The TOR-EIN2 axis mediates nuclear signalling to modulate plant growth.

The evolutionarily conserved target of rapamycin (TOR) kinase acts as a master regulator that coordinates cell proliferation and growth by integrating nutrient, energy, hormone and stress signals in all eukaryotes1,2. Research has focused mainly on TOR-regulated translation, but how TOR orchestrates the global transcriptional network remains unclear. Here we identify ethylene-insensitive protein 2 (EIN2), a central integrator3-5 that shuttles between the cytoplasm and the nucleus, as a direct substrate of TOR in Arabidopsis thaliana. Glucose-activated TOR kinase directly phosphorylates EIN2 to prevent its nuclear localization. Notably, the rapid global transcriptional reprogramming that is directed by glucose-TOR signalling is largely compromised in the ein2-5 mutant, and EIN2 negatively regulates the expression of a wide range of target genes of glucose-activated TOR that are involved in DNA replication, cell wall and lipid synthesis and various secondary metabolic pathways. Chemical, cellular and genetic analyses reveal that cell elongation and proliferation processes that are controlled by the glucose-TOR-EIN2 axis are decoupled from canonical ethylene-CTR1-EIN2 signalling, and mediated by different phosphorylation sites. Our findings reveal a molecular mechanism by which a central signalling hub is shared but differentially modulated by diverse signalling pathways using distinct phosphorylation codes that can be specified by upstream protein kinases.

STABILIZED1 Modulates Pre-mRNA Splicing for Thermotolerance.

High-temperature stress often leads to differential RNA splicing, thus accumulating different types and/or amounts of mature mRNAs in eukaryotic cells. However, regulatory mechanisms underlying plant precursor mRNA (pre-mRNA) splicing in the environmental stress conditions remain elusive. Herein, we describe that a U5-snRNP-interacting protein homolog STABILIZED1 (STA1) has pre-mRNA splicing activity for heat-inducible transcripts including HEAT STRESS TRANSCRIPTION FACTORs and various HEAT SHOCK PROTEINs for the establishment of heat stress tolerance in Arabidopsis (Arabidopsis thaliana). Our cell-based splicing reporter assay demonstrated STA1 acts on pre-mRNA splicing for specific subsets of stress-related genes. Cellular reconstitution of heat-inducible transcription cascades supported the view that STA1-dependent pre-mRNA splicing plays a role in DREB2A-dependent HSFA3 expression for heat-responsive gene expression. Further genetic analysis with a loss-of-function mutant sta1-1, STA1-expressing transgenic plants in Col background, and STA1-expressing transgenic plants in the sta1-1 background verified that STA1 is essential in expression of necessary genes including HSFA3 for two-step heat stress tolerance in plants. However, constitutive overexpression of the cDNA version of HSFA3 in the sta1-1 background is unable to execute plant heat stress tolerance in sta1-1 Consistently our global target analysis of STA1 showed that its splicing activity modulates a rather broad range of gene expression in response to heat treatment. The findings of this study reveal that heat-inducible STA1 activity for pre-mRNA splicing serves as a molecular regulatory mechanism underlying the plant stress tolerance to high-temperature stress.

The effects of a multicomponent intervention program on clinical outcomes associated with falls in healthy older adults.

BACKGROUND: Multicomponent intervention programs have been shown to be effective in reducing risk factors associated with falls, but the primary target population of these interventions is often low-functioning older adults. AIMS: The purpose of this study was to investigate the effectiveness of a multicomponent intervention program focusing on balance and muscle strength for independently functioning community-dwelling older adults. METHODS: Fifty-three independently functioning older adults, aged 80.09 ± 6.62 years, participated in a group exercise class (conducted 2 times/week for 8 weeks) emphasizing balance. Outcome measures were balance performance using the Fullerton Advanced Balance (FAB) scale and muscle strength using the Senior Fitness Test (SFT). RESULTS: The intervention improved balance (P < 0.001), and older adults who were classified as having high fall risks based on the FAB scores at pre-testing improved more than older adults who were classified as having low fall risks (P = 0.017). As a result, 22 participants transitioned from a high fall risk group at pre-testing to a low fall risk group at post-testing (P < 0.001). The intervention also enhanced both upper and lower muscle extremity strength based on SFT results (P < 0.001) regardless of participants' classification of fall risk status. CONCLUSIONS AND DISCUSSION: The multicomponent intervention conducted two times per week for 8 weeks was effective in improving balance and enhancing muscle strength of independently functioning older adults. The results underscore the importance of providing fall prevention interventions to healthy older adults, a population often not a target of balance interventions.

PHABULOSA controls the quiescent center-independent root meristem activities in Arabidopsis thaliana.

Plant growth depends on stem cell niches in meristems. In the root apical meristem, the quiescent center (QC) cells form a niche together with the surrounding stem cells. Stem cells produce daughter cells that are displaced into a transit-amplifying (TA) domain of the root meristem. TA cells divide several times to provide cells for growth. SHORTROOT (SHR) and SCARECROW (SCR) are key regulators of the stem cell niche. Cytokinin controls TA cell activities in a dose-dependent manner. Although the regulatory programs in each compartment of the root meristem have been identified, it is still unclear how they coordinate one another. Here, we investigate how PHABULOSA (PHB), under the posttranscriptional control of SHR and SCR, regulates TA cell activities. The root meristem and growth defects in shr or scr mutants were significantly recovered in the shr phb or scr phb double mutant, respectively. This rescue in root growth occurs in the absence of a QC. Conversely, when the modified PHB, which is highly resistant to microRNA, was expressed throughout the stele of the wild-type root meristem, root growth became very similar to that observed in the shr; however, the identity of the QC was unaffected. Interestingly, a moderate increase in PHB resulted in a root meristem phenotype similar to that observed following the application of high levels of cytokinin. Our protoplast assay and transgenic approach using ARR10 suggest that the depletion of TA cells by high PHB in the stele occurs via the repression of B-ARR activities. This regulatory mechanism seems to help to maintain the cytokinin homeostasis in the meristem. Taken together, our study suggests that PHB can dynamically regulate TA cell activities in a QC-independent manner, and that the SHR-PHB pathway enables a robust root growth system by coordinating the stem cell niche and TA domain.

Regulatory functions of evolutionarily conserved AN1/A20-like Zinc finger family proteins in Arabidopsis stress responses under high temperature.

AN1/A20-like Zinc finger family proteins are evolutionarily conserved regulatory components in eukaryotic signaling circuits. In Arabidopsis thaliana, the AN1/A20 Zinc finger family is encoded as 14 members in the genome and collectively referred to as stress-associated proteins (SAPs). Here we described AtSAP5 localized to the nucleus, and played a role in heat-responsive gene regulation together with MBF1c. Seedling survival assay of sap5 and mbf1c demonstrated consistent effects of AtSAP5 and MBF1C in response to two-step heat treatment, supporting their function in heat stress tolerance. Our findings yield an insight in A20/AN1-like Zinc finger protein AtSAP5 functions in plant adaptability under high temperature.

Regulatory role of BOTRYTIS INDUCED KINASE1 in ETHYLENE INSENSITIVE3-dependent gene expression in Arabidopsis.

KEY MESSAGE: Arabidopsis BIK1 negatively regulates EIN3-depedent gene expression as an immediate cellular response. BIK1 localizes to the plasma membrane and its autophosphorylation and kinase activity involves in EIN3 repression. BOTRYTIS INDUCED KINASE1 (BIK1) is a multifunctional receptor-like kinase that involves in ethylene-mediated plant defense signaling. The loss of function BIK1 becomes insensitive to ethylene, but it still accumulates a higher level of ETHYLENE INSENSITIVE3 (EIN3) that serves as the key transcription activator in ethylene signaling. To unequivocally elucidate BIK1 function on EIN3 regulation in ethylene signaling, we took a combined approach of transient expression assay and stable expression analysis of BIK1. In our cell-based functional assay BIK1 destabilized EIN3 and down-regulated EIN3-dependent transcription. Membrane localization and autophosphorylation of BIK1 were required for full repression of EIN3 function, but its kinase activity potential compromised such regulatory action. Consistently, the analysis of transgenic plants verified BIK1 function on EIN3 repression. Our findings have clarified that autophosphorylated BIK1 in the plasma membrane negatively regulates EIN3-dependent gene expression. Thus, ethylene insensitivity in bik1 appears to be an indirect or a feedback long-term response.

Inverse modulation of the energy sensor Snf1-related protein kinase 1 on hypoxia adaptation and salt stress tolerance in Arabidopsis thaliana.

Terrestrial plants are exposed to complex stresses of high salt-induced abscisic acid (ABA) and submergence-induced hypoxia when seawater floods fields. Many studies have investigated plant responses to individual stress conditions, but not so much for coupled or sequentially imposed stresses. We examined molecular regulatory mechanisms of gene expression underlying the cellular responses involved in crosstalk between salt and hypoxia stresses. Salt/ABA- and AtMYC2-dependent induction of a synthetic ABA-responsive element and the native RD22 promoters were utilized in our cell-based functional assays. Such promoter-based reporter induction was largely inhibited by hypoxia and hypoxia-inducible AKIN10 activity. Biochemical analyses showed that AKIN10 negatively modulates AtMYC2 protein accumulation via proteasome activity upon AKIN10 kinase activity-dependent protein modification. Further genetic analysis using transgenic plants expressing AKIN10 provided evidence that AKIN10 activity undermined AtMYC2-dependent salt tolerance. Our findings unravel a novel molecular interaction between the key signalling constituents leading crosstalk between salt and hypoxia stresses in Arabidopsis thaliana under the detrimental condition of submergence in saltwater.

Dual control of nuclear EIN3 by bifurcate MAPK cascades in C2H4 signalling.

A principal question in MAP kinase (MAPK/MPK) cascade signalling is how similar components dictate different specificity in the information-processing machineries from yeast to humans and plants. In Arabidopsis, how MPK3/6 modulates distinct outputs in diverse signal transduction pathways remains elusive. By combining systematic cellular and genetic screens, here we uncover a previously unexpected MKK9-MPK3/MPK6 cascade promoting ethylene-insensitive 3 (EIN3)-mediated transcription in ethylene signalling. The mkk9 mutant exhibits a broad spectrum of moderate ethylene-insensitive phenotypes, and translocated MKK9 governs nuclear signalling downstream of receptors. Breaking a linear model and conventional MAPK signalling, ethylene inactivates the negative regulator constitutive triple response 1 (CTR1, a Raf-like MAPK kinase kinase (MAPKKK)) to activate the positive MKK9-MPK3/6 cascade. The bifurcate and antagonistic CTR1 and MKK9 pathways are both critical in determining ethylene-signalling specificity through two MAPK phosphorylation sites with opposite effects on EIN3 stability. The results suggest a new paradigm for linking intertwined MAPK cascades to control quantitative responses and specificity in signalling networks.

Signaling role of fructose mediated by FINS1/FBP in Arabidopsis thaliana.

Sugars are evolutionarily conserved signaling molecules that regulate the growth and development of both unicellular and multicellular organisms. As sugar-producing photosynthetic organisms, plants utilize glucose as one of their major signaling molecules. However, the details of other sugar signaling molecules and their regulatory factors have remained elusive, due to the complexity of the metabolite and hormone interactions that control physiological and developmental programs in plants. We combined information from a gain-of-function cell-based screen and a loss-of-function reverse-genetic analysis to demonstrate that fructose acts as a signaling molecule in Arabidopsis thaliana. Fructose signaling induced seedling developmental arrest and interacted with plant stress hormone signaling in a manner similar to that of glucose. For fructose signaling responses, the plant glucose sensor HEXOKINASE1 (HXK1) was dispensable, while FRUCTOSE INSENSITIVE1 (FINS1), a putative FRUCTOSE-1,6-BISPHOSPHATASE, played a crucial role. Interestingly, FINS1 function in fructose signaling appeared to be independent of its catalytic activity in sugar metabolism. Genetic analysis further indicated that FINS1-dependent fructose signaling may act downstream of the abscisic acid pathway, in spite of the fact that HXK1-dependent glucose signaling works upstream of hormone synthesis. Our findings revealed that multiple layers of controls by fructose, glucose, and abscisic acid finely tune the plant autotrophic transition and modulate early seedling establishment after seed germination.

A 12-Week, Single-Centre, Randomised, Double-Blind, Placebo-Controlled, Parallel-Design Clinical Trial for the Evaluation of the Efficacy and Safety of Lactiplantibacillus plantarum SKO-001 in Reducing Body Fat.

There is growing evidence linking gut microbiota to overall health, including obesity risk and associated diseases. Lactiplantibacillus plantarum SKO-001, a probiotic strain isolated from Angelica gigas, has been reported to reduce obesity by controlling the gut microbiome. In this double-blind, randomised clinical trial, we aimed to evaluate the efficacy and safety of SKO-001 in reducing body fat. We included 100 participants randomised into SKO-001 or placebo groups (1:1) for 12 weeks. Dual-energy X-ray absorptiometry was used to objectively evaluate body fat reduction. Body fat percentage (p = 0.016), body fat mass (p = 0.02), low-density lipoprotein-cholesterol levels (p = 0.025), and adiponectin levels (p = 0.023) were lower in the SKO-001 group than in the placebo group after 12 weeks of SKO-001 consumption. In the SKO-001 group, the subcutaneous fat area (p = 0.003), total cholesterol levels (p = 0.003), and leptin levels (p = 0.014) significantly decreased after 12 weeks of SKO-001 consumption compared with baseline values. Additionally, SKO-001 did not cause any severe adverse reactions. In conclusion, SKO-001 is safe and effective for reducing body fat and has the potential for further clinical testing in humans.

Regulatory functions of SnRK1 in stress-responsive gene expression and in plant growth and development.

Sucrose-nonfermentation1-related protein kinase1 (SnRK1) is an evolutionarily conserved energy sensor protein that regulates gene expression in response to energy depletion in plants. Efforts to elucidate the functions and mechanisms of this protein kinase are hampered, however, by inherent growth defects of snrk1-null mutant plants. To overcome these limitations and study SnRK1 functions in vivo, we applied a method combining transient expression in leaf mesophyll protoplasts and stable expression in transgenic plants. We found that both rice (Oryza sativa) and Arabidopsis (Arabidopsis thaliana) SnRK1 activities critically influence stress-inducible gene expression and the induction of stress tolerance. Genetic, molecular, and chromatin immunoprecipitation analyses further revealed that the nuclear SnRK1 modulated target gene transcription in a submergence-dependent manner. From early seedling development through late senescence, SnRK1 activities appeared to modulate developmental processes in the plants. Our findings offer insight into the regulatory functions of plant SnRK1 in stress-responsive gene regulation and in plant growth and development throughout the life cycle.

Cell fate in the Arabidopsis root epidermis is determined by competition between WEREWOLF and CAPRICE.

The root hair and nonhair cells in the Arabidopsis (Arabidopsis thaliana) root epidermis are specified by a suite of transcriptional regulators. Two of these are WEREWOLF (WER) and CAPRICE (CPC), which encode MYB transcription factors that are required for promoting the nonhair cell fate and the hair cell fate, respectively. However, the precise function and relationship between these transcriptional regulators have not been fully defined experimentally. Here, we examine these issues by misexpressing the WER gene using the GAL4-upstream activation sequence transactivation system. We find that WER overexpression in the Arabidopsis root tip is sufficient to cause epidermal cells to adopt the nonhair cell fate through direct induction of GLABRA2 (GL2) gene expression. We also show that GLABRA3 (GL3) and ENHANCER OF GLABRA3 (EGL3), two closely related bHLH proteins, are required for the action of the overexpressed WER and that WER interacts with these bHLHs in plant cells. Furthermore, we find that CPC suppresses the WER overexpression phenotype quantitatively. These results show that WER acts together with GL3/EGL3 to induce GL2 expression and that WER and CPC compete with one another to define cell fates in the Arabidopsis root epidermis.

Preparing Undergraduate Students for Summer Research Experiences and Graduate School Applications in a Pandemic Environment: Development and Implementation of Online Modules.

Engaging students in research is a high impact practice that improves student retention and persistence in behavioral and biomedical sciences and engineering. The California State University Long Beach (CSULB) Building Infrastructure Leading to Diversity (BUILD) Program offers an intensive research training experience to undergraduate students from a wide range of health-related disciplines. The goal of this program is to provide students with research skills, psychosocial resources, and graduate school application guidance that will make them competitive for Ph.D. programs. With the COVID-19 pandemic forcing the campus closure of many universities, including CSULB, our student training had to transition from in-person training to online training. This paper discusses the development and implementation of a series of eight online modules for guiding students through the application process for summer research experiences and graduate schools. Overall, the BUILD trainees were positive about the online modules. Specifically, they indicated that the modules were useful, informative, easy to access/use, good use of their time, and a good supplemental activity to their learning community activities. Most trainees indicated that they preferred the modules to be implemented in a hybrid format, where the students can view the modules on their own first and then have an opportunity to engage in in-person/synchronous online discussions.

Formation of protein-rich coatings around lipid droplets using the electrostatic deposition method.

The purpose of this study was to determine whether protein-rich coatings could be formed around lipid droplets using an electrostatic deposition method. These coatings were assembled using two methods: (i) beta-lactoglobulin was adsorbed to beta-lactoglobulin-pectin-coated lipid droplets; (ii) beta-lactoglobulin-pectin complexes were adsorbed to beta-lactoglobulin-coated lipid droplets. Composite particles, consisting of lipid droplets with protein-rich biopolymer coatings, could be formed using both approaches (e.g., at pH 4, the protein surface load could be increased from 3 to 59 mg m(-2)). These composite particles could be made small (d < 500 nm) and relatively stable to gravitational separation at certain protein concentrations. Nevertheless, aggregation and sedimentation occurred at sufficiently high protein concentrations because of charge neutralization. The composite particles remained stable after they were heated above the thermal denaturation temperature of the globular proteins at pH 4. When the heated composite particles were adjusted to pH 7, where beta-lactoglobulin and pectin are both negatively charged, some of the pectin and beta-lactoglobulin became detached from the droplet surfaces but the protein surface load was still higher than in a nontreated sample. These composite particles may be useful for increasing the protein concentration in biopolymer coatings surrounding lipid droplets, which potentially has practical applications in the food industry (e.g., in protecting omega-3 oils from oxidation or in developing natural weighting agents).

Assembled protein nanoparticles in food or nutrition applications.

Proteins are one of the essential components of nutritional food materials and an excellent source for food-grade nanomaterials. This review focuses on select examples of nanoparticles assembled naturally, found in food-relevant materials, major approaches in assembling nanoscale structure from proteins, and general applications of protein nanoparticles in food or nutrition. Animal-sourced casein and non-animal grain storage proteins and legume storage proteins are discussed in terms of their structural assemblies. Protein solubility is a key factor in assembling protein nanoparticles with desired functional properties. Desolvation is the most common technique to prepare protein nanoparticles for insoluble proteins. Well-hydrated protein assemblies have been extensively studied through electrostatic complexes, assembled with fatty acid and starch, reassembled protein structure, and nanogels. These protein-based nanoparticles have been utilized for filler materials of films, encapsulation of bioactive molecules, and stabilization of emulsions. Most studies exploiting protein-based nanoparticles have focused on developing technologies in extraction of proteins from sources and assembly of nanoparticles in different environmental conditions.

Contributions of protein and milled chitin extracted from domestic cricket powder to emulsion stabilization.

Interfacial and emulsifying properties of fractionated cricket powder were assessed to identify whether emulsification properties originate from protein or chitin particles. Fractions extracted in alkaline water, containing high protein and mineral contents, increased the surface pressure of heptane-water interfaces with near-saturation equilibrium surface pressure of 31 mN/m. Dynamic surface pressure profiles indicated adsorption of protein clusters to the interface. Emulsification capacity of protein fraction was 50% greater than that of the source cricket flour, although oil-in-water emulsions prepared with 1-2% (w/w) protein fraction formed a cream layer within one day of storage. Emulsified layers persisted for up to 20 days, and light scattering measurements described a stable population with surface-volume-mean diameter of approximately 3 µm. Chitin-rich fractions milled to a particle size of 0.5-200 µm contributed negligible surface pressure, and its emulsification capacity was 5% of the value for the source cricket flour. Emulsions prepared with chitin-rich fractions coexisted with an unstable precipitate layer comprising 60% of the added solid, which was attributed to larger particles with poor emulsifying capability. Stable chitin-stabilized emulsion phases were resistant to creaming, yet volume-mean droplet diameter surpassed 50 µm within 24 h of storage. Both protein and chitin fractions have emulsifying capabilities but would require further processing or secondary additives to achieve desirable storage stability.

Quantitative approach to realization of ultrasonic grain refinement of Al-7Si-2Cu-1Mg alloy.

Ultrasonic melt treatment (UST) was applied to Al-7Si-2Cu-1Mg melt at various temperatures of 620, 650, 700 and 785 °C. MgAl2O4 particles which were often found to be densely populated along oxide films, became effectively dispersed and well-wetted by UST. Transmission electron microscopy work combined with crystallography analysis clearly indicates that MgAl2O4 particles can act as α-Al nucleation site with the aid of UST. However, with UST, grain refinement occurred only at temperature of 620 °C and the grain size increased from 97 to 351 µm with increase of melt temperature to 785 °C for UST. In quantitative analysis of grain size and MgAl2O4 particle diameter, it was found that ultrasonic de-agglomeration decreased mean particle size of the MgAl2O4 particles, significantly reducing size from 1.2 to 0.4 µm when temperature increased from 620 to 785 °C. Such a size reduction with increased number of MgAl2O4 particles does not always guarantee grain refinement. Thus, in this work, detailed condition for achieving grain refinement by UST is discussed based on quantitative measurement. Furthermore, we tried to suggest the most valid grain refinement mechanism among the known mechanisms by investigation of the relationship between grain size and particle size with variation of melt temperature.

STABILIZED1 as a heat stress-specific splicing factor in Arabidopsis thaliana.

To overcome high temperature stress, plants have developed transcriptional cascades which express a large amount of chaperone proteins called heat shock proteins (HSPs). In our recent publication, we reported that STABILIZED1, as an U5-snRNP-interacting protein, is involved in the splicing of heat shock factor (HSF) and HSP transcripts during high temperature stress. This indicates that not only transcriptional regulation, but also post-transcriptional regulation by STA1, is essential for the full activation of HSF-HSP cascades and for thermotolerance. Here, we observed that the splicing of HSP transcripts was induced independent of STA1 at room temperature after heat acclimation, indicating that STA1 acts as a high temperature-specific splicing factor for the splicing of HSP transcripts. Our findings suggest the molecular mechanism for how HSF and HSP transcripts are spliced well under high temperature stress that blocks the splicing of overall transcripts.

Role of impaired vision during dual-task walking in young and older adults.

While cognitive-motor interference in dual-task activities is well established, it is still unknown how such interference is influenced by concurrent visual challenges. Nineteen community-dwelling healthy, cognitively intact, older adults (Mean±SD=71.45±1.25years, 6 males) and nineteen young adults (Mean±SD=22.25±0.68years, 4 males) performed a cognitive-single-task (serial subtraction by 3), a walking-single-task and a cognitive-walking-dual-task under normal, blurred and peripheral-vision-loss conditions (artificially imposed using goggles). Gait parameters and the number of correct responses were measured. Dual task costs for both walking and cognition were computed. Results showed that higher walking cost was seen with impaired vision (p=0.05) and with older adults (p=0.03); greater cognitive cost was seen with impaired vision (p=0.01), but no difference in cognitive cost was seen between young and older adults. Thus, when faced with impaired vision, both young and older adults appear to allocate less attention to cognition than to walking, and thus prioritize walking. Future work should explore whether dual-task training under visual challenge could reduce cognitive-motor interference and reduce fall risks in older adults.

Regulatory Functions of Cellular Energy Sensor SNF1-Related Kinase1 for Leaf Senescence Delay through ETHYLENE- INSENSITIVE3 Repression.

Aging of living organisms is governed by intrinsic developmental programs, of which progression is often under the regulation of their cellular energy status. For example, calorie restriction is known to slow down aging of heterotrophic organisms from yeasts to mammals. In autotrophic plants cellular energy deprivation by perturbation of photosynthesis or sugar metabolism is also shown to induce senescence delay. However, the underlying molecular and biochemical mechanisms remain elusive. Our plant cell-based functional and biochemical assays have demonstrated that SNF1-RELATED KINASE1 (SnRK1) directly interacts, phosphorylates, and destabilizes the key transcription factor ETHYLENE INSENSITIVE3 (EIN3) in senescence-promoting hormone ethylene signaling. Combining chemical manipulation and genetic validation using extended loss-of-function mutants and gain-of-function transgenic lines, we further revealed that a SnRK1 elicitor, 3-(3,4-dichlorophenyl)-1,1-dimethylurea enables to slow down senescence-associated leaf degreening through the regulation of EIN3 in Arabidopsis. Our findings enlighten that an evolutionary conserved cellular energy sensor SnRK1 plays a role in fine-tuning of organ senescence progression to avoid sudden death during the last step of leaf growth and development.