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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., (© 2022. The Author(s).)

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.

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., (© The Author(s) 2019. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

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 β-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.

Quick suberin-based healing after wounding played a protective role for plant to prevent further damage. In this study, the stimulative effect of exogenous abscisic acid (ABA) on wound suberization in postharvest kiwifruit was evaluated through suberin staining with toluidine blue O as well as the determination of suberin phenolics and aliphatics in wound tissue. Furthermore, to reveal the regulatory involvement of ABA in wound suberization, comparative quantitative proteomics and transcriptomics analyses based on iTRAQ and qRT-PCR technique were performed. In proteomics levels, a total of 95 protein species consistently showed differential abundance between ABA and control, including 29 down-regulated and 66 up-regulated protein species. The Kyoto Encyclopedia of Genes and Genomes (KEGG) with protein-protein interaction analyses revealed that ABA mainly affected the antioxidant system, phenylpropanoid metabolism and lipid metabolism associated with wound suberization. Based on the data of proteomics analysis, the differential expressions of genes encoding 11 selected protein species were confirmed by qRT-PCR analyses. GSH-Px, MDHAR, SOD, APX, POD, PAL, CCR, PPO, CYP86B1, DGGT and KCS11 were likely to be the key enzymes that involved the response of ABA to stimulate wound suberization by mediating the antioxidant system, phenylpropanoid metabolism and lipid metabolism., Biological Significance: Kiwifruit is susceptible to physical injury causing postharvest deterioration during harvest, transportation and storage. Therefore, quick healing is important for maintaining the postharvest quality of injured fruit. This work elucidated the potential role of ABA and the proteomic mechanism of its regulation in wound suberization of postharvest kiwifruit., (Copyright © 2017 Elsevier B.V. All rights reserved.)

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 3 days 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. Wounds on Chinese yam (Dioscorea opposita) tubers can ocurr during harvesting and handling, and rapid suberisation is required to prevent 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. Wound-induced abscisic acid and jasmonic acid are likely to regulate the biosynthesis, transport and polymerisation of suberin monomers by activating relevant gene expressions, which might be involved in suberisation of cells in wound area. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

• Abscisic acid (ABA) application accelerated wound healing of Chinese yam tubers. • ABA raised the contents of suberin polyphenolics (SPP), lignin and total phenols. • ABA raised gene expressions, contents and activities of enzymes in SPP formation. Water loss, nutrient elution and pathogen attacks caused by mechanical wounding are the primary sources of postharvest loss of fruit and vegetables. Continuously increasing information has unraveled that abscisic acid (ABA) can accelerate wound healing of fruit and vegetables via stimulating suberin polyphenolics (SPP) biosynthesis, whereas the effect of ABA in wound healing of Chinese yam tubers remains unclear. The treatment of ABA and fluridone (FLD, an inhibitor of ABA biosynthesis) on postharvest wound healing of yam tubers was investigated in this study. The results showed that ABA treatment facilitated wound healing process of yam tubers, representing by the reduced weight loss and relative electrolyte leakage (REL). ABA enhanced the expression levels of phenylalanine ammonialyase (PAL), cinnamic acid 4-hydroxylase (C4H) and 4-coumarate-CoA ligase (4CL), as well as the corresponding enzyme contents and activities. Correspondingly, the contents of major monomers such as ferulic acid, caffeic acid and p -coumaric acid also increased. Simultaneously, ABA could improve the expression of peroxidase (POD), the enzyme content and activity, as well as hydrogen peroxide (H 2 O 2) production, which might contribute to the polymerization of SPP and lignin. Inversely, inhibition of ABA production by FLD application displayed an opposite effect to ABA treatment. These results demonstrate that application of ABA stimulated wound healing of Chinese yam tubers via improving the gene expressions of relevant enzymes, their contents and activities, so that maintain high quality of postharvest yam tubers. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

• AchLOX3 and AchOPR3 were involved in JA biosynthesis of kiwifruit. • ABA-induced AchABF1-1 was a nucleus-localized transcription factor. • AchABF1-1 activated AchLOX3 and AchOPR3 expression by binding to their promoters. • ABA promoted SPP formation by the induction of JA biosynthesis. Mechanical damage is a central limiting factor in the maintenance of the postharvest fruit quality during harvest, transportation, and processing. In this study, wounded kiwifruit (Actinidia chinensis cv. Xuxiang) were respectively treated with abscisic acid (ABA), fluridone (FLD, an inhibitor of ABA biosynthesis), methyl jasmonate (MeJA), diethyldithiocarbamic acid (DIECA, an inhibitor of JA biosynthesis), and ABA+DIECA to investigate the potential interaction between ABA and JA in the process of wound healing. The results showed that blocking JA biosynthesis significantly reduced the positive effect of ABA on the accumulation of suberin polyphenolic (SPP). Besides, transiently overexpressed the AchLOX3 and AchOPR3 in tobacco (Nicotiana benthamiana) leaves confirmed that these two genes respectively encoded lipoxygenase and 12-oxo-phytodienoic acid reductase involved in JA biosynthesis. Notably, the positive regulation of transcription factor (TF) AchABF1-1 on AchLOX3 and AchOPR3 was identified. ABA-induced AchABF-1 located in the nucleus and activated the transcriptional expression of AchLOX3 and AchOPR3 through binding to their promoters. These results were further confirmed in the tobacco leaves overexpressing AchABF1-1 in which AchABF1-1 promoted NbLOX and NbOPR expression with the increases of JA content. In addition, exogenous ABA increased endogenous JA content, and enhanced LOX and OPR activities as well as their related gene expression, while FLD reduced these indices. These results suggested that the promotion effect of JA on the SPP accumulation was enhanced through the promotion of JA biosynthesis by ABA-responsive transcription factor AchABF1-1 during the wound healing process of kiwifruit. [ABSTRACT FROM AUTHOR]

• AchnCYP86A1 and AchnFAR were involved in wound suberinzation of kiwifruit. • AcnMYB4 repressed AchnCYP86A1 and AchnFAR expression by binding to gene promoter. • ABA induced the AchnCYP86A1 and AchnFAR expression by inhibition of AchnMYB4. Suberin biosynthesis involves a large number of genes, and many of them are induced by abscisic acid (ABA). However, the regulation of transcription factor (TF) in response to ABA on suberin biosynthetic genes in kiwifruit has been unexplored. In this study, two genes, AchnCYP86A1 and AchnFAR respectively encoding a fatty acid ω-hydroxylase and fatty acyl-CoA reductase involved in suberin monomer biosynthesis were demonstrated in transient overexpressed tobacco (Nicotiana benthamiana). Notably, the negative regulation of AchnMYB4 on AchnCYP86A1 and AchnFAR was identified. AchnMYB4 could repress AchnCYP86A1 and AchnFAR transcript by directly binding to the gene promoter in yeast one-hybrid and dual-luciferase assays. These results were further confirmed in transient overexpressed tobacco leaves in that AchnMYB4 significantly down-regulated suberin biosynthetic genes including CYP86A1 , FAR2 and FAR3 , and reduced accumulation of ω-hydroxyacids and primary alcohols. Moreover, exogenous ABA could induce the expression of AchnCYP86A1 and AchnFAR , and the accumulation of corresponding suberin monomers by inhibition of AchnMYB4 in wound-tissue of kiwifruit. However, fluridone (an inhibitor of ABA biosynthesis) was found to counter the inductive effects of ABA. Taken together, the results suggest that ABA activates AchnCYP86A1 and AchnFAR to promote suberin monomers biosynthesis by inhibiting AchnMYB4. [ABSTRACT FROM AUTHOR]

Wound-induced suberization is an essentially protective healing process for fruit to reduce water loss and avoid infecting. However, cognate mechanisms that regulate this process are little known. To expand our knowledge of suberization induced by wounding, a wound-healing investigation together with metabolite profiling study was conducted in postharvest kiwifruit ( Actinidia deliciosa ). The development of suberization in wounded fruit was demonstrated by autofluorescence observation and toluidine blue staining at 1–4 day (d) after wounding. Activities of phenylalanine ammonia-lyase (PAL), cinnamyl-alcohol dehydrogenase (CAD) and peroxidase (POD) in wound-healing tissue were enhanced by abscisic acid (ABA). The constituent analysis of suberin including polyphenolics (SPP) and polyaliphatics (SPA) proved that exogenous ABA increased the content levels of total phenols, total flavonoids and alkanes, alkenes, alcohols, alkane acids, olefine acids, esters, glycerides and vitamin E in wound-healing tissue. Results suggested that ABA stimulated suberization through the activation of PAL, CAD and POD to accelerate wound-healing of wounded kiwifruit. [ABSTRACT FROM AUTHOR]