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FaPAO5 regulates Spm/Spd levels as a signaling during strawberry fruit ripening.

ABSTRACT: Polyamines are important for non-climacteric fruit ripening according to an analysis of the model plant strawberry. However, the molecular mechanism underlying the polyamine accumulation during ripening has not been fully elucidated. In this study, an examination of our proteome data related to strawberry fruit ripening revealed a putative polyamine oxidase 5, FaPAO5, which was localized in the cytoplasm and nucleus. Additionally, FaPAO5 expression levels as well as the abundance of the encoded protein continually decreased during ripening. Inhibiting FaPAO5 expression by RNAi promoted Spd, Spm, and ABA accumulation while inhibited H2O2 production, which ultimately enhanced ripening as evidenced by the ripening-related events and corresponding gene expression changes. The opposite effects were observed in FaPAO5-overexpressing transgenic fruits. Analyses of the binding affinity and enzymatic activity of FaPAO5 with Spm, Spd, and Put uncovered a special role for FaPAO5 in the terminal catabolism of Spm and Spd, with a K d of 0.21 and 0.29 µM, respectively. Moreover, FaPAO5 expression was inhibited by ABA and promoted by Spd and Spm. Furthermore, the RNA-seq analysis of RNAi and control fruits via differentially expressed genes (DEGs) indicated the six most enriched pathways among the differentially expressed genes were related to sugar, abscisic acid, ethylene, auxin, gibberellin, and Ca2+. Among four putative PAO genes in the strawberry genome, only FaPAO5 was confirmed to influence fruit ripening. In conclusion, FaPAO5 is a negative regulator of strawberry fruit ripening and modulates Spm/Spd levels as a signaling event, in which ABA plays a central role.

SUBMITTER: Mo A

PROVIDER: S-EPMC7189608 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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FaPAO5 regulates Spm/Spd levels as a signaling during strawberry fruit ripening.

Mo Aowai A Xu Tian T Bai Qian Q Shen Yaunyue Y Gao Fan F Guo Jiaxuan J

Plant direct 20200429 5

Polyamines are important for non-climacteric fruit ripening according to an analysis of the model plant strawberry. However, the molecular mechanism underlying the polyamine accumulation during ripening has not been fully elucidated. In this study, an examination of our proteome data related to strawberry fruit ripening revealed a putative polyamine oxidase 5, FaPAO5, which was localized in the cytoplasm and nucleus. Additionally, <i>FaPAO5</i> expression levels as well as the abundance of the e ...[more]

PMID: 32355906

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Project description:Auxin response factors (ARFs) are transcription factors that play important roles in plants. ARF2 is a member of the ARF family and participates in many plant growth and developmental processes. However, the role of ARF2 in strawberry fruit quality remains unclear. In this study, FveARF2 was isolated from the woodland strawberry 'Ruegen' using reverse transcription-polymerase chain reaction (RT-PCR), which showed that FveARF2 expression levels were higher in the stem than in other organs of the 'Ruegen' strawberry. Moreover, FaARF2 was higher in the white fruit stage of cultivated strawberry fruit than in other stage. Subcellular localization analysis showed that FveARF2 is located in the nucleus, while transcriptional activation assays showed that FveARF2 inhibited transcription in yeast. Silencing FveARF2 in cultivated strawberry fruit revealed earlier coloration and higher soluble solid, sugar, and anthocyanin content in the transgenic fruit than in the control fruit, overexpression of FveARF2 in strawberry fruit delayed ripening and lower soluble solid, sugar, and anthocyanin content compared to the control fruit. Gene expression analysis indicated that the transcription levels of the fruit ripening genes FaSUT1, FaOMT, and FaCHS increased in FveARF2-RNAi fruit and decreased in FveARF2-OE fruit, when compared with the control. Furthermore, yeast one-hybrid (Y1H) and GUS activity experiments showed that FveARF2 can directly bind to the AuxRE (TGTCTC) element in the FaSUT1, FaOMT, and FaCHS promoters in vitro and in vivo. Potassium ion supplementation improved the quality of strawberry fruit, while silencing FveARF2 increased potassium ion content in transgenic fruit. The Y1H and GUS activity experiments also confirmed that FveARF2 could directly bind to the promoter of FveKT12, a potassium transporter gene, and inhibited its expression. Taken together, we found that FveARF2 can negatively regulate strawberry fruit ripening and quality, which provides new insight for further study of the molecular mechanism of strawberry fruit ripening.

Project description:Light signaling and plant hormones, particularly ethylene and auxins, have been identified as important regulators of carotenoid biosynthesis during tomato fruit ripening. However, whether and how the light and hormonal signaling cascades crosstalk to control this metabolic route remain poorly elucidated. Here, the potential involvement of ethylene and auxins in the light-mediated regulation of tomato fruit carotenogenesis was investigated by comparing the impacts of light treatments and the light-hyperresponsive high pigment-2 (hp2) mutation on both carotenoid synthesis and hormonal signaling. Under either light or dark conditions, the overaccumulation of carotenoids in hp2 ripening fruits was associated with disturbed ethylene production, increased expression of genes encoding master regulators of ripening and higher ethylene sensitivity and signaling output. The increased ethylene sensitivity observed in hp2 fruits was associated with the differential expression of genes encoding ethylene receptors and downstream signaling transduction elements, including the downregulation of the transcription factor ETHYLENE RESPONSE FACTOR.E4, a repressor of carotenoid synthesis. Accordingly, treatments with exogenous ethylene promoted carotenoid biosynthetic genes more intensively in hp2 than in wild-type fruits. Moreover, the loss of HP2 function drastically altered auxin signaling in tomato fruits, resulting in higher activation of the auxin-responsive promoter DR5, severe down-regulation of AUXIN/INDOLE-3-ACETIC ACID (Aux/IAA) genes and altered accumulation of AUXIN RESPONSE FACTOR (ARF) transcripts. Both tomato ARF2 paralogues (Sl-ARF2a and SlARF2b) were up-regulated in hp2 fruits, which agrees with the promotive roles played by these ARFs in tomato fruit ripening and carotenoid biosynthesis. Among the genes differentially expressed in hp2 fruits, the additive effect of light treatment and loss of HP2 function was particularly evident for those encoding carotenoid biosynthetic enzymes, ethylene-related transcription factors, Aux/IAAs and ARFs. Altogether, the data uncover the involvement of ethylene and auxin as part of the light signaling cascades controlling tomato fruit metabolism and provide a new link between light signaling, plant hormone sensitivity and carotenoid metabolism in ripening fruits.

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FaPAO5 regulates Spm/Spd levels as a signaling during strawberry fruit ripening.

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FaPAO5 regulates Spm/Spd levels as a signaling during strawberry fruit ripening.

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