Influences of two transport strategies on AMPK-mediated metabolism and flesh quality of shrimp (Litopenaeus vannamei).

BACKGROUND: Water-free transportation (WFT), as a novel strategy for express delivery of live shrimp (Litopenaeus vannamei), was developed recently. However, air exposure during this transportation arouses a series of abiotic stress to the shrimp. In the present study, the influences of WFT stress on glycolysis and lipolysis metabolism and meat quality (umami flavor and drip loss) were investigated in comparison with conventional water transportation (WT). RESULTS: The results showed that type II muscle fibers with the feature of anaerobic metabolism were dominated in shrimp flesh. In addition, the increments of intracellular Ca2+ was detected in WFT and WT, which then activated the AMP-activated protein kinase pathway and promoted the consumption of glycogen, as well as the accumulation of lactate and lipolysis, under the enzymolysis of hexokinase, pyruvate kinase, lactate dehydrogenase and adipose triglyceride lipase. Glycogen glycolyzed to latate. Meanwhile, ATP degraded along with glycolysis resulting in the generation of ATP-related adenosine phosphates such as inosine monophosphate with umami flavor and phosphoric acid. More remarkable (P < 0.05) physiological changes (except lactate dehydrogenase and lactate) were observed in WFT compared to WT. Additionally, the fatty acid profile also slightly changed. CONCLUSION: The transport stress induced significant energy metabolism changes of shrimp flesh and therefore effected the flesh quality. The intensifications of freshness (K-value) of shrimp flesh were detected as a result of ATP degradation, which were more pronounced after WFT. However, the drip loss of shrimp flesh was more significantly increased (P < 0.05) after WFT compared to WT. © 2023 Society of Chemical Industry.

Tandem Mass Tag Proteomics Provides Insights into the Underlying Mechanism of Flesh Quality Degradation of Litopenaeus vannamei during Refrigerated Waterless Transport at 12 °C.

Refrigerated waterless transport at 12 °C of live shrimp (Litopenaeus vannamei) causes flesh quality deterioration, and the underlying mechanism remains unknown. Herein, proteomics and bioinformatics analyses were used to elucidate the molecular mechanism of flesh quality changes. The result showed that 33 and 44 of the differentially abundant proteins (DAPs) were, respectively, identified in the acute cold (AC) group and the combined stress of acute cold and waterless duration (AC+WD) group, which were mostly involved in the metabolism processes and cellular structure of animal tissues, and notably enriched in biological pathways such as lysosome, glycolysis/gluconeogenesis, and focal adhesion. Furthermore, the changes in color and texture properties were closely associated with tubulin, gelsolin, laminin, trypsin-1, dipeptidyl peptidase, triosephosphate isomerase, and aldehyde dehydrogenase. Therefore, these DAPs could be used as potential biomarkers to monitor the deterioration of shrimp flesh quality during refrigerated waterless transportation.

Wounding induces suberin deposition, relevant gene expressions and changes of endogenous phytohormones in Chinese yam (Dioscorea opposita) tubers.

Wounds on Chinese yam (Dioscorea opposita ) tubers can ocurr during harvest and handling, and rapid suberisation of the wound is required to prevent pathogenic infection and desiccation. However, little is known about the causal relationship among suberin deposition, relevant gene expressions and endogenous phytohormones levels in response to wounding. In this study, the effect of wounding on phytohormones levels and the expression profiles of specific genes involved in wound-induced suberisation were determined. Wounding rapidly increased the expression levels of genes, including PAL , C4H , 4CL , POD , KCSs , FARs , CYP86A1 , CYP86B1 , GPATs , ABCGs and GELPs , which likely involved in the biosynthesis, transport and polymerisation of suberin monomers, ultimately leading to suberin deposition. Wounding induced phenolics biosynthesis and being polymerised into suberin poly(phenolics) (SPP) in advance of suberin poly(aliphatics) (SPA) accumulation. Specifically, rapid expression of genes (e.g. PAL , C4H , 4CL , POD ) associated with the biosynthesis and polymerisation of phenolics, in consistent with SPP accumulation 3days after wounding, followed by the massive accumulation of SPA and relevant gene expressions (e.g. KCSs , FARs , CYP86A1 /B1 , GPATs , ABCGs , GELPs ). Additionally, wound-induced abscisic acid (ABA) and jasmonic acid (JA) consistently correlated with suberin deposition and relevant gene expressions indicating that they might play a central role in regulating wound suberisation in yam tubers.

Impacts of long-time transportation on whiteleg shrimp (Penaeus vannamei) muscle quality and underlying biochemical mechanisms.

BACKGROUND: Shrimp is widely consumed around the world. Since muscle is the primary edible component of shrimp, muscle quality (particularly texture) has a direct impact on the economic value of shrimp products. However, reports on the shrimp muscle quality influenced by transportation are rather limited, and the underlying mechanism remains unknown. RESULTS: During the simulated transportation, the water pH and total ammonia-nitrogen content and un-ionized ammonia contents were elevated. Furthermore, reductions in shrimp muscle water-holding capacity, hardness, and shear value with intensive myofibrillar protein degradation were detected. Simulated transportation decreased the pH and glycogen content of shrimp muscle while increasing lactic dehydrogenase activity and lactate content, resulting in an elevated level of free calcium ions and increased µ-calpain and general proteolytic activities. Water exchange could improve the water quality and reduce the mortality of shrimp during transportation, as well as decrease muscle textural softening by alleviating these stress responses. CONCLUSIONS: Maintaining water quality and, in particular, reducing ammonia are critical to improving shrimp survival and muscle quality during live transportation. This study is of great significance for the better maintenance of the textural properties of shrimp meat. © 2023 Society of Chemical Industry.

Participation of FaTRAB1 Transcription Factor in the Regulation of FaMADS1 Involved in ABA-Dependent Ripening of Strawberry Fruit.

Abscisic acid (ABA) plays a crucial role in regulating the ripening of non-climacteric strawberry fruit. In the present study, ABA was confirmed to promote strawberry ripening and induce the down-regulation of FaMADS1. The transient silence of FaMADS1 in strawberries promoted fruit ripening and induced the content of anthocyanin and soluble pectin but reduced firmness and protopectin through a tobacco rattle virus-induced gene silencing technique. In parallel with the accelerated ripening, the genes were significantly induced in the transiently modified fruit, including anthocyanin-related PAL6, C4H, 4CL, DFR, and UFGT, softening-related PL and XTH, and aroma-related QR and AAT2. In addition, the interaction between FaMADS1 and ABA-related transcription factors was researched. Yeast one-hybrid analysis indicated that the FaMADS1 promoter could interact with FaABI5-5, FaTRAB1, and FaABI5. Furthermore, dual-luciferase assay suggested that FaTRAB1 could actively bind with the FaMADS1 promoter, resulting in the decreased expression of FaMADS1. In brief, these results suggest that the ABA-dependent ripening of strawberry fruit was probably inhibited through inhibiting FaMADS1 expression by the active binding of transcript FaTRAB1 with the FaMADS1 promoter.

Physicochemical and functional properties of the protein-starch interaction in Chinese yam.

Protein-starch interaction has an important impact on the properties of starchy foods rich in protein, but the contribution of the interaction to Chinese yam still remains unclear. This study aimed to characterize the physicochemical and functional properties related to the possible interaction between starch and protein in Chinese yam. Differential scanning calorimetry and rapid viscosity analyzer results revealed that the gelatinization temperature increased in protein and starch cross-linked powder, while the peak viscosity and the setback viscosity decreased. The swelling power and solubility at 80°C and 95°C decreased with increasing protein ratio in the powder. In vitro starch digestibility test indicated that a high protein ratio could rapidly reduce digestible starch, but increase both slowly digestible starch and resistant starch. Protein could act as the physical barrier toward starch against heating and digestion to exert the influence on starch properties. Fourier transform infrared spectroscopy test revealed the interaction between protein and starch. These results revealed the role of protein-starch interaction and provided beneficial information for the utilization of Chinese yam.

The spatial distribution and migration of three typical fungicides in postharvest satsuma mandarin (Citrus unshiu Marc.) fruit.

Fungicides are often used to extend the storage time of postharvest satsuma mandarin fruit. In recent years, fungicide residue has become an issue of food safety. This study aimed to investigate the distribution and migration of three typical fungicides (imazalil, prochloraz, thiophanate-methyl) in postharvest satsuma mandarins using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Three fungicides could quickly penetrate satsuma mandarins and their gradient concentrations of residues in the fruit were: carpopodium > mesocarp > epicarp > pulp. However, the residues of three fungicides in the edible pulp were obviously lower than the maximum residue limit (MRL = 5.0 mg/kg in China). Residues of the three fungicides decreased in epicarp and carpopodium but increased in mesocarp and pulp during storage. Fungicides could quickly penetrate the fruit, settling primarily in the carpopodium but little in the pulp. Both epicarp and carpopodium were the breakthrough pathways for the fungicides entering the fruit, while epicarp was the main route for the penetration of fungicides. These findings shed new information on the behavior of fungicides and the safety issue of satsuma mandarins.

Exogenous ABA promotes aroma biosynthesis of postharvest kiwifruit after low-temperature storage.

MAIN CONCLUSION: Exogenous ABA played a positive role in the accumulation and biosynthesis of aroma components of postharvest kiwifruit after low-temperature storage, especially the esters production during ripening. Low-temperature storage (LTS) generally affects the aroma formation associated with the decrease in aroma quality in kiwifruit. In this work, abscisic acid (ABA) treatment after LTS increased the production of aroma components in postharvest kiwifruit and enhanced the related enzyme activity, especially alcohol acyltransferase (AAT), branched amino acid transaminase (BCAT) and hydroperoxide lyase (HPL). Corresponding to the enzyme activity, the gene expression of AchnAAT, AchnADH, AchnBCAT and AchnHPL was significantly up-regulated by ABA. The principal component analysis further illustrated the differences in aroma components between ABA and the control. The positive correlation of aroma accumulation with the expression levels of AchnPDC and AchnLOX and the enzyme activities of BCAT and pyruvate decarboxylase (PDC) was also revealed by correlation analysis. In addition, the promoter sequences of the key genes involved in aroma biosynthesis contained multiple cis-elements (ABRE and G-box) of ABA-responsive proteins. Combining the transcriptome sequencing data, the promoting role of ABA signaling in the regulation of aroma biosynthesis of postharvest kiwifruit after LTS was discussed. This study would provide a reference for improving aroma quality of postharvest kiwifruit after LTS, as well the molecular mechanism of kiwifruit aroma fading after LTS.

Transcriptional regulation of KCS gene by bZIP29 and MYB70 transcription factors during ABA-stimulated wound suberization of kiwifruit (Actinidia deliciosa).

BACKGROUND: Our previous study has demonstrated that the transcription of AchnKCS involved in suberin biosynthesis was up-regulated by exogenous abscisic acid (ABA) during the wound suberization of kiwifruit, but the regulatory mechanism has not been fully elucidated. RESULTS: Through subcellular localization analysis in this work, AchnbZIP29 and AchnMYB70 transcription factors were observed to be localized in the nucleus. Yeast one-hybrid and dual-luciferase assay proved the transcriptional activation of AchnMYB70 and transcriptional suppression of AchnbZIP29 on AchnKCS promoter. Furthermore, the transcription level of AchnMYB70 was enhanced by ABA during wound suberization of kiwifruit, but AchnbZIP29 transcription was reduced by ABA. CONCLUSIONS: Therefore, it was believed that ABA enhanced the transcriptional activation of AchnMYB70 on AchnKCS by increasing AchnMYB70 expression. On the contrary, ABA relieved the inhibitory effect of AchnbZIP29 on transcription of AchnKCS by inhibiting AchnbZIP29 expression. These results gave further insight into the molecular regulatory network of ABA in wound suberization of kiwifruit.

Abscisic acid stimulates wound suberisation in kiwifruit (Actinidia chinensis) by regulating the production of jasmonic acid, cytokinin and auxin.

Wounding induces a cascade of correlative physiological responses that lead to the repair of damaged tissue. In this study, the effect of wounding on suberin, endogenous hormones and their metabolic genes expression was observed during the wound healing of kiwifruit (Actinidia chinensis Planch.). In addition, the role of abscisic acid (ABA) in wound suberisation was investigated by analysing the coordinated regulation between ABA and other hormones. The wound healing process in kiwifruit could be divided into two stages including: (1) initial accumulation of suberin polyphenolic (SPP) and long carbon chain suberin polyaliphatic monomers (LSPA) before 24h; and (2) massive synthesis of SPP and very long carbon chain suberin polyaliphatic monomers (VLSPA) after 24h. ABA content rapidly increased and induced the jasmonic acid (JA) biosynthesis at the early stage of wound healing. ABA level gradually decreased with the expression of AchCYP707A genes, while the contents of trans-zeatin (t-ZT) and indole-3-acetic acid (IAA) steadily increased at the late stage of wound healing. Exogenous ABA stimulated JA and suberin monomers accumulation, but suppressed both t-ZT and IAA biosynthesis. The role of ABA in wound healing of kiwifruit might be involved in the coordination of both JA-mediated suberin monomers biosynthesis and t-ZT- and IAA-mediated formation of suberised cells via an interaction mechanism.

Heat shock protein 70 (HSP70) promotes air exposure tolerance of Litopenaeus vannamei by preventing hemocyte apoptosis.

Brief pretreatment of cold shock at 13 °C for 3 min proved to be an inducer of heat shock protein 70 (HSP70) and improved stress tolerance as a molecular chaperone. With the improvement of air exposure tolerance, HSP70 in shrimp hemocytes was upregulated in mRNA and protein levels after cold shock. Both HSP70 RNA interference (RNAi) gene knockdown and recombinant HSP70 (rHSP70) injection were successfully established in order to investigate the role of HSP70 in response to air exposure stress. Shrimp receiving rHSP70 showed an improved survival rate (80%) with no significant difference (p > 0.05) compared to cold shock treated shrimp (control, 90%) under air exposure, but the survival rate of HSP70-knockdown shrimp was significantly lower (62%, p < 0.05). Reactive oxygen species (ROS) content, relative expression of cytochrome c, caspase-3 activity, and apoptosis rate in hemocytes of HSP70 enriched shrimp (i.e., cold shock and rHSP70 injection) were significantly lower (p < 0.05) than HSP70-knockdown shrimp. Results suggested that HSP70 could be induced by cold shock and contributed to improve the tolerance of shrimp suffering air exposure by blocking the apoptosis pathway through scavenging intracellular ROS, inhibiting cytochrome c expression, inhibiting release from mitochondria, and inactivating caspase-3. This work updates the understanding of cold shock mechanism in water-free transportation of aquatic animals.

Exogenous 24-epibrassinolide activates detoxification enzymes to promote degradation of boscalid in cherry tomatoes.

BACKGROUND: Boscalid is often used to extend the storage time of postharvest cherry tomato. Pesticide residue has become an issue of food safety. This study sought to investigate the spatial distribution of boscalid residue in cherry tomato fruits and to determine the effect of 24-epibrassinolide (EBR) in promoting boscalid degradation. RESULTS: Boscalid could quickly penetrate into cherry tomatoes, but mainly remained in the peel. The migration of boscalid from the peel into the core was a time-consuming and complex process during storage. After 72 h, boscalid residues in the pulp and the core began to accumulate gradually. The exogenous application of EBR activated peroxidase, glutathione reductase and glutathione S-transferase, and effectively promoted the degradation of boscalid by a maximum decrease of 44.8% in peel, 54.0% in pulp and 71.2% in core. CONCLUSION: As one of the common pesticides, boscalid had a strong ability to enter the cherry tomato and thus become a potential risk for public consumption. Therefore, rational use of pesticides is recommended. The results of this study indicate that the possible risk of boscalid residue could be alleviated by EBR pretreatment through activating detoxification enzymes. © 2020 Society of Chemical Industry.

Physicochemical and structural properties of low-amylose Chinese yam (Dioscorea opposita Thunb.) starches.

Nuoshanyao (NSY), Tiegunshanyao (TSY) and Huaishanyao (HSY) are the main cultivars of Chinese yam (Dioscorea opposita Thunb.) widely grown in China. The composition, physicochemical properties, morphology, and thermal properties of the starches from these cultivars were investigated in this study. NSY starch (17.0%) was much lower in amylose content than other cultivars (33.4-34.5%). The average particle diameter of the starches ranged from 25.83 to 28.93 µm. Weight-average molecular weights (Mw) and number-average molecular weights (Mn) ranged from 1.29 to 1.84 × 105 g/mol and 5.93 to 8.36 × 104 g/mol, respectively. NSY starch had higher gelatinization temperature (71.5 °C), enthalpy (14.14 J/g), peak viscosity (8590 cP) and swelling power (12.0%) than TSY and HSY. X-ray diffraction (XRD) and Fourier transform infrared (FTIR) tests indicated that Chinese yam starches had CB-type crystalline structure with crystallinity ranging from 21.91% to 27.08% and a short-range ordered structure. To, Tp, ΔH, peak viscosity and swelling power at 95 °C were significantly correlated to amylose content. The low-amylose NSY starch was found to have high gelatinization temperature, enthalpy, peak viscosity and swelling power. These specific physicochemical and structural properties indicated the industrial potential of low-amylose yam starch.

MYB41, MYB107, and MYC2 promote ABA-mediated primary fatty alcohol accumulation via activation of AchnFAR in wound suberization in kiwifruit.

Wound damage triggers the accumulation of abscisic acid (ABA), which induces the expression of a large number of genes involved in wound suberization in plants. Fatty acyl-CoA reductase (FAR) catalyzes the generation of primary fatty alcohols by the reduction of fatty acids in suberin biosynthesis. However, the regulatory effects of transcription factors (TFs) on AchnFAR in response to ABA are unexplored. In this study, kiwifruit AchnFAR displayed a biological function analogous to that of FAR in transiently overexpressed tobacco (Nicotiana benthamiana) leaves. The positive role of TFs, including AchnMYB41, AchnMYB107, and AchnMYC2, in the regulation of AchnFAR was identified. The three TFs could individually bind to the AchnFAR promoter to activate gene transcription in yeast one-hybrid and dual-luciferase assays. Transient overexpression of TFs in tobacco leaves resulted in the upregulation of aliphatic synthesis genes (including FAR) and the increase in aliphatics, including primary alcohols, α,ω-diacids, ω-hydroxyacids, and fatty acids. Moreover, exogenous ABA treatment elevated TF-mediated AchnFAR expression and the accumulation of primary alcohols. Conversely, fluridone, an inhibitor of ABA biosynthesis, suppressed the expression of AchnFAR and TF genes and reduced the formation of primary alcohols. The results indicate that AchnMYB41, AchnMYB107, and AchnMYC2 activate AchnFAR transcription to promote ABA-mediated primary alcohol formation in wound suberization in kiwifruit.

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.

ABF2 and MYB transcription factors regulate feruloyl transferase FHT involved in ABA-mediated wound suberization of kiwifruit.

Suberin is a cell-wall biopolymer with aliphatic and aromatic domains that is synthesized in the wound tissues of plants in order to restrict water loss and pathogen infection. ω-hydroxyacid/fatty alcohol hydroxycinnamoyl transferase (FHT) is required for cross-linking of the aliphatic and aromatic domains. ABA is known to play a positive role in suberin biosynthesis but it is not known how it interacts with FHT. In this study, the kiwifruit (Actinidia chinensis) AchnFHT gene was isolated and was found to be localized in the cytosol. Transient overexpression of AchnFHT in leaves of Nicotiana benthamiana induced massive production of ferulate, ω-hydroxyacids, and primary alcohols, consistent with the in vitro ability of AchnFHT to catalyse acyl-transfer from feruloyl-CoA to ω-hydroxypalmitic acid and 1-tetradecanol. A regulatory function of four TFs (AchnABF2, AchnMYB4, AchnMYB41, and AchnMYB107) on AchnFHT was identified. These TFs localized in the nucleus and directly interacted with the AchnFHT promoter in yeast one-hybrid assays. Dual-luciferase analysis indicated that AchnABF2, AchnMYB41, and AchnMYB107 activated the AchnFHT promoter while AchnMYB4 repressed it. These findings were supported by the results of transient overexpression in N. benthamiana, in which AchnABF2, AchnMYB41, and AchnMYB107 induced expression of suberin biosynthesis genes (including FHT) and accumulation of suberin monomers, whilst AchnMYB4 had the opposite effect. Exogenous ABA induced the expression of AchnABF2, AchnMYB41, AchnMYB107, and AchnFHT and induced suberin monomer formation, but it inhibited AchnMYB4 expression. In addition, fluridone (an inhibitor of ABA biosynthesis) was found to counter the inductive effects of ABA. Activation of suberin monomer biosynthesis by AchnFHT was therefore controlled in a coordinated way by both repression of AchnMYB4 and promotion of AchnABF2, AchnMYB41, and AchnMYB107.

Evaluation of immunological response in shrimp Penaeus vannamei submitted to low temperature and air exposure.

The objective of this study is to explore the immune response of the shrimp Penaeus vannamei to low temperature and air exposure during the mimic waterless transportation. Shrimp were cold shocked at 13 °C for 3 min, then exposed to air at 15 °C for 12 h, and finally revived in water at 25 °C. Hemocyte structure remained intact with only slight distortions of some organelles and nuclear membrane under the stress. Phenoloxidase (PO), lysozyme (Lys) and gamma-glutamyl transferase (GGT) activities, glutamine (Gln) content and relative mRNA expressions of prophenoloxidase (proPO), ß-1,3-glucan binding protein (LGBP), ferrin (Fer) and glucose regulated protein 78 (GRP 78) increased and reached peak levels after 3 h-9 h of air exposure, and then decreased to relatively stable levels in the prolonged period of air exposure. The total hemocyte count (THC) and gene expressions of proPO, Fer and LGBP at the end of waterless storage were significantly lower (p < 0.05) than those in fresh shrimp while no significant difference (p ≥ 0.05) was found between revived and fresh shrimp in PO, Lys, GGT activities, Gln content and gene expression level of GRP 78. Of all the hemocytes, the percentage of semi granular cells (SGC) and granular cells (GC) significantly decreased after 6-9 h of storage, while hyaline cells (HC) dramatically increased after 9 h of storage to compensate the loss of SGC and GC. Results suggested that the low temperature and air exposure caused significant immunological response to the shrimp, but the damages to the immune system were partly reparable.

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.

Three Transcription Activators of ABA Signaling Positively Regulate Suberin Monomer Synthesis by Activating Cytochrome P450 CYP86A1 in Kiwifruit.

Wound attack stimulates accumulation of abscisic acid (ABA) that activates a number of genes associated with wound suberization of plants. Cytochrome P450 fatty acid ω-hydroxylase CYP86A1 catalyzes ω-hydroxylation of fatty acids to form the ω-functionalized monomers that play a pivotal role in suberin synthesis. However, the transcriptional regulation of ABA signaling on AchnCYP86A1 has not been characterized in kiwifruit. In this study, AchnCYP86A1, a kiwifruit homolog of Arabidopsis AtCYP86A1, was isolated. AchnCYP86A1-overexpressed N. benthamiana leaves displayed that the AchnCYP86A1 functioned as a fatty acid ω-hydroxylase associated with synthesis of suberin monomer. The regulatory function of three transcription factors (TFs, including AchnMYC2, AchnMYB41 and AchnMYB107) on AchnCYP86A1 was identified. All the three TFs were localized in nucleus and could individually interact with AchnCYP86A1 promoter to activate gene expression in yeast one-hybrid and dual-luciferase assays. The findings were further demonstrated in transient overexpressed N. benthamiana, in which all TFs notably elevated the expression of aliphatic synthesis genes including CYP86A1 and the accumulation of ω-hydroxyacids, α, ω-diacids, fatty acids and primary alcohols. Moreover, exogenous ABA induced the expression of AchnMYC2, AchnMYB41 and AchnMYB107 that promoted AchnCYP86A1 involving in suberin monomer formation. Contrary to the inductive effects of ABA, however, fluridone (an inhibitor of ABA biosynthesis) inhibited the three TFs expression and suberin monomer formation. These results indicate that AchnMYC2, AchnMYB41 and AchnMYB107 positively regulate suberin monomer synthesis by activating AchnCYP86A1 promoter in response to ABA.