Ethylene Response Factor Research Articles

Overexpression of galactinol synthase 1 from Solanum commersonii (ScGolS1) confers freezing tolerance in transgenic potato

Potato (Solanum tuberosum L.) is the fourth largest food crop in the world. Low temperatures cause serious damage to potato plants every year, and freezing tolerance has become a hot spot in potato research. Galactinol synthase (GolS) is a key enzyme in the synthesis of raffinose family oligosaccharides (RFOs), and plays an important role in the response of plants to abiotic stress. In this study, the ScGolS1 gene from Solanum commersonii was cloned and introduced into the S. tuberosum cultivars ‘Atlantic’ and ‘Desiree’ via Agrobacterium-mediated transformation. Phenotyping assays showed that overexpression of ScGolS1 could significantly improve freezing tolerance in transgenic potato plants. Further physiological and biochemical experiments showed that the transgenic lines had lower relative conductivity, malondialdehyde content, and 3,3′-diaminobenzidine staining and a higher plant survival rate compared with wild type (WT) under cold stress. Moreover, the C-repeat binding factors (CBF1, CBF2 and CBF3), the downstream cold-responsive genes COR413 and COR47, and the ethylene-responsive factor (ERF) transcription factor genes ERF3, ERF4 and ERF6, which function in the ethylene signaling pathway, were all induced by freezing treatment and expressed at higher levels in the ScGolS1 overexpression lines compared with WT. Besides, the expression of some genes such as MIPS, STS and RS from the RFO metabolic pathway was up-regulated under cold stress, resulting in changes in the content of some soluble sugars. This indicated that ScGolS1 overexpression altered the sugar composition and enhanced freezing tolerance in transgenic potato by inducing the ethylene and CBF signaling pathways. These results provided theoretical support and genetic resources for freezing tolerance breeding in potato.

GsERF1 enhances Arabidopsis thaliana aluminum tolerance through an ethylene-mediated pathway

Ethylene response factor (ERF) transcription factors constitute a subfamily of the AP2/ERF superfamily in plants and play multiple roles in plant growth and development as well as in stress responses. In this study, the GsERF1 gene from the wild soybean BW69 line (an Al-resistant Glycine soja line) was rapidly induced in response to aluminum stress. Quantitative real-time PCR (qRT–PCR) analysis showed that the GsERF1 gene maintained a constitutive expression pattern and was induced in soybean in response to aluminum stress, with increased amounts of transcripts detected in the roots. The putative GsERF1 protein, which contains an AP2 domain, was located in the nucleus and maintained transactivation activity. In addition, under AlCl3 treatment, GsERF1 overexpression increased the relative growth rate of the roots of Arabidopsis and weakened the hematoxylin staining of hairy roots. Ethylene synthesis-related genes such as ACS4, ACS5 and ACS6 were upregulated in GsERF1 transgenic lines compared with the wild type under AlCl3 treatment. Furthermore, the expression levels of stress/ABA-responsive marker genes, including ABI1, ABI2, ABI4, ABI5 and RD29B, in the GsERF1 transgenic lines were affected by AlCl3 treatment, unlike those in the wild type. Taken together, the results indicated that overexpression of GsERF1 may enhance aluminum tolerance of Arabidopsis through an ethylene-mediated pathway and/or ABA signaling pathway, the findings of which lay a foundation for breeding soybean plants tolerant to aluminum stress.

In silico Comparisons of the Ethylene Response Factor 1 (ERF1) Gene Between Malaysian Wild Banana (Musa acuminata ssp. malaccensis) and Pisang Klutuk Wulung (Musa balbisiana)

Musa balbisiana (B genome) has been observed to have a higher tolerance of biotic and abiotic stresses than Musa acuminata (A genome). Ethylene Response Factor 1 (ERF1) is a gene activator for pathogenesis-related proteins (PR proteins) such as basic chitinases and beta-1,3-glucanase. There are numerous ERF1 gene studies about Oryza sativa, but information about the banana ERF1 gene, especially in the B genome (Musa balbisiana “Pisang Klutuk Wulung”), has still not been explored thoroughly. Using annotated genomic data in an A genome (Musa acuminata ssp. malaccensis) and genomic data in a B genome (Musa balbisiana “Pisang Klutuk Wulung”), research on the ERF1 gene can be conducted at the gene sequences and amino acid sequences levels. The Musa acuminata (A genome) ERF1 gene nucleotide sequence was retrieved from the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The Musa balbisiana (B genome) ERF1 gene nucleotide sequence was identified with the nucleotide Basic Local Alignment Search Tool (BLASTn) using an A genome ERF1 gene sequence as a query. Both ERF1 gene nucleotide sequences and amino acid sequences in the A and B genomes were annotated and compared. Seven annotated genome ERF1 gene sequences from the A and B genomes were identified with the probability that these genes were actively transcribed in cell activity. ERF1 gene comparisons between the A and B genomes showed that nucleotide composition, gene structure, gene position in each respective chromosome, ERF clusterization, identified motif, and amino acid composition in each of the identified motifs have similar characteristics.

The CaPti1-CaERF3 module positively regulates resistance of Capsicum annuum to bacterial wilt disease by coupling enhanced immunity and dehydration tolerance.

Bacterial wilt, a severe disease involving vascular system blockade, is caused by Ralstonia solanacearum. Although both plant immunity and dehydration tolerance might contribute to disease resistance, whether and how they are related remains unclear. Herein, we showed that immunity against R. solanacearum and dehydration tolerance are coupled and regulated by the CaPti1-CaERF3 module. CaPti1 and CaERF3 are members of the serine/threonine protein kinase and ethylene-responsive factor families, respectively. Expression profiling revealed that CaPti1 and CaERF3 were upregulated by R. solanacearum inoculation, dehydration stress, and exogenously applied abscisic acid (ABA). They in turn phenocopied each other in promoting resistance of pepper (Capsicum annuum) to bacterial wilt not only by activating salicylic acid-dependent CaPR1, but also by activating dehydration tolerance-related CaOSM1 and CaOSR1 and inducing stomatal closure to reduce water loss in an ABA signaling-dependent manner. Our yeast two hybrid assay showed that CaERF3 interacted with CaPti1, which was confirmed using co-immunoprecipitation, bimolecular fluorescence complementation, and pull-down assays. Chromatin immunoprecipitation and electrophoretic mobility shift assays showed that upon R. solanacearum inoculation, CaPR1, CaOSM1, and CaOSR1 were directly targeted and positively regulated by CaERF3 and potentiated by CaPti1. Additionally, our data indicated that the CaPti1-CaERF3 complex might act downstream of ABA signaling, as exogenously applied ABA did not alter regulation of stomatal aperture by the CaPti1-CaERF3 module. Importantly, the CaPti1-CaERF3 module positively affected pepper growth and the response to dehydration stress. Collectively, the results suggested that immunity and dehydration tolerance are coupled and positively regulated by CaPti1-CaERF3 in pepper plants to enhance resistance against R. solanacearum.

Multilayered synergistic regulation of phytoalexin biosynthesis by ethylene, jasmonate, and MAPK signaling pathways in Arabidopsis.

Camalexin, an indolic antimicrobial metabolite, is the major phytoalexin in Arabidopsis thaliana, and plays a crucial role in pathogen resistance. Our previous studies revealed that the Arabidopsis mitogen-activated protein kinases MPK3 and MPK6 positively regulate pathogen-induced camalexin biosynthesis via phosphoactivating the transcription factor WRKY33. Here, we report that the ethylene and jasmonate (JA) pathways act synergistically with the MPK3/MPK6-WRKY33 module at multiple levels to induce camalexin biosynthesis in Arabidopsis upon pathogen infection. The ETHYLENE RESPONSE FACTOR1 (ERF1) transcription factor integrates the ethylene and JA pathways to induce camalexin biosynthesis via directly upregulating camalexin biosynthetic genes. ERF1 also interacts with and depends on WRKY33 to upregulate camalexin biosynthetic genes, indicating that ERF1 and WRKY33 form transcriptional complexes to cooperatively activate camalexin biosynthetic genes, thereby mediating the synergy of ethylene/JA and MPK3/MPK6 signaling pathways to induce camalexin biosynthesis. Moreover, as an integrator of the ethylene and JA pathways, ERF1 also acts as a substrate of MPK3/MPK6, which phosphorylate ERF1 to increase its transactivation activity and therefore further cooperate with the ethylene/JA pathways to induce camalexin biosynthesis. Taken together, our data reveal the multilayered synergistic regulation of camalexin biosynthesis by ethylene, JA, and MPK3/MPK6 signaling pathways via ERF1 and WRKY33 transcription factors in Arabidopsis.

Phosphorylation of an ethylene response factor by MPK3/MPK6 mediates negative feedback regulation of pathogen-induced ethylene biosynthesis in Arabidopsis

Plants under pathogen attack produce high levels of the gaseous phytohormone ethylene to induce plant defense responses via the ethylene signaling pathway. The 1-aminocyclopropane-1-carboxylate synthase (ACS) is a critical rate-limiting enzyme of ethylene biosynthesis. Transcriptional and post-translational upregulation of ACS2 and ACS6 by the mitogen-activated protein kinases MPK3 and MPK6 are previously shown to be crucial for pathogen-induced ethylene biosynthesis in Arabidopsis. Here, we report that the fungal pathogen Botrytis cinerea-induced ethylene biosynthesis in Arabidopsis is under the negative feedback regulation by ethylene signaling pathway. The ethylene response factor ERF1A is further found to act downstream of ethylene signaling to negatively regulate the B. cinerea-induced ethylene biosynthesis via indirectly suppressing the expression of ACS2 and ACS6. Interestingly, ERF1A is shown to also upregulate defensin genes directly and therefore promote Arabidopsis resistance to B. cinerea. Furthermore, ERF1A is identified to be a substrate of MPK3 and MPK6, which phosphoactivate ERF1A to enhance its functions in suppressing ethylene biosynthesis and inducing defensin gene expression. Taken together, our data reveal that ERF1A and its phosphorylation by MPK3/MPK6 not only mediate the negative-feedback regulation of the B. cinerea-induced ethylene biosynthesis, but also upregulate defensin gene expression to increase Arabidopsis resistance to B. cinerea.

The OsEIL1-OsERF115-target gene regulatory module controls grain size and weight in rice.

SummaryGrain size is one of the essential determinants of rice yield. Our previous studies revealed that ethylene plays an important role in grain‐size control; however, the precise mechanism remains to be determined. Here, we report that the ethylene response factor OsERF115 functions as a key downstream regulator for ethylene‐mediated grain development. OsERF115 encodes an AP2/ERF‐type transcriptional factor that is specifically expressed in young spikelets and developing caryopses. Overexpression of OsERF115 significantly increases grain length, width, thickness and weight by promoting longitudinal elongation and transverse division of spikelet hull cells, as well as enhancing grain‐filling activity, whereas its knockout mutations lead to the opposite effects, suggesting that OsERF115 positively regulates grain size and weight. OsERF115 transcription is strongly induced by ethylene, and OsEIL1 directly binds to the promoter to activate its expression. OsERF115 acts as a transcriptional repressor to directly or indirectly modulate a set of grain‐size genes during spikelet growth and endosperm development. Importantly, haplotype analysis reveals that the SNP variations in the EIN3‐binding sites of OsERF115 promoter are significantly associated with the OsERF115 expression levels and grain weight, suggesting that natural variations in the OsERF115 promoter contribute to grain‐size diversity. In addition, the OsERF115 orthologues are identified only in grass species, implying a conserved and unique role in the grain development of cereal crops. Our results provide insights into the molecular mechanism of ethylene‐mediated grain‐size control and a potential strategy based on the OsEIL1‐OsERF115‐target gene regulatory module for genetic improvement of rice yield.

Ethylene and polyamines form a negative feedback loop to regulate peach fruit ripening via the transcription factor PpeERF113 by regulating the expression of PpePAO1

Ethylene and polyamines (PAs) have antagonistic roles during fruit ripening. PAs affect both ethylene biosynthetic and signal transduction pathways, but the regulatory effects of ethylene on PA metabolism during fruit ripening have remained largely unknown. Here, we found that ethylene enhanced PA catabolism by regulating expression of a peach PA oxidase gene, PpePAO1. Knocking out PpePAO1 delayed peach fruit ripening, whereas PpePAO1 overexpression promoted tomato fruit ripening. Both in vitro and in vivo assays demonstrated that an ethylene response factor (ERF), PpeERF113, activated PpePAO1 by direct binding to the promoter. PA levels were significantly increased and ripening was delayed in PpeERF113 knockdown peach fruit. Correspondingly, PA levels were significantly decreased when PpeERF113 was overexpressed in peach. PpeERF113 was also found to be upregulated in PpePAO1 knockdowns, indicating that PA catabolism has a regulatory function in ethylene signal transduction. Together, our results suggest that ethylene accelerates peach fruit ripening in part by promoting PA catabolism through PpeERF113-mediated regulation of PpePAO1 expression. To our knowledge, this is the first report related to the regulatory mechanism of ethylene in PA catabolism during fruit ripening. This study contributes new understanding of the crosstalk between ethylene and PAs during fruit ripening.

UHPLC-MS/MS and QRT-PCR profiling of PGP agents and Rhizobium spp. of induced phytohormones for growth promotion in mungbean (var. Co4)

In present study, five potential strains with different plant growth promotion (PGP) characteristics were used. By considering various PGP properties of different bacterial strains, several treatments based on various combinations were developed and studied on mungbean (var. Co4). The quantification of the phytohormones was performed on ultrahigh-performance liquid chromatograph coupled to heated electrospray ionization tandem mass spectrometry (UHPLC/HESI-MS/MS). Indole 3-acetic acid (IAA) and Indole 3-butyric acid (IBA) were quantified in positive ionization mode while Gibberellic acid (GA3) and salicylic acid (SA) were quantified in negative ionization mode. Among all the treatments two penta combinations of consortia 1 (Rhizobium + Azospirillum + Pseudomonas + Bacillus spp. + Bacillus licheniformis) and consortia 2 (Rhizobium + Azotobacter + Pseudomonas + Bacillus spp. + Bacillus licheniformis) were found most effective. Higher amount of IAA (1.043 μg g−1), IBA (0.036 μg g−1), GA3 (1.999 μg g−1) and SA (0.098 μg g−1) Fresh weight (FW) were found in treated adolescent root tissues of consortia 2 as compared to consortia 1. Moreover, transcriptional level of the plant hormones were 2–4 fold higher in the relative gene expression study of three genes: ARF (Auxin responsive factors), ERF-IF (Ethylene-responsive Initiation Factors) and GAI (Gibberellic-Acid Insensitive) in consortia 2, on the 15th, 30th and 45th day using quantitative real time-Polymerase chain reaction (qRT-PCR). Furthermore, Yield attributing characters like, the number of nodules plant−1, number of pods plant−1, weight of nodule and seed yield plant−1 were also increased as compared to the control. As a result, the current research elucidated that penta combinations consortium of Rhizobium sp. and rhizobacteria can be developed as a single delivery system biofertilizer for enhancing mungbean productivity.

Investigating the role of the ERF104 gene in stress tolerance in model plant Arabidopsis suecica using CRISPR

ETHYLENE RESPOSNE FACTORS are highly conserved genes that have been found to be involved in stress‐related hormone signaling in plants. When these genes are over‐expressed, they can have negative impacts on plant health, leading to inhibited growth and leaf senescence under stressful conditions. Could knocking down the function of an ERF improve plant response in the face of climate change related stressors such as soil salinity? A CRISPR gene knockdown will be performed on ETHYLENE RESPONSE FACTOR 104 in Arabidopsis suecica. A. suecica is an allopolyploid plant which has shown differential expression of ERF104 in genetically different accessions. A CRISPR gene knockout of ERF104 will be performed on two different accessions of A. suecica, which will then be genotyped to look for signs of genetic mutation. Phenotypic observations of mutation will also be performed as the plants grow and are exposed to stressful conditions. Upon successful completion of the experiment, I expect to see that the ERF104gene is no longer being overexpressed in A. suecica, which should result in the plant showing fewer signs of stress when grown on salinity plates.

MtEFD and MtEFD2: Two transcription factors with distinct neofunctionalization in symbiotic nodule development.

Rhizobium-legume nitrogen-fixing symbiosis involves the formation of a specific organ, the root nodule, which provides bacteria with the proper cellular environment for atmospheric nitrogen fixation. Coordinated differentiation of plant and bacterial cells is an essential step of nodule development, for which few transcriptional regulators have been characterized. Medicago truncatula ETHYLENE RESPONSE FACTOR REQUIRED FOR NODULE DIFFERENTIATION (MtEFD) encodes an APETALA2/ETHYLENE RESPONSIVE FACTOR (ERF) transcription factor, the mutation of which leads to both hypernodulation and severe defects in nodule development. MtEFD positively controls a negative regulator of cytokinin signaling, the RESPONSE REGULATOR 4 (MtRR4) gene. Here we showed that that the Mtefd-1 mutation affects both plant and bacterial endoreduplication in nodules, as well as the expression of hundreds of genes in young and mature nodules, upstream of known regulators of symbiotic differentiation. MtRR4 expressed with the MtEFD promoter complemented Mtefd-1 hypernodulation but not the nodule differentiation phenotype. Unexpectedly, a nonlegume homolog of MtEFD, AtERF003 in Arabidopsis (Arabidopsis thaliana), could efficiently complement both phenotypes of Mtefd-1, in contrast to the MtEFD paralog MtEFD2 expressed in the root and nodule meristematic zone. A domain swap experiment showed that MtEFD2 differs from MtEFD by its C-terminal fraction outside the DNA binding domain. Furthermore, clustered regularly interspaced short palindromic repeats-CRISPR associated protein 9 (CRISPR-Cas9)mutagenesis of MtEFD2 led to a reduction in the number of nodules formed in Mtefd-1, with downregulation of a set of genes, including notably NUCLEAR FACTOR-YA1 (MtNF-YA1) and MtNF-YB16, which are essential for nodule meristem establishment. We, therefore, conclude that nitrogen-fixing symbiosis recruited two proteins originally expressed in roots, MtEFD and MtEFD2, with distinct functions and neofunctionalization processes for each of them.

Transcriptome Meta-Analysis Associated Targeting Hub Genes and Pathways of Drought and Salt Stress Responses in Cotton (Gossypium hirsutum): A Network Biology Approach.

Abiotic stress tolerance is an intricate feature controlled through several genes and networks in the plant system. In abiotic stress, salt, and drought are well known to limit cotton productivity. Transcriptomics meta-analysis has arisen as a robust method to unravel the stress-responsive molecular network in crops. In order to understand drought and salt stress tolerance mechanisms, a meta-analysis of transcriptome studies is crucial. To confront these issues, here, we have given details of genes and networks associated with significant differential expression in response to salt and drought stress. The key regulatory hub genes of drought and salt stress conditions have notable associations with functional drought and salt stress-responsive (DSSR) genes. In the network study, nodulation signaling pathways 2 (NSP2), Dehydration-responsive element1 D (DRE1D), ethylene response factor (ERF61), cycling DOF factor 1 (CDF1), and tubby like protein 3 (TLP3) genes in drought and tubby like protein 1 (TLP1), thaumatin-like proteins (TLP), ethylene-responsive transcription factor ERF109 (EF109), ETS-Related transcription Factor (ELF4), and Arabidopsis thaliana homeodomain leucine-zipper gene (ATHB7) genes in salt showed the significant putative functions and pathways related to providing tolerance against drought and salt stress conditions along with the significant expression values. These outcomes provide potential candidate genes for further in-depth functional studies in cotton, which could be useful for the selection of an improved genotype of Gossypium hirsutum against drought and salt stress conditions.

Silicon enhances the submergence tolerance of rice by regulating quiescence strategy and alleviating oxidative damage

The safety of rice production under submergence is one of the research hotspots worldwide. Although the effects of silicon (Si) on enhancing plant stress tolerance have been widely investigated, the underlying mechanisms mediated by Si under submergence remains poorly understood. In this study, wild type (WT) and Si-defective mutant (lsi1) rice were chosen to investigate the mechanisms of Si-mediated rice resistance to submergence. Our results showed that Si addition effectively mitigated oxidative damages under submergence by reducing the content of hydrogen peroxide (H2O2) and superoxide (O2.-) in WT rice plants. Moreover, Si treatment increased rice yield by 21.5% for WT rice under submergence. The application of Si significantly inhibited the elongation and internode length in WT rice under submergence, through the synergistic regulation of endogenous hormones ethylene (ET), gibberellic acid (GA) and jasmonic acid (JA). Further investigation showed that the ethylene-responsive factor (ERF) SUB1A gene was expressed under submergence in WT and lsi1 rice, but Si addition did not influence the expression of SUB1A. Interestingly, exogenous Si down-regulated the relative expression levels of Si transporter genes Lsi1 and Lsi2 in WT rice roots by 51.7% and 48.0%, respectively. However, the physiological characteristics and genes expression of lsi1 rice were not affected by Si application regardless of submergence. The present study indicated that Si enhances the submergence tolerance and reduce the adverse effects of yield loss through the removal of reactive oxygen species and the adjustment of quiescence strategy.

Roles of ERF2 in apple fruit cuticular wax synthesis

Ethylene response factors 2 (ERF2) belongs to the AP2/ERF transcription factor family and plays key roles in ethylene signal transduction. The roles of MdERF2 in apple (Malus × domestica) cuticular wax synthesis regulated by ethylene were studied in this work. Result shows that MdERF2 overexpression (ERF2-OE) up-regulated the expression levels of wax-related synthetic genes long-chain acyl-CoA synthetase 2 (MdLACS2), Wax synthase1 (MdWSD1), Eceriferum 4 (MdCER4) and Eceriferum 6 (MdCER6), while has no effect on the transcription of Eceriferum 1 (MdCER1), and shows different regulation effects on the expression levels of mid-chain alkane hydroxylase 1 (MdMAH1) between calli and fruit. Furthermore, different MdERF2 expression levels changed the apparent structure of the apple cuticular wax and positively regulated the total wax accumulation. ERF2-OE also increased alkane and alcohol production but decreased acid and ester contents. Thus, ethylene regulated the synthesis of apple cuticular wax at the transcriptional level through MdERF2, thereby altering the wax structure and composition.

Involvement of ethylene in glutamate-mediated tomato fruit resistance to Alternaria alternata

Recent progress showed the potential of glutamate to regulate stress responses by affecting phytohormones homeostasis, but much remains to be explored. To decipher the role of ethylene in glutamate-induced resistance to Alternaria alternata, ethephon application, ethylene perception inhibitor (1-MCP) experiment and ethylene measurement were pursued. Concurrently, temporal alteration in ethylene biosynthesis and signaling pathways were transcriptionally analyzed in tomato fruit. Our data exhibited that ethylene conferred tomato fruit susceptibility rather than resistance to Alternaria alternata. The enhanced resistance in glutamate treatment was related to decreased ethylene production during the early stages of interaction, accompanied by lower expression levels of SlACS1, SlACS2 and SlACO1 relative to the control. Interestingly, 1-MCP induced amounts of resistance similar to that observed in glutamate treatment, while ethylene receptors (SlETR3 and SlETR4), mitogen-activated protein kinase SlMPK3 and ethylene response factor SlERF1 were down-regulated by glutamate during infection. Collectively, antagonizing the ethylene biosynthesis or signaling might play a pivotal role in glutamate-induced resistance to the necrotrophic pathogen Alternaria alternata.

MicroRNA candidate miRcand137 in apple is induced by Botryosphaeria dothidea for impairing host defense.

MicroRNA (miRNA)-mediated gene silencing is a master gene regulatory pathway in plant-pathogen interactions. The differential accumulation of miRNAs among plant varieties alters the expression of target genes, affecting plant defense responses and causing resistance differences among varieties. Botryosphaeria dothidea is an important phytopathogenic fungus of apple (Malus domestica). Malus hupehensis (Pamp.) Rehder, a wild apple species, is highly resistant, whereas the apple cultivar "Fuji" is highly susceptible. Here, we identified a 22-nt miRNA candidate named miRcand137 that compromises host resistance to B. dothidea infection and whose processing was affected by precursor sequence variation between M. hupehensis and "Fuji." miRcand137 guides the direct cleavage of and produced target-derived secondary siRNA against Ethylene response factor 14 (ERF14), a transcriptional activator of pathogenesis-related homologs that confers disease resistance to apple. We showed that miRcand137 acts as an inhibitor of apple immunity by compromising ERF14-mediated anti-fungal defense and revealed a negative association between miRcand137 expression and B. dothidea sensitivity in both resistant and susceptible apples. Furthermore, MIRCAND137 was transcriptionally activated by the invading fungi but not by the fungal elicitor, implying B. dothidea induced host miRcand137 as an infection strategy. We propose that the inefficient miRcand137 processing in M. hupehensis decreased pathogen-initiated miRcand137 accumulation, leading to higher resistance against B. dothidea.

Untangling the role of ethylene beyond fruit development and ripening: A physiological and molecular perspective focused on the Monilinia-peach interaction

It is already well known that ethylene plays a crucial role in peach fruit growth and ripening, by triggering an unset of biochemical and physiological changes that finally make the fruit attractive for consumption. This said, ethylene is not only responsible for fruit ripening but, in conjunction with other hormones, or key compounds (ROS, polyamines, etc.) is involved in the plant response to numerous abiotic stresses (drought, salt and heat tolerance) as well as the plant/fruit response against certain pathogens. Among peaches, one of the most devastating pathogens is the brown rot causing fungus Monilinia spp. that can affect the fruit both on the field or postharvest. Nonetheless scarce information exists regarding the Monilinia-peach interaction from a physiological and molecular perspective. In this sense, recent studies point out to the importance of ethylene during such interaction, which seems to be dependent on the fruit developmental stage and also on the Monilinia species or even the strain's virulence. Why the fruit or the fungus reacts different to distinct Monilinia species or strains and why such reaction depends on the fruit physiological stage is, however, still elusive. Accordingly, this review aims to shed light on the role of ethylene, alone or through a complex cross-talk with other compounds, not only during peach development and ripening but also during the Monilinia-peach interaction. Based on the available literature, it is clear that not only ethylene biosynthesis but ethylene signaling and the activation of ethylene response factors via ROS may play an essential role during this specific host-pathogen interaction.

PpERF/ABR1 functions as an activator to regulate PpPG expression resulting in fruit softening during storage in peach (Prunus persica)

Fruit softening is a complex process that accelerates postharvest spoilage and shortens the shelf life of fruit. The precise mechanisms underlying fruit softening remain elusive, especially in terms of genetic function. Here, we analyzed in deep the RNA-Seq data of ‘Hongli’ (readily softens after harvest) and ‘Baili’ (slowly softens after harvest) peach fruit. We identified polygalacturonase (PpPG) gene in the ‘plum3’ module as a potential fruit softening biomarker in peach through weighted gene co-expression network analysis (WGCNA). Through transcription factor-target gene regulatory network analysis and molecular approaches, we identified an ethylene response factor (ERF) transcript factor, PpERF/ABR1, as a transcriptional activator that binds directly to the promoter of PpPG. Moreover, transient overexpression of PpERF/ABR1 also enhanced PpPG expression, thereby resulting in fruit softening. These results provide a theoretical basis for the molecular mechanism of ERF in the transcription regulation of cell wall-related genes during fruit softening.

Genome-Wide Expression and Physiological Profiling of Pearl Millet Genotype Reveal the Biological Pathways and Various Gene Clusters Underlying Salt Resistance.

Pearl millet (Pennisetum glaucum L.) is a vital staple food and an important cereal crop used as food, feed, and forage. It can withstand heat and drought due to the presence of some unique genes; however, the mechanism of salt stress has been missing in pearl millet until now. Therefore, we conducted a comparative transcriptome profiling to reveal the differentially expressed transcripts (DETs) associated with salt stress in pearl millet at different time points, such as 1, 3, and 7 h, of salt treatment. The physiological results suggested that salt stress significantly increased proline, malondialdehyde (MDA) content, and hydrogen peroxide (H2O2) in pearl millet at 1, 3, and 7 h of salt treatment. In addition, pearl millet plants regulated the activities of superoxide dismutase, catalase, and peroxidase to lessen the impact of salinity. The transcriptomic results depicted that salt stress upregulated and downregulated the expression of various transcripts involved in different metabolic functions. At 1 and 7 h of salt treatment, most of the transcripts were highly upregulated as compared to the 3 h treatment. Moreover, among commonly enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, the mitogen-activated protein kinase (MAPK) signaling pathway and peroxisome pathway were significantly enriched. The DETs related to hormone signaling (auxins, ethylene, gibberellin, and abscisic acid), kinases, protein modifications, and degradation were also identified, depicting the possible role of hormones and kinases to enhance plant tolerance against salt stress. Furthermore, the transcription factors, such as ethylene-responsive element binding factors (ERF), basic helix-loop-helix (bHLH), HMG box-containing protein (HBP), MADS, myeloblastosis (MYB), and WRKY, were predicted to significantly regulate different transcripts involved in salt stress responses at three different time points. Overall, this study will provide new insights to better understand the salt stress regulation mechanisms in pearl millet to improve its resistance against salinity and to identify new transcripts that control these mechanisms in other cereals.

Hydrogen sulphide (H2 S) in the hidden half: Role in root growth, stress signalling and rhizospheric interactions.

Apart from nitric oxide (NO) and carbon monoxide (CO), hydrogen sulphide (H2 S) has emerged as a potential gasotransmitter that has regulatory roles in root differentiation, proliferation and stress signalling. H2 S metabolism in plants exhibits spatio-temporal differences that are intimately associated with sulphide signalling in the cytosol and other subcellular components, e.g. chloroplast and mitochondria. H2 S biosynthesis in plant organs uses both enzymatic and non-enzymatic pathways. H2 S generation in roots and aerial organs is modulated by developmental phase and changes in environmental stimuli. H2 S has an influential role in root development and in the nodulation process. Studies have revealed that H2 S is a part of the auxin and NO signalling pathways in roots, which induce lateral root formation. At the molecular level, exogenous application of H2 S regulates expression of several transcription factors, viz. LBD (Lateral organ Boundaries Domain), MYB (myeloblastosis) and AP2/ERF (Apetala 2/ Ethylene Response Factor), which stimulate upregulation of PpLBD16 (Lateral organ boundaries domain 16), thereby significantly increasing the number of lateral roots. Concomitantly, H2 S acts as a crucial signalling molecule in roots during various abiotic stresses, e.g. drought, salinity heavy metals (HMs), etc., and augments stress tolerance in plants. Interestingly, extensive crosstalk exists between H2 S, NO, ABA, calcium and ethylene during stress, which escalate plant defence and regulate plant growth and productivity. Hence, the present review will elaborate the role of H2 S in root development, stress alleviation, legume-Rhizobium symbiosis and rhizosphere signalling. The review also examines the mechanism of H2 S-mediated abiotic stress mitigation and cross-talk with other signaling molecules.