How the Ethylene Biosynthesis Pathway of Semi-Halophytes Is Modified with Prolonged Salinity Stress Occurrence?

The mechanism of ethylene (ET)-regulated salinity stress response remains largely unexplained, especially for semi-halophytes and halophytes. Here, we present the results of the multifaceted analysis of the model semi-halophyte Mesembryanthemum crystallinum L. (common ice plant) ET biosynthesis pathway key components' response to prolonged (14 days) salinity stress. Transcriptomic analysis revealed that the expression of 3280 ice plant genes was altered during 14-day long salinity (0.4 M NaCl) stress. A thorough analysis of differentially expressed genes (DEGs) showed that the expression of genes involved in ET biosynthesis and perception (ET receptors), the abscisic acid (ABA) catabolic process, and photosynthetic apparatus was significantly modified with prolonged stressor presence. To some point this result was supported with the expression analysis of the transcript amount (qPCR) of key ET biosynthesis pathway genes, namely ACS6 (1-aminocyclopropane-1-carboxylate synthase) and ACO1 (1-aminocyclopropane-1-carboxylate oxidase) orthologs. However, the pronounced circadian rhythm observed in the expression of both genes in unaffected (control) plants was distorted and an evident downregulation of both orthologs' was induced with prolonged salinity stress. The UPLC-MS analysis of the ET biosynthesis pathway rate-limiting semi-product, namely of 1-aminocyclopropane-1-carboxylic acid (ACC) content, confirmed the results assessed with molecular tools. The circadian rhythm of the ACC production of NaCl-treated semi-halophytes remained largely unaffected by the prolonged salinity stress episode. We speculate that the obtained results represent an image of the steady state established over the past 14 days, while during the first hours of the salinity stress response, the view could be completely different.

Shade stress triggers ethylene biosynthesis to accelerate soybean senescence and impede nitrogen remobilization.

In gramineae-soybean intercropping systems, shade stress caused by taller plants impacts soybean growth specifically during the reproductive stage. However, the effects of shade stress on soybean senescence remain largely unexplored. In this research, we applied artificial shade treatments with intensities of 75% (S75) and 50% (S50) to soybean plants at the onset of flowering to simulate the shade stress experienced by soybeans in the traditional and optimized maize-soybean intercropping systems, respectively. Compared to the normal light control, both shade treatments led to a rapid decline in the dry matter content of soybean vegetative organs and accelerated their abscission. Moreover, shade treatments triggered the degradation of chlorophyll and soluble proteins in leaves and increased the expression of genes associated with leaf senescence. Metabolic profiling further revealed that ethylene biosynthesis and signal transduction were induced by shade treatment. In addition, the examination of nitrogen content demonstrated that shade treatments impeded the remobilization of nitrogen in vegetative tissues, consequently reducing the seed nitrogen harvest. It's worth noting that these negative effects were less pronounced under the S50 treatment compared to the S75 treatment. Taken together, this research demonstrates that shade stress during the reproductive stage accelerates soybean senescence and impedes nitrogen remobilization, while optimizing the field layout to improve soybean growth light conditions could mitigate these challenges in the maize-soybean intercropping system.

The Role of Plant Hormones in the Interaction of Colletotrichum Species with Their Host Plants.

Colletotrichum is a plant pathogenic fungus which is able to infect virtually every economically important plant species. Up to now no common infection mechanism has been identified comparing different plant and Colletotrichum species. Plant hormones play a crucial role in plant-pathogen interactions regardless whether they are symbiotic or pathogenic. In this review we analyze the role of ethylene, abscisic acid, jasmonic acid, auxin and salicylic acid during Colletotrichum infections. Different Colletotrichum strains are capable of auxin production and this might contribute to virulence. In this review the role of different plant hormones in plant-Colletotrichum interactions will be discussed and thereby auxin biosynthetic pathways in Colletotrichum spp. will be proposed.

Regulation of wound ethylene biosynthesis by NAC transcription factors in kiwifruit.

BACKGROUND: The phytohormone ethylene controls many processes in plant development and acts as a key signaling molecule in response to biotic and abiotic stresses: it is rapidly induced by flooding, wounding, drought, and pathogen attack as well as during abscission and fruit ripening. In kiwifruit (Actinidia spp.), fruit ripening is characterized by two distinct phases: an early phase of system-1 ethylene biosynthesis characterized by absence of autocatalytic ethylene, followed by a late burst of autocatalytic (system-2) ethylene accompanied by aroma production and further ripening. Progress has been made in understanding the transcriptional regulation of kiwifruit fruit ripening but the regulation of system-1 ethylene biosynthesis remains largely unknown. The aim of this work is to better understand the transcriptional regulation of both systems of ethylene biosynthesis in contrasting kiwifruit organs: fruit and leaves. RESULTS: A detailed molecular study in kiwifruit (A. chinensis) revealed that ethylene biosynthesis was regulated differently between leaf and fruit after mechanical wounding. In fruit, wound ethylene biosynthesis was accompanied by transcriptional increases in 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS), ACC oxidase (ACO) and members of the NAC class of transcription factors (TFs). However, in kiwifruit leaves, wound-specific transcriptional increases were largely absent, despite a more rapid induction of ethylene production compared to fruit, suggesting that post-transcriptional control mechanisms in kiwifruit leaves are more important. One ACS member, AcACS1, appears to fulfil a dominant double role; controlling both fruit wound (system-1) and autocatalytic ripening (system-2) ethylene biosynthesis. In kiwifruit, transcriptional regulation of both system-1 and -2 ethylene in fruit appears to be controlled by temporal up-regulation of four NAC (NAM, ATAF1/2, CUC2) TFs (AcNAC1-4) that induce AcACS1 expression by directly binding to the AcACS1 promoter as shown using gel-shift (EMSA) and by activation of the AcACS1 promoter in planta as shown by gene activation assays combined with promoter deletion analysis. CONCLUSIONS: Our results indicate that in kiwifruit the NAC TFs AcNAC2-4 regulate both system-1 and -2 ethylene biosynthesis in fruit during wounding and ripening through control of AcACS1 expression levels but not in leaves where post-transcriptional/translational regulatory mechanisms may prevail.

A guanidine-degrading enzyme controls genomic stability of ethylene-producing cyanobacteria.

Recent studies have revealed the prevalence and biological significance of guanidine metabolism in nature. However, the metabolic pathways used by microbes to degrade guanidine or mitigate its toxicity have not been widely studied. Here, via comparative proteomics and subsequent experimental validation, we demonstrate that Sll1077, previously annotated as an agmatinase enzyme in the model cyanobacterium Synechocystis sp. PCC 6803, is more likely a guanidinase as it can break down guanidine rather than agmatine into urea and ammonium. The model cyanobacterium Synechococcus elongatus PCC 7942 strain engineered to express the bacterial ethylene-forming enzyme (EFE) exhibits unstable ethylene production due to toxicity and genomic instability induced by accumulation of the EFE-byproduct guanidine. Co-expression of EFE and Sll1077 significantly enhances genomic stability and enables the resulting strain to achieve sustained high-level ethylene production. These findings expand our knowledge of natural guanidine degradation pathways and demonstrate their biotechnological application to support ethylene bioproduction.

Verticillium dahliae secretory effector PevD1 induces leaf senescence by promoting ORE1-mediated ethylene biosynthesis.

Leaf senescence, the final stage of leaf development, is influenced by numerous internal and environmental signals. However, how biotic stresses such as pathogen infection regulate leaf senescence remains largely unclear. In this study, we found that the premature leaf senescence in Arabidopsis caused by the soil-borne vascular fungus Verticillium dahliae was impaired by disruption of a protein elicitor from V. dahliae 1 named PevD1. Constitutive or inducible overexpression of PevD1 accelerated Arabidopsis leaf senescence. Interestingly, a senescence-associated NAC transcription factor, ORE1, was targeted by PevD1. PevD1 could interact with and stabilize ORE1 protein by disrupting its interaction with the RING-type ubiquitin E3 ligase NLA. Mutation of ORE1 suppressed the premature senescence caused by overexpressing PevD1, whereas overexpression of ORE1 or PevD1 led to enhanced ethylene production and thereby leaf senescence. We showed that ORE1 directly binds the promoter of ACS6 and promotes its expression for mediating PevD1-induced ethylene biosynthesis. Loss-of-function of ACSs could suppress V. dahliae-induced leaf senescence in ORE1-overexpressing plants. Furthermore, we found thatPevD1 also interacts with Gossypium hirsutum ORE1 (GhORE1) and that virus-induced gene silencing of GhORE1 delays V. dahliae-triggered leaf senescence in cotton, indicating a possibly conserved mechanism in plants. Taken together, these results suggest that V. dahliae induces leaf senescence by secreting the effector PevD1 to manipulate the ORE1-ACS6 cascade, providing new insights into biotic stress-induced senescence in plants.

Salicylic acid modulates ACS, NHX1, sos1 and HKT1;2 expression to regulate ethylene overproduction and Na+ ions toxicity that leads to improved physiological status and enhanced salinity stress tolerance in tomato plants cv. Pusa Ruby.

Tomato is an important crop for its high nutritional and medicinal properties. The role of salicylic acid (SA) in 1-aminocyclopropane-1-carboxylate synthase (ACS), sodium-hydrogen exchanger (NHX1), salt overly sensitive 1 (sos1) and high-affinity K+ transporter (HKT1;2) transcripts, and ACS enzyme activity and ethylene (ET) production, and growth and physiological attributes was evaluated in tomato cv. Pusa Ruby under salinity stress. Thirty days-old seedlings treated with 0 mM NaCl, 250 mM NaCl, 250 mM NaCl plus 100 µM SA were assessed for different growth and physiological parameters at 45 DAS. Results showed ACS, NHX1, sos1 and HKT1;2 transcripts were significantly changed in SA treated plants. The ACS enzyme activity and ET content were considerably decreased in SA treated plants. Shoot length (SL), root length (RL), number of leaves (NL), leaf area per plant (LA), shoot fresh weight (SFW) and root fresh weight (RFW) were also improved under SA treatment. Conversely, the electrolyte leakage and sodium ion (Na+) content were significantly reduced in SA treated plants. In addition, the endogenous proline and potassium ion (K+) content, and K+/Na+ ratio were considerably increased under SA treatment. Likewise, antioxidant enzymes (SOD, CAT, APX and GR) profile were better in SA treated plant. The present findings suggest that SA reverse the negative effects of salinity stress and stress induced ET production by modulating ACS, NHX, sos1 and HKT1;2 transcript level, and improving various growth and physiological parameters, and antioxidants enzymes profile. This will contribute to a better understanding of salinity stress tolerance mechanisms of tomato plants involving SA and ET cross talk and ions homeostasis to develop more tolerant plant.

The regulation of 1-methylcyclopropene treatment on the subfamily genes expression of ethylene response factors in peaches during storage.

BACKGROUND: Ethylene response factors (ERFs) perform diverse functions in fruit development, ripening and senescence. However, the effects of postharvest treatments on ERF genes have not been widely investigated due to the lack of peach ERF genomic information. The aim of this study was to investigate the ERF genes' expression of freshly harvested peach during storage after 1-methylcyclopropene (1-MCP) treatment. METHODS: 10 µL L-1 1-MCP was used to fumigate peaches. Treated peaches and control peaches were stored at 20°C for 9 days. Fruit firmness, ethylene production and the transcript abundance of ERFs were evaluated during storage. RESULTS: 127 AP2/ERF genes were identified genome using RNA-sequencing (RNA-seq). Expression profiles of 39 ERF genes were considered at day 0, 3, 5 and 7. Results showed that 1-MCP inhibited some ERF genes' expression (e.g., Prupe.5G117800), some genes were generally up-regulated responding to 1-MCP (e.g., Prupe.6G039700), while the other ERF genes displayed no significant difference between the two groups (e.g., Prupe.1G130300). CONCLUSIONS: These data revealed that peach ERF genes perform diverse functions during fruit growth, ripening and senescence. The different responses of ERF genes to postharvest 1-MCP treatment may be useful to understand the roles of ethylene and ERF genes in controlling technological aspects of postharvest peach conservation.

Distinct Functions of Ethylene and ACC in the Basal Land Plant Marchantia polymorpha.

Ethylene is a gaseous phytohormone involved in various physiological processes, including fruit ripening, senescence, root hair development and stress responses. Recent genomics studies have suggested that most homologous genes of ethylene biosynthesis and signaling are conserved from algae to angiosperms, whereas the function and biosynthesis of ethylene remain unknown in basal plants. Here, we examined the physiological effects of ethylene, an ethylene precursor, 1-aminocyclopropane-1-carboxylic acid (ACC) and an inhibitor of ethylene perception, silver thiosulfate (STS), in a basal land plant, Marchantia polymorpha. M. polymorpha plants biosynthesized ethylene, and treatment with high concentrations of ACC slightly promoted ethylene production. ACC remarkably suppressed the growth of thalli (vegetative organs) and rhizoids (root-hair-like cells), whereas exogenous ethylene slightly promoted thallus growth. STS suppressed thallus growth and induced ectopic rhizoid formation on the dorsal surface of thalli. Thus, ACC and ethylene have different effects on the vegetative growth of M. polymorpha. We generated single and double mutants of ACC synthase-like (ACSL) genes, MpACSL1 and MpACSL2. The mutants did not show obvious defects in thallus growth, ACC content and ethylene production, indicating that MpACSL genes are not essential for the vegetative growth and biosynthesis of ACC and ethylene. Gene expression analysis suggested the involvement of MpACSL1 and MpACSL2 in stress responses. Collectively, our results imply ethylene-independent function of ACC and the absence of ACC-mediated ethylene biosynthesis in M. polymorpha.

Promotion of tomato growth by the volatiles produced by the hypovirulent strain QT5-19 of the plant gray mold fungus Botrytis cinerea.

Our previous study identified a hypovirulent strain QT5-19 of Botrytis cinerea, the causal agent of the plant gray mold disease, and found that QT5-19 can produce volatile organic compounds (VOCs) with high antifungal activity and high control efficacy against B. cinerea. However, impact of the QT5-19 VOCs on plant growth remains unknown. This study was conducted to investigate the impact of the QT5-19 VOCs on tomato growth, and to elucidate the mechanisms for the plant growth-promoting (PGP) activity of the QT5-19 VOCs. Results showed that compared to the control treatment, the QT5-19 VOCs significantly (P < 0.05) promoted tomato growth, and the PGP activity of the QT5-19 VOCs acted in dose- and time-dependent manners. Results also showed that the values of photosynthetic assimilation, stomatal conductance and transpiration, water use efficiency and chlorophyll content in the treatments of the QT5-19 VOCs were significantly (P < 0.05) higher than the corresponding values in the control treatment. The QT5-19 VOCs up-regulated expression of the genes for expansins (EXP2, EXP9 and EXP18), IAA (SlIAA1, SlIAA3 and SlIAA9), cytokinins (SlCKX1) and gibberellins in leaves and/or roots, whereas down-regulated expression of the gene ACO1 for ethylene in both organs. Moreover, enhanced accumulation of auxins and decreased accumulation of ethylene were observed in tomato roots in the treatment of the QT5-19 VOCs, compared to the control treatment. These results suggest that the QT5-19 VOCs probably promote tomato growth through improving photosynthesis and biosynthesis of expansins and IAA, and reducing ethylene biosynthesis. This study suggests that QT5-19 is a versatile biocontrol control agent.

Auxin-Induced SaARF4 Downregulates SaACO4 to Inhibit Lateral Root Formation in Sedum alfredii Hance.

Lateral root (LR) formation promotes plant resistance, whereas high-level ethylene induced by abiotic stress will inhibit LR emergence. Considering that local auxin accumulation is a precondition for LR generation, auxin-induced genes inhibiting ethylene synthesis may thus be important for LR development. Here, we found that auxin response factor 4 (SaARF4) in Sedum alfredii Hance could be induced by auxin. The overexpression of SaARF4 decreased the LR number and reduced the vessel diameters. Meanwhile, the auxin distribution mode was altered in the root tips and PIN expression was also decreased in the overexpressed lines compared with the wild-type (WT) plants. The overexpression of SaARF4 could reduce ethylene synthesis, and thus, the repression of ethylene production decreased the LR number of WT and reduced PIN expression in the roots. Furthermore, the quantitative real-time PCR, chromatin immunoprecipitation sequencing, yeast one-hybrid, and dual-luciferase assay results showed that SaARF4 could bind the promoter of 1-aminocyclopropane-1-carboxylate oxidase 4 (SaACO4), associated with ethylene biosynthesis, and could downregulate its expression. Therefore, we concluded that SaARF4 induced by auxin can inhibit ethylene biosynthesis by repressing SaACO4 expression, and this process may affect auxin transport to delay LR development.

Gene expression patterns in shoots of Camelina sativa with enhanced salinity tolerance provided by plant growth promoting bacteria producing 1-aminocyclopropane-1-carboxylate deaminase or expression of the corresponding acdS gene.

Growth of plants in soil inoculated with plant growth promoting bacteria (PGPB) producing 1-aminocyclopropane-1-carboxylate (ACC) deaminase or expression of the corresponding acdS gene in transgenic lines reduces the decline in shoot length, shoot weight and photosynthetic capacity triggered by salt stress in Camelina sativa. Reducing the levels of ethylene attenuated the salt stress response as inferred from decreases in the expression of genes involved in development, senescence, chlorosis and leaf abscission that are highly induced by salt to levels that may otherwise have a negative effect on plant growth and productivity. Growing plants in soil treated with Pseudomonas migulae 8R6 negatively affected ethylene signaling, auxin and JA biosynthesis and signalling, but had a positive effect on the regulation of genes involved in GA signaling. In plants expressing acdS, the expression of the genes involved in auxin signalling was positively affected, while the expression of genes involved in cytokinin degradation and ethylene biosynthesis were negatively affected. Moreover, fine-tuning of ABA signaling appears to result from the application of ACC deaminase in response to salt treatment. Moderate expression of acdS under the control of the root specific rolD promoter or growing plants in soil treated with P. migulae 8R6 were more effective in reducing the expression of the genes involved in ethylene production and/or signaling than expression of acdS under the more active Cauliflower Mosaic Virus 35S promoter.

Dimerization of the ETO1 family proteins plays a crucial role in regulating ethylene biosynthesis in Arabidopsis thaliana.

ETHYLENE OVERPRODUCER1 (ETO1), ETO1-LIKE1 (EOL1), and EOL2 are members of the Broad complex, Tramtrack, Bric-a-brac (BTB) protein family that collectively regulate type-2 1-aminocyclopropane-1-carboxylic acid synthase (ACS) activity in Arabidopsis thaliana. Although ETO1 and EOL1/EOL2 encode structurally related proteins, genetic studies suggest that they do not play an equivalent role in regulating ethylene biosynthesis. The mechanistic details underlying the genetic analysis remain elusive. In this study, we reveal that ETO1 collaborates with EOL1/2 to play a key role in the regulation of type-2 ACS activity via protein-protein interactions. ETO1, EOL1, and EOL2 exhibit overlapping but distinct tissue-specific expression patterns. Nevertheless, neither EOL1 nor EOL2 can fully complement the eto1 phenotype under control of the ETO1 promoter, which suggests differential functions of ETO1 and EOL1/EOL2. ETO1 forms homodimers with itself and heterodimers with EOLs. Furthermore, CULLIN3 (CUL3) interacts preferentially with ETO1. The BTB domain of ETO1 is sufficient for interaction with CUL3 and is required for homodimerization. However, domain-swapping analysis in transgenic Arabidopsis suggests that the BTB domain of ETO1 is essential but not sufficient for a full spectrum of ETO1 function. The missense mutation in eto1-5 generates a substitution of phenylalanine with an isoleucine in ETO1F466I that impairs its dimerization and interaction with EOLs but does not affect binding to CUL3 or ACS5. Overexpression of ETO1F466I in Arabidopsis results in a constitutive triple response phenotype in dark-grown seedlings. Our findings reveal the mechanistic role of protein-protein interactions of ETO1 and EOL1/EOL2 that is crucial for their biological function in ethylene biosynthesis.

Glutathione and ethylene biosynthesis reveal that the glume and lemma have better tolerance to water deficit in wheat.

As senescence progresses, the sensitivity of wheat organs to plant hormones during the grain-filling stages cannot be ignored. Especially under water deficit situation, non-leaf organs (spikes) have better photosynthesis and drought-tolerance traits than flag leaves. However, the mechanism of ethylene synthesis in wheat organs under water deficit remains unclear. We have studied the influence of water deficit in wheat flag leaves and spike bracts on photosynthetic parameters and on the expression of key enzymes involved in the ethylene biosynthesis pathway during the late grain-filling stages. More stable chlorophyll content (Chl), maximum PSII quantum yield (Fv/Fm), nonphotochemical quenching (NPQ) and maximal efficiency of PSII photochemistry under light adaptation (Fv'/Fm') were observed in the spike bracts than that in the flag leaves during the late grain-filling stages. In addition, the activity of glutathione reductase (GR), γ-glutamylcysteine synthetase (γ-ECS), 1-aminocyclopropane-1-carboxylic (ACC) acid synthase (ACS), and ACC oxidase (ACO) induced ethylene synthesis and influenced plant growth. Further analysis of genes encoding cysteine-ethylene related proteins (γ-ECS, GR, ACO, ACS1, and ASC2) demonstrated that ear organs and flag leaves exhibited different expression patterns. These findings will facilitate future investigations of the regulatory senescence response mechanisms of cysteine interaction with ethylene in wheat under conditions of drought stress.

Integrated analysis of lncRNA and mRNA transcriptomes reveals the potential regulatory role of lncRNA in kiwifruit ripening and softening.

Kiwifruit has gained increasing attention worldwide for its unique flavor and high nutritional value. Rapid softening after harvest greatly shortens its shelf-life and reduces the commercial value. Therefore, it is imperative and urgent to identify and clarify its softening mechanism. This study aimed to analyze and compare the long noncoding RNA (lncRNA) and mRNA expression patterns in ABA-treated (ABA) and room temperature (RT)-stored fruits with those in freshly harvested fruits (CK) as control. A total of 697 differentially expressed genes (DEGs) and 81 differentially expressed lncRNAs (DELs) were identified while comparing ABA with CK, and 458 DEGs and 143 DELs were detected while comparing RT with CK. The Kyoto Encyclopedia of Genes and Genomes analysis of the identified DEGs and the target genes of DELs revealed that genes involved in starch and sucrose metabolism, brassinosteroid biosynthesis, plant hormone signal transduction, and flavonoid biosynthesis accounted for a large part. The co-localization networks, including 38 DEGs and 31 DELs in ABA vs. CK, and 25 DEGs and 25 DELs in RT vs. CK, were also performed. Genes related to fruit ripening, such as genes encoding ß-galactosidase, mannan endo-1,4-ß-mannosidase, pectinesterase/pectinesterase inhibitor, and NAC transcription factor, were present in the co-localization network, suggesting that lncRNAs were involved in regulating kiwifruit ripening. Notably, several ethylene biosynthesis- and signaling-related genes, including one 1-aminocyclopropane-1-carboxylic acid oxidase gene and three ethylene response factor genes, were found in the co-localization network of ABA vs. CK, suggesting that the promoting effect of ABA on ethylene biosynthesis and fruit softening might be embodied by increasing the expression of these lncRNAs. These results may help understand the regulatory mechanism of lncRNAs in ripening and ABA-induced fruit softening of kiwifruit.

Endogenous Polyamines and Ethylene Biosynthesis in Relation to Germination of Osmoprimed Brassica napus Seeds under Salt Stress.

Currently, seed priming is reported as an efficient and low-cost approach to increase crop yield, which could not only promote seed germination and improve plant growth state but also increase abiotic stress tolerance. Salinity represents one of the most significant abiotic stresses that alters multiple processes in plants. The accumulation of polyamines (PAs) in response to salt stress is one of the most remarkable plant metabolic responses. This paper examined the effect of osmopriming on endogenous polyamine metabolism at the germination and early seedling development of Brassica napus in relation to salinity tolerance. Free, conjugated and bound polyamines were analyzed, and changes in their accumulation were discussed with literature data. The most remarkable differences between the corresponding osmoprimed and unprimed seeds were visible in the free (spermine) and conjugated (putrescine, spermidine) fractions. The arginine decarboxylase pathway seems to be responsible for the accumulation of PAs in primed seeds. The obvious impact of seed priming on tyramine accumulation was also demonstrated. Moreover, the level of ethylene increased considerably in seedlings issued from primed seeds exposed to salt stress. It can be concluded that the polyamines are involved in creating the beneficial effect of osmopriming on germination and early growth of Brassica napus seedlings under saline conditions through moderate changes in their biosynthesis and accumulation.

Melatonin treatment reduces ethylene production and maintains fruit quality in apple during postharvest storage.

The effects of treatment with melatonin on ripening of 'Fuji' apples during storage at 1 °C for 56 d were investigated. The apples were harvested at the commercial ripening stage and treated with 1 mmol L-1 melatonin. Compared with the control, melatonin treated apples had significant reduced ethylene production (28 d-56 d) and weight loss (14 d-56 d) during storage (p < 0.05). Also, the melatonin treatment maintained better apple skin structure throughout storage. The reduced ethylene production was regulated by the decreased expressions of MdACO1, MdACS1, MdAP2.4 and MdERF109, based on RNA-Seq analysis, which was validated using qRT-PCR analysis. Moreover, the activity of 3 enzymes, including peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT), were significantly increased in melatonin treated fruit (p < 0.05). Taken together, this study highlights the inhibitory effects of melatonin in ethylene biosynthesis and factors influencing postharvest quality in apple.

Balancing of hormonal biosynthesis and catabolism pathways, a strategy to ameliorate the negative effects of heat stress on reproductive growth.

In pea (Pisum sativum L.), moderate heat stress during early flowering/fruit set increased seed/ovule abortion, and concomitantly produced fruits with reduced ovary (pericarp) length, and fewer seeds at maturity. Plant hormonal networks coordinate seed and pericarp growth and development. To determine if these hormonal networks are modulated in response to heat stress, we analyzed the gene expression patterns and associated these patterns with precursors, and bioactive and inactive metabolites of the auxin, gibberellin (GA), abscisic acid (ABA), and ethylene biosynthesis/catabolism pathways in young developing seeds and pericarps of non-stressed and 4-day heat-stressed fruits. Our data suggest that within the developing seeds heat stress decreased bioactive GA levels reducing GA growth-related processes, and that increased ethylene levels may have promoted this inhibitory response. In contrast, heat stress increased auxin biosynthesis gene expression and auxin levels in the seeds and pericarps, and seed ABA levels, both effects can increase seed sink strength. We hypothesize that seeds with higher auxin- and ABA-induced sink strength and adequate bioactive GA levels will set and continue to grow, while the seeds with lower sink strength (low auxin, ABA, and GA levels) will become more sensitive to heat stress-induced ethylene leading to ovule/seed abortion.

Identification of Novel Molecular Regulators Modulating Ethylene Biosynthesis Using EMS-Based Genetic Screening.

The gaseous hormone ethylene regulates a diverse range of plant development and stress responses. Ethylene biosynthesis is tightly regulated by the transcriptional and posttranscriptional regulation of ethylene biosynthetic enzymes. ACC synthase (ACS) is the rate-limiting enzyme that controls the speed of ethylene biosynthesis in plant tissues, thus serving as a primary target for biotic and abiotic stresses to modulate ethylene production. Despite the critical role of ACS in ethylene biosynthesis, only a few regulatory components regulating ACS stability or ACS transcript levels have been identified and characterized. Here we show a genetic approach for identifying novel regulatory components in ethylene biosynthesis by screening EMS-mutagenized Arabidopsis seeds.

Targeted mutagenesis of POLYAMINE OXIDASE 5 that negatively regulates mesocotyl elongation enables the generation of direct-seeding rice with improved grain yield.

Under conditions of labor or resource scarcity, direct seeding, rather than transplantation, is the preferred mode of rice (Oryza sativa) cultivation. This approach requires varieties that exhibit uniform seedling emergence. Mesocotyl elongation (ME), the main driver of rapid emergence of rice seedlings from soil, is enhanced by darkness and inhibited by light. Plant polyamine oxidases (PAOs) oxidize polyamines (PAs) and release H2O2. Here, we established that OsPAO5 expression in rice seedlings is increased in the presence of light and inhibited by darkness. To determine its role in ME, we created OsPAO5 mutants using CRISPR/Cas9. Compared with the wild type, pao5 mutants had longer mesocotyls, released less H2O2, and synthesized more ethylene. The mutant seedlings emerged at a higher and more uniform rate, indicating their potential for use in direct seeding. Nucleotide polymorphism analysis revealed that an SNP (PAO5-578G/A) located 578 bp upstream of the OsPAO5 start codon alters its expression, and was selected during rice mesocotyl domestication. The PAO5-578G genotype conferring a long mesocotyl mainly exists in wild rice, most Aus accessions, and some Geng (Japonica) accessions. Intriguingly, knocking out OsPAO5 can remarkably increase the grain weight, grain number, and yield potential. In summary, we developed a novel strategy to obtain elite rice with higher emergence vigor and yield potential, which can be conveniently and widely used to breed varieties of direct-seeding rice.