Spatiotemporal feedforward between PKM2 tetramers and mTORC1 prompts mTORC1 activation.

Most mammalian cells couple glucose availability to anabolic processes via the mTORC1 pathway. However, the mechanism by which fluctuations in glucose availability are rapidly translated into mTORC1 signals remains elusive. Here, we show that cells rapidly respond to changes in glucose availability through the spatial coupling of mTORC1 and tetramers of the key glycolytic enzyme pyruvate kinase M2 (PKM2) on lysosomal surfaces in the late G1/S phases. The lysosomal localization of PKM2 tetramers enables rapid increases in local ATP concentrations around lysosomes to activate mTORC1, while bypassing the need to elevate global ATP levels in the entire cell. In essence, this spatial coupling establishes a feedforward loop to enable mTORC1 to rapidly sense and respond to changes in glucose availability. We further demonstrate that this mechanism ensures robust cell proliferation upon fluctuating glucose availability. Thus, we present mechanistic insights into the rapid response of the mTORC1 pathway to changes in glucose availability. The underlying mechanism may be applicable to the control of other cellular processes.

MaMYB4 Recruits Histone Deacetylase MaHDA2 and Modulates the Expression of ω-3 Fatty Acid Desaturase Genes during Cold Stress Response in Banana Fruit.

Linoleic acid (LA; C18:2) and α-linolenic acid (ALA; C18:3) are two essential unsaturated fatty acids that play indispensable roles in maintaining membrane integrity in cold stress, and ω-3 fatty acid desaturases (FADs) are responsible for the transformation of LA into ALA. However, how this process is regulated at transcriptional and posttranscriptional levels remains largely unknown. In this study, an MYB transcription factor, MaMYB4, of a banana fruit was identified and found to target several ω-3 MaFADs, including MaFAD3-1, MaFAD3-3, MaFAD3-4 and MaFAD3-7, and repress their transcription. Intriguingly, the acetylation levels of histones H3 and H4 in the promoters of ω-3 MaFADs were elevated in response to cold stress, which was correlated with the enhancement in the transcription levels of ω-3 MaFADs and the ratio of ALA/LA. Moreover, a histone deacetylase MaHDA2 physically interacted with MaMYB4, thereby leading to the enhanced MaMYB4-mediated transcriptional repression of ω-3 MaFADs. Collectively, these data demonstrate that MaMYB4 might recruit MaHDA2 to repress the transcription of ω-3 MaFADs by affecting their acetylation levels, thus modulating fatty acid biosynthesis. Our findings provided new molecular insights into the regulatory mechanisms of fatty acid biosynthesis in cold stress in fruits.

A New Regulatory Network Controls Chilling Injury in Peach Fruit by γ-Aminobutyric Acid.

The control of chilling injury in peach fruit by a new regulator network, that exogenous γ-aminobutyric acid (GABA) regulates the metabolisms of polyamines (PAs), the GABA shunt, and proline, is still unclear. This study found that GABA induced an increase in the expression of PpADC and PpODC and a decrease in the expression of PpPAO expression, resulting in the accumulation of PAs. There was also an increase in the expression of PpGAD, which improved GABA content, and an increase in the expression of PpP5CS and PpOAT, which improved proline content. The correlation analysis showed that an increase in PpADC/PpP5CS expression was closely associated with the accumulation of putrescine and that the synergistic increase in the expression of PpODC and PpGAD/PpP5CS/PpOAT was closely related to the accumulation of spermine, proline, and GABA induced by GABA. Importantly, arginine and PpADC played a key role in putrescine accumulation, whereas ornithine and PpODC/PpOAT played a crucial role in the synergistic accumulation of spermine, proline, and GABA induced by GABA. This study provides new information on GABA-induced cold tolerance in peach fruit.

NAC-mediated membrane lipid remodeling negatively regulates fruit cold tolerance.

Low temperatures are known to destroy cell membranes' structural integrity by affecting the remodeling of their phospholipids. Fruits stored at low temperature are prone to chilling injury, characterized by discoloration, absence of ripening, surface pitting, growth inhibition, flavor loss, decay, and wilting. Phosphatidic acid, a vital second-messenger lipid in plants, is known to accumulate in response to different kinds of stress stimuli. However, the regulatory mechanism of its production from the degradation of phospholipids remains poorly understood. We identified two cold-responsive NAC (NAM/ATAF1/CUC2) transcription factors from bananas, namely, MaNAC25 and MaNAC28, which negatively regulated cold tolerance in banana fruits by upregulating the expression of phospholipid degradation genes in banana fruits. Furthermore, MaNAC25 and MaNAC28 formed a positive feedback loop to induce phospholipid degradation and produce phosphatidic acid. In contrast, ethylene directly inhibited the degradation of phospholipids in banana and transgenic tomato fruits. In addition, ethylene reduced the activity of MaNAC25 and MaNAC28, thereby inhibiting phospholipid degradation. To conclude, NAC-mediated membrane lipid remodeling negatively regulates the cold tolerance of banana and transgenic tomato fruits.

Membrane lipid metabolism influences chilling injury during cold storage of peach fruit.

Peach fruit is prone to chilling injury (CI) during long-term low temperature (LT) storage, and develops protective mechanisms against LT stress. This study revealed that 4 °C storage induced the occurrence of CI in peach fruit by promoting the expression of membrane lipid metabolism genes and phosphatidic acid (PA) accumulation, and stimulated a protective mechanism for peach fruit against LT by increasing diacylglycerol (DAG), triacylglycerol (TAG) and several phosphatidylcholine (PC) components in the later storage stage. In contrast, 0 °C delayed the occurrence of CI in peach fruit by delaying the degradation of phospholipids, upregulation of fatty acid desaturase (FAD) and the process of fatty acid unsaturation, and maintaining higher levels of PC and PE. Results from this study provide new information on the mechanism of CI in peach fruit, and lay the foundation for the transcriptional regulation mechanism of CI and cold tolerance in peach fruit mediated by membrane lipid metabolism.

Heterodimerization of MaTCP proteins modulates the transcription of MaXTH10/11 genes during banana fruit ripening.

The biological processes involved in banana fruit ripening are extremely complex and modulated by a number of genes such as transcription factors (TFs). Although TFs like MADS, ERF and NAC are implicated in controlling banana ripening, little is known about other TFs such as TCP in this process. In this work, 25 MaTCPs named MaTCP1 to MaTCP25 were characterized from our previously reported transcriptomes related to banana ripening. Expression analysis revealed that these MaTCPs displayed differential expression patterns during the progression of banana ripening. Particularly, MaTCP5, MaTCP19 and MaTCP20 were ethylene-inducible and nuclear-localized, with MaTCP5 and MaTCP20 acting as transcriptional activators while MaTCP19 being a transcriptional inhibitor. Moreover, MaTCP5 and MaTCP20 promoted the transcription of MaXTH10/11 that may play a role in fruit softening during banana ripening, whereas MaTCP19 repressed their transcription, by directly binding to their promoters. Importantly, protein-protein interaction assays demonstrated that MaTCP20 physically interacts with MaTCP5 and MaTCP19 to form heterodimers in vitro and in vivo, and these protein complexes affects their transcriptional activities in regulating the target genes. Taken together, our results provide an overview of the interactions between MaTCPs in controlling the ripening-associated genes and lay a foundation for further investigation of MaTCP gene family in regulating banana fruit ripening.

Accumulation of carotenoids and expression of carotenogenic genes in peach fruit.

To understand better the regulatory mechanism of the carotenoid accumulation, the expression profile of relevant carotenoid genes and metabolites were compared between two peach cultivars with different colors during fruit development. Meanwhile, the change pattern of carotenoid content and expression of carotenoid metabolic genes in peaches after harvest in response to blue light were also investigated. As compared to the yellow fleshed-cultivar 'Jinli', lower carotenoid levels were observed in skin and pulp in white peach cultivar 'Hujing', which might be explained by differentially expression of PpCCD4 gene. With respect to 'Jinli', the carotenoid accumulation during fruit development in fruit skin was partially linked with the transcriptional regulation of PpFPPS, PpGGPS, PpLCYB and PpCHYB. However, in the pulp, the accumulation might be also associated with the increased transcriptions of PpPDS, along with the above four genes. Blue light treatment induced carotenoid accumulation in 'Jinli' peaches during storage. In addition, the treated-fruit displayed higher expression of all the eight genes analysed with a lesser extent on PpCCD4, which suggested that the much more increased carotenoid synthesis rate could result in the higher carotenoid content in blue light-treated fruit. The results presented herein contribute to further elucidating the regulatory mechanism of carotenoid accumulation in peach fruit.

Exogenous Melatonin Treatment Increases Chilling Tolerance and Induces Defense Response in Harvested Peach Fruit during Cold Storage.

The effect of exogenous melatonin on chilling injury in peach fruit after harvest was investigated. To explore the optimum concentration of melatonin for chilling tolerance induction, peach fruit were treated with 50, 100, or 200 µM melatonin for 120 min and then stored for 28 days at 4 °C. The results showed that application of melatonin at 100 µM was most effective in reducing chilling injury of peach fruit after harvest. Peaches treated with melatonin at this concentration displayed higher levels of extractable juice rate and total soluble solids than the non-treated peaches. In addition, melatonin treatment enhanced expression of PpADC, PpODC, and PpGAD and consequently increased polyamines and γ-aminobutyric acid (GABA) contents. Meanwhile, the upregulated transcripts of PpADC and PpODC and inhibited PpPDH expression resulted in the higher proline content in melatonin-treated fruit compared to the control fruit. Our results revealed that melatonin treatment may be a useful technique to alleviate chilling injury in cold-stored peach fruit. The chilling tolerance of harvested peaches induced by melatonin treatment is associated with higher levels of polyamine, GABA, and proline. These data provided here are the first protective evidence of exogenous melatonin in harvested horticultural products in response to direct chilling stress.