Quantitative proteomic analysis of tomato fruit ripening behavior in response to exogenous abscisic acid.

BACKGROUND: To determine how abscisic acid (ABA) affects tomato fruit ripening at the protein level, mature green cherry tomato fruit were treated with ABA, nordihydroguaiaretic acid (NDGA) or sterile water (control, CK). The proteomes of treated fruit were analyzed and quantified using tandem mass tags (TMTs) at 7 days after treatment, and the gene transcription abundances of differently expressed proteins (DEPs) were validated with quantitative real-time polymerase chain reaction. RESULTS: Postharvest tomato fruit underwent faster color transformation and ripening than the CK when treated with ABA. In total, 6310 proteins were identified among the CK and treatment groups, of which 5359 were quantified. Using a change threshold of 1.2 or 0.83 times, 1081 DEPs were identified. Among them, 127 were upregulated and 127 were downregulated in the ABA versus CK comparison group. According to KEGG and protein-protein interaction network analyses, the ABA-regulated DEPs were primarily concentrated in the photosynthesis system and sugar metabolism pathways, and 102 DEPs associated with phytohormones biosynthesis and signal transduction, pigment synthesis and metabolism, cell wall metabolism, photosynthesis, redox reactions, allergens and defense responses were identified in the ABA versus CK and NDGA versus CK comparison groups. CONCLUSION: ABA affects tomato fruit ripening at the protein level to some extent. The results of this study provided comprehensive insights and data for further research on the regulatory mechanism of ABA in tomato fruit ripening. © 2023 Society of Chemical Industry.

Effects of Exogenous Abscisic Acid on Bioactive Components and Antioxidant Capacity of Postharvest Tomato during Ripening.

Abscisic acid (ABA) is a phytohormone which is involved in the regulation of tomato ripening. In this research, the effects of exogenous ABA on the bioactive components and antioxidant capacity of the tomato during postharvest ripening were evaluated. Mature green cherry tomatoes were infiltrated with either ABA (1.0 mM) or deionized water (control) and stored in the dark for 15 days at 20 °C with 90% relative humidity. Fruit colour, firmness, total phenolic and flavonoid contents, phenolic compounds, lycopene, ascorbic acid, enzymatic activities, and antioxidant capacity, as well as the expression of major genes related to phenolic compounds, were periodically monitored. The results revealed that exogenous ABA accelerated the accumulations of total phenolic and flavonoid contents; mostly increased the contents of detected phenolic compounds; enhanced FRAP and DPPH activity; and promoted the activities of PAL, POD, PPO, CAT, and APX during tomato ripening. Meanwhile, the expressions of the major genes (PAL1, C4H, 4CL2, CHS2, F3H, and FLS) involved in the phenylpropanoid pathway were up-regulated (1.13- to 26.95-fold) in the tomato during the first seven days after treatment. These findings indicated that ABA promoted the accumulation of bioactive components and the antioxidant capacity via the regulation of gene expression during tomato ripening.

Suppression of Cell Wall Degrading Enzymes and their Encoding Genes in Button Mushrooms (Agaricus bisporus) by CaCl2 and Citric Acid.

Fresh button mushrooms (Agaricus bisporus) were harvested and treated with a solution of 1.5% CaCl2 + 0.5% citric acid and stored for 16 days at 12 °C. The effects of this treatment on firmness, weight, color, cell wall compositions (cellulose and chitin) and cell wall degrading enzymes (cel1ulase, beta-1, 3 glucanase, chitinase and phenylalanine ammonialyase) were investigated during post-harvest storage. The expressions of major genes (Cel1, Glu1, Chi1 and PAL1) involved in cell wall degradation during post-harvest storage were also monitored. The results revealed that the post-harvest chemical treatment maintained better firmness, weight, color and inhibited cellulase, beta-1, 3 glucanase, chitinase and phenylalanine ammonialyase activities. These findings showed that the down-regulation of cell wall degrading enzymes is a possible mechanism that delays the softening of button mushrooms by the application of combined chemical treatment.

Transcriptome profiling of postharvest strawberry fruit in response to exogenous auxin and abscisic acid.

MAIN CONCLUSION: Auxin and abscisic acid regulate strawberry fruit ripening and senescence through cross-talk of their signal transduction pathways that further modulate the structural genes related to physico-chemical properties of fruit. The physiological and transcriptomic changes in harvested strawberry fruits in responses to IAA, ABA and their combination were analyzed. Exogenous IAA delayed the ripening process of strawberries after harvest while ABA promoted the postharvest ripening. However, treatment with a combination of IAA and ABA did not slow down nor accelerate the postharvest ripening in the strawberry fruits. At the molecular level, exogenous IAA up regulated the expressions of genes related to IAA signaling, including AUX/IAA, ARF, TOPLESS and genes encoding E3 ubiquitin protein ligase and annexin, and down regulated genes related to pectin depolymerization, cell wall degradation, sucrose and anthocyanin biosyntheses. In contrast, exogenous ABA induced genes related to fruit softening, and genes involved in signaling pathways including SKP1, HSPs, CK2, and SRG1. Comparison of transcriptomes in responses to individual treatments with IAA or ABA or the combination revealed that there were cooperative and antagonistic actions between IAA and ABA in fruit. However, 17% of the differentially expressed unigenes in response to the combination of IAA and ABA were unique and were not found in those unigenes responding to either IAA or ABA alone. The analyses also found that receptor-like kinases and ubiquitin ligases responded to both IAA and ABA, which seemed to play a pivotal role in both hormones' signaling pathways and thus might be the cross-talk points of both hormones.

Involvement of three annexin genes in the ripening of strawberry fruit regulated by phytohormone and calcium signal transduction.

KEY MESSAGE: Three annexin genes may be involved in the ripening progress of strawberry fruit. Phytohormones and calcium regulate the expressions of three annexin genes during strawberry fruit ripening. Plant annexins are multi-functional membrane- and Ca(2+)-binding proteins that are involved in various developmental progresses and stress responses. Three annexins FaAnn5a, FaAnn5b and FaAnn8 cDNA obtained from strawberry fruit encode amino acid sequences of approximately 35 kDa containing four annexin repeats, Ca(2+)-binding site, GTP-binding motif, peroxidase residue, and conserved amino acid residues of tryptophan, arginine and cysteine. During fruit development, the transcript levels of FaAnn5a and FaAnn5b increased while FaAnn5b declined after 3/4R stage. The expression patterns of annexins suggested their potential roles in strawberry fruit development and ripening. Expressions of annexin genes were also highly correlated with hormone levels. In addition, exogenous abscisic acid (ABA) enhanced the expressions of FaAnn5a and FaAnn8 while exogenous auxin (IAA) retarded it. However, both ABA and IAA promoted the transcript levels of FaAnn5b, indicating the independent regulation of annexins in fruit likely due to multi-functions of their large family. The responses of annexin genes to exogenous ABA and IAA inhibitors verified the involvement of annexins in plant hormone signaling. Besides, calcium restrained the expressions of FaAnn5s (FaAnn5a and FaAnn5b) but promoted the expression of FaAnn8. Effects of calcium and ethylene glycol tetraacetic acid (EGTA) on the transcript levels of annexins confirmed that calcium likely mediated hormone signal transduction pathways, which helped to elucidate the mechanism of calcium in fruit ripening. Therefore, FaAnn5s and FaAnn8 might be involved in plant hormones' regulation in the development and ripening of strawberry fruit through calcium signaling in the downstream.

Alleviation of chilling injury in postharvest tomato fruit by preconditioning with ultraviolet irradiation.

BACKGROUND: Tomato fruit is usually stored at low temperatures for delayed ripening and extended shelf life. However, tomato fruit is susceptible to chilling injury when exposed to low temperatures. In this study, the potential effects of preconditioning with UV-C or UV-B irradiation on chilling injury of postharvest tomato fruit were investigated. RESULTS: Mature-green tomato fruit were exposed to 4 kJ m(-2) UV-C or 20 kJ m(-2) UV-B irradiation and stored for 20 days at 2 °C and subsequently 10 days at 20 °C. UV irradiation was effective in reducing chilling injury index and delaying ethylene peak. Furthermore, UV irradiation preserved storage quality as manifested by reduced weight loss, better retention of firmness, and higher contents of total soluble solids, soluble protein and soluble sugar during subsequent storage at 20 °C. UV-C irradiation significantly delayed the development of the red colour after 10 days of storage at 20 °C. On the other hand, UV irradiation decreased total phenolics content and antioxidant capacity, suggesting possibly reduced stress response to low temperature resulted from enhanced physiological adaptation by UV preconditioning. CONCLUSION: Our results suggest that preconditioning with UV-C or UV-B irradiation in appropriate doses had a positive effect on alleviating chilling injury in postharvest tomato fruit.

Integrated application of nitric oxide and modified atmosphere packaging to improve quality retention of button mushroom (Agaricus bisporus).

Button mushrooms (Agaricus bisporus) were dipped for 10min in different concentrations (0.5, 1, and 2mM) of 2,2'-(hydroxynitrosohydrazino)-bisethanamine (DETANO), a nitric oxide donor, then packed in biorientated polypropylene (BOPP) bags, heat sealed and stored at 4°C for 16days (d). Mushroom weight loss, firmness, colour, percent open caps, total phenolics, ascorbic acid and H2O2 contents, superoxide anion (O2(-)) production rate and activities of polyphenol oxidase (PPO), superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) were measured. The results indicate that treatment with 1mM DETANO maintained a high level of firmness, delayed browning and cap opening, promoted the accumulation of phenolics, ascorbic acid and reduced the increases in both O2(-) production rate and H2O2 content. Furthermore, NO inhibited the activity of PPO, and increased the antioxidant enzymes activities of CAT, SOD and APX throughout storage period. Thus it was observed that application of NO in combination with modified atmosphere packaging (MAP) can extend the storage life of button mushroom up to 12d.

Structure and composition changes in the cell wall in relation to texture of shiitake mushrooms (Lentinula edodes) stored in modified atmosphere packaging.

BACKGROUND: Firmness in shiitake mushroom (Lentinula edodes) is an important textural attribute affecting consumer attitudes toward freshness and quality. In this study, the effects of modified atmosphere packaging (MAP) treatments on structure and composition changes in cell walls in relation to the texture of mushrooms were investigated. RESULTS: Shiitake mushrooms were packaged in low density polyethylene bags with no holes (M(0)), two microholes (M(1)), four macroholes (M(2)), stored at 4 degrees C for 16 days with non-wrapped mushrooms as control. Control mushrooms showed the highest firmness value due to significant increase of cellulose and chitin. All three MAP treatments reduced losses of protein and polysaccharides; the M(2) treatment can best preserve the original texture while mushrooms in M(0) became soft and deteriorated, possibly due to higher CO(2) accumulation, lower cellulose and chitin content. Transmission electron microscopy performed on caps at harvest and after 16 days indicated that disintegration of plasmalemma had been alleviated by M(2) treatment, leading to better preservation of the cell wall. CONCLUSION: Our results suggest that differences in firmness of shiitake mushrooms during storage may be due to differences in cellulose and chitin concentrations. M(2) treatment may be a useful way of maintaining shiitake mushrooms texture during storage at 4 degrees C.

Accumulation of lignin and malondialdehyde in relation to quality changes of button mushrooms (Agaricus bisporus) stored in modified atmosphere.

Whole fresh button mushrooms (Agaricus bisporus) were stored in unsealed bags in two types of modified atmosphere packaging (MAP), namely, active and passive, at 4 °C. The packaging film was 0.04 mm low-density polyethylene and the gas composition of the active MAP was 12% O(2), 2.5% CO(2) and 85.5% N(2). Firmness of Mushroom showed a positive correlation with the accumulation of lignin in the tissue. On the other hand, changes of malondialdehyde content with storage time were proportional to the evolution of browning. The button mushrooms in control treatment developed severe browning at the end of the 15-day storage, while the mushrooms of both active and passive MAP treatments browned slightly. MAP treatments could not inhibit phenylalanine ammonia lyase activity; however, it can reduce the lignification process by the inhibition of peroxidase (POD) activity and the accumulation of lignin. Correlation between the cinnamyl alcohol dehydrogenase activity and lignin accumulation was not obvious. Our results suggested that an increase in the firmness of mushrooms during senescence may be a consequence of tissue lignification, a process associated with increase in POD activity. Both active and passive MAPs were useful for the conservation of tenderness and whiteness in button mushrooms.

Characterization of a chitin-glucan complex from the fruiting body of Termitomyces albuminosus (Berk.) Heim.

Chitin-glucan complex (CGC), the main component of fungal cell wall, is reported to have wide applications in the food and pharmaceutical industries because of its physical and physiological activities. In this study, CGC was extracted from the fruiting body of Termitomyces albuminosus (Berk.) Heim with the treatments of deproteination, demineralization and depigmentation to obtain a yield of 13.46%, and its properties were investigated. The results indicated that CGC from T. albuminosus contained glucan and chitin in a molar ratio of 46:54, with very low contents of proteins and inorganic salts. The chitin in CGC was in the α-form, with crystallinity index of 64.81% and degree of acetylation of 65.40%. The surface morphology of CGC was dense and firm with no nanofibers and nanopores as observed by scanning electron microscopy, and the peak degradation temperature was determined to be 314.88 °C. This study suggested that CGC from T. albuminosus was promising to be an alternative source of crustacean chitinous products in the industry of food, medicine, waste water treatment and so on in the future.

Isolation, molecular characterization and antioxidant activity of a water-soluble polysaccharide extracted from the fruiting body of Termitornyces albuminosus (Berk.) Heim.

A water-soluble polysaccharide WSP1 was extracted from the fruiting body of Termitornyces albuminosus. Its molecular weight, monosaccharide composition and molecular structure were determined by GPC, GC-MS, UV spectroscopy, FT-IR spectroscopy, methylation analysis, NMR (1D and 2D) and AFM. Moreover, the antioxidant activity of WSP1 was evaluated in vitro by the tests of reducing power, scavenging ability on DPPH radical and hydroxyl radical, and chelating ability on ferrous ion. The results indicated that the molecular weight of WSP1 was 9 kDa, and it was mainly composed of fucose and galactose in a molar ratio of 1:3.09. Based on monosaccharide composition, methylation analysis and NMR, the possible repeating unit of WSP1 was presented as follows: →2-α-l-Fucp-1→ (6-α-d-Galp-1)3→. The antioxidant assay revealed that, in the concentration range tested in this experiment, WSP1 had strong scavenging ability on DPPH radical, suggesting that WSP1 could be potentially used as a powerful radical scavenger.

Positive Regulation of the Transcription of AchnKCS by a bZIP Transcription Factor in Response to ABA-Stimulated Suberization of Kiwifruit.

Wound-induced suberization is an essentially protective healing process for wounded fruit to reduce water loss and microbial infection. It has been demonstrated that abscisic acid (ABA) could promote wound suberization, but the molecular mechanism of ABA regulation remains little known. In this study, the transcript level of Achn030011 (designated as AchnKCS), coding a ß-ketoacyl-coenzyme A synthase (KCS) involved in suberin biosynthesis, was found to be significantly upregulated by ABA in wounded kiwifruit. A bZIP transcription factor (Achn270881), a possible downstream transcription factor in the ABA signaling pathway, was screened and designated as AchnbZIP12 according to its homology with related Arabidopsis transcription factors. A yeast one-hybrid assay demonstrated that AchnbZIP12 could interact with the AchnKCS promoter. Furthermore, significant trans-activation of AchnbZIP12 on AchnKCS was verified. The transcript level of AchnbZIP12 was also upregulated upon treatment with ABA. These results imply that AchnbZIP12 acts as a positive regulator in ABA-mediated AchnKCS transcription during wound suberization of kiwifruit.

Integrated analysis of high-throughput sequencing data shows abscisic acid-responsive genes and miRNAs in strawberry receptacle fruit ripening.

The perception and signal transduction of the plant hormone abscisic acid (ABA) are crucial for strawberry fruit ripening, but the underlying mechanism of how ABA regulates ripening-related genes has not been well understood. By employing high-throughput sequencing technology, we comprehensively analyzed transcriptomic and miRNA expression profiles simultaneously in ABA- and nordihydroguaiaretic acid (NDGA, an ABA biosynthesis blocker)-treated strawberry fruits with temporal resolution. The results revealed that ABA regulated many genes in different pathways, including hormone signal transduction and the biosynthesis of secondary metabolites. Transcription factor genes belonging to WRKY and heat shock factor (HSF) families might play key roles in regulating the expression of ABA inducible genes, whereas the KNOTTED1-like homeobox protein and Squamosa Promoter-Binding-like protein 18 might be responsible for ABA-downregulated genes. Additionally, 20 known and six novel differentially expressed miRNAs might be important regulators that assist ABA in regulating target genes that are involved in versatile physiological processes, such as hormone balance regulation, pigments formation and cell wall degradation. Furthermore, degradome analysis showed that one novel miRNA, Fa_novel6, could degrade its target gene HERCULES1, which likely contributed to fruit size determination during strawberry ripening. These results expanded our understanding of how ABA drives the strawberry fruit ripening process as well as the role of miRNAs in this process.

[Application of Vis/NIR diffuse reflectance spectroscopy to the detection and identification of transgenic tomato leaf].

The feasibility of Vis/NIR spectroscopy technique for rapid and non-invasive detection of transgenic tomato leaves from conventional ones was investigated by means of spectral diffuse reflectance mode. A total of 68 samples (38 transgenic ones and 30 non-transgenic ones) were used for classification. The calibration and validation results were analyzed via discriminant analysis (DA) and partial least squares (PLS) discriminant method using TQ 6.2. 1 quantitative software. Models based on the different spectral pre-processing methods (multiplicative signal correction (MSC), first and second derivative) were compared. It was found that the classification accuracy using DA was higher than that using PLS and the best results were gained by using spectra after MSC with InGaAs detector and the classification accuracy was 89.7% (accuracy of 86.8% for transgenic samples and 93.3% for non-transgenic ones). The results show that Vis-NIR diffuse reflectance spectroscopy technique is a feasible and fast method for non-invasive detection of transgenic and non-transgenic tomato leaves.

SlAREB1 transcriptional activation of NOR is involved in abscisic acid-modulated ethylene biosynthesis during tomato fruit ripening.

Many studies have shown that abscisic acid (ABA) regulates climacteric fruits ripening by inducing ethylene production. Nevertheless, the key components involved in the crosstalk between these two phytohormones in controlling fruit ripening remain unknown. SlAREB1, a downstream transcription factor in ABA signaling pathway, has been reported to mediate ABA signaling that regulates tomato ripening through induction of ethylene biosynthetic genes. NOR, a member of NAC domain family, was proved to act upstream of ethylene and essential for ripening- and ethylene-associated genes expression. Here, we found that the expression of SlAREB1 and NOR are both ABA-inducible, and SlAREB1 transcription reaches the peak level prior to NOR during the ripening process. Yeast one-hybrid (Y1H), electrophoretic mobility shift assay (EMSA) and dual luciferase assay indicated NOR as a novel direct target of SlAREB1. Transient over-expression of SlAREB1 in tomato fruits results in elevated expression of NOR as well as a number of downstream ethylene biosynthetic genes including SlACS2, SlACS4 and SlACO1, suggesting that SlAREB1 can mediate ABA signal to activate NOR transcription and ultimately promote ethylene synthesis. Based on these data, we present a model suggesting that the SlAREB1-NOR regulation is a crucial node modulating ABA-regulated ethylene biosynthesis during tomato fruit ripening.

Contribution of abscisic acid to aromatic volatiles in cherry tomato (Solanum lycopersicum L.) fruit during postharvest ripening.

Fruit aroma development depends on ripening. Abscisic acid (ABA) has been reported to be involved in the regulation of tomato fruit ripening. In the present study, the effects of exogenous ABA on aromatic volatiles in tomato fruit during postharvest ripening were studied. The results showed that exogenous ABA accelerated color development and ethylene production as well as the accumulation of carotenoids, total phenolics and linoleic acid in tomato fruit during ripening. Moreover, exogenous ABA increased the accumulation of volatile compounds such as 1-peten-3-one (2.06-fold), ß-damascenone (1.64-fold), benzaldehyde (3.29-fold) and benzyl cyanide (4.15-fold); induced the expression of key genes implicated in the biosynthesis pathways of aromatic volatiles, including TomloxC, HPL, ADH2, LeCCD1B and SlBCAT1 (the values of the log2 fold changes ranged from -3.02 to 2.97); and promoted the activities of lipoxygenase (LOX), hydroperoxide lyase (HPL) and alcohol dehydrogenase (ADH). In addition, the results of promoter analyses revealed that cis-acting elements involved in ABA responsiveness (ABREs) exist in 8 of the 12 key genes involved in volatile biosynthesis, suggesting that ABA potentially affects aromatic volatile emissions via the regulation of gene expression profiles.

Quantification of chlorophyll content and classification of nontransgenic and transgenic tomato leaves using visible/near-infrared diffuse reflectance spectroscopy.

Visible/near-infrared (vis/NIR) spectroscopy combined with multivariate analysis was used to quantify chlorophyll content in tomato leaves and classify tomato leaves with different genes. In this study, transgenic tomato leaves with antisense LeETR1 (n = 106) and their parent nontransgenic ones (n = 102) were measured in vis/NIR diffuse reflectance mode. Quantification of chlorophyll content was achieved by partial least-squares regression with a cross-validation prediction error equal to 2.87. Partial least-squares discriminant analysis was performed to classify leaves. The results show that differences between transgenic and nontransgenic tomato leaves do exist, and excellent classification can be obtained after optimizing spectral pretreatment. The classification accuracy can reach to 100% using the derivative of spectral data in the full and partial wavenumber range. These results demonstrate that vis/NIR spectroscopy together with chemometrics techniques could be used to quantify chlorophyll content and differentiate tomato leaves with different genes, which offers the benefit of avoiding time-consuming, costly, and laborious chemical and sensory analysis.

Characteristics of transgenic tomatoes antisensed for the ethylene receptor genes LeETR1 [corrected] and LeETR2 [corrected].

Two stable transformed lines containing antisense LeETR1 [corrected] or LeETR2 [corrected] sequences and their hybridized line were investigated to determine the effect of LeETR1 [corrected] and LeETR2 [corrected] specificity in the ethylene receptor family in tomato (Lycopersicon esculentum Mill.) on ethylene signaling. The transgenic line ale1 containing antisense LeETR1 [corrected] displayed shorter length of seedling grown in the dark and adult plant in the light, severe epinastic petiole, and accelerated abscission of petiole explant and senescence of flower explant, compared with its wild type B1. The transgenic line ale2 containing antisense LeETR2 [corrected] also exhibited shorter hypocotyls and slightly accelerated abscission. The phenotypes of cross line dale of LeETR1 [corrected] and LeETR2 [corrected] were close to ale1 in many aspects. These results suggested that LeETR1 [corrected] probably plays a relatively important role in ethylene signaling of tomato growth and development.

Contribution of polyamines metabolism and GABA shunt to chilling tolerance induced by nitric oxide in cold-stored banana fruit.

Effect of exogenous nitric oxide (NO) on polyamines (PAs) catabolism, γ-aminobutyric acid (GABA) shunt, proline accumulation and chilling injury of banana fruit under cold storage was investigated. Banana fruit treated with NO sustained lower chilling injury index than the control. Notably elevated nitric oxide synthetase activity and endogenous NO level were observed in NO-treated banana fruit. PAs contents in treated fruit were significantly higher than control fruit, due to the elevated activities of arginine decarboxylase and ornithine decarboxylase. NO treatment increased the activities of diamine oxidase, polyamine oxidase and glutamate decarboxylase, while reduced GABA transaminase activity to lower levels compared with control fruit, which resulted the accumulation of GABA. Besides, NO treatment upregulated proline content and significantly enhanced the ornithine aminotransferase activity. These results indicated that the chilling tolerance induced by NO treatment might be ascribed to the enhanced catabolism of PAs, GABA and proline.

Comprehensive Analysis of ABA Effects on Ethylene Biosynthesis and Signaling during Tomato Fruit Ripening.

ABA has been widely acknowledged to regulate ethylene biosynthesis and signaling during fruit ripening, but the molecular mechanism underlying the interaction between these two hormones are largely unexplored. In the present study, exogenous ABA treatment obviously promoted fruit ripening as well as ethylene emission, whereas NDGA (Nordihydroguaiaretic acid, an inhibitor of ABA biosynthesis) application showed the opposite biological effects. Combined RNA-seq with time-course RT-PCR analysis, our study not only helped to illustrate how ABA regulated itself at the transcription level, but also revealed that ABA can facilitate ethylene production and response probably by regulating some crucial genes such as LeACS4, LeACO1, GR and LeETR6. In addition, investigation on the fruits treated with 1-MCP immediately after ABA exposure revealed that ethylene might be essential for the induction of ABA biosynthesis and signaling at the onset of fruit ripening. Furthermore, some specific transcription factors (TFs) known as regulators of ethylene synthesis and sensibility (e.g. MADS-RIN, TAGL1, CNR and NOR) were also observed to be ABA responsive, which implied that ABA influenced ethylene action possibly through the regulation of these TFs expression. Our comprehensive physiological and molecular-level analysis shed light on the mechanism of cross-talk between ABA and ethylene during the process of tomato fruit ripening.