Phospholipase Dα1 Acts as a Negative Regulator of High Mg2+-Induced Leaf Senescence in Arabidopsis

Magnesium (Mg2+) is a macronutrient involved in essential cellular processes. Its deficiency or excess is a stress factor for plants, seriously affecting their growth and development and therefore, its accurate regulation is essential. Recently, we discovered that phospholipase Dα1 (PLDα1) activity is vital in the stress response to high-magnesium conditions in Arabidopsis roots. This study shows that PLDα1 acts as a negative regulator of high-Mg2+-induced leaf senescence in Arabidopsis. The level of phosphatidic acid produced by PLDα1 and the amount of PLDα1 in the leaves increase in plants treated with high Mg2+. A knockout mutant of PLDα1 (pldα1-1), exhibits premature leaf senescence under high-Mg2+ conditions. In pldα1-1 plants, higher accumulation of abscisic and jasmonic acid (JA) and impaired magnesium, potassium and phosphate homeostasis were observed under high-Mg2+ conditions. High Mg2+ also led to an increase of starch and proline content in Arabidopsis plants. While the starch content was higher in pldα1-1 plants, proline content was significantly lower in pldα1-1 compared with wild type plants. Our results show that PLDα1 is essential for Arabidopsis plants to cope with the pleiotropic effects of high-Mg2+ stress and delay the leaf senescence.

PcGCE is a potent elicitor of defense responses in aspen

Using microbial enzymes in transgenesis is a powerful means to introduce new functionalities in plants. Glucuronoyl esterase (GCE) is a microbial enzyme hydrolyzing the ester bond between lignin and 4-O-methyl-α-D-glucuronic acid present as a side chain of glucuronoxylan. This bond mediates lignin-carbohydrate complex (LCC) formation, considered as crucial factor of lignocellulose recalcitrance to saccharification. Previous studies showed that hybrid aspen (Populus tremula L. x tremuloides Michx.) constitutively expressing Phanerochaete carnosa Burt GCE (PcGCE) had better efficiency of cellulose-to-glucose conversion but were stunned and had lower cellulose content indicating that more studies are needed to design strategy for deployment of this enzyme in planta. Here we report that the transgenic plants exhibit premature leaf senescence, increased accumulation of calcium oxalate crystals, tyloses and necrotic lesions and have strongly activated immune defense reactions as revealed by their altered profiles of transcriptomes, metabolomes and hormones in the leaves. To elucidate if these effects are triggered by damage-associated molecular patterns (DAMPs) or by PcGCE protein perceived as a pathogen-associated molecular pattern (PAMP), we ectopically expressed in aspen an enzymatically inactive PcGCES217A. The mutated PcGCE induced similar growth retardation, leaf necrosis and premature senescence as the active one, providing evidence that PcGCE protein is recognized as PAMP. Transcriptomics analysis of young expanding leaves of 35S:PcGCE plants identified several candidates for receptors of PcGCE, which were not expressed in developing wood tissues. Grafting experiments showed that PcGCE transcripts are not cell-to-cell mobile and that leaves augment systemic responses. In agreement, expressing PcGCE in developing wood by using the wood-specific promoter (WP), avoided all off-target effects. Moreover, WP:PcGCE lines grew better than control plants providing evidence that this strategy can be used in transgenic crops dedicated for biorefinery.

Colored Shade Nets can Relieve Abnormal Fruit Softening and Premature Leaf Senescence of “Jumeigui” Grapes during Ripening Under Greenhouse Conditions

When grapes reach maturity, they usually experience extremely high-temperature periods in southern China, causing premature leaf senescence, abnormal fruit softening, and fruiting period shortening. Their quality and production efficiency are also severely affected. ‘Jumeigui’ grapes were examined in terms of fruit quality and leaf senescence under shading treatments; green, blue, black, and gray aluminum foil nets were used for shading, and their spectra were measured. At the same density, shade net color significantly affected cooling and shading efficiencies, and gray net had the best light transmission and cooling effect. Shading treatment significantly alleviated abnormal grape softness during hot periods. Total soluble solids (TSS) content and grape coloration were affected under gray, blue, and green shade nets. TSS exceeded 18% under gray, blue, and green nets, meeting the requirements of first-class high-quality fruit. However, peel coloration was not notably affected under gray and blue shade nets, while the non-shading treatment produced clear heat-stress damage, especially on the edges of old leaves. The net photosynthetic rate of the bottom five old leaves under the non-shading treatment was significantly lower than that under the shading treatment, indicating that high light and heat caused premature leaf senescence. In summary, colored shade nets can reduce the temperature and light in the greenhouse, while alleviating premature senescence of perennial grape plants. However, the quality of the grapes treated using black shade nets was poor; superior quality was achieved using gray and blue shade nets. These results can be applied in future cultivation facilities during high-temperature periods.

Integration of Transcriptomic And Proteomic Analyses Reveals Several Levels of Metabolic Regulation In The Excess Starch And Early Senescent Leaf Mutant lses1 In Rice

Background: The normal metabolism of transitory starch in leaves plays an important role in ensuring photosynthesis, delaying senescence and maintaining high yield in crops. OsCKI1 (casein kinase I1) plays crucial regulatory roles in multiple important physiological processes, including root development, hormonal signaling and low temperature-treatment adaptive growth in rice; however, its potential role in regulating temporary starch metabolism or premature leaf senescence remains unclear. To reveal the molecular regulatory mechanism of OsCKI1 in rice leaves, physiological, transcriptomic and proteomic analyses of leaves of the mutant lses1 (leaf starch excess and senescence 1), allelic to osckI1, and its wild-type variety (WT) were performed. Results: Phenotypic identification and physiological measurements showed that the lses1 mutant exhibited starch excess in the leaves and an obvious leaf tip withering phenotype as well as high ROS and MDA contents, low chlorophyll content and protective enzyme activities compared to WT. Transcriptomic and proteomic analyses showed that the correlations of most genes at the transcription and translation levels were limited. However, the changes of several important genes related to carbohydrate metabolism and apoptosis at the mRNA and protein levels were consistent. The protein-protein interaction (PPI) network might play accessory roles in promoting premature senescence of lses1 leaves. Comprehensive transcriptomic and proteomic analysis indicated that multiple key genes/proteins related to starch and sugar metabolism, apoptosis and ABA signaling exhibited significant differential expression. Abnormal increase in temporary starch was highly correlated with the expression of starch biosynthesis-related genes, which might be the main factor that causes premature leaf senescence and changes in multiple metabolic levels in leaves of lses1. In addition, significant up regulation of four proteins associated with ABA accumulation and signaling were detected in the lses1 mutant, suggesting that ABA may involve in multiple metabolic regulation via LSES1/OsCKI1 and the formation of mutant phenotype in lses1 leaves.Conclusion: The current study established the high correlation between the changes in physiological characteristics and mRNA and protein expression profiles in lses1 leaves, and emphasized the positive effect of excessive starch on accelerating premature leaf senescence. The expression patterns of genes/proteins related to starch biosynthesis and ABA signaling were analyzed via transcriptomes and proteomes, which provided a novel direction and research basis for the subsequent exploration of the regulation mechanism of temporary starch and apoptosis via LSES1/OsCKI1 in rice.

Phospholipase Dα1 acts as a negative regulator of high Mg2+-induced leaf senescence in Arabidopsis

Magnesium is a macronutrient involved in essential cellular processes. Its deficiency or excess is a stress factor for plants, seriously affecting their growth and development and therefore, its accurate regulation is essential. Recently, we discovered that phospholipase Dα1 (PLDα1) activity is vital in the stress response to high-magnesium conditions in Arabidopsis roots. This study shows that PLDα1 acts as a negative regulator of high-Mg2+-induced leaf senescence in Arabidopsis. The level of phosphatidic acid produced by PLDα1 and the amount of PLDα1 in the leaves increase in plants treated with high Mg2+. A knockout mutant of PLDα1 (plda1-1), exhibits premature leaf senescence under high-Mg2+ conditions. In pldα1-1 plants, higher accumulation of abscisic and jasmonic acid and impaired magnesium, potassium and phosphate homeostasis were observed under high-Mg2+ conditions. High Mg2+ also led to an increase of starch and proline content in Arabidopsis plants. While the starch content was higher in plda1-1 plants, proline content was significantly lower in plda1-1 compared with WT. Our results show that PLDα1 is essential for Arabidopsis plants to cope with the pleiotropic effects of high-Mg2+ stress and delay the leaf senescence.

PREMATURE SENESCENCE LEAF 50 Promotes Heat Stress Tolerance in Rice (Oryza sativa L.)

Background Heat stress is a major environmental factor that could induce premature leaf senescence in plants. So far, a few rice premature senescent leaf mutants have been reported to involve in heat tolerance. Findings We identified a premature senescence leaf 50 (psl50) mutant that exhibited a higher heat susceptibility with decreased survival rate, over-accumulated hydrogen peroxide (H2O2) content and increased cell death under heat stress compared with the wild-type. The causal gene PREMATURE SENESCENCE LEAF 50 (PSL50) was isolated by using initial map-based resequencing (IMBR) approach, and we found that PSL50 promoted heat tolerance probably by acting as a modulator of H2O2 signaling in response to heat stress in rice (Oryza sativa L.). Conclusions PSL50 negatively regulates heat-induced premature leaf senescence in rice.

Low Light/Darkness as Stressors of Multifactor-Induced Senescence in Rice Plants

Leaf senescence, as an integral part of the final development stage for plants, primarily remobilizes nutrients from the sources to the sinks in response to different stressors. The premature senescence of leaves is a critical challenge that causes significant economic losses in terms of crop yields. Although low light causes losses of up to 50% and affects rice yield and quality, its regulatory mechanisms remain poorly elucidated. Darkness-mediated premature leaf senescence is a well-studied stressor. It initiates the expression of senescence-associated genes (SAGs), which have been implicated in chlorophyll breakdown and degradation. The molecular and biochemical regulatory mechanisms of premature leaf senescence show significant levels of redundant biomass in complex pathways. Thus, clarifying the regulatory mechanisms of low-light/dark-induced senescence may be conducive to developing strategies for rice crop improvement. This review describes the recent molecular regulatory mechanisms associated with low-light response and dark-induced senescence (DIS), and their effects on plastid signaling and photosynthesis-mediated processes, chloroplast and protein degradation, as well as hormonal and transcriptional regulation in rice.

OsNBL1, a Multi-Organelle Localized Protein, Plays Essential Roles in Rice Senescence, Disease Resistance, and Salt Tolerance

Background Plant senescence is a complicated process involving multiple regulations, such as temperature, light, reactive oxygen species (ROS), endogenous hormone levels, and diseases. Although many such genes have been characterized to understand the process of leaf senescence, there still remain many unknowns, and many more genes need to be characterized. Results We identified a rice mutant nbl1 with a premature leaf senescence phenotype. The causative gene, OsNBL1, encodes a small protein with 94 amino acids, which is conserved in monocot, as well as dicot plants. Disruption of OsNBL1 resulted in accelerated dark-induced leaf senescence, accompanied by a reduction in chlorophyll content and up-regulation of several senescence-associated genes. Notably, the nbl1 mutant was more susceptible to rice blast and bacterial blight but more tolerant to sodium chloride. Several salt-induced genes, including HAK1, HAK5, and three SNAC genes, were also up-regulated in the nbl1 mutant. Additionally, the nbl1 mutant was more sensitive to salicylic acid. Plants overexpressing OsNBL1 showed delayed dark-induced senescence, consistent with a higher chlorophyll content compared to wild-type plants. However, the overexpression plants were indistinguishable from the wild-types for resistance to the rice blast disease. OsNBL1 is a multi-organelle localized protein and interacts with OsClpP6, which is associated with senescence. Conclusions We described a novel leaf senescence mutant nbl1 in rice. It is showed that OsNBL1, a multi-organelle localized protein which interacts with a plastidic caseinolytic protease OsClpP6, is essential for controlling leaf senescence, disease resistance, and salt tolerance.

Strigolactone and Ethylene Inhibitor Suppressing Dark-induced Leaf Senescence in Perennial Ryegrass Involving Transcriptional Downregulation of Chlorophyll Degradation

Accelerated or premature leaf senescence induced by dark conditions could be associated with chlorophyll degradation and regulated by hormones. To study the effects of strigolactone (SL) on dark-induced leaf senescence and to examine the interaction effects of SL and ethylene on regulating dark-induced leaf senescence, plants of perennial ryegrass (Lolium perenne) exposed to darkness for 8 days were treated with a synthetic SL analogue (GR24), aminoethoxyvinyl glycine [AVG (an ethylene biosynthesis inhibitor)], or SL and AVG by foliar spray. Chlorophyll content, photochemical efficiency, electrolyte leakage, and ethylene production were measured. Expressions of genes associated with leaf senescence, SL biosynthesis and signaling, ethylene biosynthesis and signaling, and chlorophyll biosynthesis and degradation were determined. Foliar application of GR24 promoted leaf senescence in perennial ryegrass grown in darkness, and the intensity of action increased with the GR24 concentration. SL-accelerated leaf senescence was associated with the downregulation of four chlorophyll biosynthesis-associated genes and upregulation of four chlorophyll degradation-associated genes. AVG had functions counteractive to SL, suppressing dark-induced leaf senescence by downregulating chlorophyll degradation genes and SL synthesis genes. Our results suggested that SL and ethylene interactively regulated leaf senescence, mainly by controlling chlorophyll degradation induced by darkness in perennial ryegrass.

Premature Senescence Leaf 50 Promotes Heat Stress Tolerance in Rice (Oryza Sativa L.)

BackgroundHeat stress is a major environmental factor that could induce premature leaf senescence in plants. So far, few rice premature senescent leaf mutants have been reported to involve in heat tolerance.FindingsWe identified a premature senescence leaf 50 (psl50) mutant that exhibited a higher heat susceptibility with decreased survival rate, over-accumulated hydrogen peroxide (H2O2) content and increased cell death under heat stress compared with the wild-type. The causal gene PREMATURE SENESCENCE LEAF 50 (PSL50) was isolated by using initial map-based resequencing (IMBR) approach, and we found that PSL50 promoted heat tolerance probably by acting as a modulator of H2O2 signaling in response to heat stress in rice (Oryza sativa L.).ConclusionsPSL50 negatively regulates heat-induced premature leaf senescence in rice.