionary period of interaction with plants in which it behaved as a plant pathogen until plants developed a defense system to limit its colonization through a defense response mediated by SA.

Rhodopseudomonas palustris PSB-06 Induces Plant Defense and Suppresses the Transmission of Tomato Chlorosis Virus by Bemisia tabaci MED

Tomato chlorosis virus (ToCV) is an RNA virus and a member of the Closteroviridae, Crinivirus, that is primarily vectored by Bemisia tabaci MED (B. tabaci MED). An outbreak of ToCV causes destructive damage to tomato plants and other solanaceous vegetables. Currently, ToCV has not been effectively controlled. Rhodopseudomonas palustris PSB-06 is a novel biological agent that is effective at controlling the tobacco mosaic virus (TMV). In this study, we investigated the role of PSB-06 in ToCV-infected tomato plants, and we studied the effects of PSB-06 on plant defense and plant photosynthetic pathways. Furthermore, the effect of PSB-06 on the acquisition and transmission of B. tabaci MED was determined. The results showed that compared with water-treated tomato plants, the contents of jasmonic acid increased, and the activities of catalase, peroxidase and superoxide dismutase increased significantly in tomato plants treated with PSB-06. The relative expression of genes involved in chlorophyll development, chlorophyll metabolism and photosynthesis also increased significantly. Simultaneously, treatment with PSB-06 reduced the acquisition and transmission of B. tabaci MED. We verified the hypothesis that PSB-06 is effective at controlling ToCV by promoting plant defense responses and reducing the amount of ToCV in tomato plants. We also confirmed the ability of B. tabaci MED to transmit ToCV. This study should help to control B. tabaci MED and reduce the spread of ToCV.

Transcriptomic analysis provides insight into defensive strategies in response to continuous cropping in strawberry (Fragaria × ananassa Duch.) plants

BackgroundStrawberries are an important economic fruit crop world-wide. In strawberry cultivation, continuous cropping (CC) can seriously threaten yield and quality. However, our understanding of the gene expression changes in response to CC and during subsequent defense processes is limited. In this study, we analyzed the impact of CC on the transcriptome of strawberry roots using RNA-Seq technology to elucidate the effect of CC and the subsequent molecular changes.ResultsWe found that CC significantly affects the growth of strawberry plants. The transcriptome analysis identified 136 differentially expressed genes (DEGs), including 49 up-regulated and 87 down-regulated DEGs. A Gene Ontology (GO) analysis indicated that the up-regulated DEGs were mainly assigned to defense-related GO terms, and most down-regulated DEGs were assigned to nutrient-related GO terms. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the responsive DEGs were classified in a large number of important biological pathways, such as phenylalanine metabolism, starch and sucrose metabolism, phenylpropanoid biosynthesis, glutathione metabolism and plant-pathogen interaction. We also found that four WRKY transcription factors and three peroxidase genes involved in plant defense pathways were up-regulated in the roots of strawberry plants subjected to CC.ConclusionSeveral unigenes involved in plant defense processes, such as CNGCs, WRKY transcription factors, PR1, and peroxidase genes with highly variable expression levels between non-CC and CC treatments may be involved in the regulation of CC in strawberry. These results indicate that strawberry roots reallocate development resources to defense mechanisms in response to CC. This study will further deepen our understanding of the fundamental regulatory mechanisms of strawberry resource reallocation in response to CC.

Histone modification and chromatin remodeling in plant response to pathogens.

As sessile organisms, plants are constantly exposed to changing environments frequently under diverse stresses. Invasion by pathogens, including virus, bacterial and fungal infections, can severely impede plant growth and development, causing important yield loss and thus challenging food/feed security worldwide. During evolution, plants have adapted complex systems, including coordinated global gene expression networks, to defend against pathogen attacks. In recent years, growing evidences indicate that pathogen infections can trigger local and global epigenetic changes that reprogram the transcription of plant defense genes, which in turn helps plants to fight against pathogens. Here, we summarize up plant defense pathways and epigenetic mechanisms and we review in depth current knowledge’s about histone modifications and chromatin-remodeling factors found in the epigenetic regulation of plant response to biotic stresses. It is anticipated that epigenetic mechanisms may be explorable in the design of tools to generate stress-resistant plant varieties.

Negative Interactions Balance Growth and Defense in Plants Confronted with Herbivores or Pathogens.

Plants have evolved a series of defensive mechanisms against pathogens and herbivores, but the defense response always leads to decreases in growth or reproduction, which has serious implications for agricultural production. Growth and defense are negatively regulated not only through metabolic consumption but also through the antagonism of different phytohormones, such as jasmonic acid (JA) and salicylic acid (SA). Meanwhile, plants can limit the expression of defensive metabolites to reduce the costs of defense by producing constitutive defenses such as glandular trichomes or latex and accumulating specific metabolites, determining the activation of plant defense or the maintenance of plant growth. Interestingly, plant defense pathways might be prepared in advance which may be transmitted to descendants. Plants can also use external organisms to protect themselves, thus minimizing the costs of defense. In addition, plant relatives exhibit cooperation to deal with pathogens and herbivores, which is also a way to regulate growth and defense.

Overexpression of the salicylic acid binding protein 2 (SABP2) from tobacco enhances tolerance against Huanglongbing in transgenic citrus.

Overexpression of the salicylic acid binding protein 2 (SABP2) gene from Tobacco results in enhanced tolerance to Huanglongbing (HLB; citrus greening disease) in transgenic sweet oranges. Huanglongbing (HLB), the most destructive citrus disease, is caused by Candidatus Liberibacter asiaticus (CaLas). Currently, no cure for this disease exists, and all commercially planted cultivars are highly susceptible. Salicylic Acid Binding Protein 2 (SABP2) is a well-characterized protein essential for establishing systemic acquired resistance (SAR) in tobacco.The constitutive over expression of SABP2 from tobacco (NtSABP2) in 'Hamlin' sweet orangeresulted in the production of several transgenic lines with variable transcript levels. Transient expression ofthe NtSABP2-EGFP fusion protein in Nicotiana benthamiana plants demonstrated thatNtSABP2was cytosolicin its subcellular localization. In a long-term fieldstudy, we identified a SABP2 transgenic linewithsignificantly reduced HLB symptoms that maintained aconsistently low CaLas titer. Transcriptome analysis of this selectedtransgenic line demonstrated upregulation of several genes related to plant defense and SAR pathways. Genes, such as NPR family genes and those coding for monooxygenases and lipoxygenases, were upregulated in the35S-NtSABP2 overexpressing line and might be candidates for incorporation into our citrus improvement program.

Using crystallographic 'failures' to identify artificial and endogenous inhibitors for plant growth and defence pathways

Using crystallographic 'failures' to identify artificial and endogenous inhibitors for plant growth and defence pathways

The downside of metabolic diversity: Postingestive rearrangements by specialized insects

Deploying toxins in complex mixtures is thought to be advantageous and is observed during antagonistic interactions in nature. Toxin mixtures are widely utilized in medicine and pest control, as they are thought to slow the evolution of detoxification counterresponses in the targeted organisms. Here we show that caterpillars rearrange key constituents of two distinct plant defense pathways to postingestively disable the defensive properties of both pathways. Specifically, phenolic esters of quinic acid, chlorogenic acids (CAs), potent herbivore and ultraviolet (UV) defenses, are reesterified to decorate particular sugars of 17-hydroxygeranyllinalool diterpene glycosides (HGL-DTGs) and prevent their respective anti–herbivore defense functions. This was discovered through the employment of comparative metabolomics of the leaves of Nicotiana attenuata and the frass of this native tobacco’s specialist herbivore, Manduca sexta larvae. Feeding caterpillars on leaves of transgenic plants abrogated in each of the two pathways, separately and together, revealed that one of the fully characterized frass conjugates, caffeoylated HGL-DTG, originated from ingested CA and HGL-DTGs and that both had negative effects on the defensive function of the other compound class, as revealed by rates of larval mass gain. This negative defensive synergy was further explored in 183 N. attenuata natural accessions, which revealed a strong negative covariance between the two defense pathways. Further mapping analyses in a biparental recombinant inbred line (RIL) population imputed quantitative trait loci (QTLs) for the two pathways at distinct genomic locations. The postingestive repurposing of defense metabolism constituents reveals a downside of deploying toxins in mixtures, a downside which plants in nature have evolved to counter.

β-Glucan and its nanocomposites in sustainable agriculture and environment: an overviewof mechanisms and applications.

β-Glucan is an eco-friendly, biodegradable, and economical biopolymer with important roles for acquiring adaptations to mitigate climate change in crop plants. β-Glucan plays a crucial role in the activation of functional plant innate immune system by triggering the downward signaling cascade/s, resulting in the accumulation of different pathogenesis-related proteins (PR-proteins), reactive oxygen species (ROS), antioxidant defense enzymes, Ca2+-influx as well as activation of mitogen-activated protein kinase (MAPK) pathway. Recent experimental studies have shown that β-glucan recognition is mediated by co-receptor LysMPRR (lysin motif pattern recognition receptor)-CERK1 (chitin elicitor receptor kinase 1), LYK4, and LYK5 (LysM-containing receptor-like kinase), as well as different receptor systems in plants that could be plant species-specific and/or age and/or tissue-dependent. Transgenic overexpression of β-glucanase, chitinase, and/or in combination with other PR-proteins like cationic peroxidase, AP24,thaumatin-likeprotein 1 (TLP-1) has also been achieved for improving plant disease resistance in crop plants, but the transgenic methods have some ethical and environmental concerns. In this regard, elicitation of plant immunity using biopolymer like β-glucan and chitosan offers an economical, safe, and publicly acceptable method. The β-glucan and chitosan nanocomposites have proven to be useful for the activation of plant defense pathways and to enhance plant response/systemic acquired resistance (SAR) against broad types of plant pathogens and mitigating multiple stresses under the changing climate conditions.

Oviposition by a Specialist Herbivore Increases Susceptibility of Canola to Herbivory by a Generalist Herbivore.

Oviposition by specialist herbivores can alter the suitability of the host plant to subsequent infestation by other herbivores. In this study, we tested the effect of previous oviposition on canola, Brassica napus L., by a Brassica specialist, the diamondback moth (DBM), Plutella xylostella (L.) (Lepidoptera: Plutellidae), on subsequent herbivory by the generalist feeder, the bertha armyworm (BAW), Mamestra configurata Walker (Lepidoptera: Noctuidae). The effect of DBM oviposition on subsequent BAW oviposition and larval feeding was tested in no-choice and choice experiments. Oviposition of BAW was not altered by DBM eggs on canola plants, however, BAW had increased larval feeding on plants with DBM eggs. These results suggest that oviposition by a specialist herbivore increased the susceptibility of the host plant to generalist herbivory. In a preliminary experiment, salicylic acid, jasmonic acid, and its conjugates were not altered by DBM oviposition on canola, however, further experimentation is needed to determine if oviposition affects expression of plant defense pathways and other plant traits.

Rhizosphere bacteria induce programmed cell death defence genes and signalling in chilli pepper.

To understand how beneficial bacteria assist chilli plants (Capsicum annuum) in defence against biotrophic or hemibiotrophic pathogens. We quantified marker genes of plant defence pathways in Phytophthora capsici-infected chilli pepper treated with anti-oomycete plant growth-promoting rhizobacteria, Bacillus amyloliquefaciens, Bacillus velezensis and Acinetobacter sp. Plants displayed strong resistance, and the pathogen load in the roots was significantly lower in infected plants treated with bacterial biocontrol agents at all time points tested (1, 2 and 7days after pathogen inoculation, p<0.05). Gene expression profiling revealed that P. capsici infection in the absence of beneficial bacteria led to the upregulation of a wide array of defence genes. The addition of biocontrol bacteria modulated defence by further enhancing genes involved in programmed cell death, such as CaLOX1, CaPAL1, CaChitIV and CaPTI1, while suppressing others CaLRR1, a negative regulator of cell death. Our results suggest that the bacteria exerted a combined effect by directly antagonizing the pathogen and enhancing the expression of key plant defence genes, including those involved in cell death, causing resistance at early stages of infection by this hemibiotrophic pathogen.

Cloning the barley nec3 disease lesion mimic mutant using complementation by sequencing.

Disease lesion mimic (DLM) or necrotic mutants display necrotic lesions in the absence of pathogen infections. They can show improved resistance to some pathogens and their molecular dissection can contribute to revealing components of plant defense pathways. Although forward-genetics strategies to find genes causal to mutant phenotypes are available in crops, these strategies require the production of experimental cross populations, mutagenesis, or gene editing and are time- and resource-consuming or may have to deal with regulated plant materials. In this study, we described a collection of 34 DLM mutants in barley (Hordeum vulgare L.) and applied a novel method called complementation by sequencing (CBS), which enables the identification of the gene responsible for a mutant phenotype given the availability of two or more chemically mutagenized individuals showing the same phenotype. Complementation by sequencing relies on the feasibility to obtain all induced mutations present in chemical mutants and on the low probability that different individuals share the same mutated genes. By CBS, we identified a cytochrome P450 CYP71P1 gene as responsible for orange blotch DLM mutants, including the historical barley nec3 locus. By comparative phylogenetic analysis we showed that CYP71P1 gene family emerged early in angiosperm evolution but has been recurrently lost in some lineages including Arabidopsis thaliana (L.) Heynh. Complementation by sequencing is a straightforward cost-effective approach to clone genes controlling phenotypes in a chemically mutagenized collection. The TILLMore (TM) collection will be instrumental for understanding the molecular basis of DLM phenotypes and to contribute knowledge about mechanisms of host-pathogen interaction.

RNA Sequencing of Arabidopsis thaliana Seedlings after Non-Thermal Plasma-Seed Treatment Reveals Upregulation in Plant Stress and Defense Pathways

Not all agricultural practices are sustainable; however, non-thermal plasma treatment of seeds may be an eco-friendly alternative to improve macroscopic plant growth parameters. Despite the numerous successful results of plasma-seed treatments reported in the literature, there is a large gap in our understanding of how non-thermal plasma treatments affect seeds, especially due to the plethora of physical, chemical, and biological variables. This study uses RNA sequencing to characterize the changes in gene transcription in Arabidopsis thaliana (L.) Heynh. seeds 6 days after exposure to surface dielectric barrier discharge plasma treatment. Here, we provide an overview of all pathways that are differentially expressed where few genes are upregulated and many genes are downregulated. Our results reveal that plasma treatment time is a parameter that can activate different pathways in plant defense. An 80 s treatment upregulates the glucosinolate pathway, a defense response to insects and herbivores to deter feeding, whereas a shorter treatment of 60 s upregulates the phenylpropanoid pathway, which reinforces the cell wall with lignin and produces antimicrobial compounds, a defense response to bacterial or fungal plant pathogens. It seems that plasma elicits a wounding response from the seed in addition to redox changes. This suggests that plasma treatment can be potentially applied in agriculture to protect plants against abiotic and biotic stresses without discharging residues into the environment.