Gene Expression In Plants Research Articles

An enhancer-promoter-transcription factor module orchestrates plant immune homeostasis by constraining camalexin biosynthesis

Effective plant defense against pathogens relies on highly coordinated regulation of immune gene expression. Enhancers, as cis-regulatory elements, are indispensable determinants of dynamic gene regulation, but the molecular functions in plant immunity are not well understood. In this study, we identified a novel enhancer, CORE PATTERN-INDUCED ENHANCER 35 (CPIE35), which is rapidly activated upon pathogenic elicitation and negatively regulates anti-fungal resistance through modulating WRKY15 expression. During immune activation, CPIE35 activates the transcription of WRKY15 by forming chromatin loops with the promoter of WRKY15 in a WRKY18/40/60-, WRKY33-, and MYC2-dependent manner. WRKY15 directly binds to the promoters of PAD3 and GSTU4, suppressing their expression and leading to the reduced camalexin synthesis and resistance. Interestingly, CPIE35 region is evolutionarily conserved among Brassicaceae plants, and the CPIE35-WRKY15 module exerts similar functions in Brassica napus to negatively regulate anti-fungal resistance. Our work reveals the “enhancer-promoter-transcription factor” regulatory mechanism in maintenance of immune homeostasis, highlighting the importance and conserved role of enhancers in fine-tuning immune gene expression in plants.

Combined Transcriptomic and Metabolomic Analyses Reveal the Mechanisms by Which the Interaction Between Sulfur and Nitrogen Affects Garlic Yield and Quality

Nitrogen and sulfur are essential macronutrients in plant growth and development, and their interaction profoundly influences gene expression, metabolic activities, and adaptability in plants, directly affecting plant growth and yield. Garlic (Allium sativum L.) is a crop of significant economic and medicinal value. However, despite the critical role of the nitrogen–sulfur interaction in garlic’s adaptability, yield, and quality, the specific mechanisms underlying these effects remain unclear. In this study, transcriptomic and metabolomic analyses were employed to investigate the effects of combined sulfur and nitrogen application on garlic bulb tissues. The results show that the combined application of sulfur and nitrogen significantly increased the diameter and weight of garlic bulbs by 14.96% and 35.47%, respectively. The content of alliin increased by 28.48%, while the levels of abscisic acid (ABA), jasmonic acid (JA), salicylic acid (SA), and gibberellin (GA) increased by 15.82%, 12.94%, 32.34%, and 48.13%, respectively. Additionally, the activities of alliinase, superoxide dismutase (SOD), and catalase (CAT) were enhanced by 7.93%, 4.48%, and 19.74%, respectively. Moreover, the application of sulfur and nitrogen significantly reduced the malondialdehyde (MDA) content and peroxidase (POD) activity in garlic bulbs by 29.66% and 9.42%, respectively, thereby improving garlic’s adaptability and growth potential. Transcriptomic analysis revealed differentially expressed genes in several key pathways, including plant hormone signal transduction, RNA degradation, glutathione metabolism, amino acid biosynthesis, and glycerophospholipid metabolism. Metabolomic analysis identified 80 differentially abundant metabolites primarily consisting of amino acids, indole carboxylic acids, and fatty acids. The integrated transcriptomic and metabolomic analyses highlighted the pivotal roles of glutathione metabolism, glycerophospholipid metabolism, and amino acid biosynthesis pathways in the synergistic effects of sulfur and nitrogen. This study not only provides critical scientific evidence for understanding the mechanisms underlying the nitrogen–sulfur interaction’s impact on the yield and quality of garlic but also offers a scientific basis for optimizing nutrient management strategies to enhance garlic yield and quality.

The response of DNA methyltransferase and demethylase genes to abiotic stresses in tomato seedling

DNA methylation plays an important role in regulating plant growth, development and gene expression. However, less is known about the response of DNA methyltransferase and demethylase genes to various stresses. In this study, the effects of abiotic stresses on DNA methylation gene expression patterns in tomato seedlings were investigated. Results showed that most tomato DNA methyltransferase and demethylase genes contained stress-related elements. The expression of SlDML1 was significantly induced by cadmium (Cd) and sodium chloride (NaCl) stresses. SlDML2 was more sensitive and reached its maximum value under polyethylene (PEG) stress at 24 h. The expression of SlMET3L was repressed to varying degrees under Cd, NaCl and PEG stresses at 48 h. However, 5-aza-2′-deoxycytidine (5-azadC) treatment decreased the Cd and PEG stress tolerance by down-regulating the expression of DNA methyltransferase except for the SlMET3L, and up-regulating the expression levels of SlDML2, SlDML3 and SlDML4, cadmium transporters (SlHMA5, SlCAX3, and SlACC3) and osmoregulators (SlDREB, SlLEA and SlHSP70). Whereas 5-azadC treatment alleviated the salt stress through up-regulating DNA methyltransferase gene expression, and down-regulating the expression level of SlDML1, SlDML3, and SlDML4, SlHKT1, SlNHX1, and SlSOS1. Collectively, 5-azadC impaired Cd and PEG stress tolerance and enhanced salt stress tolerance by regulating the expression of methylation-related and stress-related genes in tomato seedlings. These results may provide useful information for further analysing function and evolution of DNA methylation methyltransferase and demethylase genes in tomato under stress conditions.

A history-dependent integrase recorder of plant gene expression with single-cell resolution

During development, most cells experience a progressive restriction of fate that ultimately results in a fully differentiated mature state. Understanding more about the gene expression patterns that underlie developmental programs can inform engineering efforts for new or optimized forms. Here, we present a four-state integrase-based recorder of gene expression history and demonstrate its use in tracking gene expression events in Arabidopsis thaliana in two developmental contexts: lateral root initiation and stomatal differentiation. The recorder uses two serine integrases to mediate sequential DNA recombination events, resulting in step-wise, history-dependent switching between expression of fluorescent reporters. By using promoters that express at different times along each of the two differentiation pathways to drive integrase expression, we tie fluorescent status to an ordered progression of gene expression along the developmental trajectory. In one snapshot of a mature tissue, our recorder is able to reveal past gene expression with single cell resolution. In this way, we are able to capture heterogeneity in stomatal development, confirming the existence of two alternate paths of differentiation.

Integrating Biofertilizers with Organic Fertilizers Enhances Photosynthetic Efficiency and Upregulates Chlorophyll-Related Gene Expression in Rice

Biofertilizers offer a sustainable method for improving rice growth and productivity, yet their effects on the interaction between plant growth, photosynthetic activity, and gene expression remain under-researched. This study examines how biofertilizer influences rice physiology, focusing on photosynthetic regulation and expression of chlorophyll-related genes. Eight fertilizer treatments were applied: control (CNT), biofertilizer (BF), deactivated biofertilizer (DABF), rice straw (RS), rice straw with biofertilizer (RS+BF), organic fertilizer (OF), organic fertilizer with biofertilizer (OF+BF), and inorganic fertilizer (IOF). Plant height, tiller number, SPAD, NDVI, chlorophyll content, and photosynthesis rates were measured, while gene expression analysis was conducted using RT-qPCR. The OF+BF treatment produced the most significant results, leading to a 31% increase in plant height, a 135% increase in tiller number, and a 42% increase in chlorophyll content (SPAD values) compared to the control. Additionally, OF+BF enhanced photosynthetic efficiency by 74%, with the highest net photosynthetic rate of 48.23 μmol CO2 m−2 s−1. Gene expression analysis revealed that OF+BF upregulated key photosynthesis-related genes, such as OsChlD and OsCHLM, showing a 70% and 90% increase in expression. These findings highlight the potential of integrating biofertilizers with organic fertilizers to sustainably boost rice growth and productivity, contributing to global food security and climate change mitigation.

The effect of spermidine supplementation on polyamine metabolism and Fusarium infection in Linum usitatissimum

Flax (Linum usitatissimum) is a valuable industrial crop in temperate climate zones. However, the prevalence of fungal diseases, particularly those caused by Fusarium oxysporum sp. linii, poses a substantial threat, leading to significant crop losses and diminished interest in flax cultivation. In this study, flax plants were treated with spermidine and subsequently infected with Fusarium oxysporum to investigate multiple aspects: (1) the uptake of exogenous spermidine by the plants, (2) the impact of this uptake on the levels of other polyamines and the expression of polyamine biosynthesis genes, and (3) the effects of fungal infection on these parameters. The results demonstrated that flax plants effectively absorb spermidine, leading to substantial changes in the levels of polyamines and the expression of related genes in both roots and shoots. Notably, spermidine treatment resulted in significant alterations in the levels of putrescine and spermine, as well as in the transcript levels of key genes involved in polyamine biosynthesis. Moreover, Fusarium oxysporum infection itself induced changes in polyamine content and gene expression, which varied between root and shoot tissues. When spermidine-treated plants were infected with Fusarium oxysporum, a more complex response was observed, characterized by modifications in polyamine levels and gene expression that suggest an enhanced defense mechanism against the pathogen. These findings indicate that polyamine treatment not only affects the infection process but also modulates the plant's internal polyamine metabolism and gene expression, contributing to an improved resistance to fungal attack. This study highlights the potential of spermidine as a protective agent in flax and provides a foundation for further exploration of polyamine-related defense mechanisms.

Genome-Wide Identification and Expression Analysis of the REF Genes in 17 Species.

Natural rubber production currently relies heavily on a single species, Hevea brasiliensis, underscoring the urgent need to identify alternative sources to alleviate the strain on natural rubber production. The rubber elongation factor (REF) and small rubber particle protein (SRPP), both members of the REF/SRPP gene family, are crucial for natural rubber biosynthesis. However, research on the REF gene has predominantly focused on H. brasiliensis and Taraxacum kok-saghyz. We conducted a comprehensive genome-wide identification and characterization of the REF gene, identifying 87 REF protein sequences across 17 plants species. We observed a significant increase in the copy numbers and expression of REF genes in rubber-producing plants. Notably, in H. brasiliensis, T. kok-saghyz, Eucommia ulmoides, Lactuca sativa, and other rubber-yielding species, the number of REF genes has markedly increased. Furthermore, some REF genes in H. brasiliensis form a distinct clade in phylogenetic analyses and exhibit differences in conserved motif arrangements and tertiary protein structures compared to other REF genes. These findings suggest that REF genes in rubber-producing plants may have undergone independent evolution, leading to changes in copy number and structure. These alterations could contribute to the production of natural rubber in these species. The results of this study provide a scientific basis for further research into the mechanisms of rubber production in plants and for identifying potential rubber-producing species.

From grass to yeast; functional insights from heterologous expression of LfHKT2;1 in ion regulation

Saccharomyces cervisceae mutants lacking the major potassium trasnporters trk1 and trk2 show hypersensitivity to aminoglycoside antibiotic hygromycin (hyg). This study demonstrates that expression of the inward K+ transporter LfHKT2;1 can suppress this hygromycin sensitivity in the trk1, trk2 double mutant strain. Growth complementation was performed on solid yeast peptone dextrose (YPD) media supplemented with hygromycin B. Both, wild type and trk1 trk2 yeast strains exhibited growth inhibition in the presence of hygromycin. However the potassium uptake-deficient trk1 trk2 strain (control) showed complete growth arrest when exposed to hygromycin, while expression of LfHKT2;1 resulted in growth recovery. Increased sodium concentrations caused cellular toxicity in the trk1 trk2 strain, which was exacerbated by the addition of hygromycin to the media. The hypersensitivity of trk1 trk2 mutant yeast cells expressing LfHKT2;1 to sodium suggested the presence of an additional sodium uptake system on the membrane, which was further confirmed by transient GFP expression assays. These results provide conclusive evidence that heterologous expression of LfHKT2;1 confers both sodium and K+ uptake capabilities in hygromycin supplemented YPD media, thereby rescuing the growth of K+ transport-deficient S. cerevisiae mutants. This highlights the potential of plant gene expression in yeast as a valuable tool for studying ion transport mechanisms and gene function under stress conditions.

Machine learning-aided microRNA discovery for olive oil quality.

MicroRNAs (miRNAs) are key regulators of gene expression in plants, influencing various biological processes such as oil quality and seed development. Although, our knowledge about miRNAs in olive (Olea europaea L.) is progressing, with several miRNAs being identified in previous studies, but most of these reported miRNAs have been predicted without the aid of a reference genome, primarily due to limited genome accessibility at the time. However, significant knowledge gaps still need to be improved in this area. This study addresses the complexities of miRNA detection in olive, using a high quality reference genome and a combination of genomics and machine learning-based methods. By leveraging random forest and support vector machine algorithms, we successfully identified 56 novel miRNAs in olive, surpassing the limitations of conventional homology-based methods. Our subsequent analysis revealed that some of these miRNAs are implicated in the regulation of key genes involved in oil quality. Within the context of oil biosynthesis pathways, the novel miRNA Oeu124369 regulates fatty acid biosynthesis by targeting acetyl-CoA acyltransferase 1 and palmitoyl-protein thioesterase, thereby influencing the production of acetyl-CoA and palmitic acid, respectively. These findings underscore the power of machine learning in unraveling the complex miRNA regulatory network in olive and provide a high quality miRNA resource for future research aimed at improving olive oil production by exploring the target genes of the identified miRNAs to understand their role and their biological processes.

Hormonal signaling regulates photosynthetic function of alfalfa (Medicago sativa L.) under NaHCO3 stress

The physiological and molecular mechanisms underlying salt-alkali tolerance in Medicago sativa are of significant importance for the development of animal husbandry on salt-alkali lands and the restoration of vegetation in such areas. This study utilized salt-alkali tolerance Medicago sativa 'Zhaodong' (ZD) and salt-alkali sensitive variety M. sativa 'Zhongmu No.1′ (ZM) as materials. Physiological analyses, transcriptomic sequencing, and hormone-targeted metabolomics techniques were employed to investigate the differential responses of the two alfalfa varieties to NaHCO3 stress in terms of morphology, photosynthetic functionality, and oxidative damage indicators. Additionally, weighted gene co-expression network analysis (WGCNA) was utilized to elucidate key mechanisms underlying salt-alkali tolerance in alfalfa. The results indicate that NaHCO3 stress leads to photosynthetic inhibition and oxidative damage in alfalfa leaves. Under NaHCO3 stress, PSI in alfalfa leaves exhibits higher stability compared to PSII. The salt-alkali tolerance alfalfa variety ZD demonstrates stronger tolerance compared to the salt-alkali sensitive variety ZM. Furthermore, differentially expressed genes (DEGs) between the two varieties under NaHCO3 stress are primarily enriched in KEGG pathways such as chlorophyll synthesis, photosynthesis, carbon fixation, and plant hormone synthesis and signaling. Weighted gene co-expression network analysis (WGCNA) was conducted based on physiological and transcriptomic data. Most differentially expressed genes (DEGs) in the top two modules with the highest correlation to physiological indicators such as photosynthesis are enriched in hormone synthesis and signal transduction pathways. Additionally, key transcription factors involved in hormone signal transduction were identified within these modules, such as MYC2 and ABI5, which regulate jasmonic acid (JA) and abscisic acid (ABA) signaling, respectively. These findings suggest that plant hormone signaling may play a critical role in regulating salt-alkali tolerance in alfalfa. Further analysis was conducted on plant hormone levels and gene expression involved in biosynthesis and signal transduction processes. The results indicate that NaHCO3 stress leads to significant accumulation of ABA and JA content in alfalfa leaves. The biosynthesis and signal transduction pathways of ABA and JA are activated under NaHCO3 stress. Additionally, the salt-alkali tolerance alfalfa variety ZD exhibits a more sensitive response to ABA and JA signals compared to ZM. Salicylic acid (SA) shows a positive response to NaHCO3 stress only in the ZD variety, which may be one of the key reasons for its stronger salt-alkali tolerance. Under NaHCO3 stress, overall growth-promoting hormones (IAA, GA, CK) are downregulated in ZD but upregulated in ZM, indicating that the salt-alkali tolerance alfalfa variety ZD mainly regulates the balance between growth and resistance by modulating the ratio of growth-promoting and stress-related hormones in response to NaHCO3 stress. This study reveals that hormone signaling plays a key role in regulating the photosynthetic function of alfalfa in response to salt-alkali stress, which provides theoretical basis and clues for the molecular breeding of alfalfa for salt-alkali tolerance.

Análisis de la expresión del gen TRI5 de Fusarium graminearum en coleóptilos de trigo y maíz.

The necrotrophic fungus F. graminearum is responsible for causing Fusarium head blight in wheat and ear rotin maize. This fungal infection leads to reduced crop yields and contamination of the crops with mycotoxinslike trichothecene deoxynivalenol (DON), which can cause poisoning in animals and humans. The productionof DON is facilitated by a set of genes, with the TRI5 gene encoding trichodiene synthase being a key player.While some factors that can activate the TRI5 gene in wheat are known, it's unclear whether this gene isactivated when the fungus infects wheat or maize coleoptiles. This study reveals that using ammonium sulfateas a nitrogen source induces the expression of the TRI5 gene in minimal media. In wheat coleoptiles, F.graminearum forms infection structures at 5 days post-infection (dpi), where the induction of the TRI5 geneoccurs. On the other hand, there is no TRI5 gene induction in maize coleoptiles. Finally, the TRI5 geneexpression in plants was confirmed using semi-quantitative PCR, validating the induction in wheat coleoptilesat 5 dpi.

Comparative Analysis of the Codon Usage Pattern in the Chloroplast Genomes of Gnetales Species

Codon usage bias refers to the preferential use of synonymous codons, a widespread phenomenon found in bacteria, plants, and animals. Codon bias varies among species, families, and groups within kingdoms and between genes within an organism. Codon usage bias (CUB) analysis sheds light on the evolutionary dynamics of various species and optimizes targeted gene expression in heterologous host plants. As a significant order of gymnosperms, species within Gnetales possess extremely high ecological and pharmaceutical values. However, comprehensive analyses of CUB within the chloroplast genomes of Gnetales species remain unexplored. A systematic analysis was conducted to elucidate the codon usage patterns in 13 diverse Gnetales species based on the chloroplast genomes. Our results revealed that chloroplast coding sequences (cp CDSs) in 13 Gnetales species display a marked preference for AT bases and A/T-ending codons. A total of 20 predominantly high-frequency codons and between 2 and 7 optimal codons were identified across these species. The findings from the ENC-plot, PR2-plot, and neutrality analyses suggested that both mutation pressure and natural selection exert influence on the codon bias in these 13 Gnetales species, with natural selection emerging as the predominant influence. Correspondence analysis (COA) demonstrated variation in the codon usage patterns among the Gnetales species and indicated mutation pressure is another factor that could impact CUB. Additionally, our research identified a positive correlation between the measure of idiosyncratic codon usage level of conservatism (MILC) and synonymous codon usage order (SCUO) values, indicative of CUB’s potential influence on gene expression. The comparative analysis concerning codon usage frequencies among the 13 Gnetales species and 4 model organisms revealed that Saccharomyces cerevisiae and Nicotiana tabacum were the optimal exogenous expression hosts. Furthermore, the cluster and phylogenetic analyses illustrated distinct patterns of differentiation, implying that codons, even with weak or neutral preferences, could affect the evolutionary trajectories of these species. Our results reveal the characteristics of codon usage patterns and contribute to an enhanced comprehension of evolutionary mechanisms in Gnetales species.

Assessing Cold Plasma's Impact on Banana Growth and Fusarium Wilt Control.

Bananas (Musa spp.), which serve millions of people worldwide, face a serious threat from Fusarium wilt (FW) disease caused by Fusarium oxysporum f. sp. cubense (Foc). Developing disease-resistant varieties particularly through breeding is challenging due to banana's seedless nature (parthenocarpic). As an alternative, cold plasma (CP) technology, has the potential to be used for crop improvement. Our study demonstrates a favourable impact of CP on the growth performance of banana (Berangan cultivar, AAA) in terms of height, leaf number and stem diameter. CP-treated plants also displayed delayed disease progression as well as lower disease severity indicated by slightly lower value of leaf symptoms index and rhizome discoloration index compared to the control plants. Additionally, quantitative real-time polymerase chain reaction analysis revealed differential expression of several defence (PR1, WRKY22, PAL, and CEBiP) and growth (Cytochrome P450, NAC68, and CAT) related genes in CP-treated plants, particularly in conjunction with Foc infection. These findings shed light on the potential use of CP in managing FW in banana and offer insights into possible mechanism behind improved traits.

Bacillus siamensis orchestrates plant gene reprogramming and rhizosphere microbiome reshaping to bolster maize crop performance

AbstractPlant growth‐promoting rhizobacteria (e.g., Bacillus) confer health benefits to the host. Bacillus siamensis has been used to control plant disease in fruits and vegetables, but its potential effects on crop performance remain unclear. This study investigated changes in the growth, root transcriptomic profile, and rhizosp here microbiome of maize plants (Zea may L.) following inoculation with B. siamensis 33. All inoculated plants grew better than non‐inoculated controls in pots, as exemplified by 24.0% and 6.8% increase in plant height and stem diameter, respectively (p < 0.01). Shoot dry weight also increased by 20.6% upon inoculation, accompanied by 46.9% increase in root dry weight (p < 0.01). Transcriptomic analysis revealed that inoculation induced (2.16‐ to 5.53‐fold) upregulated expression of genes related to auxin signal transduction in maize. The expression of genes involved in phenylpropanoid biosynthesis, glutathione metabolism, and plant defense‐related signal (e.g., jasmonic acid and salicylic acid) transduction was (2.06‐ to 11.30‐fold) upregulated in inoculated plants. Upon inoculation, bacterial α‐diversity trended higher and potentially beneficial taxa (e.g., Sphingobium, Flavihumibacter, and Parasegetibacter) were enriched in the rhizosphere, likely a result of enhanced root exudation of specific metabolites (e.g., flavonoids) and subsequent recruitment of beneficial microbial taxa. Potential microbial functions associated with nitrogen cycling and pollutant degradation were stimulated by inoculation. These findings indicate that B. siamensis 33 mediates the reprogramming of plant gene expression and reshaping of rhizosphere microbiome structure to bolster maize crop performance.

Design and Characterization of a Transcriptional Repression Toolkit for Plants.

Regulation of gene expression is essential for all life. Tools to manipulate the gene expression level have therefore proven to be very valuable in efforts to engineer biological systems. However, there are few well-characterized genetic parts that reduce gene expression in plants, commonly known as transcriptional repressors. We characterized the repression activity of a library consisting of repression motifs from approximately 25% of the members of the largest known family of repressors. Combining sequence information with our trans-regulatory function data, we next generated a library of synthetic transcriptional repression motifs with function predicted in advance. After characterizing our synthetic library, we demonstrated not only that many of our synthetic constructs were functional as repressors but also that our advance predictions of repression strength were better than random guesses. Finally, we assessed the functionality of known transcriptional repression motifs from a wide range of eukaryotes. Our study represents the largest plant repressor motif library experimentally characterized to date, providing unique opportunities for tuning transcription in plants.

Combined transcriptome and metabolome analysis identifies triterpenoid-induced defense responses in Myzus persicae Sülzer-infested peach.

Piercing/sucking insects such as green peach aphid (GPA) (Myzus persicae) cause direct damage by obtaining phloem nutrients and indirect damage by spreading plant viruses. To investigate the response of peach trees (Prunus persica) to aphids, the leaf transcriptome and metabolome of two genotypes with different sensitivities to GPA were studied. The gene expression of aphid-susceptible plants infested with aphids was similar to that of control plants, whereas the gene expression of aphid-resistant plants infested with aphids showed strong induced changes in gene expression compared with control plants. Furthermore, gene transcripts in defense-related pathways, including plant-pathogen interaction, MAPK signaling, and several metabolic pathways, were strongly enriched upon aphid infestation. Untargeted secondary metabolite profiling confirmed that aphid infestation induced larger changes in aphid-resistant than in aphid-susceptible peaches. Consistent with transcriptomic alterations, nine triterpenoids showed highly significant GPA-induced accumulation in aphid-resistant peaches, whereas triterpenoid abundance remained predominantly unchanged or undetected in aphid-susceptible peaches. Furthermore, some types of transcription factors (including WRKYs, ERFs, and NACs) were strongly induced upon GPA infestation in aphid-resistant, but not in aphid-susceptible peaches. These results suggested that the accumulation of specialized triterpenoids and the corresponding pathway transcripts may play a key role in peach GPA resistance.

Analysis of Plant and Fungal Transcripts from Resistant and Susceptible Phenotypes of Leptospermum scoparium Challenged by Austropuccinia psidii.

Austropuccinia psidii is the causal pathogen of myrtle rust disease of Myrtaceae. To gain understanding of the initial infection process, gene expression in germinating A. psidii urediniospores and in Leptospermum scoparium-inoculated leaves were investigated via analyses of RNA sequencing samples taken 24 and 48 h postinoculation (hpi). Principal component analyses of transformed transcript count data revealed differential gene expression between the uninoculated L. scoparium control plants that correlated with the three plant leaf resistance phenotypes (immunity, hypersensitive response, and susceptibility). Gene expression in the immune resistant plants did not significantly change in response to fungal inoculation, whereas susceptible plants showed differential expression of genes in response to fungal challenge. A putative disease resistance gene, jg24539.t1, was identified in the L. scoparium hypersensitive response phenotype family. Expression of this gene may be associated with the phenotype and could be important for further understanding the plant hypersensitive response to A. psidii challenge. Differential expression of pathogen genes was found between samples taken 24 and 48 hpi, but there were no significant differences in pathogen gene expression that were associated with the three different plant leaf resistance phenotypes. There was a significant decrease in the abundance of fungal transcripts encoding three putative effectors and a putative carbohydrate-active enzyme between 24 and 48 hpi, suggesting that the encoded proteins are important during the initial phase of infection. These transcripts, or their translated proteins, may be potential targets to impede the early phases of fungal infection by this wide-host-range obligate biotrophic basidiomycete.

Rhizosphere microbiomes derived from vermicompost alter gene expression and regulatory pathways in tomato (Solanum lycopersicum, L.).

The gut microbiome of worms from composting facilities potentially harbors organisms that are beneficial to plant growth and development. In this experiment, we sought to examine the potential impacts of rhizosphere microbiomes derived from Eisenia fetida worm castings (i.e. vermicompost) on tomato (Solanum lycopersicum, L.) plant growth and physiology. Our experiment consisted of a greenhouse trial lasting 17weeks total in which tomato plants were grown with one of three inoculant treatments: a microbial inoculant created from vermicompost (V), a microbial inoculant created from sterilized vermicompost (SV), and a no-compost control inoculant (C). We hypothesized that living microbiomes from the vermicompost inoculant treatment would enhance host plant growth and gene expression profiles compared to plants grown in sterile and control treatments. Our data showed that bacterial community composition was significantly altered in tomato rhizospheres, but fungal community composition was highly variable in each treatment. Plant phenotypes that were significantly enhanced in the vermicompost and sterile vermicompost treatments, compared to the control, included aboveground biomass and foliar δ15N nitrogen. RNA sequencing revealed distinct gene expression changes in the vermicompost treatment, including upregulation of nutrient transporter genes such as Solyc06g074995 (high affinity nitrate transporter), which exhibited a 250.2-fold increase in expression in the vermicompost treatment compared to both the sterile vermicompost and control treatments. The plant transcriptome data suggest that rhizosphere microbiomes derived from vermicompost can influence tomato gene expression and growth-related regulatory pathways, which highlights the value of RNA sequencing in uncovering molecular responses in plant microbiome studies.

Transcriptional responses of durum wheat to chronic chromium exposure reveal candidate proteins involved in metal detoxification and compartmentalization

Chromium phytotoxicity results in relevant alterations to plant physiology, gene expression, and genomic DNA methylation at a transgenerational level. Herein, transcriptional responses of durum wheat (Triticum turgidum L.) to chronic chromium exposure were explored in roots and leaves by RNA-seq approach. Plants grown all the time in a hydroponic system supplemented with 2.5 and 10 µM hexavalent chromium were compared to unstressful control plants, assessing biomass and seed yield analyses after senescence. Then, transcriptomic analysis was performed with these plants kept under 10 µM chromium 50 days after the onset of exposure. The chromium concentrations used were considered the lowest dose sufficient to alter gene expression without impeding plant development, while the sampling time reflected the effects in the pre-harvest phase and long-lasting defense mechanisms. Root and leaf samples from plants kept under 10 µM chromium stress and from unstressful control plants were analyzed, generating 12 RNA-seq libraries. In total, 965 and 810 transcripts were found to be differentially expressed, respectively, in roots and leaves in response to 10 µM chromium stress. In roots, transcriptional changes were noted in the primary and secondary metabolism, redox homeostasis, protein modification, solute transport, nutrient uptake, and external stimuli responses. Meanwhile, the transcriptional changes in leaves were primarily found in the secondary metabolism, hormone-related pathways, chromatin modifications, cell division, protein modification and homeostasis, solute transport, and nutrient uptake. In particular, the metal uptake and translocation pathways were studied with greater emphasis to identify key proteins involved in chromium transport and compartmentalization. Furthermore, several genes involved in the biosynthesis of malate-derived organic acids, trace metal transport/detoxification/chelation, and vacuolar compartmentalization were linked to primary defense responses, and some of them were also associated with two putative gene clusters. Therefore, these genes and gene clusters are suggested as valuable biotechnological targets for future proof-of-concept studies aimed at genetic engineering of durum wheat to improve plant tolerance to chromium exposure.

New evidence of the timing of arsenic accumulation and expression of arsenic-response genes in field-grown Pteris vittata plants under different arsenic concentrations

The timing and efficiency of arsenic (As) accumulation is crucial for using the hyperaccumulator P. vittata in remediation of As-contaminated soils. In this study, through an innovative microXRF-based approach, using a new “pinna powder” sampling method, we monitored As accumulation over time in fronds of individual P. vittata plants grown in the greenhouse and in the field on two natural soils, with high (750 mg/kg) and moderate (58.4 mg/kg) As concentrations. Results, validated by multivariant statistical analysis show that the peak of As occurs on both soils at 45/60 days and at 100/120 days in greenhouse and field grown plants, respectively. Furthermore, in field trials, the timing of As accumulation in both soils was similar during the first autumn-winter and the second spring-summer phytoextraction cycle. After the two cycles, soil As content was reduced by 70.4% in the high-As soil and 26.4% in the moderate one. Moreover, candidate genes involved in As hyperaccumulation -the arsenite antiporter PvACR3, the As (V)-reductases Pv2.5-8 and the organic cation transporter PvOCT4- are expressed in response to As in field-grown plants with similar kinetics in both soils. In conclusion, we established by this innovative technique, the timing of maximum As accumulation that is linked to the intrinsic hyperaccumulation mechanism and represents a highly powerful tool to set up the duration of phytoextraction cycles.