Exogenous BR delayed peach fruit softening by inhibiting pectin degradation enzyme genes.

Peach fruit deteriorates and senesces rapidly when stored at room temperature. Brassinosteroids (BRs) play an important role in regulating plant growth and development and maintaining fruit quality. However, little information is available on the effect of BRs on the senescence of harvested peach fruit. In this study, different concentrations of BR were used to treat 'Hongniang' peach fruit, and the results showed that 10 µM BR was the most beneficial concentration to delay the senescence of peach fruits. BR treatment delayed the decrease of fruit firmness, the release of ethylene, the increase in water-soluble pectin (WSP) and ionic-soluble pectin (ISP) content and the decrease in covalently bound pectin (CBP) content, inhibited the activities of pectin degradation enzymes, and inhibited the gene expression of PpPME1/3, PpPG, PpARF2, and PpGAL2/16. In addition, BR treatment also inhibited the expression of PpBES1-5/6. Cis-acting regulatory element analysis of pectin degradation enzyme promoters showed that many of them contained BES1 binding elements. All the above results showed that BR treatment had a positive effect on delaying the senescence of peach fruit and prolonging its storage period.

Low temperature inhibits pectin degradation by PpCBFs to prolong peach storage time.

Low-temperature storage is a widely used method for peach fruit storage. However, the impact of PpCBFs on pectin degradation during low-temperature storage is unclear. As such, in this study, we stored the melting-flesh peach cultivar "Fuli" at low temperature (LT, 6°C) and room temperature (RT, 25°C) to determine the effect of different temperatures on its physiological and biochemical changes. Low-temperature storage can inhibit the softening of "Fuli" peaches by maintaining the stability of the cell wall. It was found that the contents of water-soluble pectin and ionic-soluble pectin in peach fruit stored at RT were higher than those stored at LT. The enzyme activities of polygalacturonase (PG), pectate lyase (PL), and pectin methylesterase (PME) were all inhibited by LT. The expressions of PpPME3, PpPL2, and PpPG were closely related to fruit firmness, but PpCBF2 and PpCBF3 showed higher expression levels at LT than RT. The promoters of PpPL2 and PpPG contain the DER motif, which suggested that PpCBF2 and PpCBF3 might negatively regulate their expression by directly binding to their promoters. These results indicated that LT may maintain firmness by activating PpCBFs to repress pectin-degradation-related enzyme genes during storage.

Self-Supervised Monocular Depth Estimation With Self-Perceptual Anomaly Handling.

It is attractive to extract plausible 3-D information from a single 2-D image, and self-supervised learning has shown impressive potential in this field. However, when only monocular videos are available as training data, moving objects at similar speeds to the camera can disturb the reprojection process during training. Existing methods filter out some moving pixels by comparing pixelwise photometric error, but the illumination inconsistency between frames leads to incomplete filtering. In addition, existing methods calculate photometric error within local windows, which leads to the fact that even if an anomalous pixel is masked out, it can still implicitly disturb the reprojection process, as long as it is in the local neighborhood of a nonanomalous pixel. Moreover, the ill-posed nature of monocular depth estimation makes the same scene correspond to multiple plausible depth maps, which damages the robustness of the model. In order to alleviate the above problems, we propose: 1) a self-reprojection mask to further filter out moving objects while avoiding illumination inconsistency; 2) a self-statistical mask method to prevent the filtered anomalous pixels from implicitly disturbing the reprojection; and 3) a self-distillation augmentation consistency loss to reduce the impact of ill-posed nature of monocular depth estimation. Our method shows superior performance on the KITTI dataset, especially when evaluating only the depth of potential moving objects.

The Malus domestica metal tolerance protein MdMTP11.1 was involved in the detoxification of excess manganese in Arabidopsis thaliana.

Ion homeostasis is maintained in plant cells by specialized transporters. However, functional studies on Mn transporters in apple trees have not been reported. MdMTP11.1, which encodes a putative Mn-MTP transporter in Malus domestica, was expressed highly in leaves and induced by Mn stress. Subcellular localization analysis of the MdMTP11.1-GFP fusion protein indicated that MdMTP11.1 was targeted to the Golgi. Meanwhile, overexpression of MdMTP11.1 in Arabidopsis thaliana conferred increased resistance to plants under toxic Mn levels, as evidenced by increased biomass of whole plant and length of primary root. Analysis of Mn bioaccumulation indicated that overexpression of MdMTP11.1 effectively reduced the content of Mn in every subcellular component and chemical forms when the plants were subjected with Mn stress. The majority of Mn of action were bound to cell wall and combined with un-dissolved phosphate. Besides, contents of malondialdehyde (MDA), proline and hydrogen peroxide (H2O2) were significantly lower, while content of chlorophyll and activities of CAT, SOD, POD and APX were significantly higher in MdMTP11.1-over-expressing plants compared with that in wild type plants under Mn stress. Taken together, these results suggest that MdMTP11.1 is a Mn specific transporter localized to the Golgi can maintain the phenotype, reduce the Mn accumulation and alleviate damage of oxidative stress, conferring the positive role of Mn tolerance.

Genome-wide characterization of SINA E3 ubiquitin ligase family members and their expression profiles in response to various abiotic stresses and hormones in kiwifruit.

SINA (Seven in absentia) proteins in the subtype of E3 ubiquitin ligase family have important functions in regulating the growth and development as well as in response to abiotic and biotic stresses in plants. However, the characteristics and possible functions of SINA family proteins in kiwifruit are not studied. In this research, a total number of 11 AcSINA genes in the kiwifruit genome were identified. Chromosome location and multiple sequence alignment analyses indicated that they were unevenly distributed on 10 chromosomes and all contained the typical N-terminal RING domain and C-terminal SINA domain. Phylogenetic, gene structure and collinear relationship analyses revealed that they were highly conserved with the same gene structure, and have gone through segmental duplication events. Expression pattern analyses demonstrated that all AcSINAs were ubiquitously expressed in roots, stems and leaves, and were responsive to different abiotic and plant hormone treatments with overlapped but distinct expression patterns. Further yeast two-hybrid and Arabidopsis transformation analyses demonstrated most AcSINAs interacted with itself or other AcSINA members to form homo- or heterodimers, and ectopic expression of AcSINA2 in Arabidopsis led to hypersensitive growth phenotype of transgenic seedlings to ABA treatment. Our results reveal that AcSINAs take part in the response to various abiotic stresses and hormones, and provide important information for the functional elucidation of AcSINAs in vine fruit plants.

Low temperatures inhibit the pectin degradation of 'Docteur Jules Guyot' pear (Pyrus communis L.).

The most remarkable characteristic of European pears is extremely perishable and difficult to store after postharvest softening. Low-temperature storage is one of the most commonly used methods to prolong the shelf life of European pears. However, the regulatory mechanism of the low-temperature delay of the softening of European pears is still unclear. In this study, the fruit firmness, pectin polysaccharide content, pectin-degrading enzyme activity, and pectin degradation gene expression of 'Docteur Jules Guyot' pears under low temperature (LT) and room temperature (RT) were analyzed. It was found that water-soluble pectin (WSP) was significantly negatively correlated with fruit flesh firmness, and the activities of several pectin-degrading enzymes were inhibited under LT storage conditions. In addition, it was also found that the gene expression patterns of PcPME2, PcPME3, PcPG1, PcPG2, PcPL, PcGAL1, PcGAL2, PcGAL4, and PcARF1 were inhibited by LT. The C-repeat binding factors PcCBF1 and PcCBF2 were also inhibited by long-term LT storage. Correlation analysis showed that the expression of PcCBFs was positively correlated with pectin-degradation enzyme genes, and we found that the promoters of many pectin-degradation enzyme genes contain the CRT/DRE motif, which CBF can directly bind. Therefore, it is speculated that long-term low-temperature conditions inhibit pectin degradation through PcCBFs.

High Throughput Omnidirectional Printing of Tubular Microstructures from Elastomeric Polymers.

Bioelastomers are extensively used in biomedical applications due to their desirable mechanical strength, tunable properties, and chemical versatility; however, three-dimensional (3D) printing bioelastomers into microscale structures has proven elusive. Herein, a high throughput omnidirectional printing approach via coaxial extrusion is described that fabricates perfusable elastomeric microtubes of unprecedently small inner diameter (350-550 µm) and wall thickness (40-60 µm). The versatility of this approach is shown through the printing of two different polymeric elastomers, followed by photocrosslinking and removal of the fugitive inner phase. Designed experiments are used to tune the microtube dimensions and stiffness to match that of native ex vivo rat vasculature. This approach affords the fabrication of multiple biomimetic shapes resembling cochlea and kidney glomerulus and affords facile, high-throughput generation of perfusable structures that can be seeded with endothelial cells for biomedical applications. Post-printing laser micromachining is performed to generate micro-sized holes (520 µm) in the tube wall to tune microstructure permeability. Importantly, for organ-on-a-chip applications, the described approach takes only 3.6 min to print microtubes (without microholes) over an entire 96-well plate device, in contrast to comparable hole-free structures that take between 1.5 and 6.5 days to fabricate using a manual 3D stamping approach.

Metabolome and Transcriptome Analyses of Anthocyanin Accumulation Mechanisms Reveal Metabolite Variations and Key Candidate Genes Involved in the Pigmentation of Prunus tomentosa Thunb. Cherry Fruit.

Prunus tomentosa Thunb. has excellent nutritional, economic, and ornamental values with different fruit color. The red coloration of fruit is determined by anthocyanin pigmentation, which is an attractive trait for consumers. However, the mechanisms underlying fruit color formation in the P. tomentosa cherry are not well understood. In this research, the pigmentation patterns in red-color P. tomentosa (RP) fruit and white-color P. tomentosa (WP) were evaluated. Anthocyanin content in matured RP fruit was significantly abundant compared with WP fruit. Metabolomic profiling revealed that pelargonidin 3-O-glucoside, cyanidin 3-O-rutinoside, and pelargonidin 3-O-rutinoside were the predominant anthocyanin compounds in the RP fruit, while, WP fruit had less anthocyanin compositions and lower level. Then, integrative analyses of transcriptome and metabolome identified 285 significant differentially expressed genes (DEGs) closely related to anthocyanin differentially expressed metabolites (DEMs). Among them, nine genes were involved in anthocyanin biosynthesis, transport and degradation pathway, including four biosynthesis genes (PtPAL1, PtDFR, PtANS, and PtUFGT), two transport genes (PtGST11, PtABC10), and three degradation genes (PtPOD1, PtPOD16, PtPOD73). Transcriptome data and real-time PCR showed that the transcript levels of biosynthesis and transport genes were significantly higher in RP than in WP, especially PtANS, PtUFGT, and PtGST11, suggesting they may play key roles in red-colored fruit formation. Meanwhile, the degradation-related genes PtPOD1/16/73 took on exactly opposite trend, suggesting their potential effects on anthocyanin degradation. These results provide novel insights into color patterns formation mechanisms of cherries fruit, and the candidate key genes identified in anthocyanin biosynthesis, transport and degradation may provide a valuable resource for cherry breeding research in future.

Heterologous expression of ISU1 gene from Fragaria vesca enhances plant tolerance to Fe depletion in Arabidopsis.

Iron-sulfur (Fe-S) cluster assembly genes play important roles in plant growth and development. However, their biological function in fruit crops is still unknown, especially in strawberry. In this study, Fe depletion significantly inhibited the growth, photosynthesis, Fe accumulation level and the enzyme activity of Fe-S proteins of aconitase (ACO), nitrate reductase (NiR) and succinate dehydrogenase (SDH) in strawberry seedlings. In addition, 40 Fe-S cluster assembly genes were isolated from strawberry, which were significantly varied among different tissues/organs and were differentially responded to Fe depletion in different tissue parts. In total, 79% of the responsive genes were up-regulated in shoots, while 65% of the responsive genes were down-regulated in roots under Fe depletion. Moreover, the expression level of ISU1 was the highest in strawberry tissues, especially in young fruits, and over-expression of ISU1 gene in Arabidopsis significantly enhanced the Fe accumulation, leaf total chlorophyll, ACO and SDH activities in transgenic lines, and strengthened plant tolerance to Fe depletion. This study provides gene resources to elucidate the molecular mechanisms of Fe-S cluster assembly in strawberry, and lays a theoretical foundation to reveal Fe nutrition and metabolism in Rosaceae fruits.

Self-Supervised Monocular Depth Estimation With Multiscale Perception.

Extracting 3D information from a single optical image is very attractive. Recently emerging self-supervised methods can learn depth representations without using ground truth depth maps as training data by transforming the depth prediction task into an image synthesis task. However, existing methods rely on a differentiable bilinear sampler for image synthesis, which results in each pixel in a synthetic image being derived from only four pixels in the source image and causes each pixel in the depth map to perceive only a few pixels in the source image. In addition, when calculating the photometric error between a synthetic image and its corresponding target image, existing methods only consider the photometric error within a small neighborhood of each single pixel and therefore ignore correlations between larger areas, which causes the model to tend to fall into the local optima for small patches. In order to extend the perceptual area of the depth map over the source image, we propose a novel multi-scale method that downsamples the predicted depth map and performs image synthesis at different resolutions, which enables each pixel in the depth map to perceive more pixels in the source image and improves the performance of the model. As for the locality of photometric error, we propose a structural similarity (SSIM) pyramid loss to allow the model to sense the difference between images in multiple areas of different sizes. Experimental results show that our method achieves superior performance on both outdoor and indoor benchmarks.

High-quality genome assembly of 'Cuiguan' pear (Pyrus pyrifolia) as a reference genome for identifying regulatory genes and epigenetic modifications responsible for bud dormancy.

Dormancy-associated MADS-box (DAM) genes serve as crucial regulators of the endodormancy cycle in rosaceous plants. Although pear DAM genes have been identified previously, the lack of a high-quality reference genome and techniques to study gene function have prevented accurate genome-wide analysis and functional verification of such genes. Additionally, the contribution of other genes to the regulation of endodormancy release remains poorly understood. In this study, a high-quality genome assembly for 'Cuiguan' pear (Pyrus pyrifolia), which is a leading cultivar with a low chilling requirement cultivated in China, was constructed using PacBio and Hi-C technologies. Using this genome sequence, we revealed that pear DAM genes were tandemly clustered on Chr8 and Chr15 and were differentially expressed in the buds between 'Cuiguan' and the high-chilling-requirement cultivar 'Suli' during the dormancy cycle. Using a virus-induced gene silencing system, we determined the repressive effects of DAM genes on bud break. Several novel genes potentially involved in the regulation of endodormancy release were identified by RNA sequencing and H3K4me3 chromatin immunoprecipitation sequencing analyses of 'Suli' buds during artificial chilling using the new reference genome. Our findings enrich the knowledge of the regulatory mechanism underlying endodormancy release and chilling requirements and provide a foundation for the practical regulation of dormancy release in fruit trees as an adaptation to climate change.

Alternative splicing of the dormancy-associated MADS-box transcription factor gene PpDAM1 is associated with flower bud dormancy in 'Dangshansu' pear (Pyrus pyrifolia white pear group).

Alternative splicing (AS) plays a crucial role in plant growth, development and response to various environmental changes. However, whether alternative splicing of MADS-box transcription factors contributes to the flower bud dormancy process in fruit trees still remains unknown. In this work, the AS profile of genes in the dormant flower buds of 'Dangshansu' pear tree were examined. A total number of 3661 alternatively spliced genes were identified, and three mRNA isoforms of the dormancy associated MADS box (DAM) gene, PpDAM1, derived by alternative splicing, designated as PpDAM1.1, PpDAM1.2 and PpDAM1.3, were characterized. Bimolecular fluorescence complementation (BiFC) analysis indicated that AS of PpDAM1 didn't affect the nucleus localization and homo-/heterodimerization of PpDAM1.1, PpDAM1.2 and PpDAM1.3 proteins, but disturbed the translocation of PpDAM1.1/PpDAM1.1, PpDAM1.3/PpDAM1.3, PpDAM1.1/PpDAM1.3, and PpDAM1.2/PpDAM1.3 dimers to the nucleus. Constitutive expression of PpDAM1.2, but not PpDAM1.1 and PpDAM1.3, in Arabidopsis retarded the growth and development of transgenic plants. Further comparative expression analyses of PpDAM1.1, PpDAM1.2 and PpDAM1.3 in the flower buds of 'Dangshansu' and a less dormant pear cultivar, 'Cuiguan', exhibited that the expression of all the three isoforms in 'Dangshansu' were significantly higher than in 'Cuiguan', especially PpDAM1.2, which showed a predominantly higher expression than PpDAM1.1 and PpDAM1.3 in both cultivars. Our results suggest that alternative splicing of PpDAM1 could play a crucial role in pear flower bud dormancy process.

Downregulation of long non-coding RNA LINC-PINT serves as a diagnostic and prognostic biomarker in patients with non-small cell lung cancer.

Long non-coding RNAs (lncRNAs) play an important role in gene regulation. Several lncRNAs have been demonstrated to be associated with the diagnosis and prognosis of non-small cell lung cancer (NSCLC). The present study aimed to investigate the role of lncRNA long intragenic non-protein-coding RNA p53-induced transcript (LINC-PINT) in NSCLC to identify a novel non-invasive biomarker for the diagnosis and prognosis of patients with NSCLC. Reverse transcription-quantitative PCR analysis was performed to detect LINC-PINT expression in the tissue and serum samples of patients with NSCLC. The diagnostic and prognostic values of LINC-PINT were assessed via the receiver operating characteristic curve, and Kaplan-Meier and Cox regression analyses, respectively. The results demonstrated that LINC-PINT expression was significantly downregulated in NSCLC serum samples and tissues. In addition, serum LINC-PINT exhibited diagnostic value in patients with NSCLC, and may be used to predict prognosis. Furthermore, aberrant LINC-PINT expression in tumor tissues was significantly associated with lymph node metastasis, tumor size, differentiation and TNM stage. Taken together, the results of the present study suggest that lncRNA LINC-PINT may be an independent diagnostic and prognostic biomarker in NSCLC.

The Chloroplastic Small Heat Shock Protein Gene KvHSP26 Is Induced by Various Abiotic Stresses in Kosteletzkya virginica.

Small heat shock proteins (sHSPs) are a group of chaperone proteins existed in all organisms. The functions of sHSPs in heat and abiotic stress responses in many glycophyte plants have been studied. However, their possible roles in halophyte plants are still largely known. In this work, a putative sHSP gene KvHSP26 was cloned from K. virginica. Bioinformatics analyses revealed that KvHSP26 encoded a chloroplastic protein with the typical features of sHSPs. Amino acid sequence alignment and phylogenetic analysis demonstrated that KvHSP26 shared 30%-77% homology with other sHSPs from Arabidopsis, cotton, durian, salvia, and soybean. Quantitative real-time PCR (qPCR) assays exhibited that KvHSP26 was constitutively expressed in different tissues such as leaves, stems, and roots, with a relatively higher expression in leaves. Furthermore, expression of KvHSP26 was strongly induced by salt, heat, osmotic stress, and ABA in K. virginica. All these results suggest that KvHSP26 encodes a new sHSP, which is involved in multiple abiotic stress responses in K. virginica, and it has a great potential to be used as a candidate gene for the breeding of plants with improved tolerances to various abiotic stresses.

Inhibition and enhancement of linear and nonlinear optical effects by conical phase front shaping for femtosecond laser material processing.

The emergence of high-powered femtosecond lasers presents the opportunity for large volume processing inside of transparent materials, wherein a myriad of nonlinear optical and aberration effects typically convolves to distort the focused beam shape. In this paper, convex and concave conical phase fronts were imposed on femtosecond laser beams and focussed into wide-bandgap glass to generate a vortex beam with tuneable Gaussian-Bessel features offset from the focal plane. The influence of Kerr lensing, plasma defocussing, and surface aberration on the conical phase front shaping were examined over low to high pulse energy delivery and for shallow to deep processing tested to 2.5 mm focussing depth. By isolating the underlying processes, the results demonstrate how conical beams can systematically manipulate the degree of nonlinear interaction and surface aberration to facilitate a controllable inhibition or enhancement of Kerr lensing, plasma defocussing, and surface aberration effects. In this way, long and uniform filament tracks have been generated over shallow to deep focussing by harnessing surface aberration and conical beam shaping without the destabilizing Kerr lensing and plasma defocussing effects. A facile means for compressing and stretching of the focal interaction volume is presented for controlling the three-dimensional micro- and nano-structuring of transparent materials.

Compensating deep focusing distortion for femtosecond laser inscription of low-loss optical waveguides.

Various beam shaping approaches were examined to counter the negative influence of surface aberration arising when inscribing optical waveguides deeply inside of glass with a femtosecond laser. Aberration correction was found unable to completely recover the low-loss waveguide properties, prompting a comprehensive examination of waveguides formed with focused Gaussian-Bessel beams. Diverging conical phase fronts are presented as a hybrid means of partial aberration correction to improve insertion loss and a new, to the best of our knowledge, means of asymmetric beam shaping. In this way, low-loss waveguides are presented over shallow to deep writing depth (2.8 mm) where morphological and modal properties could be further tuned with conical phase front.

Aberrant expression of miR-4728 in patients with non-small cell lung cancer and its regulatory effects on tumor progression in tumor cells.

Non-small cell lung cancer (NSCLC) is a common malignant tumor with poor prognosis and an increasing number of cases. MicroRNA (miR)-4728 is related with the progression of various types of cancer, and is dysregulated in NSCLC, which indicates that miR-4728 may serve as a biomarker for NSCLC. The present study aimed to investigate the clinical significance of miR-4728 in NSCLC diagnosis and prognosis, and to explore the biological function of miR-4728 in NSCLC progression. Serum and tissue samples were collected from 122 patients with NSCLC. By conducting reverse transcription-quantitative PCR, the Cell Counting Kit-8 assay and Transwell assays, the expression of miR-4728 and its effect on NSCLC cell proliferation, migration and invasion were investigated. The diagnostic value of miR-4728 was evaluated by plotting a receiver operating characteristic curve, and Kaplan-Meier and Cox regression analyses were conducted to assess the prognostic value of miR-4728. miR-4728 was significantly downregulated in NSCLC serum and tissue samples compared with healthy controls, with a relatively high diagnostic accuracy and ability to predict poor overall survival time in patients with NSCLC. By conducting gain- and loss-of-function experiments, the results indicated that miR-4728 knockdown significantly promoted NSCLC cell proliferation, migration and invasion compared with the inhibitor negative control (NC) group. By contrast, miR-4728 overexpression displayed the opposite effect on NSCLC cell proliferation, migration and invasion. The present study indicated that miR-4728 was downregulated in NSCLC and may serve as a candidate diagnostic and prognostic biomarker. NSCLC cell proliferation, migration and invasion were inhibited by miR-4728 overexpression compared with the mimic NC group, which suggested that miR-4728 may serve as a therapeutic target for NSCLC.

MicroRNA-374b-5p Functions as a Tumor Suppressor in Non-Small Cell Lung Cancer by Targeting FOXP1 and Predicts Prognosis of Cancer Patients.

BACKGROUND: Lung cancer remains the most frequent malignancy worldwide with increasing morbidity and mortality. This study aimed to assess the expression of microRNA-374b-5p (miR-374b-5p) in tissues and cell lines of non-small cell lung cancer (NSCLC) and to evaluate the prognostic value of miR-374b-5p as well as its biological function in tumor progression. MATERIALS AND METHODS: Expression of miR-374b-5p in NSCLC patients and cells was estimated using quantitative real-time PCR. The prognostic value of miR-374b-5p was evaluated using Kaplan-Meier method and Cox regression analysis. Gain-of-function and loss-of-function cell experiments were performed to examine the effects of miR-374b-5p on NSCLC cell proliferation, migration and invasion. A luciferase activity assay was used to confirm the target gene of miR-374b-5p. RESULTS: miR-374b-5p expression levels were decreased in tumorous tissues and cell lines compared with the normal tissues or cells (P < 0.05). The expression of miR-374b-5p was associated with the patients' tumor size, lymph node metastasis and TNM stage (all P < 0.05). Patients with low miR-374b-5p expression have a shorter survival time (log-rank P = 0.001), and the downregulated expression of miR-374b-5p was determined to be an independent prognostic indicator of NSCLC. In NSCLC cells, the overexpression of miR-374b-5p could inhibit NSCLC cell proliferation, migration and invasion and could directly target FOXP1. CONCLUSION: This study found that the decreased miR-374b-5p predicts poor prognosis of NSCLC, and the upregulation of miR-374b-5p can inhibit NSCLC cell proliferation, migration and invasion. The data obtained from this study provide a novel candidate prognostic biomarker and a potential therapeutic target for NSCLC.

Transcriptome co-expression network analysis identifies key genes and regulators of ripening kiwifruit ester biosynthesis.

BACKGROUND: Aroma is an important organoleptic quality for fruit and has a large influence on consumer preference. Kiwifruit esters undergo rapid and substantial changes contributing to the flavor during fruit ripening. Part of enzymes and their coding genes have been indicated potential candidates for flavor-related esters synthesis. However, there still exist obvious gaps in the biosynthetic pathways of esters and the mechanisms regulating ester biosynthesis in kiwifruit remain unknown. RESULTS: Using gas chromatography-mass spectrometry (GC-MS), volatile compounds of kiwifruit were quantified in response to ethylene (ETH, 100 µl/l, 24 h, 20 °C) and 1-methylcyclopropene (1-MCP, 1 µl/l, 24 h, 20 °C). The results indicated that esters showed the most substantial changes enhanced by ethylene and were inhibited by 1-MCP. Correlations between RNA-seq results and concentrations of esters, constructed using Weighted Gene Co-Expression Network Analysis (WGCNA) indicated that three structural genes (fatty acid desaturase, AdFAD1; aldehyde dehydrogenase, AdALDH2; alcohol acyltransferase, AdAT17) had similar expression patterns that paralled the changes in total ester content, and AdFAD1 transcripts exhibited the highest correlation. In order to search for potential regulators for ester biosynthesis, 14 previously reported ethylene-responsive transcription factors (TFs) were included in the correlation analysis with esters and their biosynthetic genes. Using dual-luciferase assay, the in vivo regulatory activities of TFs on ester biosynthetic gene promoters were investigated and the results indicated that AdNAC5 and AdDof4 (DNA binding with one finger) trans-activated and trans-suppressed the AdFAD1 promoter. CONCLUSIONS: The present study advanced the molecular basis of ripening-related ester biosynthesis in kiwifruit by identifying three biosynthetic related genes AdFAD1, AdALDH2 and AdAT17 by transcriptome analysis, and highlighted the function of two TFs by transactivation studies.