Regulation Of Ethylene Biosynthesis Research Articles

New insights into the regulation of ethylene biosynthesis during leaf senescence in Arabidopsis.

New insights into the regulation of ethylene biosynthesis during leaf senescence in Arabidopsis.

Carbon dioxide sensitization delays the postharvest ripening and fatty acids composition of Capsicum fruit by regulating ethylene biosynthesis, malic acid and reactive oxygen species metabolism.

The online version contains supplementary material available at 10.1007/s12298-024-01471-4.

Casein kinase 1 AELs promote senescence by enhancing ethylene biosynthesis through phosphorylating WRKY22 transcription factor.

Leaf senescence is a complex process regulated by developmental and environmental factors, and plays a pivotal role in the development and life cycle of higher plants. Casein kinase 1 (CK1) is a highly conserved serine/threonine protein kinase in eukaryotes and functions in various cellular processes including cell proliferation, light signaling and hormone effects of plants. However, the biological function of CK1 in plant senescence remains unclear. Through systemic genetic and biochemical studies, we here characterized the function of Arabidopsis EL1-like (AEL), a CK1, in promoting leaf senescence by stimulating ethylene biosynthesis through phosphorylating transcription factor WRKY22. Seedlings lacking or overexpressing AELs presented delayed or accelerated leaf senescence, respectively. AELs interact with and phosphorylate WRKY22 at Thr57, Thr60 and Ser69 residues to enhance whose transactivation activity. Being consistent, increased or suppressed phosphorylation of WRKY22 resulted in the promoted or delayed leaf senescence. WRKY22 directly binds to promoter region and stimulates the transcription of 1-amino-cyclopropane-1-carboxylate synthase 7 gene to promote ethylene level and hence leaf senescence. Our studies demonstrated the crucial role of AEL-mediated phosphorylation in regulating ethylene biosynthesis and promoting leaf senescence by enhancing WRKY22 transactivation activity, which helps to elucidate the fine-controlled ethylene biosynthesis and regulatory network of leaf senescence.

The Aux/IAA Factor PbIAA.C3 Positively Regulates Ethylene Biosynthesis During Pear Fruit Ripening by Activating the PbACS1b Transcription

The Aux/IAA Factor PbIAA.C3 Positively Regulates Ethylene Biosynthesis During Pear Fruit Ripening by Activating the PbACS1b Transcription

The Ethylene Biosynthetic Enzymes, 1-Aminocyclopropane-1-Carboxylate (ACC) Synthase (ACS) and ACC Oxidase (ACO): The Less Explored Players in Abiotic Stress Tolerance.

Ethylene is an essential plant hormone, critical in various physiological processes. These processes include seed germination, leaf senescence, fruit ripening, and the plant's response to environmental stressors. Ethylene biosynthesis is tightly regulated by two key enzymes, namely 1-aminocyclopropane-1-carboxylate synthase (ACS) and 1-aminocyclopropane-1-carboxylate oxidase (ACO). Initially, the prevailing hypothesis suggested that ACS is the limiting factor in the ethylene biosynthesis pathway. Nevertheless, accumulating evidence from various studies has demonstrated that ACO, under specific circumstances, acts as the rate-limiting enzyme in ethylene production. Under normal developmental processes, ACS and ACO collaborate to maintain balanced ethylene production, ensuring proper plant growth and physiology. However, under abiotic stress conditions, such as drought, salinity, extreme temperatures, or pathogen attack, the regulation of ethylene biosynthesis becomes critical for plants' survival. This review highlights the structural characteristics and examines the transcriptional, post-transcriptional, and post-translational regulation of ACS and ACO and their role under abiotic stress conditions. Reviews on the role of ethylene signaling in abiotic stress adaptation are available. However, a review delineating the role of ACS and ACO in abiotic stress acclimation is unavailable. Exploring how particular ACS and ACO isoforms contribute to a specific plant's response to various abiotic stresses and understanding how they are regulated can guide the development of focused strategies. These strategies aim to enhance a plant's ability to cope with environmental challenges more effectively.

The transcription factor HSFA7b controls thermomemory at the shoot apical meristem by regulating ethylene biosynthesis and signaling in Arabidopsis

The shoot apical meristem (SAM) is responsible for overall shoot growth by generating all aboveground structures. Recent research has revealed that the SAM displays an autonomous heat stress (HS) memory of a previous non-lethal HS event. Considering the importance of the SAM for plant growth, it is essential to determine how its thermomemory is mechanistically controlled. Here, we report that HEAT SHOCK TRANSCRIPTION FACTOR A7b (HSFA7b) plays a crucial role in this process in Arabidopsis, as the absence of functional HSFA7b results in the temporal suppression of SAM activity after thermopriming. We found that HSFA7b directly regulates ethylene response at the SAM by binding to the promoter of the key ethylene signaling gene ETHYLENE-INSENSITIVE 3 to establish thermotolerance. Moreover, we demonstrated that HSFA7b regulates the expression of ETHYLENE OVERPRODUCER 1 (ETO1) and ETO1-LIKE 1, both of which encode ethylene biosynthesis repressors, thereby ensuring ethylene homeostasis at the SAM. Taken together, these results reveal a crucial and tissue-specific role for HSFA7b in thermomemory at the Arabidopsis SAM.

A peach ethylene response factor PpERF61 is involved in fruit ripening by modulating ripening-related genes and PpSEP1

Fruit ripening is a highly coordinated process at the transcriptional level, requiring a hierarchical regulatory network formed by transcription factor and ripening-related genes. MADS-box transcription factors are core regulators that play a key function in the ripening process of fruit, targeting a large amount of downstream ripening-related genes. However, the upstream regulators of MADS-box transcription factors are poorly understood. In this study, we identified an ethylene response factor PpERF61 by yeast one-hybrid screening with the promoter of PpSEP1 (MADS-RIN homologs). The expression pattern of PpERF61 was closely associated with ethylene biosynthesis and fruit softening across six peach cultivars. The transient overexpression and VIGS assay showed that overexpression or silencing of PpERF61 in peach promoted or inhibited fruit softening, while ethylene production was increased or decreased, respectively, demonstrating that PpERF61 has effect on ethylene generation and fruit softening during peach ripening. As a nuclear-localized transcription factor, PpERF61 activated the ripening-related genes transcription, including PpACO1, PpACS1, PpPL1, and PpPL15, by binding directly to their promoters. Interestingly, PpERF61 acts as upstream of PpSEP1 and binds directly to the PpSEP1 promoter, thereby forming PpERF61-PpSEP1 module to promote the PpPG2 and PpPG3 transcription. Based on these findings, we conclude that PpERF61 positively regulates ethylene biosynthesis and fruit softening by directly activating ripening-related genes or activating the PpERF61-PpSEP1 signaling cascade. Collectively, these results enrich knowledge of the complex cascade network for peach fruit ripening.

MaMADS1-MaNAC083 transcriptional regulatory cascade regulates ethylene biosynthesis during banana fruit ripening.

The hormone ethylene is crucial in the regulation of ripening in climacteric fruit, such as bananas. The transcriptional regulation of ethylene biosynthesis throughout banana fruit ripening has received much study, but the cascaded transcriptional machinery of upstream transcriptional regulators implicated in the ethylene biosynthesis pathway is still poorly understood. Here we report that ethylene biosynthesis genes, including MaACS1, MaACO1, MaACO4, MaACO5, and MaACO8, were upregulated in ripening bananas. NAC (NAM, ATAF, CUC) transcription factor, MaNAC083, a ripening and ethylene-inhibited gene, was discovered as a potential binding protein to the MaACS1 promoter by yeast one-hybrid screening. Further in vitro and in vivo experiments indicated that MaNAC083 bound directly to promoters of the five ethylene biosynthesis genes, thereby transcriptionally repressing their expression, which was further verified by transient overexpression experiments, where ethylene production was inhibited through MaNAC083-modulated transcriptional repression of ethylene biosynthesis genes in banana fruits. Strikingly, MaMADS1, a ripening-induced MADS (MCM1, AGAMOUS, DEFICIENS, SRF4) transcription factor, was found to directly repress the expression of MaNAC083, inhibiting trans-repression of MaNAC083 to ethylene biosynthesis genes, thereby attenuating MaNAC083-repressed ethylene production in bananas. These findings collectively illustrated the mechanistic basis of a MaMADS1-MaNAC083-MaACS1/MaACOs regulatory cascade controlling ethylene biosynthesis during banana fruit ripening. These findings increase our knowledge of the transcriptional regulatory mechanisms of ethylene biosynthesis at the transcriptional level and are expected to help develop molecular approaches to control ripening and improve fruit storability.

GhXB38D represses cotton fibre elongation through ubiquitination of ethylene biosynthesis enzymes GhACS4 and GhACO1.

Ethylene plays an essential role in the development of cotton fibres. Ethylene biosynthesis in plants is elaborately regulated by the activities of key enzymes, 1-aminocyclopropane-1-carboxylate oxidase (ACO) and 1-aminocyclopropane-1-carboxylate synthase (ACS); however, the potential mechanism of post-translational modification of ACO and ACS to control ethylene synthesis in cotton fibres remains unclear. Here, we identify an E3 ubiquitin ligase, GhXB38D, that regulates ethylene biosynthesis during fibre elongation in cotton. GhXB38D gene is highly expressed in cotton fibres during the rapid elongation stage. Suppressing GhXB38D expression in cotton significantly enhanced fibre elongation and length, accompanied by the up-regulation of genes associated with ethylene signalling and fibre elongation. We demonstrated that GhXB38D interacts with the ethylene biosynthesis enzymes GhACS4 and GhACO1 in elongating fibres and specifically mediates their ubiquitination and degradation. The inhibition of GhXB38D gene expression increased the stability of GhACS4 and GhACO1 proteins in cotton fibres and ovules, resulting in an elevated concentration of ethylene. Our findings highlight the role of GhXB38D as a regulator of ethylene synthesis by ubiquitinating ACS4 and ACO1 proteins and modulating their stability. GhXB38D acts as a negative regulator of fibre elongation and serves as a potential target for enhancing cotton fibre yield and quality through gene editing strategy.

MaNAC029 modulates ethylene biosynthesis and fruit quality and undergoes MaXB3-mediated proteasomal degradation during banana ripening

IntroductionsEthylene regulates ripening by activating various metabolic pathways that controlcolor, aroma, flavor, texture, and consequently, the quality of fruits. However, the modulation of ethylene biosynthesis and quality formation during banana fruit ripening remains unclear. ObjectivesThe present study aimed to identify the regulatory module that regulates ethylene and fruit quality-related metabolisms during banana fruit ripening. MethodsWe used RNA-seq to compare unripe and ripe banana fruits and identified a ripening-induced NAC transcription factor, MaNAC029. We further performed DNA affinity purification sequencing to identify the MaNAC029′s target genes involved in ethylene biosynthesis and fruit quality formation, and electrophoretic mobility shift assay, chromatin immunoprecipitation with real-time polymerase chain reaction and dual luciferase assays to explore the underlying regulatory mechanisms. Immunoprecipitation combined with mass spectrometry, yeast two-hybrid assay, and bimolecular fluorescence complementation assay were used to screen and verify the proteins interacting with MaNAC029. Finally, the function of MaNAC029 and its interacting protein associated with ethylene biosynthesis and quality formation was verified through transient overexpression experiments in banana fruits. ResultsThe study identified a nucleus-localized, ripening-induced NAC transcription factor MaNAC029. It transcriptionally activated genes associated with ethylene biosynthesis and a variety of cellular metabolisms related to fruit quality formation (cell wall degradation, starch degradation, aroma compound synthesis, and chlorophyll catabolism) by directly modulating their promoter activity during ripening. Overexpression of MaNAC029 in banana fruits activated ethylene biosynthesis and accelerated fruit ripening and quality formation. Notably, the E3 ligase MaXB3 interacted with and ubiquitinated MaNAC029 protein, facilitating MaNAC029 proteasomal degradation. Consistent with this finding, MaXB3 overexpression attenuated MaNAC029-enhanced ethylene biosynthesis and quality formation. ConclusionOur findings demonstrate that a MaXB3-MaNAC029 module regulates ethylene biosynthesis and a series of cellular metabolisms related to fruit quality formation during banana ripening. These results expand the understanding of the transcriptional and post-translational mechanisms of fruit ripening and quality formation.

Comparative transcriptome analysis of the climacteric of apple fruit uncovers the involvement of transcription factors affecting ethylene biosynthesis

Apple (Malus domestica) fruit generally undergoes a climacteric. During its ripening process, there is a peak in ethylene release and its firmness simultaneously decreases. Although more in-depth research into the mechanism of climacteric-type fruit ripening is being carried out, some aspects remain unclear. In this study, we compared the transcriptomes of 0-Pre and 15-Post (pre- and post-climacteric fruit), and 15-Post and 15-MCP [fruit treated with 1-MCP (1-methylcyclopropene)]. Various transcription factors, such as MADS-box, ERF, NAC, Dof and SHF were identified among the DEGs (differential gene expressions). Furthermore, these transcription factors were selected for further validation analysis by qRT-PCR. Moreover, yeast one hybrid (Y1H), β-glucuronidase (GUS) transactivation assay and dual-luciferase reporter assay showed that MdAGL30, MdAGL104, MdERF008, MdNAC71, MdDof1.2, MdHSFB2a and MdHSFB3 bound to MdACS1 promoter and directly regulated its transcription, thereby regulating ethylene biosynthesis in apple fruit. Our results provide useful information and new insights for research on apple fruit ripening.

WRKY29 transcription factor regulates ethylene biosynthesis and response in arabidopsis

The gaseous phytohormone ethylene participates in a lot of physiological processes in plants. 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS, EC 4.4.1.14) and the ACC oxidase (ACO, EC 1.14.17.4) are key enzymes in ethylene biosynthesis. However, how ACSs and ACOs are regulated at the transcriptional level is largely unknown. In the present study, we showed that an Arabidopsis (Arabidopsis thaliana) WRKY-type transcription factor (TF), WRKY29 positively regulated the expression of ACS5, ACS6, ACS8, ACS11 and ACO5 genes and thus promoted basal ethylene production. WRKY29 protein was localized in nuclei and was a transcriptional activator. Overexpression of WRKY29 caused pleiotropic effect on plant growth, development and showed obvious response even without ACC treatment. Inducible overexpression of WRKY29 also reduced primary root elongation and lateral root growth. A triple response assay of overexpression and mutant seedlings of WRKY29 showed that overexpression seedlings had shorter hypocotyls than the transgenic GFP (Green Fluorescence Protein) control, while mutants had no difference from wild-type. A qRT-PCR assay demonstrated that expression of multiple ACSs and ACO5 was up-regulated in WRKY29 overexpression plants. A transactivation assay through dual luciferase reporter system confirmed the regulation of promoters of ACS5, ACS6, ACS8, ACS11 and ACO5 by WRKY29. Both in vivo chromatin immunoprecipitation (ChIP)- quantitative PCR and in vitro electrophoretic mobility shift assay (EMSA) revealed that WRKY29 directly bound to the promoter regions of its target genes. Taken together, these results suggest that WRKY29 is a novel TF positively regulating ethylene production by modulating the expression of ACS and ACO genes.

Abscisic Acid: Role in Fruit Development and Ripening.

Abscisic acid (ABA) is a plant growth regulator known for its functions, especially in seed maturation, seed dormancy, adaptive responses to biotic and abiotic stresses, and leaf and bud abscission. ABA activity is governed by multiple regulatory pathways that control ABA biosynthesis, signal transduction, and transport. The transport of the ABA signaling molecule occurs from the shoot (site of synthesis) to the fruit (site of action), where ABA receptors decode information as fruit maturation begins and is significantly promoted. The maximum amount of ABA is exported by the phloem from developing fruits during seed formation and initiation of fruit expansion. In the later stages of fruit ripening, ABA export from the phloem decreases significantly, leading to an accumulation of ABA in ripening fruit. Fruit growth, ripening, and senescence are under the control of ABA, and the mechanisms governing these processes are still unfolding. During the fruit ripening phase, interactions between ABA and ethylene are found in both climacteric and non-climacteric fruits. It is clear that ABA regulates ethylene biosynthesis and signaling during fruit ripening, but the molecular mechanism controlling the interaction between ABA and ethylene has not yet been discovered. The effects of ABA and ethylene on fruit ripening are synergistic, and the interaction of ABA with other plant hormones is an essential determinant of fruit growth and ripening. Reaction and biosynthetic mechanisms, signal transduction, and recognition of ABA receptors in fruits need to be elucidated by a more thorough study to understand the role of ABA in fruit ripening. Genetic modifications of ABA signaling can be used in commercial applications to increase fruit yield and quality. This review discusses the mechanism of ABA biosynthesis, its translocation, and signaling pathways, as well as the recent findings on ABA function in fruit development and ripening.

SnRK2 subfamily I protein kinases regulate ethylene biosynthesis by phosphorylating HB transcription factors to induce ACO1 expression in apple.

Summary Ethylene (ETH) controls climacteric fruit ripening and can be triggered by osmotic stress. However, the mechanism regulating ETH biosynthesis during fruit ripening and under osmotic stress is largely unknown in apple (Malus domestica).Here, we explored the roles of SnRK2 protein kinases in ETH biosynthesis related to fruit ripening and osmoregulation. We identified the substrates of MdSnRK2‐I using phosphorylation analysis techniques. Finally, we identified the MdSnRK2‐I‐mediated signaling pathway for ETH biosynthesis related to fruit ripening and osmoregulation.The activity of two MdSnRK2‐I members, MdSnRK2.4 and MdSnRK2.9, was significantly upregulated during ripening or following mannitol treatment. Overexpression of MdSnRK2‐I increased ETH biosynthesis under normal and osmotic conditions in apple fruit. MdSnRK2‐I phosphorylated the transcription factors MdHB1 and MdHB2 to enhance their protein stability and transcriptional activity on MdACO1. MdSnRK2‐I also interacted with MdACS1 and increased its protein stability through two phosphorylation sites. The increased MdACO1 expression and MdACS1 protein stability resulted in higher ETH production in apple fruit. In addition, heterologous expression of MdSnRK2‐I or manipulation of SlSnRK2‐I expression in tomato (Solanum lycopersicum) fruit altered fruit ripening and ETH biosynthesis.We established that MdSnRK2‐I functions in fruit ripening and osmoregulation, and identified the MdSnRK2‐I‐mediated signaling pathway controlling ETH biosynthesis.

Reduction of ethylene biosynthesis in sugarcane induces growth and investment in the non-enzymatic antioxidant apparatus.

Lower ethylene production in sugarcane results in plants with higher stature, expression of growth-promoting genes, higher photosynthetic rate, and increased antioxidant compounds. The hormone ethylene is involved in critical processes in sugarcane, such as the growth and accumulation of sucrose. The lack of mutants for ethylene biosynthesis or signaling genes makes it difficult to understand the role of this phytohormone throughout sugarcane development. This study aimed to evaluate the physiology and development of sugarcane plants with low ethylene production. To achieve this goal, we used RNA interference to silence three genes, ScACS1, ScACS2, and ScACS3, encoding 1-aminocyclopropane-1-carboxylic acid synthases (ACS), responsible for a limiting step of the ethylene biosynthesis pathway. Sugarcane plants with reduced ethylene levels presented increased growth, faster germination of lateral gems, and activation of non-enzymatic antioxidant mechanisms. We observed an augmentation in the expression of ScACO5, which encodes the final enzyme regulating ethylene biosynthesis, and ScERF1, encoding a transcription factor, linked to the ethylene response. The increase in plant height was correlated with higher expression of ScPIF3, ScPIF4, and ScPIF5, which encode for transcription factors related to growth induction. Interestingly, there was also an increase in the expression of the ScGAI gene, which encodes a DELLA protein, a growth repressor. The final content of sucrose in the stems was not affected by the low levels of ethylene, although the rate of CO2 assimilation was reduced. This study reports for the first time the impacts of low endogenous production of ethylene in sugarcane and provides helpful insights on the molecular mechanisms behind ethylene responses.

PuWRKY31 affects ethylene production in response to sucrose signal in pear fruit.

The ripening of climacteric fruits is mainly controlled by the plant hormone ethylene. The regulatory effect of sucrose on ethylene biosynthesis in fruits remains unclear. Here we examined ethylene production in two Ussurian pear (Pyrus ussuriensis) varieties, ‘Nanguo’ (NG) pear and its bud sport variety (BNG), which has a higher sucrose content. We found that the peak of ethylene release occurred earlier in BNG fruit than in NG fruit during ripening. The expression of the transcription factor PuWRKY31 was higher in BNG fruit than in NG fruit, and was induced by sucrose treatment. Furthermore, PuWRKY31 bound to the promoters of ethylene biosynthetic genes and upregulated their transcription. Our findings suggest a mechanism by which sucrose regulates ethylene biosynthesis in pears.

Regulation of ethylene biosynthesis and signal transduction by nitric oxide leading to resistance against Alternaria alternata in Hami melon.

Hami melons are tasty and nutritive, but susceptibility to the fungus Alternaria alternata is one of the main problems leading to the postharvest loss of this fruit. The purpose of this research was to evaluate the effectiveness of nitric oxide (NO) on regulation of ethylene biosynthesis as well as signal transduction against black spot disease caused by A. alternata in the Hami melon. Nitric oxide reduced the growth of lesion diameter and lesion depth in melons inoculated with A. alternata. Ethylene production was significantly inhibited by NO, which was supported by the reduction of 1-aminocyclopropene-1-carboxylate (ACC) synthase (ACS) activity and the deferment of ACC content and ACC oxidase (ACO) activity. Nitric oxide treatment also significantly regulated the expression of four ethylene biosynthesis genes CmACS1, CmACS2, CmACO1, and CmACO2, and eight signal ethylene transduction genes CmETR1, CmETR2, CmCTR1, CmEIN2, CmEIL1, CmEBF1, CmERF1B and CmERF2. The modes of NO regulating these genes can be divided into five categories: promotion (CmEIN2, and CmEIL1), delay (CmACS1, CmETR2, CmCTR1 and CmERF2), up-regulation (CmETR1, CmEBF1 and CmERF1B), down-regulation (CmACS2), and first inhibition and then induction (CmACO1 and CmACO2). The NO treatment enhanced the postharvest disease resistance of Hami melon attacked by A. alternata, possibly by postponing ethylene biosynthesis and signal transduction. © 2021 Society of Chemical Industry.

Delay of ripening and senescence in mango fruit by 6-benzylaminopurine is associated with inhibition of ethylene biosynthesis and membrane lipid catabolism

6-Benzylaminopurine (6-BA), a synthetic cytokinin compound, plays a vital role in modulating diverse physiological processes in plants and is widely applied in agriculture. This study aims to investigate the effects of 6-BA on ripening and senescence and its possible mechanisms in harvested ‘Guifei’ mango fruit. The results exhibited that the application of 0.2 g L−1 6-BA to mango fruit inhibited the changes in firmness, flesh color, soluble solids content (SSC), titratable acidity (TA) and contents of carotenoid and chlorophyll, while retarded the climacteric process of respiration and ethylene. The delay of ethylene production due to 6-BA application was correlated with suppressed activities of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase (ACS) and ACC oxidase (ACO) in mango fruit. 6-BA treatment also reduced the accumulation of reactive oxygen species (ROS) (·O2- and H2O2) and malonaldehyde (MDA) and maintained a lower membrane permeability. Moreover, 6-BA treatment inhibited the activities of phospholipase D (PLD) and lipoxygenase (LOX) and downregulated the relative expression of MiPLD and MiLOX, which resulted in reduced degradation of phosphatidylcholine (PC) to phosphatidic acid (PA) and limited lipid peroxidation. The results suggest that 6-BA could delay ripening and senescence in mango fruit through regulating ethylene biosynthesis, ROS production and membrane lipid metabolism.

A tomato NAC transcription factor, SlNAM1, positively regulates ethylene biosynthesis and the onset of tomato fruit ripening.

Fruit ripening in tomato (Solanum lycopersicum) is the result of selective expression of ripening-related genes, which are regulated by transcription factors (TFs). The NAC (NAM, ATAF1/2, and CUC2) TF family is one of the largest families of plant-specific TFs and members are involved in a variety of plant physiological activities, including fruit ripening. Fruit ripening-associated NAC TFs studied in tomato to date include NAC-NOR (non-ripening), SlNOR-like1 (non-ripening like1), SlNAC1, and SlNAC4. Considering the large number of NAC genes in the tomato genome, there is little information about the possible roles of other NAC members in fruit ripening, and research on their target genes is lacking. In this study, we characterize SlNAM1, a NAC TF, which positively regulates the initiation of tomato fruit ripening via its regulation of ethylene biosynthesis. The onset of fruit ripening in slnam1-deficient mutants created by CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9) technology was delayed, whereas fruit ripening in OE-SlNAM1 lines was accelerated compared with the wild type. The results of RNA-sequencing (RNA-seq) and promoter analysis suggested that SlNAM1 directly binds to the promoters of two key ethylene biosynthesis genes (1-aminocyclopropane-1-carboxylate synthase: SlACS2 and SlACS4) and activates their expression. This hypothesis was confirmed by electrophoretic mobility shift assays and dual-luciferase reporter assay. Our findings provide insights into the mechanisms of ethylene production and enrich understanding of the tomato fruit ripening regulatory network.

The B’ζ subunit of protein phosphatase 2A negatively regulates ethylene signaling in Arabidopsis

Ethylene is a major plant hormone that regulates plant growth, development, and defense responses to biotic and abiotic stresses. The major pieces of the ethylene signaling pathway have been put together, although several details still need to be elucidated. For instance, the phosphorylation and dephosphorylation processes controlling the ethylene responses are poorly understood and need to be further explored. The type 2A protein phosphatase (PP2A) was suggested to play an important role in the regulation of ethylene biosynthesis, where the A1 subunit of PP2A was shown to be involved in the regulation of the rate-limiting enzyme of the ethylene biosynthetic pathway. However, whether other subunits of PP2A play roles in the ethylene signal transduction pathway is yet to be answered. In this study, we demonstrate that a B subunit, PP2A-B’ζ, positively regulates plant germination and seedling development, as a pp2a-b’ζ mutant is very sensitive to ethylene treatment. Furthermore, PP2A-B’ζ interacts with and stabilizes the kinase CTR1 (Constitutive Triple Response 1), a key enzyme in the ethylene signal transduction pathway, and like CTR1, PP2A-B’ζ negatively regulates ethylene signaling in plants.