Plant Pathogen Defense Research Articles

Deciphering the genomic regions governing major agronomic traits and blast resistance using genome wide association mapping in finger millet

Finger millet (Eleusine coracana L.) is climate resilient minor millet of Asia and Africa with wide adaptation and unparallel nutritional profile. To date, genomic resources available in finger millet are scanty and genetic control of agronomic traits remains elusive. Here, a collection of eco-geographically diverse 186 genotypes was quantified for variation in 13 agronomic traits and reaction to blast to identify marker-trait associations (MTAs) using genotyping-by-sequencing (GBS) and genome-wide association study (GWAS). GBS generated 2977 high quality single nucleotide polymorphism (SNPs) markers and identified three subpopulations with varying admixture levels. General linear and mixed model approaches of GWAS to correct for population structure and genetic relatedness identified 132 common MTAs for agronomic traits across the years. The phenotypic variance explained by the makers varied from 4.8% (TP692389-flag leaf width) to 20% (TP714446-green fodder weight). Of these, 26 MTAs showed homology with candidate genes having role in plant growth, development and photosynthesis in the genomes of foxtail millet, rice, maize, wheat and barley. We also found 4 common MTAs for neck blast resistance, which explained 5.9–15.1% phenotypic variance. Three MTAs for neck blast resistance showed orthologues in related genera having putative functions in pathogen defense in plants. The results of this work lay a foundation for understanding the genetic architecture of agronomic traits and blast resistance in finger millet and provide a framework for genomics assisted breeding.

Exogenous Application of a Plant Elicitor Induces Volatile Emission in Wheat and Enhances the Attraction of an Aphid Parasitoid Aphidius gifuensis.

It is well known that plant elicitors can induce plant defense against pests. The herbivore-induced plant volatile (HIPV) methyl salicylate (MeSA), as a signaling hormone involved in plant pathogen defense, is used to recruit natural enemies to protect wheat and other crops. However, the defense mechanism remains largely unknown. Here, the headspace volatiles of wheat plants were collected and analyzed by gas chromatography-mass spectrometry (GC-MS), gas chromatography with electroantennographic detection (GC-EAD) and principal component analysis (PCA). The results showed that exogenous application of MeSA induced qualitative and quantitative changes in the volatiles emitted from wheat plants, and these changes were mainly related to Carveol, Linalool, m-Diethyl-benzene, p-Cymene, Nonanal, D-limonene and 6-methyl-5-Hepten-2-one. Then, the electroantennogram (EAG) and Y-tube bioassay were performed to test the physiological and behavioral responses of Aphidius gifuensis Ashmesd to the active volatile compounds (p-Cymene, m-Diethyl-benzene, Carveol) that identified by using GC-EAD. The female A. gifuensis showed strong physiological responses to 1 μg/μL p-Cymene and 1 μg/μL m-Diethyl-benzene. Moreover, a mixture blend was more attractive to female A. gifuensis than a single compound. These findings suggested that MeSA could induce wheat plant indirect defense against wheat aphids through attracting parasitoid in the wheat agro-ecosystem.

Comprehensive analysis of bZIP gene family and function of RcbZIP17 on Botrytis resistance in rose (Rosa chinensis)

Rose is an important ornamental plant and a commercial source of natural fragrances and essential oils. However, gray mold is the most serious disease of rose during cultivation and post-harvest. In plants, the bZIP gene family is one of the important families of transcription factors that regulate many biological processes, including pathogen defense, light response, and stress signaling. Although there were reports that bZIPs play a key role in plant pathogen defense, there have been no comprehensive studies or functional analysis of the bZIP gene family in rose (Rosa sp.) yet. In this study, a genome-wide analysis of the bZIP family gene (RcbZIPs) in rose was carried out to investigate their phylogenetic relationships, gene structure, chromosome localizations, collinearity, and the function of RcbZIP17. In the rose genome, 48 RcbZIP genes were identified and divided into 11 subgroups by phylogenetic relationships. Moreover, gene structure analysis agreed with phylogenetic tree grouping. The RcbZIPs were unevenly distributed on all seven chromosomes. The gene duplication events indicated that segmental duplication of the entire genome may play a key role in gene duplication. A Ka/Ks analysis showed that they mainly underwent purifying selection with limited functional differentiation. Gene expression analysis showed that 16 RcbZIPs were significantly differential expressed by Botrytis cinerea infection. It is speculated that these RcbZIPs are candidate genes for regulating the response of rose to gray mold infection. According to virus-induced gene silencing (VIGS) and overexpression (OE) studies, RcbZIP17 was implicated in the resistance of rose to gray mold. This study provides useful information for further study on the function of the bZIP gene family in rose.

Silencing of a Cotton Actin-Binding Protein GhWLIM1C Decreases Resistance against Verticillium dahliae Infection.

LIM proteins are widely spread in various types of plant cells and play diversely crucial cellular roles through actin cytoskeleton assembly and gene expression regulation. Till now, it has not been clear whether LIM proteins function in plant pathogen defense. In this study, we characterized a LIM protein, GhWLIM1C, in upland cotton (Gossypium hirsutum). We found that GhWLIM1C could bind and bundle the actin cytoskeleton, and it contains two LIM domains (LIM1 and LIM2). Both the two domains could bind directly to the actin filaments. Moreover, the LIM2 domain additionally bundles the actin cytoskeleton, indicating that it possesses a different biochemical activity than LIM1. The expression of GhWLIM1C responds to the infection of the cotton fungal pathogen Verticillium dahliae (V. dahliae). Silencing of GhWLIM1C decreased cotton resistance to V. dahliae. These may be associated with the down regulated plant defense response, including the PR genes expression and ROS accumulation in the infected cotton plants. In all, these results provide new evidence that a plant LIM protein functions in plant pathogen resistance and the assembly of the actin cytoskeleton are closely related to the triggering of the plant defense response.

Genome-Wide Analysis of the Peptidase M24 Superfamily in Triticum aestivum Demonstrates That TaM24-9 Is Involved in Abiotic Stress Response.

The peptidase M24 (Metallopeptidase 24, M24) superfamily is essential for plant growth, stress response, and pathogen defense. At present, there are few systematic reports on the identification and classification of members of the peptidase M24 proteins superfamily in wheat. In this work, we identified 53 putative candidate TaM24 genes. According to the protein sequences characteristics, these members can be roughly divided into three subfamilies: I, II, III. Most TaM24 genes are complex with multiple exons, and the motifs are relatively conserved in each sub-group. Through chromosome mapping analysis, we found that the 53 genes were unevenly distributed on 19 wheat chromosomes (except 3A and 3D), of which 68% were in triads. Analysis of gene duplication events showed that 62% of TaM24 genes in wheat came from fragment duplication events, and there were no tandem duplication events to amplify genes. Analysis of the promoter sequences of TaM24 genes revealed that cis-acting elements were rich in response elements to drought, osmotic stress, ABA, and MeJA. We also studied the expression of TaM24 in wheat tissues at developmental stages and abiotic stress. Then we selected TaM24-9 as the target for further analysis. The results showed that TaM24-9 genes strengthened the drought and salt tolerance of plants. Overall, our analysis showed that members of the peptidase M24 genes may participate in the abiotic stress response and provided potential gene resources for improving wheat resistance.

Photoperiod Stress in Arabidopsis thaliana Induces a Transcriptional Response Resembling That of Pathogen Infection.

Plants are exposed to regular diurnal rhythms of light and dark. Changes in the photoperiod by the prolongation of the light period cause photoperiod stress in short day-adapted Arabidopsis thaliana. Here, we report on the transcriptional response to photoperiod stress of wild-type A. thaliana and photoperiod stress-sensitive cytokinin signaling and clock mutants and identify a core set of photoperiod stress-responsive genes. Photoperiod stress caused altered expression of numerous reactive oxygen species (ROS)-related genes. Photoperiod stress-sensitive mutants displayed similar, but stronger transcriptomic changes than wild-type plants. The alterations showed a strong overlap with those occurring in response to ozone stress, pathogen attack and flagellin peptide (flg22)-induced PAMP triggered immunity (PTI), which have in common the induction of an apoplastic oxidative burst. Interestingly, photoperiod stress triggers transcriptional changes in jasmonic acid (JA) and salicylic acid (SA) biosynthesis and signaling and results in increased JA, SA and camalexin levels. These responses are typically observed after pathogen infections. Consequently, photoperiod stress increased the resistance of Arabidopsis plants to a subsequent infection by Pseudomonas syringae pv. tomato DC3000. In summary, we show that photoperiod stress causes transcriptional reprogramming resembling plant pathogen defense responses and induces systemic acquired resistance (SAR) in the absence of a pathogen.

BoPEP4, a C-Terminally Encoded Plant Elicitor Peptide from Broccoli, Plays a Role in Salinity Stress Tolerance.

Plant peptide hormones play various roles in plant development, pathogen defense and abiotic stress tolerance. Plant elicitor peptides (Peps) are a type of damage-associated molecular pattern (DAMP) derived from precursor protein PROPEPs. In this study, we identified nine PROPEP genes in the broccoli genome. qRT-PCR analysis indicated that the expression levels of BoPROPEPs were induced by NaCl, ABA, heat, SA and P. syringae DC3000 treatments. In order to study the functions of Peps in salinity stress response, we synthesized BoPep4 peptide, the precursor gene of which, BoPROPEP4, was significantly responsive to NaCl treatment, and carried out a salinity stress assay by exogenous application of BoPep4 in broccoli sprouts. The results showed that the application of 100 nM BoPep4 enhanced tolerance to 200 mM NaCl in broccoli by reducing the Na+/K+ ratio and promoting accumulation of wax and cutin in leaves. Further RNA-seq analysis identified 663 differentially expressed genes (DGEs) under combined treatment with BoPep4 and NaCl compared with NaCl treatment, as well as 1776 genes differentially expressed specifically upon BoPep4 and NaCl treatment. GO and KEGG analyses of these DEGs indicated that most genes were enriched in auxin and ABA signal transduction, as well as wax and cutin biosynthesis. Collectively, this study shows that there was crosstalk between peptide hormone BoPep4 signaling and some well-established signaling pathways under salinity stress in broccoli sprouts, which implies an essential function of BoPep4 in salinity stress defense.

Small RNA and Degradome Sequencing Reveal Important MicroRNA Function in Nicotiana tabacum Response to Bemisia tabaci.

MicroRNAs (miRNAs), a class of small non-coding regulatory RNAs, are key molecules in many biological and metabolic processes of plant growth, development and stress response via targeting mRNAs. The phloem-feeding insect whitefly Bemisia tabaci (Hemiptera, Aleyrodidae) is a serious pest that causes devastating harm to agricultural production worldwide. However, the function of host miRNAs in the response to whitefly infestation remains unclear. Here, we sequenced the small RNA and degradome of tobacco (Nicotiana tabacum L.), after and before infestation by B. tabaci. We identified 1291 miRNAs belonging to 138 miRNA families including 706 known miRNAs and 585 novel miRNAs. A total of 47 miRNAs were differentially expressed, of which 30 were upregulated and 17 were downregulated by whitefly exposure. Then, computational analysis showed that the target genes of differential miRNAs were involved in R gene regulation, plant innate immunity, plant pathogen defense, the plant hormone signal pathway and abiotic stress tolerance. Furthermore, degradome analysis demonstrated that 253 mRNAs were cleaved by 66 miRNAs. Among them, the targets cleaved by upregulated miR6025, miR160, miR171, miR166 and miR168 are consistent with our prediction, suggesting that pathogen-related miRNAs may function in plant defense against whitefly. Moreover, our results show that plant miRNA response and miRNA-mediated post-transcriptional regulation for phloem-feeding insect infestation are similar to pathogen invasion. Our study provides additional data to further elucidate how host plants respond and defend the phloem-feeding insects.

Mycorrhizal symbiosis and phosphorus supply determine interactions among plants with contrasting nutrient‐acquisition strategies

Abstract Highly diverse plant communities growing on nutrient‐impoverished soils are test beds for theories on species coexistence. Here, neighbouring mycorrhizal and non‐mycorrhizal plants compete for limited phosphorus. The impact of below‐ground interactions on community dynamics is underexplored. We used an experimental approach to investigate effects of inoculation with arbuscular mycorrhizal (AM) fungi and a phosphorus supply gradient on competitive and facilitative interactions among mixed assemblages of woody plants in microcosms. The plant species, one cluster root‐forming (CR) species and four AM species, are native to jarrah forest that grows on nutrient‐impoverished soils in south‐western Australia. We measured plant growth in microcosms, with and without inoculation with the AM fungus Rhizophagus irregularis, and across a gradient of P supply: 0, 9, 27 and 243 mg P per kg of soil. Our data show evidence of plant–plant facilitation at low P supply and competition at high P supply. Growth of the CR species, Hakea undulata, was highest in microcosms with 0P and without AM inoculation. One AM species, Bossiaea aquifolium, also performed better at lower P levels, possibly benefitting from P mobilised by H. undulata. The other three AM species, one strongly obligates, performed better at higher P levels. Data for Acacia celastrifolia suggested it was facultatively mycotropic, and because there was no correlation between AM colonisation and the relative inoculum effect, we suggest positive effects of AM inoculation at 9P might be due to benefits other than P acquisition, such as pathogen defence. Benefit of AM inoculation diminished for three of four mycorrhizal species at the highest P level as we had predicted. The fourth species, Eucalyptus marginata (jarrah), had higher growth in microcosms that were not inoculated with AM, perhaps because the species benefits more from ectomycorrhizas. Synthesis. Our experimental data suggest spatial heterogeneity of soil P, coupled with a diversity of nutrient‐acquisition strategies, and plasticity among plant–plant and plant–AM fungi interactions, contributes to plant species coexistence in the nutrient‐impoverished jarrah forest. Our research highlights the importance of below‐ground mechanisms for understanding factors determining community structure including a potential role of AM fungi in plant pathogen defence.

Identification of miRNAs and Their Targets Involved in Flower and Fruit Development across Domesticated and Wild Capsicum Species.

MicroRNAs (miRNAs) are regulators of the post-transcription stage of gene activity documented to play central roles in flower and fruit development in model plant species. However, little is known about their roles and differences in domesticated and wild Capsicum species. In this study, we used high-throughput sequencing to analyze the miRNA content at three developmental stages (flower, small fruit, and middle fruit) from two cultivated (C. baccatum and C. annuum) and two wild (C. chacoense and C. eximium) pepper species. This analysis revealed 22 known and 27 novel miRNAs differentially expressed across species and tissues. A number of stage- and species-specific miRNAs were identified, and Gene Ontology terms were assigned to 138 genes targeted by the miRNAs. Most Gene Ontology terms were for the categories “genetic information processing”, “signaling and cellular processes”, “amino acid metabolism”, and “carbohydrate metabolism”. Enriched KEGG analysis revealed the pathways amino acids, sugar and nucleotide metabolism, starch and sucrose metabolism, and fructose-mannose metabolism among the principal ones regulated by miRNAs during pepper fruit ripening. We predicted miRNA–target gene interactions regulating flowering time and fruit development, including miR156/157 with SPL genes, miR159 with GaMYB proteins, miR160 with ARF genes, miR172 with AP2-like transcription factors, and miR408 with CLAVATA1 gene across the different Capsicum species. In addition, novel miRNAs play an important role in regulating interactions potentially controlling plant pathogen defense and fruit quality via fructokinase, alpha-L-arabinofuranosidase, and aromatic and neutral amino acid transporter. Overall, the small RNA-sequencing results from this study represent valuable information that provides a solid foundation for uncovering the miRNA-mediated mechanisms of flower and fruit development between domesticated and wild Capsicum species.

Guard cell redox proteomics reveals a role of lipid transfer protein in plant defense

Redox-based post-translational modifications (PTMs) involving protein cysteine residues as redox sensors are important to various physiological processes. However, little is known about redox-sensitive proteins in guard cells and their functions in stomatal immunity. In this study, we applied an integrative protein labeling method cysTMTRAQ, and identified guard cell proteins that were altered by thiol redox PTMs in response to a bacterial flagellin peptide flg22. In total, eight, seven and 20 potential redox-responsive proteins were identified in guard cells treated with flg22 for 15, 30 and 60 min, respectively. The proteins fall into several functional groups including photosynthesis, lipid binding, oxidation-reduction, and defense. Among the proteins, a lipid transfer protein (LTP)-II was confirmed to be redox-responsive and involved in plant resistance to Pseudomonas syringe pv. tomato DC3000. This study not only creates an inventory of potential redox-sensitive proteins in flg22 signal transduction in guard cells, but also highlights the biological relevance of the lipid transfer protein in plant defense against bacterial pathogens. SignificanceProtein redox modifications play important roles in many physiological processes. However, redox proteomics has rarely been studied in plant single cell-types. In this study, isobaric tandem mass tag-based redox proteomics technology was applied to discover redox-sensitive proteins and corresponding cysteine residues in guard cell response to a bacterial flagellin peptide flg22. Many redox-responsive proteins related to photosynthesis, lipid binding, oxidation-reduction, and defense were identified. Using reverse genetics and biochemical analyses, a lipid transfer protein was functionally characterized to be involved in plant defense against pathogens. The study highlights the utility of redox proteomics in discovering new proteins and redox modifications in important stomatal guard cell functions. Furthermore, detailed functional characterization demonstrates the biological relevance of the redox-responsive lipid transfer protein in plant pathogen defense.

In Vivo Imaging of Plant Oxygen Levels.

Oxygen is essential for multicellular aerobic life due to its central role in energy metabolism. The availability of oxygen can drop below the level to sustain oxidative phosphorylation when plants are flooded, posing a severe threat to survival. However, under non-stressful conditions, the internal oxygen concentration of most plant tissue is not in equilibrium with the environment, which is attributed to cellular respiration and diffusion constrains imposed by O2 barriers and bulky tissue. This is exemplified by the observations of steep oxygen gradients in roots, fruits, tubers, anthers and meristems. To adapt to a varying availability of oxygen, plants sense O2 via the conditional proteolysis of transcriptional regulators. This mechanism acts to switch oxidative metabolism to anaerobic fermentation, but it was also shown to play a role in plant development and pathogen defense. To investigate how dynamic and spatial distribution of O2 impacts on these processes, accurate mapping of its concentration in plants is essential. Physical oxygen sensors have been employed for decades to profile internal oxygen concentrations in plants, while genetically encoded oxygen biosensors have only recently started to see use. Driven by the critical role of hypoxia in human pathology and development, several novel oxygen-sensing devices have also been characterized in cell lines and animal model organisms. This review aims to provide an overview of available oxygen biosensors and to discuss their potential application to image oxygen levels in plants.

A novel sweetpotato RING-H2 type E3 ubiquitin ligase gene IbATL38 enhances salt tolerance in transgenic Arabidopsis

Arabidopsis Toxicos en Levadura (ATL) proteins compose a subfamily of E3 ubiquitin ligases and play major roles in regulating plant growth, cold, drought, oxidative stresses response and pathogen defense in plants. However, the role in enhancing salt tolerance has not been reported to date. Here, we cloned a novel RING-H2 type E3 ubiquitin ligase gene, named IbATL38, from sweetpotato cultivar Lushu 3. This gene was highly expressed in the leaves of sweetpotato and strongly induced by NaCl and abscisic acid (ABA). This IbATL38 was localized to nucleus and plasm membrane and possessed E3 ubiquitin ligase activity. Overexpression of IbATL38 in Arabidopsis significantly enhanced salt tolerance, along with inducible expression of a series of stress-responsive genes and prominently decrease of H2O2 content. These results suggest that IbATL38 as a novel E3 ubiquitin ligase may play an important role in salt stress response.

High-throughput sequencing analysis of the rhizosphere arbuscular mycorrhizal fungi (AMF) community composition associated with Ferula sinkiangensis

BackgroundFerula sinkiangensis is an increasingly endangered medicinal plant. Arbuscular mycorrhiza fungi (AMF) are symbiotic microorganisms that live in the soil wherein they enhance nutrient uptake, stress resistance, and pathogen defense in host plants. While such AMF have the potential to contribute to the cultivation of Ferula sinkiangensis, the composition of AMF communities associated with Ferula sinkiangensis and the relationship between these fungi and other pertinent abiotic factors still remains to be clarified.ResultsHerein, we collected rhizosphere and surrounding soil samples at a range of depths (0–20, 20–40, and 40–60 cm) and a range of slope positions (bottom, middle, top). These samples were then subjected to analyses of soil physicochemical properties and high-throughput sequencing (Illumina MiSeq). We determined that Glomus and Diversispora species were highly enriched in all samples. We further found that AMF diversity and richness varied significantly as a function of slope position, with this variation primarily being tied to differences in relative Glomus and Diversispora abundance. In contrast, no significant relationship was observed between soil depth and overall AMF composition, although some AMF species were found to be sensitive to soil depth. Many factors significantly affected AMF community composition, including organic matter content, total nitrogen, total potassium, ammonium nitrogen, nitrate nitrogen, available potassium, total dissolvable salt levels, pH, soil water content, and slope position. We further determined that Shannon diversity index values in these communities were positively correlated with total phosphorus, nitrate-nitrogen levels, and pH values (P < 0.05), whereas total phosphorus, total dissolvable salt levels, and pH were positively correlated with Chao1 values (P < 0.05).ConclusionIn summary, our data revealed that Glomus and Diversispora are key AMF genera found within Ferula sinkiangensis rhizosphere soil. These fungi are closely associated with specific environmental and soil physicochemical properties, and these soil sample properties also differed significantly as a function of slope position (P < 0.05). Together, our results provide new insights regarding the relationship between AMF species and Ferula sinkiangensis, offering a theoretical basis for further studies of their development.

Role of brassinosteroids in mitigating abiotic stresses in plants

Plants require extrinsic factors like air, water, light, nutrition, etc. for the regulation of their growth and development. Similarly, phytohormones are equally important for plants as intrinsic factors. Phytohormones are active molecules vital for various aspects in growth and development starting from embryogenesis, plant-pathogen defense and organ size regulation to reproductive development. These hormones also play an active role in mediating defense response against biotic and abiotic stresses in plants. According to estimates, a substantial loss in agricultural yields leading to concerns on food security worldwide has been reported due to abiotic stresses like salinity, extreme temperatures, drought, etc. To cope up with harsh stress conditions, plants develop certain altered growth patterns and physiological processes. Among various groups of phytohormones produced by plants, those which are based on isoprenoid origin are quite important in safeguarding plants against environmental stress. Brassinosteroids are one of the novel groups of plant hormones of isoprenoid origin. Due to its remarkable growth supporting property, these are regarded as phytohormones with pleiotropic effects. They dominate miscellaneous physiological activities such as growth, development, rhizogenesis, seed germination, senescence and most importantly abiotic stress tolerance in plants. In general, when a plant perceives a signal produced by stress, then it triggers a cascade mechanism of signal transduction with plant growth regulators, acting as alphatransducers.

The Sweetpotato BTB-TAZ Protein Gene, IbBT4, Enhances Drought Tolerance in Transgenic Arabidopsis.

BTB-TAZ (BT)-domain proteins regulate plant development and pathogen defense. However, their roles in resistance to abiotic stresses remain largely unknown. In this study, we found that the sweetpotato BT protein-encoding gene IbBT4 significantly enhanced the drought tolerance of Arabidopsis. IbBT4 expression was induced by PEG6000, H2O2 and brassinosteroids (BRs). The IbBT4-overexpressing Arabidopsis seeds presented higher germination rates and longer roots in comparison with those of WT under 200 mM mannitol stress. Under drought stress the transgenic Arabidopsis plants exhibited significantly increased survival rates and BR and proline contents and decreased water loss rates, MDA content and reactive oxygen species (ROS) levels. IbBT4 overexpression upregulated the BR signaling pathway and proline biosynthesis genes and activated the ROS-scavenging system under drought stress. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays revealed that the IbBT4 protein interacts with BR-ENHANCED EXPRESSION 2 (BEE2). Taken together, these results indicate that the IbBT4 gene provides drought tolerance by enhancing both the BR signaling pathway and proline biosynthesis and further activating the ROS-scavenging system in transgenic Arabidopsis.

Cellular Stress Upregulates Indole Signaling Metabolites in Escherichia coli

Escherichia coli broadly colonize the intestinal tract of humans and produce a variety of small molecule signals. However, many of these small molecules remain unknown. Here, we describe a family of widely distributed bacterial metabolites termed the "indolokines." In E.coli, the indolokines are upregulated in response to a redox stressor via aspC and tyrB transaminases. Although indolokine 1 represents a previously unreported metabolite, four of the indolokines (2-5) were previously shown to be derived from indole-3-carbonyl nitrile (ICN) in the plant pathogen defense response. We show that the indolokines are produced in a convergent evolutionary manner relative to plants, enhance E.coli persister cell formation, outperform ICN protection in an Arabidopsis thaliana-Pseudomonas syringae infection model, trigger a hallmark plant innate immune response, and activate distinct immunological responses in primary human tissues. Our molecular studies link a family of cellular stress-induced metabolites to defensive responses across bacteria, plants, and humans.

Exogenous sodium diethyldithiocarbamate, a Jasmonic acid biosynthesis inhibitor, induced resistance to powdery mildew in wheat.

Jasmonic acid (JA) is an important plant hormone associated with plant–pathogen defense. To study the role of JA in plant–fungal interactions, we applied a JA biosynthesis inhibitor, sodium diethyldithiocarbamate (DIECA), on wheat leaves. Our results showed that application of 10 mM DIECA 0–2 days before inoculation effectively induced resistance to powdery mildew (Bgt) in wheat. Transcriptome analysis identified 364 up‐regulated and 68 down‐regulated differentially expressed genes (DEGs) in DIECA‐treated leaves compared with water‐treated leaves. Gene ontology (GO) enrichment analysis of the DEGs revealed important GO terms and pathways, in particular, response to growth hormones, activity of glutathione metabolism (e.g., glutathione transferase activity), oxalate oxidase, and chitinase activity. Gene annotaion revealed that some pathogenesis‐related (PR) genes, such as PR1.1, PR1, PR10, PR4a, Chitinase 8, beta‐1,3‐glucanase, RPM1, RGA2, and HSP70, were induced by DIECA treatment. DIECA reduced JA and auxin (IAA) levels, while increased brassinosteroid, glutathione, and ROS lesions in wheat leaves, which corroborated with the transcriptional changes. Our results suggest that DIECA can be applied to increase plant immunity and reduce the severity of Bgt disease in wheat fields.

Computational identification of conserved microRNAs and functional annotation of their target genes in Citrus limon: Identification of microRNA in Citrus limon

Citrus limon (lemon) is an important fruit crop of Rutaceae family with high nutritive value. The microRNAs (miRNAs) are a class of small RNAs, approximately 22 nucleotides long. They are evolutionary conserved and function at post-transcriptional level to regulate the expressivity of genes in both plants and animals. They have crucial roles in different biological and metabolic processes like growth and development, pathogen defense, abiotic and biotic stress response in plants. Computational analysis of Citrus limon expressed sequence tags (ESTs) resulted in the identification of one conserved microRNA (clm-miR5658). The secondary structure of newly identified microRNA was generated based on minimal folding energy (MFE= −50.60 kcal/mol) and minimal folding energy index (MFEI= 0.81). Phylogenetic analysis of miR5658 family revealed a close association of clm-miR5658 with bra-miR5658. Further, a total of 14 potential target genes of the putative miRNA were identified and functionally annotated. The findings of this study will contribute to understand the role of miRNAs in lemon and expected to lay foundation for further research on lemon miRNAs.

Apple ethylene response factor MdERF11 confers resistance to fungal pathogen Botryosphaeria dothidea

Ethylene response factor (ERF) is a plant-specific transcription factor involved in many biological processes including root formation, hypocotyl elongation, fruit ripening, organ senescence and stress responses, as well as fruit quality formation. However, its underlying mechanism in plant pathogen defense against Botryosphaeria dothidea (B. dothidea) remains poorly understood. Here, we isolate MdERF11, an apple nucleus-localized ERF transcription factor, from apple cultivar ‘Royal Gala’. qRT-PCR assays show that the expression of MdERF11 is significantly induced in apple fruits after B. dothidea infection. Overexpression of MdERF11 gene in apple calli significantly increases the resistance to B.dothidea infection, while silencing MdERF11 in apple calli results in reduced resistance. Ectopic expression of MdERF11 in Arabidopsis also exhibits enhanced resistance to B. dothidea infection compared to that of wild type. Infections in apple calli and Arabidopsis leaves by B. dothidea respectively cause an increase in endogenous levels of salicylic acid (SA) followed by induction of SA synthesis-related and signaling-related gene expression. Taken together, these findings illustrate a potential mechanism by which MdERF11 elevates plant pathogen defense against B. dothidea by regulating SA synthesis pathway.