Ethylene Response Pathway Research Articles

Chromosome-scale genome assembly clarifies the mechanism of flooding tolerance and evolutionary history of Myricaria laxiflora

Myricaria laxiflora, belonging to the Tamaricaceae family, is a valuable plant resource naturally occurring along the riverbanks of the Yangtze River Basin. In this study, we constructed a chromosome-level M. laxiflora genome which was 1.34 Gb in size, with 12 chromosomes and a contig N50 of 11.69 Mb. In total, 23,719 protein-coding genes were discovered across the M. laxiflora genome. Phylogenetic investigation indicated that Tamaricaceae and Polygonaceae diverged between 65 and 81 million years ago (Mya). M. laxiflora underwent one whole genome duplication (WGD) between 19.74 and 24.60 Mya, and experienced a long terminal repeat (LTR) insertion around 0.15 Mya. These genomic changes may be related to environmental upheavals (such as the uplift of the Qinghai-Tibet Plateau) experienced by M. laxiflora during its evolution, and also may have provided a foundation for its evolution and adaptation. To analyze the flooding adaption mechanism of M. laxiflora, we compared morphological, physiological, and transcriptomic changes under control and submerged conditions. Overall, M. laxiflora was found to adopt a "quiescence" strategy when completely submerged. Specifically, in response to submersion, photosynthesis weakens, growth stagnates, glycolysis is activated, and the antioxidant enzyme system is enhanced. The “ethylene response pathway” was ascertained as a likely regulator of this process, and a MlERF-VII gene (Mla11G026380) may be the key gene regulating this process. Our findings shed light on the underlying mechanisms behind the flooding resistance displayed by M. laxiflora, while also establishing a basis for the conservation of its genetic resources.

Physiological Changes and Transcriptomic Analysis throughout On-Tree Fruit Ripening Process in Persimmon (Diospyros kaki L.).

The involvement of effectors and transcriptional regulators in persimmon fruit maturation has been mostly approached by the literature under postharvest conditions. In order to elucidate the participation of these genes in the on-tree fruit maturation development, we have collected samples from seven persimmon germplasm accessions at different developmental stages until physiological maturation. This study has focused on the expression analysis of 13 genes involved in ethylene biosynthesis and response pathways, as well as the evolution of important agronomical traits such as skin colour, weight, and firmness. Results revealed different gene expression patterns, with genes up- and down-regulated during fruit development progression. A principal component analysis was performed to correlate gene expression with agronomical traits. The decreasing expression of the ethylene biosynthetic genes DkACO1, DkACO2, and DkACS2, in concordance with other sensing (DkERS1) and transduction genes (DkERF18), provides a molecular mechanism for the previously described high production of ethylene in immature detached fruits. On the other side, DkERF8 and DkERF16 are postulated to induce fruit softening and skin colour change during natural persimmon fruit ripening via DkXTH9 and DkPSY activation, respectively. This study provides valuable information for a better understanding of the ethylene signalling pathway and its regulation during on-tree fruit ripening in persimmon.

Plant microbiota controls an alternative root branching regulatory mechanism in plants

The establishment of beneficial interactions with microbes has helped plants to modulate root branching plasticity in response to environmental cues. However, how the plant microbiota harmonizes with plant roots to control their branching is unknown. Here, we show that the plant microbiota influences root branching in the model plant Arabidopsis thaliana. We define that the microbiota's ability to control some stages in root branching can be independent of the phytohormone auxin that directs lateral root development under axenic conditions. In addition, we revealed a microbiota-driven mechanism controlling lateral root development that requires the induction of ethylene response pathways. We show that the microbial effects on root branching can be relevant for plant responses to environmental stresses. Thus, we discovered a microbiota-driven regulatory pathway controlling root branching plasticity that could contribute to plant adaptation to different ecosystems.

ETHYLENE-INSENSITIVE 3-LIKE 2 regulates β-carotene and ascorbic acid accumulation in tomatoes during ripening.

ETHYLENE-INSENSITIVE 3/ETHYLENE-INSENSITIVE 3-LIKEs (EIN3/EILs) are important ethylene response factors during fruit ripening. Here, we discovered that EIL2 controls carotenoid metabolism and ascorbic acid (AsA) biosynthesis in tomato (Solanum lycopersicum). In contrast to the red fruits presented in the wild type (WT) 45 d after pollination, the fruits of CRISPR/Cas9 eil2 mutants and SlEIL2 RNA interference lines (ERIs) showed yellow or orange fruits. Correlation analysis of transcriptome and metabolome data for the ERI and WT ripe fruits revealed that SlEIL2 is involved in β-carotene and AsA accumulation. ETHYLENE RESPONSE FACTORs (ERFs) are the typical components downstream of EIN3 in the ethylene response pathway. Through a comprehensive screening of ERF family members, we determined that SlEIL2 directly regulates the expression of 4 SlERFs. Two of these, SlERF.H30 and SlERF.G6, encode proteins that participate in the regulation of LYCOPENE-β-CYCLASE 2 (SlLCYB2), encoding an enzyme that mediates the conversion of lycopene to carotene in fruits. In addition, SlEIL2 transcriptionally repressed L-GALACTOSE 1-PHOSPHATE PHOSPHATASE 3 (SlGPP3) and MYO-INOSITOL OXYGENASE 1 (SlMIOX1) expression, which resulted in a 1.62-fold increase of AsA via both the L-galactose and myoinositol pathways. Overall, we demonstrated that SlEIL2 functions in controlling β-carotene and AsA levels, providing a potential strategy for genetic engineering to improve the nutritional value and quality of tomato fruit.

DNA hypomethylation mediates flower opening and senescence in sweet osmanthus through auxin and ethylene responsive pathways

Recent studies have shown that DNA methylation is critical for fruit ripening and leaf senescence, but its effect on flowering is unclear. This study investigated the dynamic changes in DNA methylation and its regulatory role on flower opening and senescence of Osmanthus fragrans ‘Liuyejingui’ by whole-genome bisulfite sequencing and transcriptome sequencing at six developmental stages (unopened floral bud to late flower senescence, S1 to S6). In total 3222 differentially methylated regions (DMRs) were identified in senescent flowers, of which 89% were hypomethylated (hypo-DMRs), and the number increased form S1 to S6, suggesting that DNA demethylation is associated with the progression of flowering. Analysis of hypo-DMR-associated up-regulated differentially expressed genes revealed that small auxin up-regulated RNAs (OfSAURs) were the most highly significantly enriched from S3 to S5, and ethylene-responsive transcription factors (OfERFs) were strongly activated in senescent flowers. These results suggested that DNA hypomethylation was important for flower expansion and flower senescence through the auxin and ethylene response pathways. Spray application of a DNA methylation inhibitor to unopened floral buds triggered a premature opening and senescence phenotype by promoting endogenous ethylene production and carotenoid accumulation, and indicated that DNA demethylation mediated flower opening and senescence by activating the expression of OfSAURs and OfERFs. An epigenetic regulatory three-component is proposed, in which flower opening and senescence are regulated by DNA demethylation, mediating the expression of specific gene families, together with the plant hormones auxin and ethylene, to transition the flower into an opening- and senescence-competent state. Data availabilityThe genome and transcriptome sequencing data of S1-S6 in O. fragrans ‘Liuyejingui’ were under the public database of the National Center of Biotechnology Information (NCBI) BioProject under the accession number of PRJNA679852 (https://www.ncbi.nlm.nih.gov/bioproject/PRJNA679852). Whole Genome Bisulfate Sequencing and transcriptome sequencing data in DNA methylation inhibitor 5-azacytidine and ddH2O treatment have been deposited into the public database of NCBI BioProject under the accession number of PRJNA798828 and PRJNA798559.

Induced resistance to Fusarium wilt of banana caused by Tropical Race 4 in Cavendish cv Grand Naine bananas after challenging with avirulent Fusarium spp.

In the last century, Fusarium wilt of banana (FWB) destroyed the banana cultivar Gros Michel. The Cavendish cultivars saved the global banana industry, and currently they dominate global production (~50%) and the export trade (~95%). However, a new strain called Tropical Race 4 (TR4) surfaced in the late 1960’s, spread globally and greatly damages Cavendish plantations as well as manifold local varieties that are primarily grown by small holders. Presently, there is no commercially available replacement for Cavendish and hence control strategies must be developed and implemented to manage FWB. Here, we studied whether it is possible to induce resistance to TR4 by pre-inoculations with different Fusarium spp. Only pre-treatments with an avirulent Race 1 strain significantly reduced disease development of TR4 in a Cavendish genotype and this effect was stable at various nutritional and pH conditions. We then used transcriptome analysis to study the molecular basis of this response. Several genes involved in plant defence responses were up-regulated during the initial stages of individual infections with TR4 and Race 1, as well as in combined treatments. In addition, a number of genes in the ethylene and jasmonate response pathways as well as several gibberellin synthesis associated genes were induced. We observed upregulation of RGA2 like genes in all treatments. Hence, RGA2 could be a key factor involved in both R1 and TR4 resistance. The data support the hypothesis that activating resistance to Race 1 in Cavendish bananas affects TR4 development and provide a first insight of gene expression during the interaction between various Fusarium spp. and banana.

Accumulation of the gaseous hormone ethylene helps roots sense compact soil.

Soil compaction, in which soil grains are pressed together leaving less pore space for air and water, is a persistent problem in mechanized agriculture. Most plant roots fail to penetrate soil if it is too dense. One might assume that they are physically unable to penetrate the compact soil. However, new research demonstrates a more complex mechanism that requires the build-up of the volatile plant hormone ethylene in the rhizosphere1. Ethylene itself can arrest growth and, in compact soil, it is present in higher concentrations near roots due to its reduced ability to diffuse. Roots that lack the ethylene response pathway grow better through compact soil, demonstrating that it is physically possible to do so. The work suggests new levers for crop improvement in increasingly degraded soils.

Role of ethylene in effective establishment of the peanut-bradyrhizobia symbiotic interaction.

Ethylene has been implicated in nitrogen fixing symbioses in legumes, where rhizobial invasion occurs via infection threads (IT). In the symbiosis between peanut (Arachis hypogaea L.) and bradyrhizobia, the bacteria penetrate the root cortex intercellularly and IT are not formed. Little attention has been paid to the function of ethylene in the establishment of this symbiosis. The aim of this article is to evaluate whether ethylene plays a role in the development of this symbiotic interaction and the participation of Nod Factors (NF) in the regulation of ethylene signalling. Manipulation of ethylene in peanut was accomplished by application of 1-aminocyclopropane-1-carboxylic acid (ACC), which mimics applied ethylene, or AgNO3, which blocks ethylene responses. To elucidate the participation of NF in the regulation of ethylene signalling, we inoculated plants with a mutant isogenic rhizobial strain unable to produce NF and evaluated the effect of AgNO3 on gene expression of NF and ethylene responsive signalling pathways. Data revealed that ethylene perception is required for the formation of nitrogen-fixing nodules, while addition of ACC does not affect peanut symbiotic performance. This phenotypic evidence is in agreement with transcriptomic data from genes involved in symbiotic and ethylene signalling pathways. NF seem to modulate the expression of ethylene signalling genes. Unlike legumes infected through IT formation, ACC addition to peanut does not adversely affect nodulation, but ethylene perception is required for establishment of this symbiosis. Evidence for the contribution of NF to the modulation of ethylene-inducible defence gene expression is provided.

The RING E3 ligase SDIR1 destabilizes EBF1/EBF2 and modulates the ethylene response to ambient temperature fluctuations in Arabidopsis

The gaseous phytohormone ethylene mediates numerous aspects of plant growth and development as well as stress responses. The F-box proteins EIN3-binding F-box protein 1 (EBF1) and EBF2 are key components that ubiquitinate and degrade the master transcription factors ethylene insensitive 3 (EIN3) and EIN3-like 1 (EIL1) in the ethylene response pathway. Notably, EBF1 and EBF2 themselves undergo the 26S proteasome-mediated proteolysis induced by ethylene and other stress signals. However, despite their importance, little is known about the mechanisms regulating the degradation of these proteins. Here, we show that a really interesting new gene (RING)-type E3 ligase, salt- and drought-induced ring finger 1 (SDIR1), positively regulates the ethylene response and promotes the accumulation of EIN3. Further analyses indicate that SDIR1 directly interacts with EBF1/EBF2 and targets them for ubiquitination and proteasome-dependent degradation. We show that SDIR1 is required for the fine tuning of the ethylene response to ambient temperature changes by mediating temperature-induced EBF1/EBF2 degradation and EIN3 accumulation. Thus, our work demonstrates that SDIR1 functions as an important modulator of ethylene signaling in response to ambient temperature changes, thereby enabling plant adaptation under fluctuating environmental conditions.

Pleiotropic function of the SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE gene TaSPL14 in wheat plant architecture

Main conclusionThe function of SQUAMOSA PROMOTER-BINDING PROTEIN-BOX gene TaSPL14 in wheat is similar to that of OsSPL14 in rice in regulating plant height, panicle length, spikelet number, and thousand-grain weight of wheat, but differs during tiller development. TaSPL14 may regulate spike development via ethylene-response gene EIN3-LIKE 1 (TaEIL1), ETHYLENE-RESPONSIVE TRANSCRIPTION FACTOR 2.11 (TaRAP2.11), and ETHYLENE-RESPONSIVE TRANSCRIPTION FACTOR 1 (TaERF1), but not DENSE AND ERECT PANICLE 1 (TaDEP1) in wheat.The SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE gene OsSPL14 from rice is considered to be a major determinant of ideal plant architecture consisting of few unproductive tillers, more grains per spike, and high resistance of stems to lodging. However, the function of its orthologous gene, TaSPL14, in wheat is unknown. Here, we reported the functional similarities and differences between TaSPL14 and OsSPL14. Similar to OsSPL14 knock-outs in rice, wheat TaSPL14 knock-out plants exhibited decreased plant height, panicle length, spikelet number, and thousand-grain weight. In contrast to OsSPL14, however, TaSPL14 did not affect tiller number. Transcriptome analysis revealed that the expression of genes related to ethylene response was significantly decreased in young spikes of TaSPL14 knock-out lines as compared with wild type. TaSPL14 directly binds to the promoters of the ethylene-response genes TaEIL1, TaRAP2.11, and TaERF1, and promotes their expression, suggesting that TaSPL14 might regulate wheat spike development via the ethylene-response pathway. The elucidation of TaSPL14 will contribute to understanding of the molecular mechanisms that underlie wheat plant architecture.

CLA4 regulates leaf angle through multiple hormone signaling pathways in maize.

Leaf angle is an important agronomic trait in cereals and shares a close relationship with crop architecture and grain yield. Although it has been previously reported that ZmCLA4 can influence leaf angle, the underlying mechanism remains unclear. In this study, we used the Gal4-LexA/UAS system and transactivation analysis to demonstrate in maize (Zea mays) that ZmCLA4 is a transcriptional repressor that regulates leaf angle. DNA affinity purification sequencing (DAP-Seq) analysis revealed that ZmCLA4 mainly binds to promoters containing the EAR motif (CACCGGAC) as well as to two other motifs (CCGARGS and CDTCNTC) to inhibit the expression of its target genes. Further analysis of ZmCLA4 target genes indicated that ZmCLA4 functions as a hub of multiple plant hormone signaling pathways: ZmCLA4 was found to directly bind to the promoters of multiple genes including ZmARF22 and ZmIAA26 in the auxin transport pathway, ZmBZR3 in the brassinosteroid signaling pathway, two ZmWRKY genes involved in abscisic acid metabolism, ZmCYP genes (ZmCYP75B1, ZmCYP93D1) related to jasmonic acid metabolism, and ZmABI3 involved in the ethylene response pathway. Overall, our work provides deep insights into the ZmCLA4 regulatory network in controlling leaf angle in maize.

SlymiR482e‐3p mediates tomato wilt disease by modulating ethylene response pathway

<i>Sly</i>miR482e‐3p mediates tomato wilt disease by modulating ethylene response pathway

Glyoxylic acid overcomes 1-MCP-induced blockage of fruit ripening in Pyrus communis L. var. \u2018D\u2019Anjou\u2019

1-methylcyclopropene (1-MCP) in an ethylene receptor antagonist that blocks ethylene perception and downstream ripening responses in climacteric fruit imparting a longer shelf life. However, in European pear, the application of 1-MCP irreversibly obstructs the onset of system 2 ethylene production resulting in perpetually unripe fruit with undesirable quality. Application of exogenous ethylene, carbon dioxide and treatment to high temperatures is not able to reverse the blockage in ripening. We recently reported that during cold conditioning, activation of alternative oxidase (AOX) occurs pre-climacterically. In this study, we report that activation of AOX via exposure of 1-MCP treated ‘D’Anjou’ pear fruit to glyoxylic acid triggers an accelerated ripening response. Time course physiological analysis revealed that ripening is evident from decreased fruit firmness and increased internal ethylene. Transcriptomic and functional enrichment analyses revealed genes and ontologies implicated in glyoxylic acid-mediated ripening, including AOX, TCA cycle, fatty acid metabolism, amino acid metabolism, organic acid metabolism, and ethylene-responsive pathways. These observations implicate the glyoxylate cycle as a biochemical hub linking multiple metabolic pathways to stimulate ripening through an alternate mechanism. The results provide information regarding how blockage caused by 1-MCP may be circumvented at the metabolic level, thus opening avenues for consistent ripening in pear and possibly other fruit.

Ascorbate oxidation activates systemic defence against root-knot nematode Meloidogyne graminicola in rice.

Ascorbic acid (AA) is the major antioxidant buffer produced in the shoot tissue of plants. Previous studies on root-knot nematode (RKN; Meloidogyne graminicola)-infected rice (Oryza sativa) plants showed differential expression of AA-recycling genes, although their functional role was unknown. Our results confirmed increased dehydroascorbate (DHA) levels in nematode-induced root galls, while AA mutants were significantly more susceptible to nematode infection. External applications of ascorbate oxidase (AO), DHA, or reduced AA, revealed systemic effects of ascorbate oxidation on rice defence versus RKN, associated with a primed accumulation of H2O2 upon nematode infection. To confirm and further investigate these systemic effects, a transcriptome analysis was done on roots of foliar AO-treated plants, revealing activation of the ethylene (ET) response and jasmonic acid (JA) biosynthesis pathways in roots, which was confirmed by hormone measurements. Activation of these pathways by methyl-JA, or ethephon treatment can complement the susceptibility phenotype of the rice Vitamin C (vtc1) mutant. Experiments on the jasmonate signalling (jar1) mutant or using chemical JA/ET inhibitors confirm that the effects of ascorbate oxidation are dependent on both the JA and ET pathways. Collectively, our data reveal a novel pathway in which ascorbate oxidation induces systemic defence against RKNs.

Ethylene Biosynthesis Inhibition Combined with Cyanide Degradation Confer Resistance to Quinclorac in Echinochloa crus-galli var. mitis.

Echinochloa crus-galli var. mitis has rarely been reported for herbicide resistance, and no case of quinclorac resistance has been reported so far. Synthetic auxin-type herbicide quinclorac is used extensively to control rice weeds worldwide. A long history of using quinclorac in Chinese rice fields escalated the resistance in E. crus-galli var. mitis against this herbicide. Bioassays in Petri plates and pots exhibited four biotypes that evolved into resistance to quinclorac ranking as JS01-R > AH01-R > JS02-R > JX01-R from three provinces of China. Ethylene production in these biotypes was negatively correlated with resistance level and positively correlated with growth inhibition. Determination of the related ethylene response pathway exhibited resistance in biotypes that recorded a decline in 1-aminocyclopropane-1-carboxylic acid (ACC) content, ACC synthase oxidase activities, and less inducible ACS and ACO genes expressions than the susceptible biotype, suggesting that there was a positive correlation between quinclorac resistance and ethylene biosynthesis inhibition. Cyanides produced during the ethylene biosynthesis pathway mainly degraded by the activity of β-cyanoalanine synthase (β-CAS). Resistant biotypes exhibited higher β-CAS activity than the susceptible ones. Nucleotide changes were found in the EcCAS gene of resistant biotypes as compared to sensitive ones that caused three amino acid substitutions (Asn-105-Lys, Gln-195-Glu, and Gly-298-Val), resulting in alteration of enzyme structure, increased binding residues in the active site with its cofactor, and decreased binding free energy; hence, its activity was higher in resistant biotypes. Moreover, these mutations increased the structural stability of the enzyme. In view of the positive correlation between ethylene biosynthesis inhibition and cyanide degradation with resistance level, it is concluded that the alteration in ethylene response pathway or at least variation in ACC synthase and ACC oxidase enzyme activities—due to less relative expression of ACS and ACO genes and enhanced β-CAS activity, as well as mutation and increased relative expression of EcCAS gene—can be considered as a probable mechanism of quinclorac resistance in E. crus-galli var. mitis.

Light Modulates Ethylene Synthesis, Signaling, and Downstream Transcriptional Networks to Control Plant Development.

The inhibition of hypocotyl elongation by ethylene in dark-grown seedlings was the basis of elegant screens that identified ethylene-insensitive Arabidopsis mutants, which remained tall even when treated with high concentrations of ethylene. This simple approach proved invaluable for identification and molecular characterization of major players in the ethylene signaling and response pathway, including receptors and downstream signaling proteins, as well as transcription factors that mediate the extensive transcriptional remodeling observed in response to elevated ethylene. However, the dark-adapted early developmental stage used in these experiments represents only a small segment of a plant’s life cycle. After a seedling’s emergence from the soil, light signaling pathways elicit a switch in developmental programming and the hormonal circuitry that controls it. Accordingly, ethylene levels and responses diverge under these different environmental conditions. In this review, we compare and contrast ethylene synthesis, perception, and response in light and dark contexts, including the molecular mechanisms linking light responses to ethylene biology. One powerful method to identify similarities and differences in these important regulatory processes is through comparison of transcriptomic datasets resulting from manipulation of ethylene levels or signaling under varying light conditions. We performed a meta-analysis of multiple transcriptomic datasets to uncover transcriptional responses to ethylene that are both light-dependent and light-independent. We identified a core set of 139 transcripts with robust and consistent responses to elevated ethylene across three root-specific datasets. This “gold standard” group of ethylene-regulated transcripts includes mRNAs encoding numerous proteins that function in ethylene signaling and synthesis, but also reveals a number of previously uncharacterized gene products that may contribute to ethylene response phenotypes. Understanding these light-dependent differences in ethylene signaling and synthesis will provide greater insight into the roles of ethylene in growth and development across the entire plant life cycle.

CHITINASE LIKE1 Regulates Root Development of Dark-Grown Seedlings by Modulating Ethylene Biosynthesis in Arabidopsis thaliana.

The plant hormone ethylene plays a regulatory role in development in light- and dark-grown seedlings. We previously isolated a group of small-molecule compounds with a quinazolinone backbone, which were named acsinones (for ACC synthase inhibitor quinazolinones), that act as uncompetitive inhibitors of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS). Thus, the triple response phenotype, which consists of shortened hypocotyls and roots, radial swelling of hypocotyls and exaggerated curvature of apical hooks, was suppressed by acsinones in dark-grown (etiolated) ethylene overproducer (eto) seedlings. Here, we describe our isolation and characterization of an Arabidopsis revert to eto1 9 (ret9) mutant, which showed reduced sensitivity to acsinones in etiolated eto1 seedlings. Map-based cloning of RET9 revealed an amino acid substitution in CHITINASE LIKE1 (CTL1), which is required for cell wall biogenesis and stress resistance in Arabidopsis. Etiolated seedlings of ctl1ret9 showed short hypocotyls and roots, which were augmented in combination with eto1-4. Consistently, ctl1ret9 seedlings showed enhanced sensitivity to exogenous ACC to suppress primary root elongation as compared with the wild type. After introducing ctl1ret9 to mutants completely insensitive to ethylene, genetic analysis indicated that an intact ethylene response pathway is essential for the alterations in root and apical hook but not hypocotyl in etiolated ctl1ret9 seedlings. Furthermore, a mild yet significantly increased ethylene level in ctl1 mutants was related to elevated mRNA level and activity of ACC oxidase (ACO). Moreover, genes associated with ethylene biosynthesis (ACO1 and ACO2) and response (ERF1 and EDF1) were upregulated in etiolated ctl1ret9 seedlings. By characterizing a new recessive allele of CTL1, we reveal that CTL1 negatively regulates ACO activity and the ethylene response, which thus contributes to understanding a role for ethylene in root elongation in response to perturbed cell wall integrity.

AtCSLD3 and GhCSLD3 mediate root growth and cell elongation downstream of the ethylene response pathway in Arabidopsis.

CSLD3, a gene of the cellulose synthase-like D family, affects root hair elongation, but its interactions with ethylene signaling and phosphate-starvation are poorly understood. Here, we aim to understand the role of CSLD3 in the context of the ethylene signaling and phosphate starvation pathways in Arabidopsis plant growth. Therefore, we performed a comparative analysis of the csld3-1 mutant, CSLD3-overexpressing lines, and ethylene-response mutants, such as the constitutive ethylene-response mutant i-ctr1. We found that CSLD3 overexpression enhanced root and hypocotyl growth by increasing cell elongation, and that the root growth was highly sensitive to ethylene treatment (1 µM ACC), in particular under phosphate starvation. However, the CSLD3-mediated hypocotyl elongation occurred independently of the ethylene signaling pathway. Notably, the typical induction of root hair and root elongation by ethylene and phosphate-starvation was completely abolished in the csld3-1 mutant. Furthermore, i-ctr1 csld3-1 double-mutants were hairless like the csld3-1 parent, confirming that CSLD3 acts downstream of the ethylene signaling pathway during root growth. Moreover, the CSLD3 levels positively correlated with cellulose levels, indicating a role of CSLD3 in cellulose synthesis, which may explain the observed growth effects. Our results establish how CSLD3 works in the context of the ethylene signaling and phosphate-starvation pathways during root hair growth, cell elongation, and cell wall biosynthesis.

Characterization and potential of plant growth promoting rhizobacteria isolated from native Andean crops.

Bacteria isolated from soil and rhizosphere samples collected in Peru from Andean crops were tested in vitro and in vivo to determine their potential as plant growth promoters and their ability to induce systemic resistance to Alternaria alternata in tomato plants. The isolates were identified by sequencing their 16S ribosomal RNA gene. Test for phosphate solubilization, and indolacetic acid were also carried out, together with in vitro antagonism assays in dual cultures towards the plant pathogens Fusarium solani, A. alternata and Curvularia lunata. The three most promising isolates (Pa15, Ps155, Ps168) belonged to the genus Pseudomonas. Further assays were carried out with tomato plants to assess their plant protection effect towards A. alternata and as growth promoters. Inoculation of tomato seeds with all isolates significantly enhanced seed germination, plantlets emergence and plant development. Bacterial inoculation also reduce damage level caused by A. alternata. The expression levels of three tomato genes involved in the jasmonate (AOS), ethylene responsive (ERF-2) and pathogenesis related (PR-P2) pathways were determined in plants challenged with A. alternata, alone or with each bacterial isolate, respectively. Results showed that at 24h after infection, in absence of the pathogen, the expression level of the tested genes was very low. The presence of A. alternata alone and in combination with bacteria increased the transcripts of all genes. Data showed a potential of best performing isolate Ps168 to sustain tomato plants nutrition and activate defense-related genes for protection by pathogenic fungi.

Different Preclimacteric Events in Apple Cultivars with Modified Ripening Physiology

“Anna” is an early season apple cultivar exhibiting a fast softening and juiciness loss during storage, in comparison to two mid-late season cultivars “Galaxy” and “GD.” The poor storage capacity of “Anna” was correlated with high lipid oxidation-related autoluminescence, high respiration and ethylene production rates, associated with high expression of MdACO1, 2, 4, 7, and MdACS1. All cultivars at harvest responded to exogenous ethylene by enhancing ethylene production, typical of system-II. The contribution of pre-climacteric events to the poor storage capacity of “Anna” was examined by comparing respiration and ethylene production rates, response to exogenous ethylene, expression of genes responsible for ethylene biosynthesis and response, and developmental regulators in the three cultivars throughout fruit development. In contrast to the “Galaxy” and “GD,” “Anna” showed higher ethylene production and respiration rates during fruit development, and exhibited auto-stimulatory (system II-like) effect in response to exogenous ethylene. The higher ethylene production rate in “Anna” was correlated with higher expression of ethylene biosynthesis genes, MdACS3a MdACO2, 4, and 7 during early fruit development. The expression of negative regulators of ripening (AP2/ERF) and ethylene response pathway, (MdETR1,2 and MdCTR1) was lower in “Anna” in comparison to the other two cultivars throughout development and ripening. Similar pattern of gene expression was found for SQUAMOSA promoter binding protein (SBP)-box genes, including MdCNR and for MdFUL. Taken together, this study provides new understanding on pre-climacteric events in “Anna” that might affect its ripening behavior and physiology following storage.