Search Results (35)

Oyster mushrooms (Pleurotus ostreatus) are one of the most commonly grown edible mushrooms using compost, which contains high concentrations of ammonia. In this study, inoculation of the oyster mushroom culture substrate with ammonia-assimilating bacterium Enterobacter sp. B12, either before or after composting, reduced the ammonia nitrogen content, increased the total nitrogen content of the compost, and enhanced the mushroom yield. Co-cultivation with P. ostreatus mycelia on potato dextrose agar (PDA) plates containing 200 mM NH₄⁺, B12 reduced reactive oxygen species (ROS) accumulation in the mycelia and downregulated the expression of the ROS-generating enzymes NADPH oxidase A (NOXA) and the stress hormone ethylene synthase 1-aminocyclopropane-1-carboxylate oxidase (ACO). It also downregulated the expression of the ammonia-assimilating related genes in the mycelia, such as glutamate dehydrogenase (GDH), glutamate synthase (GOGAT), glutamine synthetase (GS), ammonia transporter protein (AMT), and amino acid transporter protein (AAT), while upregulating its own ammonia-assimilation genes. These findings suggest that the mechanism by which B12 promoted oyster mushroom growth was that B12 assimilated ammonia, alleviated ammonia stress, mitigated ROS accumulation in the mycelia, and supplied ammonia and amino acids to the mycelia. To our knowledge, ammonia-assimilating bacteria are a novel type of mushroom growth promoter (MGP). Full article

The plant hormone ethylene elicits crucial regulatory effects on plant growth, development, and stress resistance. As the enzyme that catalyzes the final step of ethylene biosynthesis, 1-Aminocyclopropane-1-carboxylic acid oxidase (ACO) plays a key role in precisely controlling ethylene production. However, the functional characterization of the ACO gene family in rice remains largely unexplored. In this study, we performed a phylogenetic analysis of seven OsACO genes (OsACO1–OsACO7), which were classified into three subfamilies (Types I, II, and III). The members within the same clades exhibited similar tertiary structures and conserved protein motifs. We conducted inter/intraspecies covariance assays of OsACOs to elucidate their evolutionary and duplication events. Numerous cis-acting elements identified in OsACO promoter regions are associated with development, hormonal stimuli, and environmental responses. The expression assay by RT-qPCR revealed that OsACO genes exhibited tissue-specific expression and were significantly altered under various abiotic stresses, indicating their potential involvement in these processes regulated at the transcriptional level. Additionally, we predicted candidate-targeting miRNAs and identified putative cysteine sites of S-nitrosylation (SNO) and S-sulfhydration (SSH) in OsACOs, providing insights into their post-transcriptional and post-translational regulatory mechanisms. These findings pave the way for the further investigation of OsACO functions and their potential applications in improving rice growth and stress resilience by modulating ethylene biosynthesis. Full article

The immune response in plants is regulated by several phytohormones and involves the overexpression of defense genes, including the pathogenesis-related (PR-) genes. The data reported in this paper indicate that nematodes can suppress the immune response by inhibiting the expression of defense genes. Transcripts from nine defense genes were detected by qRT-PCR in the roots of tomato plants at three and seven days post-inoculation (dpi) with living juveniles (J2s) of Meloidogyne incognita (root-knot nematodes, RKNs). All the salicylic acid (SA)-responsive genes tested (PR-1, PR-2, PR-4b, PR-5) were down-regulated in response to nematode infection. On the contrary, the expression of jasmonic acid (JA)-responsive genes, including ACO (encoding the enzyme 1-aminocyclopropane-1-carboxylic acid oxidase, which catalyzes the last step of ethylene (ET) biosynthesis) and JERF3 (Jasmonate Ethylene Response Factor 3), was unaffected by the infection. Conversely, the effect of nematode attack on the activities of the defense enzymes endoglucanase and endochitinase, encoded by PR-2 and PR-3, respectively, changed depending on the tested dpi. At 5 dpi, both enzymes were inhibited in inoculated plants compared to healthy controls. The genes encoding glutathione peroxidase (GPX) and catalase (CAT), both part of the antioxidant plant system, were highly overexpressed. Additionally, the activity of the antioxidant enzymes superoxide dismutase (SOD), CAT, and ascorbate peroxidase (APX) was enhanced in infected roots. Isoelectrofocusing of root extracts revealed novel SOD isoforms in samples from inoculated plants. Furthermore, plants were pre-treated with an array of key compounds, including hormone generators, inhibitors of SA or JA-mediated defense pathways, reactive oxygen species (ROS) scavengers and generators, inhibitors of ROS generation, and compounds that interfere with calcium-mediated metabolism. After treatments, plants were inoculated with RKNs, and nematodes were allowed to complete their life cycle. Factors of plant growth and infection level in treated plants were compared with those from untreated inoculated plants. Generally, compounds that decreased SA and/or ROS levels increased infection severity, while those that reduced JA/ET levels did not affect infection rates. ROS generators induced resistance against the pests. Compounds that silence calcium signaling by preventing its intake augmented infection symptoms. The data shown in this paper indicate that SA-mediated plant immune responses are consistently suppressed during the early stages of nematode infection, and this restriction is associated with the activation of the antioxidant ROS-scavenging system. Full article

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. Full article

Sexual differentiation is an important developmental phenomenon in cucurbits that directly affects fruit yield. The natural existence of multiple flower types in melon offers an inclusive structure for studying the molecular basis of sexual differentiation. The current study aimed to identify and characterize the molecular network involved in sex determination and female development in melon. Male and female pools separated by the F₂ segregated generation were used for sequencing. The comparative multi-omics data revealed 551 DAPs and 594 DEGs involved in multiple pathways of melon growth and development, and based on functional annotation and enrichment analysis, we summarized four biological process modules, including ethylene biosynthesis, flower organ development, plant hormone signaling, and ubiquitinated protein metabolism, that are related to female development. Furthermore, the detailed analysis of the female developmental regulatory pathway model of ethylene biosynthesis, signal transduction, and target gene regulation identified some important candidates that might have a crucial role in female development. Two CMTs ((cytosine-5)-methyltransferase), one AdoHS (adenosylhomocysteinase), four ACSs (1-aminocyclopropane-1-carboxylic acid synthase), three ACOs (ACC oxidase), two ARFs (auxin response factor), four ARPs (auxin-responsive protein), and six ERFs (Ethylene responsive factor) were identified based on various female developmental regulatory models. Our data offer new and valuable insights into female development and hold the potential to offer a deeper comprehension of sex differentiation mechanisms in melon. Full article

The main aim of this study was to understand the regulation of the biosynthesis of phytohormones as signaling molecules in the defense mechanisms of pea seedlings during the application of abiotic and biotic stress factors. It was important to identify this regulation at the molecular level in Pisum sativum L. seedlings under the influence of various concentrations of lead—i.e., a low concentration increasing plant metabolism, causing a hormetic effect, and a high dose causing a sublethal effect—and during feeding of a phytophagous insect with a piercing-sucking mouthpart—i.e., pea aphid (Acyrthosiphon pisum (Harris)). The aim of the study was to determine the expression level of genes encoding enzymes of the biosynthesis of signaling molecules such as phytohormones—i.e., jasmonates (JA/MeJA), ethylene (ET) and abscisic acid (ABA). Real-time qPCR was applied to analyze the expression of genes encoding enzymes involved in the regulation of the biosynthesis of JA/MeJA (lipoxygenase 1 (LOX1), lipoxygenase 2 (LOX2), 12-oxophytodienoate reductase 1 (OPR1) and jasmonic acid-amido synthetase (JAR1)), ET (1-aminocyclopropane-1-carboxylate synthase 3 (ACS3)) and ABA (9-cis-epoxycarotenoid dioxygenase (NCED) and aldehyde oxidase 1 (AO1)). In response to the abovementioned stress factors—i.e., abiotic and biotic stressors acting independently or simultaneously—the expression of the LOX1, LOX2, OPR1, JAR1, ACS3, NCED and AO1 genes at both sublethal and hormetic doses increased. Particularly high levels of the relative expression of the tested genes in pea seedlings growing at sublethal doses of lead and colonized by A. pisum compared to the control were noticeable. A hormetic dose of lead induced high expression levels of the JAR1, OPR1 and ACS3 genes, especially in leaves. Moreover, an increase in the concentration of phytohormones such as jasmonates (JA and MeJA) and aminococyclopropane-1-carboxylic acid (ACC)-ethylene (ET) precursor was observed. The results of this study indicate that the response of pea seedlings to lead and A. pisum aphid infestation differed greatly at both the gene expression and metabolic levels. The intensity of these defense responses depended on the organ, the metal dose and direct contact of the stress factor with the organ. Full article

Stem nematode disease can seriously reduce the yield of sweet potato (Ipomoea batatas (L.) Lam). To explore resistance mechanism to stem nematode in sweet potato, transcriptomes and metabolomes were sequenced and compared between two sweet potato cultivars, the resistant Zhenghong 22 and susceptible Longshu 9, at different times after stem nematode infection. In the transcriptional regulatory pathway, mitogen-activated protein kinase signaling was initiated in Zhenghong 22 at the early stage of infection to activate genes related to ethylene production. Stem nematode infection in Zhenghong 22 also triggered fatty acid metabolism and the activity of respiratory burst oxidase in the metabolic pathway, which further stimulated the glycolytic and shikimic pathways to provide raw materials for secondary metabolite biosynthesis. An integrated analysis of the secondary metabolic regulation pathway in the resistant cultivar Zhenghong 22 revealed the accumulation of tryptophan, phenylalanine, and tyrosine, leading to increased biosynthesis of phenylpropanoids and salicylic acid and enhanced activity of the alkaloid pathway. Stem nematode infection also activated the biosynthesis of terpenoids, abscisic acid, zeatin, indole, and brassinosteroid, resulting in improved resistance to stem nematode. Finally, analyses of the resistance regulation pathway and a weighted gene co-expression network analysis highlighted the importance of the genes itf14g17940 and itf12g18840, encoding a leucine-rich receptor-like protein and 1-aminocyclopropane-1-carboxylate synthase, respectively. These are candidate target genes for increasing the strength of the defense response. These results provide new ideas and a theoretical basis for understanding the mechanism of resistance to stem nematode in sweet potato. Full article

Cadmium (Cd) is a heavy metal that can cause damage to living organisms at different levels. Even at low concentrations, Cd can be toxic to plants, causing harm at multiple levels. As they are unable to move away from areas contaminated by Cd, plants have developed various defence mechanisms to protect themselves. Hyperaccumulators, which can accumulate and detoxify heavy metals more efficiently, are highly valued by scientists studying plant accumulation and detoxification mechanisms, as they provide a promising source of genes for developing plants suitable for phytoremediation techniques. So far, several genes have been identified as being upregulated when plants are exposed to Cd. These genes include genes encoding transcription factors such as iron-regulated transporter-like protein (ZIP), natural resistance associated macrophage protein (NRAMP) gene family, genes encoding phytochelatin synthases (PCs), superoxide dismutase (SOD) genes, heavy metal ATPase (HMA), cation diffusion facilitator gene family (CDF), Cd resistance gene family (PCR), ATP-binding cassette transporter gene family (ABC), the precursor 1-aminocyclopropane-1-carboxylic acid synthase (ACS) and precursor 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) multigene family are also influenced. Thanks to advances in omics sciences and transcriptome analysis, we are gaining more insights into the genes involved in Cd stress response. Recent studies have also shown that Cd can affect the expression of genes related to antioxidant enzymes, hormonal pathways, and energy metabolism. Full article

The 1-aminocyclopropane-1-carboxylic acid (ACC) pathway that synthesizes ethylene is shared in seed plants, fungi and probably other organisms. However, the evolutionary relationship of the key enzyme ACC oxidase (ACO) in the pathway among organisms remains unknown. Herein, we cloned, expressed and characterized five ACOs from the straw mushroom (Volvariella volvacea) and the oyster mushroom (Pleurotus ostreatus): VvACO1-4 and PoACO. The five mushroom ACOs and the previously identified AbACO of the button mushroom contained all three conserved residues that bound to Fe(II) in plant ACOs. They also had variable residues that were conserved and bound to ascorbate and bicarbonate in plant ACOs and harbored only 1–2 of the five conserved ACO motifs in plant ACOs. Particularly, VvACO2 and AbACO had only one ACO motif 2. Additionally, VvACO4 shared 44.23% sequence identity with the cyanobacterium Hapalosiphon putative functional ACO. Phylogenetic analysis showed that the functional ACOs of monocotyledonous and dicotyledonous plants co-occurred in Type I, Type II and Type III, while putative functional gymnosperm ACOs also appeared in Type III. The putative functional bacterial ACO, functional fungi and slime mold ACOs were clustered in ancestral Type IV. These results indicate that ACO motif 2, ACC and Fe(II) are essential for ACO activity. The ACOs of the other organisms may come from the horizontal transfer of fungal ACOs, which were found ordinarily in basidiomycetes. It is mostly the first case for the horizontal gene transfers from fungi to seed plants. The horizontal transfer of ACOs from fungi to plants probably facilitates the fungal-plant symbioses, plant–land colonization and further evolution to form seeds. Full article

Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) is a serious invasive pest in China. In this study, we determined whether exogenous jasmonic acid (JA) and ethylene (ET) treatments could induce resistance against F. occidentalis in faba bean plants. First, we investigated the effects of different concentrations of JA or ET alone on F. occidentalis and then assessed the effects of optimal concentrations of JA and ET combined. Our results showed that the optimal concertation of JA was 2 mmol/L and ET was 0.5 mmol/L. JA + ET mixture showed the greatest inhibitory effect in terms of oviposition and feeding. JA with ET was found to induce changes in the activities of lipoxygenase (LOX), allene oxide synthase (AOS), polyphenol oxidase (PPO), 1-aminocyclopropane 1-carboxylic acid synthase (ACS), and trypsin inhibitor (TI). This treatment also activated or inhibited the relative expression levels of LOX1, ACO2, ACS2, and AP2/ERF. Treatment of faba bean plants with JA and ET significantly prolonged F. occidentalis development and adult preoviposition period, significantly reduced per-female oviposition, and altered male longevity and offspring demographic parameters. These results indicate that JA with ET can induce defenses against the growth and development of F. occidentalis in faba bean plants. Full article

Barnyardgrass (a monocotyledon) and yerbadetajo (a dicotyledon) are the most troublesome weeds in rice fields in China. The synthetic auxin herbicide florpyrauxifen-benzyl can effectively control both weeds. The objective of this research was to clarify modes of action of florpyrauxifen-benzyl in barnyardgrass and yerbadetajo. Our results showed that yerbadetajo was more sensitive to florpyrauxifen-benzyl than barnyardgrass: the 50% growth rate inhibition in barnyardgrass and yerbadetajo was 4.14 and 0.38 g a.i. ha⁻¹, respectively. Florpyrauxifen-benzyl induced the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) and abscisic acid (ABA) in yerbadetajo within 24 h, while in barnyardgrass, the highest production occurred at 12 h and decreased at 24 h. ACC and ABA levels in yerbadetajo at 24 h of treatment were significantly higher than those in barnyardgrass at any time. There were less differentially expressed genes related to ethylene and ABA synthesis in barnyardgrass than in yerbadetajo. There were five genes induced to increase by florpyrauxifen-benzyl in barnyardgrass and eleven genes in yerbadetajo. More ACC oxidase genes (ACO) were induced in barnyardgrass and more ACC synthesis genes (ACS) in yerbadetajo, especially three ACS3 (>30 fold). We speculated that differences in gene expression caused differences in ethylene and ABA production, leading to differences in phytotoxicity. Full article

Fruits of wild tomato species show different ethylene-dependent ripening characteristics, such as variations in fruit color and whether they exhibit a climacteric or nonclimacteric ripening transition. 1-Aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS) and ACC oxidase (ACO) are key enzymes in the ethylene biosynthetic pathway encoded by multigene families. Gene duplication is a primary driver of plant diversification and angiosperm evolution. Here, interspecific variations in the molecular regulation of ethylene biosynthesis and perception during fruit ripening in domesticated and wild tomatoes were investigated. Results showed that the activated ACS genes were increased in number in red-ripe tomato fruits than in green-ripe tomato fruits; therefore, elevated dosage of ACS enzyme promoted ripening ethylene production. Results showed that the expression of three ACS isogenes ACS1A, ACS2, and ACS4, which are involved in autocatalytic ethylene production, was higher in red-ripe tomato fruits than in green-ripe tomato fruits. Elevated ACS enzyme dosage promoted ethylene production, which corresponded to the climacteric response of red-ripe tomato fruits. The data suggest that autoinhibitory ethylene production is common to all tomato species, while autocatalytic ethylene production is specific to red-ripe species. The essential regulators Non-ripening (NOR) and Ripening-Inhibitor (RIN) have experienced gene activation and overlapped with increasing ACS enzyme dosage. These complex levels of transcript regulation link higher ethylene production with spatiotemporal modulation of gene expression in red-ripe tomato species. Taken together, this study shows that bursts in ethylene production that accompany fruit color changes in red-ripe tomatoes are likely to be an evolutionary adaptation for seed dispersal. Full article

Postharvest yellowing of leafy plant is a manifestation of senescence, and melatonin (MT) is known to delay leaf senescence in some higher plants. Herein, we investigated the effect of exogenous MT treatment on postharvest pakchoi by monitoring the ethylene biosynthesis and respiratory metabolism. Results showed that exogenous MT effectively extended the shelf life, delayed leaf yellowing, minimized the alteration in Fv/Fm ratio and maintained higher integrity of chloroplast in pakchoi. There was a significant correlation between yellowing index, respiration rate and ethylene production. MT treatments greatly delayed the yellowing process of pakchoi that was associated with the reduced activity of glycolysis pathway and tricarboxylic acid cycle (TCA), increased proportion of pentose phosphate pathway (PPP) in respiratory metabolism, as manifested by the lower activity of phosphohexose isomerase (PHI), succinate dehydrogenase (SDH) and cytochrome C oxidase (COX), downregulated the expression of their corresponding genes, but enhanced the activity and expression level of 6 phosphogluconate dehydrogenase (6PGDH). MT also markedly maintain chlorophyll content by inhibiting ethylene production and action during shelf life, likely a consequence of reduced activities of 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS) and ACC oxidase (ACO), as well as the expression levels of their related genes. These results collectively indicate that melatonin alleviated leaf yellowing of postharvest pakchoi might be attributed to the suppression of the ethylene biosynthesis and respiratory metabolism, and our findings contribute to provide a good candidate measure for extending shelf life and reducing postharvest loss of pakchoi. Full article

Rice grain yield is a complex and highly variable quantitative trait consisting of several key components, including the grain weight, the effective panicles per unit area, and the grain number per panicle (GNPP). The GNPP is a significant contributor to grain yield controlled by multiple genes (QTL) and is crucial for improvement. Attempts have been made to find genes for this trait, which has always been a challenging and arduous task through conventional methods. We combined a BSA analysis, RNA profiling, and a metabolome analysis in the present study to identify new candidate genes involved in the GNPP. The F₂ population from crossing R4233 (high GNPP) and Ce679 (low GNPP) revealed a frequency distribution fitting two segregated genes. Three pools, including low, middle, and high GNPP, were constructed and a BSA analysis revealed six candidate regions spanning 5.38 Mb, containing 739 annotated genes. Further, a conjunctive analysis of BSA-Seq and RNA-Seq showed 31 differentially expressed genes (DEGs) in the candidate intervals. Subsequently, a metabolome analysis showed 1024 metabolites, with 71 significantly enriched, including 44 up and 27 downregulated in Ce679 vs. R4233. A KEGG enrichment analysis of these 31 DEGs and 71 differentially enriched metabolites (DEMs) showed two genes, Os12g0102100 and Os01g0580500, significantly enriched in the metabolic pathways’ biosynthesis of secondary metabolites, cysteine and methionine metabolism, and fatty acid biosynthesis. Os12g0102100, which encodes for the alcohol dehydrogenase superfamily and a zinc-containing protein, is a novel gene whose contribution to the GNPP is not yet elucidated. This gene coding for mitochondrial trans-2-enoyl-CoA reductase is involved in the biosynthesis of myristic acid, also known as tetradecanoic acid. The Os01g0580500 coding for the enzyme 1-aminoclopropane-1-carboxylate oxidase (OsACO7) is responsible for the final step of the ethylene biosynthesis pathway through the conversion of 1-aminocyclopropane-1-carboxylic acid (ACC) into ethylene. Unlike Os12g0102100, this gene was significantly upregulated in R4233, downregulated in Ce679, and significantly enriched in two of the three metabolite pathways. This result pointed out that these two genes are responsible for the difference in the GNPP in the two cultivars, which has never been identified. Further validation studies may disclose the physiological mechanisms through which they regulate the GNPP in rice. Full article

The transgenic tobacco (Nicotiana tabacum L.) plants with the modified levels of alternative oxidase (AOX) were used to evaluate the physiological roles of AOX in regulating nitro-oxidative stress and metabolic changes after exposing plants to hypoxia for 6 h. Under normoxia, AOX expression resulted in the decrease of nitric oxide (NO) levels and of the rate of protein S-nitrosylation, while under hypoxia, AOX overexpressors exhibited higher NO and S-nitrosylation levels than knockdowns. AOX expression was essential in avoiding hypoxia-induced superoxide and H₂O₂ levels, and this was achieved via higher activities of catalase and glutathione reductase and the reduced expression of respiratory burst oxidase homolog (Rboh) in overexpressors as compared to knockdowns. The AOX overexpressing lines accumulated less pyruvate and exhibited the increased transcript and activity levels of pyruvate decarboxylase and alcohol dehydrogenase under hypoxia. This suggests that AOX contributes to the energy state of hypoxic tissues by stimulating the increase of pyruvate flow into fermentation pathways. Ethylene biosynthesis genes encoding 1-aminocyclopropane-1-carboxylic acid (ACC) synthase, ACC oxidase, and ethylene-responsive factors (ERFs) were induced during hypoxia and correlated with AOX and NO levels. We conclude that AOX controls the interaction of NO, reactive oxygen species, and ethylene, triggering a coordinated downstream defensive response against hypoxia. Full article