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.

Specificity of H2O2 signaling in leaf senescence: is the ratio of H2O2 contents in different cellular compartments sensed in Arabidopsis plants?

Leaf senescence is an integral part of plant development and is driven by endogenous cues such as leaf or plant age. Developmental senescence aims to maximize the usage of carbon, nitrogen and mineral resources for growth and/or for the sake of the next generation. This requires efficient reallocation of the resources out of the senescing tissue into developing parts of the plant such as new leaves, fruits and seeds. However, premature senescence can be induced by severe and long-lasting biotic or abiotic stress conditions. It serves as an exit strategy to guarantee offspring in an unfavorable environment but is often combined with a trade-off in seed number and quality. In order to coordinate the very complex process of developmental senescence with environmental signals, highly organized networks and regulatory cues have to be in place. Reactive oxygen species, especially hydrogen peroxide (H2O2), are involved in senescence as well as in stress signaling. Here, we want to summarize the role of H2O2 as a signaling molecule in leaf senescence and shed more light on how specificity in signaling might be achieved. Altered hydrogen peroxide contents in specific compartments revealed a differential impact of H2O2 produced in different compartments. Arabidopsis lines with lower H2O2 levels in chloroplasts and cytoplasm point to the possibility that not the actual contents but the ratio between the two different compartments is sensed by the plant cells.

Sitosterol-mediated Antioxidant Regulation to Enhance Heat Tolerance in Creeping Bentgrass

Heat stress symptoms in cool-season plants are characterized by loss of chlorophyll (Chl) and membrane stability, as well as oxidative damage. The objectives of this study were to determine whether foliar application of β-sitosterol, a naturally occurring plant metabolite, may promote heat tolerance by suppressing heat-induced leaf senescence as indicated by the maintenance of healthy turf quality (TQ), and Chl and membrane stability; and to determine its roles in regulating antioxidant metabolism in creeping bentgrass (Agrostis stolonifera). ‘Penncross’ plants were exposed to heat stress (35/30 °C day/night) optimal temperature conditions (nonstressed control, 22/17 °C day/night) for a duration of 28 days in environment-controlled growth chambers. Plants were foliar-treated with β-sitosterol (400 µM) or water only (untreated control) before heat stress, and at 7-day intervals through 28 days of heat stress. Plants treated with β-sitosterol had significantly greater TQ and Chl content, and significantly less electrolyte leakage (EL) than untreated controls at 21 and 28 days of heat stress. Application of β-sitosterol reduced malondialdehyde (MDA) content significantly at 21 and 28 days of heat stress, and promoted the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) from 14 through 28 days of heat stress. β-Sitosterol effectively improved heat tolerance through suppression of leaf senescence in creeping bentgrass exposed to heat stress in association with the alleviation of membrane lipid peroxidation and activation of the enzymatic antioxidant system.

Direct Phenological Responses but Later Growth Stimulation upon Spring and Summer/Autumn Warming of Prunus spinosa L. in a Common Garden Environment

Future predictions of forest ecosystem responses are a challenge, as global temperatures will further rise in the coming decades at an unprecedented rate. The effect of elevated temperature on growth performance and phenology of three Prunus spinosa L. provenances (originating from Belgium, Spain, and Sweden) in a common garden environment was investigated. One-year-old seedlings were grown in greenhouse conditions and exposed to ambient and elevated temperatures in the spring (on average 5.6 °C difference) and in the late summer/autumn of 2018 (on average 1.9 °C difference), while they were kept hydrated, in a factorial design. In the following years, all plants experienced the same growing conditions. Bud burst, leaf senescence, height, and diameter growth were recorded. Height and radial growth were not affected in the year of the treatments (2018) but were enhanced the year after (2019), whereas phenological responses depended on the temperature treatments in the year of the treatments (2018) with little carry-over effects in the succeeding years. Spring warming enhanced more height growth in the succeeding year, whereas summer/autumn warming stimulated more radial growth. Spring warming advanced bud burst and shortened the leaf opening process whereas summer/autumn warming delayed leaf senescence and enlarged the duration of this phenophase. These results can help predict the putative shifts in species composition of future forests and woody landscape elements.

Integrative Analyses of Biochemical Properties and Transcriptome Reveal the Dynamic Changes in Leaf Senescence of Tobacco (Nicotiana tabacum L.)

Leaf senescence is an important process of growth and development in plant, and it is a programmed decline controlled by a series of genes. In this study, the biochemical properties and transcriptome at five maturity stages (M1∼M5) of tobacco leaves were analyzed to reveal the dynamic changes in leaf senescence of tobacco. A total of 722, 1,534, 3,723, and 6,933 genes were differentially expressed (DEG) between M1 and M2, M1 and M3, M1 and M4, and M1 and M5, respectively. Significant changes of nitrogen, sugars, and the DEGs related to metabolite accumulation were identified, suggesting the importance of energy metabolism during leaf senescence. Gene Ontology (GO) analysis found that DEGs were enriched in biosynthetic, metabolic, photosynthesis, and redox processes, and especially, the nitrogen metabolic pathways were closely related to the whole leaf senescence process (M1∼M5). All the DEGs were grouped into 12 expression profiles according to their distinct expression patterns based on Short Time-series Expression Miner (STEM) software analysis. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis found that these DEGs were enriched in pathways of carbon metabolism, starch and sucrose metabolism, nitrogen metabolism, and photosynthesis among these expression profiles. A total of 30 core genes were examined by Weight Gene Co-expression Network Analysis (WGCNA), and they appeared to play a crucial role in the regulatory of tobacco senescence. Our results provided valuable information for further functional investigation of leaf senescence in plants.

Triticum Aestivum L. cv. Guizi 1 ANS-6D Positively Regulates Leaf Senescence Through the Abscisic Acid Mediated Chlorophyll Degradation in Tobacco

Anthocyanidin synthase (ANS) is involved in the synthesis of anthocyanins, which are important phytonutrients because of their beneficial effects on human health. Here, we identified ANS-6D of purple-colored Triticum aestivum L. cv. Guizi 1 (Gz) that is involved in leaf senescence through the abscisic acid (ABA) mediated chlorophyll degradation pathway in tobacco. After characterizing the leaf-senescence phenotype in GzANS-6D overexpression (OxGzANS-6D) lines, we found that the increased anthocyanin accumulation and decreased chlorophyll content in OxGzANS-6D lines were closely correlated with the expression levels of anthocyanin synthesis-related structural genes and senescence marker genes, as well as the accumulation of reactive oxygen species. The endogenous ABA content increased and ethylene content decreased in OxGzANS-6D transgenic lines compared with wild type. Additionally, the levels of the abscisic acid-responsive transcription factors ABF1 and ABF2, as well as those of chlorophyll degradation-related genes (PAO, NYC, SGR and CHL), were significantly higher in OxGzANS-6D transgenic lines than in wild type. Furthermore, we found that GzABF1 and NtABF1 binds to the promoter of GzANS-6D, and NtABF2 binds to the promoter of NtSGR. Thus, GzANS-6D participated in leaf senescence through ABA-mediated chlorophyll degradation, and ABF1/2 play important role in GzANS-6D functions.

Identification and Characterization of an Early Leaf Senescence Gene ELS1 in Soybean

Early leaf senescence phenotype in soybean could be helpful to shorten the maturation period and prevent green stem disorder. From a high-density mutation library, we identified two early leaf senescence soybean mutant lines, els1-1 (early leaf senescence 1) and els1-2. The chlorophyll contents of both els1-1 and els1-2 were low in pre-senescent leaves. They degraded rapidly in senescent leaves, revealing that ELS1 is involved in chlorophyll biosynthesis during leaf development and chlorophyll degradation during leaf senescence. The causal mutations in els1 were identified by next-generation sequencing-based bulked segregant analysis. ELS1 encodes the ortholog of the Arabidopsis CaaX-like protease BCM1, which is localized in chloroplasts. Soybean ELS1 was highly expressed in green tissue, especially in mature leaves. The accumulation of photosystem I core proteins and light-harvesting proteins in els1 was low even in pre-senescent leaves, and their degradation was accelerated during leaf senescence. These results suggest that soybean ELS1 is involved in both chlorophyll synthesis and degradation, consistent with the findings in Arabidopsis BCM1. The gene els1, characterized by early leaf senescence and subsequent early maturation, does not affect the flowering time. Hence, the early leaf senescence trait regulated by els1 helps shorten the harvesting period because of early maturation characteristics. The els1-1 allele with weakly impaired function of ELS1 has only a small effect on agricultural traits and could contribute to practical breeding.