The PCY-SAG14 phytocyanin module regulated by PIFs and miR408 promotes dark-induced leaf senescence in Arabidopsis.

Leaf senescence is a critical process in plants and has a direct impact on many important agronomic traits. Despite decades of research on senescence-altered mutants via forward genetics and functional assessment of senescence-associated genes (SAGs) via reverse genetics, the senescence signal and the molecular mechanism that perceives and transduces the signal remain elusive. Here, using dark-induced senescence (DIS) of Arabidopsis leaf as the experimental system, we show that exogenous copper induces the senescence syndrome and transcriptomic changes in light-grown plants parallel to those in DIS. By profiling the transcriptomes and tracking the subcellular copper distribution, we found that reciprocal regulation of plastocyanin, the thylakoid lumen mobile electron carrier in the Z scheme of photosynthetic electron transport, and SAG14 and plantacyanin (PCY), a pair of interacting small blue copper proteins located on the endomembrane, is a common thread in different leaf senescence scenarios, including DIS. Genetic and molecular experiments confirmed that the PCY-SAG14 module is necessary and sufficient for promoting DIS. We also found that the PCY-SAG14 module is repressed by a conserved microRNA, miR408, which in turn is repressed by phytochrome interacting factor 3/4/5 (PIF3/4/5), the key trio of transcription factors promoting DIS. Together, these findings indicate that intracellular copper redistribution mediated by PCY-SAG14 has a regulatory role in DIS. Further deciphering the copper homeostasis mechanism and its interaction with other senescence-regulating pathways should provide insights into our understanding of the fundamental question of how plants age.

The abscisic acid-responsive element binding factors MAPKKK18 module regulates abscisic acid-induced leaf senescence in Arabidopsis.

The mitogen-activated protein kinase kinase kinase 18 (MAPKKK18) has been reported to play a role in abiotic stress priming in long-term abscisic acid (ABA) response including drought tolerance and leaf senescence. However, the upstream transcriptional regulators of MAPKKK18 remain to be determined. Here, we report ABA-responsive element binding factors (ABFs) as upstream transcription factors of MAPKKK18 expression. Mutants of abf2, abf3, abf4, and abf2abf3abf4 dramatically reduced the transcription of MAPKKK18. Our electrophoresis mobility shift assay and dual-luciferase reporter assay demonstrated that ABF2, ABF3, and ABF4 bound to ABA-responsive element cis-elements within the promoter of MAPKKK18 to transactivate its expression. Furthermore, enrichments of the promoter region of MAPKKK18 by ABF2, ABF3, and ABF4 were confirmed by in vivo chromatin immunoprecipitation coupled with quantitative PCR. In addition, we found that mutants of mapkkk18 exhibited obvious delayed leaf senescence. Moreover, a genetic study showed that overexpression of ABF2, ABF3, and ABF4 in the background of mapkkk18 mostly phenocopied the stay-green phenotype of mapkkk18 and, expression levels of five target genes of ABFs, that is, NYE1, NYE2, NYC1, PAO, and SAG29, were attenuated as a result of MAPKKK18 mutation. These findings demonstrate that ABF2, ABF3, and ABF4 act as transcription regulators of MAPKKK18 and also suggest that, at least in part, ABA acts in priming leaf senescence via ABF-induced expression of MAPKKK18.

Delayed senescence and crop performance under stress: always a functional couple?

Exposure to abiotic stresses accelerates leaf senescence in most crop plant species, thereby reducing photosynthesis and other assimilatory processes. In some cases, genotypes with delayed leaf senescence (i.e. 'stay-green') show stress resistance, particularly in cases of water deficit, and this has led to the proposal that senescence delay improves crop performance under some abiotic stresses. In this review, we summarize the evidence for increased resistance to abiotic stress, mostly water deficit, in genotypes with delayed senescence, and specifically focus on the physiological mechanisms and agronomic conditions under which the stay-green trait may ameliorate grain yield under stress.

WHIRLY proteins, multi-layer regulators linking the nucleus and organelles in developmental and stress-induced senescence of plants.

Plant senescence is an integrated programme of plant development that aims to remobilize nutrients and energy from senescing tissues to developing organs under developmental and stress-induced conditions. Upstream in the regulatory network, a small family of single-stranded DNA/RNA-binding proteins known as WHIRLYs occupy a central node, acting at multiple regulatory levels and via trans-localization between the nucleus and organelles. In this review, we summarize the current progress on the role of WHIRLY members in plant development and stress-induced senescence. WHIRLY proteins can be traced back in evolution to green algae. WHIRLY proteins trade off the balance of plant developmental senescence and stress-induced senescence through maintaining organelle genome stability via R-loop homeostasis, repressing the transcription at a configuration condition, and recruiting RNA to impact organelle RNA editing and splicing, as evidenced in several species. WHIRLY proteins also act as retrograde signal transducers between organelles and the nucleus through protein modification and stromule or vesicle trafficking. In addition, WHIRLY proteins interact with hormones, reactive oxygen species and environmental signals to orchestrate cell fate in an age-dependent manner. Finally, prospects for further research and promotion to improve crop production under environmental constraints are highlighted.

Changes in leaf lifespan, nitrogen resorption, and mean residence time of leaf nitrogen along a soil fertility gradient in an evergreen oak tree.

The ability of plants to retain nitrogen (N) for a long period of time is critical to their N use efficiency, growth, and fitness, particularly in infertile environments. The mean residence time of leaf N (MRTL) and its two determinants, leaf lifespan and N resorption efficiency (rN, the fraction of the total leaf N pool that is resorbed during leaf senescence), have been hypothesized to increase plastically with decreasing soil N fertility but this remains to be fully tested. To avoid confusion by random changes in these characteristics in a relatively narrow N fertility range, MRTL, leaf lifespan, and N resorption efficiency were measured in Quercus glauca over a broad N fertility range. In the high to moderate N fertility range, leaf lifespan and rN increased with decreasing N addition rate, and thus the MRTL increased. However, in the moderate to low N fertility range, leaf lifespan increased but rN decreased significantly, so MRTL decreased. The decrease in rN occurred because the senesced leaf N concentration was almost constant at the lower limit while the green leaf N concentration decreased in this range. The hump-shaped quadratic responses of MRTL and rN along the N fertility gradient suggest that incorrect conclusions about the response of these traits to N fertility variation may be drawn from experiments that include only a few fertility levels, and N recycling within leaf canopy alone cannot achieve efficient N use in infertile environments.

Leaf drought adaptive response in winter oilseed rape is altered at the onset of senescence: a study combining NMR relaxometry, multi-omics and microscopy.

Climate change is bringing more frequent and intense droughts, reducing overall water availability and adversely affecting crops. There is a need to improve our understanding of the tissular and cellular adaptation mechanisms that are critical for plant water conservation strategies. Here, we have used NMR relaxometry in combination with microscopy and multi-omic analysis to study the effects of progressive soil drought on winter oilseed rape (WOSR, Brassica napus L., cv. Aviso) leaves. This study reveals the structural and metabolic adjustments these leaves operate to maintain cell homeostasis. Our results are original in showing that the adaptive responses are altered in leaves at the onset of senescence, associated with changes in metabolic plasticity and mesophyll structures. Thus, long-term responses in young leaves involving osmotic adjustment were combined with the maintenance of tissue hydration and cell growth, contributing to high survival and recovery capacity. For the first time, short-term responses observed in early senescent-old leaves were associated with early drought-induced dehydration of the spongy layer. However, this dehydration was not followed by osmotic adjustment and did not allow maintenance of leaf tissue turgor. These findings open further studies on the genetic variability of drought responses related to identified short- and long-term structural and metabolic plasticity traits in Brassica species.

Arrest, Senescence and Death of Shoot Apical Stem Cells in Arabidopsis thaliana.

Shoot stem cells act as the source of the aboveground parts of flowering plants. A precise regulatory basis is required to ensure that plant stem cells show the right status during the stages of proliferation, senescence and cell death. Over the past few decades, the genetic circuits controlling stem cell fate, including the regulatory pathways of establishment, maintenance and differentiation, have been largely revealed. However, the morphological changes and molecular mechanisms of the final stages of stem cells, which are represented by senescence and cell death, have been less studied. The senescence and death of shoot stem cells are under the control of a complex series of pathways that integrate multiple internal and external signals. Given the crucial roles of shoot stem cells in influencing plant longevity and crop yields, researchers have attempted to uncover details of stem cell senescence and death. Recent studies indicate that stem cell activity arrest is controlled by the FRUITFULL-APETALA2 pathway and the plant hormones auxin and cytokinin, while the features of senescent and dead shoot apical stem cells have also been described, with dynamic changes in reactive oxygen species implicated in stem cell death. In this review, we highlight the recent breakthroughs that have enriched our understanding of senescence and cell death processes in plant stem cells.

Arabidopsis WRKY71 regulates ethylene-mediated leaf senescence by directly activating EIN2, ORE1 and ACS2 genes.

Leaf senescence is a pivotal step in the last stage of the plant life cycle and is influenced by various external and endogenous cues. A series of reports have indicated the involvement of the WRKY transcription factors in regulating leaf senescence, but the molecular mechanisms and signaling pathways remain largely unclear. Here we provide evidence demonstrating that WRKY71 acts as a positive regulator of leaf senescence in Arabidopsis. WRKY71-1D, an overexpressor of WRKY71, exhibited early leaf senescence, while wrky71-1, the WRKY71 loss-of-function mutant, displayed delayed leaf senescence. Accordingly, a set of senescence-associated genes (SAGs) were substantially elevated in WRKY71-1D but markedly decreased in wrky71-1. Chromatin immunoprecipitation assays indicated that WRKY71 can bind directly to the promoters of SAG13 and SAG201. Transcriptome analysis suggested that WRKY71 might mediate multiple cues to accelerate leaf senescence, such as abiotic stresses, dark and ethylene. WRKY71 was ethylene inducible, and treatment with the ethylene precursor 1-amino-cyclopropane-1-carboxylic acid enhanced leaf senescence in WRKY71-1D but caused only a marginal delay in leaf senescence in wrky71-1. In vitro and in vivo assays demonstrated that WRKY71 can directly regulate ETHYLENE INSENSITIVE2 (EIN2) and ORESARA1 (ORE1), genes of the ethylene signaling pathway. Consistently, leaf senescence of WRKY71-1D was obviously retarded in the ein2-5 and nac2-1 mutants. Moreover, WRKY71 was also proved to interact with ACS2 in vitro and in vivo. Treatment with AgNO3 and aminoethoxyvinylglycine and acs2-1 could greatly arrest the leaf senescence of WRKY71-1D. In conclusion, our data revealed that WRKY71 mediates ethylene signaling and synthesis to hasten leaf senescence in Arabidopsis.

Multiple phosphorylation events of the mitochondrial membrane protein TTM1 regulate cell death during senescence.

The role of mitochondria in programmed cell death (PCD) during animal growth and development is well documented, but much less is known for plants. We previously showed that the Arabidopsis thaliana triphosphate tunnel metalloenzyme (TTM) proteins TTM1 and TTM2 are tail-anchored proteins that localize in the mitochondrial outer membrane and participate in PCD during senescence and immunity, respectively. Here, we show that TTM1 is specifically involved in senescence induced by abscisic acid (ABA). Moreover, phosphorylation of TTM1 by multiple mitogen-activated protein (MAP) kinases regulates its function and turnover. A combination of proteomics and in vitro kinase assays revealed three major phosphorylation sites of TTM1 (Ser10, Ser437, and Ser490). Ser437, which is phosphorylated upon perception of senescence cues such as ABA and prolonged darkness, is phosphorylated by the MAP kinases MPK3 and MPK4, and Ser437 phosphorylation is essential for TTM1 function in senescence. These MPKs, together with three additional MAP kinases (MPK1, MPK7, and MPK6), also phosphorylate Ser10 and Ser490, marking TTM1 for protein turnover, which likely prevents uncontrolled cell death. Taken together, our results show that multiple MPKs regulate the function and turnover of the mitochondrial protein TTM1 during senescence-associated cell death, revealing a novel link between mitochondria and PCD.

Potencial dendrocronológico de tres especies de Podocarpáceas de la Cordillera de los Andes / Dendrochronological potential of three Podocarpaceae species from the Andean Cordillera

Resumen Introducción: Poco se conoce del potencial dendrocronológico de las Podocarpáceas en el trópico. Objetivo: Explorar el potencial dendrocronológico de tres especies de podocarpáceas: Retrophyllum rospigliosii, Podocarpus oleifolius y Prumnopitys harmsiana. Métodos: De plantaciones no manejadas localizadas en los Andes colombianos, se muestrearon y analizaron 88 árboles: 30 muestras de R. rospigliosii provenientes de secciones transversales, 30 y 28 muestras de P. oleifolius y P. harmsiana, respectivamente, provenientes de núcleos de madera extraídos con barreno de incrementos. Las muestras se procesaron siguiendo las técnicas dendrocronológicas estándar. Resultados: En general, las características anatómicas de los anillos de crecimiento son similares para las tres especies, con una anatomía simple de traqueidas alineadas radialmente por tratarse de coníferas. Dado que la edad conocida de la plantación coincide con el número de anillos se considera una fuerte evidencia de la frecuencia anual de su formación en R. rospigliosii y P. oleifolius, las cuales presentaron buena sincronización (cofechado) con una inter-correlación promedio de 0.55 (r-Pearson). Para P. harmsiana no fue posible concretar series de ancho de anillos de las muestras recolectadas. Las series estandarizadas de R. rospigliosii y P. oleifolius mostraron una relación con los registros instrumentales de precipitación y temperatura, indicando que estas especies pueden ser promisorias para estudios adicionales. Conclusión: La investigación dendrocronología con especies de Podocarpáceas podría realizarse exitosamente con R. rospigliosii y P. oleifolius, pero no con P. harmsiana.

Abstract Introduction: Little is known about the dendrochronological potential of Podocarpaceaes in the tropics. Objective: To explore the dendrochronological potential of three Podocarpaceae species: Retrophyllum rospigliosii, Podocarpus oleifolius, and Prumnopitys harmsiana. Methods: From a non-managed plantation in the Andean cordillera in Colombia, a total of 88 trees were analyzed: 30 samples of cross-sections of R. rospigliosii, and 30 and 28 samples of P. oleifolius and P. harmsiana, respectively, obtained with an increment borer. Samples were processed according to standard dendrochronological methods. Results: The anatomical characteristics of the growth rings of the three species are similar, with a simple conifer anatomy with radially oriented tracheids. Since the known age of the plantation coincides with the number of tree rings this is strong evidence of annual tree-ring frequency of R. rospigliosii and P. oleifolius which also showed a satisfactory cross-dating with an average inter-correlation of 0.55 (r-Pearson). For P. harmsiana, it was not possible to build a tree-ring series from the collected samples. R. rospigliosii and P. oleifolius standardized ring-width chronologies showed a relationship with the instrumental records of rainfall and temperature, indicating these species may be promising further studies. Conclusions: Dendrochronological research with Podocarpaceae species could be carried out successfully with R. rospigliosii and P. oleifolius but not with P. harmsiana.