Induced Leaf Senescence Research Articles

Genome-wide association study of image-based trait reveals the genetic architecture of dark-induced leaf senescence in rice

Abstract Darkness is often used as an effective measure to induce leaf senescence. Although many senescence-related genes in rice have been reported, the genome-wide genetic architecture underlying leaf senescence remains poorly understood. In our study, indica and japonica rice showed contrasting responses to dark-induced leaf senescence (DILS). Genome-wide association studies (GWAS) combined with transcriptomic analyses revealed 57, 97, and 48 loci involved in the regulation of the onset, progression, and ending of DILS, respectively. Haplotype analyses showed that the senescence-related loci differentially accumulated in indica and japonica accessions and functioned additively to regulate DILS. A total of 357 candidate genes were identified that are involved in various senescence-related processes such as lipid and amino acid catabolism, photosynthesis, response to reactive oxygen species, and regulation of defence response. In addition, functional analyses of candidate genes revealed that OsMYB21 positively regulates the onset of DILS, while OsSUB1B negatively regulates its progression. Thus, our results provide new insights into the genetic regulation of DILS in rice.

Unveiling the role of epigenetics in leaf senescence: a comparative study to identify different epigenetic regulations of senescence types in barley leaves

BackgroundDevelopmental leaf senescence (DLS) is an irreversible process followed by cell death. Dark-induced leaf senescence (DILS) is a reversible process that allows adaptations to changing environmental conditions. As a result of exposure to adverse environmental changes, plants have developed mechanisms that enable them to survive. One of these is the redirection of metabolism into the senescence pathway. The plant seeks to optimise resource allocation. Our research aims to demonstrate how epigenetic machinery regulates leaf senescence, including its irreversibility.ResultsIn silico analyses allowed the complex identification and characterisation of 117 genes involved in epigenetic processes in barley. These genes include those responsible for DNA methylation, post-translational histone modifications, and ATP-dependent chromatin remodelling complexes. We then performed RNAseq analysis after DILS and DLS to evaluate their expression in senescence-dependent leaf metabolism. Principal component analysis revealed that evaluated gene expression in developmental senescence was similar to controls, while induced senescence displayed a distinct profile. Western blot experiments revealed that senescence engages senescence-specific histone modification. During DILS and DLS, the methylation of histone proteins H3K4me3 and H3K9me2 increased. H3K9ac acetylation levels significantly decreased during DILS and remained unchanged during DLS.ConclusionsThe study identified different epigenetic regulations of senescence types in barley leaves. These findings are valuable for exploring epigenetic regulation of senescence-related molecular mechanisms, particularly in response to premature, induced leaf senescence. Based on the results, we suggest the presence of an epigenetically regulated molecular switch between cell survival and cell death in DILS, highlighting an epigenetically driven cell survival metabolic response.

ArNAC148 induces Acer rubrum leaf senescence by activating the transcription of the ABA receptor gene ArPYR13

Acer rubrum, an ornamental tree known for its stunning autumn colors, has an elusive molecular mechanism that governs its leaf senescence. We performed the genome-wide analysis of NAC transcription factor genes and PYRABACTIN RESISTANCE1-LIKE (PYLs) and found that ArNAC148 and ArPYL13 were significantly upregulated in senescing leaves as compared to mature leaves. Subcellular localization studies confirmed the nuclear localization of ArNAC148 and the cytoplasmic localization of ArPYL13. Electrophoretic mobility shift assay and yeast one-hybrid assay demonstrated that ArNAC148 directly binds to the promoter of ArPYL13. Luciferase reporter assays further showed that ArNAC148 activates the transcription of ArPYL13. The transient expression of ArNAC148 and ArPYL13 in tobacco leaves promoted chlorophyll degradation, increased H2O2 level, MDA contents, and electrolyte leakage in response to abscisic acid (ABA). Moreover, the virus-induced gene silencing of ArNAC148 and ArPYL13 in A. rubrum produced results that were opposite to those observed in transient expression experiments. Our findings suggest that ArNAC148 induces leaf senescence by directly activating the transcription of ArPYL13, providing insights into the ABA-mediated regulatory mechanisms governing leaf senescence in A. rubrum. This study offers new perspectives for researchers to explore the roles of NAC and PYL genes in regulating leaf senescence in woody ornamental plants.

Temperature variations impacting leaf senescence initiation pathways alter leaf fall timing patterns in northern deciduous forests

Simulating the timing of leaf fall in large scale is crucial for accurate estimation of ecosystem carbon sequestration. However, the limited understanding of leaf senescence mechanisms often impedes the accuracy of simulation and prediction. In this study, we employed the advanced process-based models to fit remote sensing-derived end dates of the growing season (EOS) across deciduous broadleaf forests in the Northern Hemisphere, and revealed the spatial pattern associated with two leaf senescence pathways (i.e., either photoperiod- or temperature- initiated leaf senescence) and their potential effects on EOS prediction. The results show that the pixel-specific optimum models effectively fitted all EOS time series. Leaf senescence in 67.6 % and 32.4 % of pixels was initiated by shortening daylength and declining temperature, respectively. Shortening daylength triggered leaf senescence occurs mainly in areas with shorter summer daylength and/or warmer autumns, whereas declining temperature induced leaf senescence appears primarily in areas with longer summer daylength and/or colder autumns. The strong dependence of leaf senescence initiation cues on local temperature conditions implies that the ongoing increase in autumn temperature has the potential to alter the leaf senescence initiation, shifting from temperature cues to photoperiod signals. This shift would occur in 26.2–49.6 % of the areas where leaf senescence is initiated by declining temperature under RCP 4.5 and 8.5 scenarios, while forest areas where leaf senescence is induced by shortening daylength may expand northward. The overall delaying of the currently predicted EOS would therefore slow down by 4.5–10.3 % under the two warming scenarios. This implies that the adaptive nature of plants will reduce the overestimation of changes in carbon exchange capacity between ecosystems and atmosphere. Our study offers novel insights into understanding the mechanism of leaf senescence and improving the estimation of autumn phenology and ecosystem carbon balance in the deciduous broadleaf forests.

Assessing the growth dynamics of alfalfa varieties (Medicago sativa cv. Bilensoy 80 and Nimet) response to varied carbon dioxide (CO2) concentrations

Rising atmospheric CO2 levels drive greenhouse effects, elevating temperatures, and diminishing water accessibility in semi-arid regions, affecting agriculture. Alfalfa contributes to climate change mitigation by sequestering carbon, enhancing soil fertility and carbon storage, reducing synthetic nitrogen fertilizer use, preventing soil erosion, supplying high-quality livestock feed, and serving as a bioenergy source. This research examined the effects of elevated CO2 levels in climate change scenarios (600, 800, and 1000 ppm, with control at 400 ppm) on two alfalfa varieties, Medicago sativa cv. Nimet and Bilensoy-80. The experiments were conducted in specialized Climate Change Simulation Greenhouses, allowing control of CO2, water, and temperature variables. Results revealed a positive relationship between higher CO2 concentrations and increased photosynthesis (P ≤ 0.001), promoting the plant growth leaf area (P ≤ 0.001), yields and both leaf (P ≤ 0.05) and stem dry biomass (P ≤ 0.001). At 1000 ppm CO2, a saturation point was reached, halting further photosynthesis. This down-regulation was linked to decreased intercellular CO2 levels, which expedited chlorophyll and breakdown and potentially induced leaf senescence. High CO2 levels led to greater biomass, as anticipated. However, total protein levels, a forage quality indicator, initially decreased with high CO2 concentrations (up to 1000 ppm) due to an inverse relationship with shoot yield. Surprisingly, the 1000 ppm CO2 concentration mitigated this protein reduction in both alfalfa varieties.

Ozone stress response of leaf BVOC emission and photosynthesis in mountain birch (Betula pubescens spp. czerepanovii) depends on leaf age.

Oxidative stress from ozone (O3) causes plants to alter their emission of biogenic volatile organic compounds (BVOC) and their photosynthetic rate. Stress reactions from O3 on birch trees can result in prohibited plant growth and lead to increased BVOC emission rates as well as changes in their compound blend to emit more monoterpenes (MT) and sesquiterpenes (SQT). BVOCs take part in atmospheric reactions such as enhancing the production of secondary organic aerosols (SOA). As the compound blend and emission rate change with O3 stress, this can influence the atmospheric conditions by affecting the production of SOA. Studying the stress responses of plants provides important information on how these reactions might change, which is vital to making better predictions of the future climate. In this study, measurements were taken to find out how the leaves of mature mountain birch trees (Betula pubescens ssp. czerepanovii) respond to different levels of elevated O3 exposure insitu depending on leaf age. We found that leaves from both early and late summers responded with induced SQT emission after exposure to 120 ppb O3. Early leaves were, however, more sensitive to increased O3 concentrations, with enhanced emission of green leaf volatiles (GLV) and tendencies of both induced leaf senescence as well as poor recovery in the photosynthetic rate between exposures. Late leaves had more stable photosynthetic rates throughout the experiment and responded less to exposure at different O3 levels.

The mechanism of low blue light-induced leaf senescence mediated by GmCRY1s in soybean

Leaf senescence is a crucial trait that has a significant impact on crop quality and yield. Previous studies have demonstrated that light is a key factor in modulating the senescence process. However, the precise mechanism by which plants sense light and control senescence remains largely unknown, particularly in crop species. In this study, we reveal that the reduction in blue light under shading conditions can efficiently induce leaf senescence in soybean. The blue light receptors GmCRY1s rather than GmCRY2s, primarily regulate leaf senescence in response to blue light signals. Our results show that GmCRY1s interact with DELLA proteins under light-activated conditions, stabilizing them and consequently suppressing the transcription of GmWRKY100 to delay senescence. Conversely, LBL reduces the interaction between GmCRY1s and the DELLA proteins, leading to their degradation and premature senescence of leaves. Our findings suggest a GmCRY1s-GmDELLAs-GmWRKY100 regulatory cascade that is involved in mediating LBL-induced leaf senescence in soybean, providing insight into the mechanism of how light signals regulate leaf senescence. Additionally, we generate GmWRKY100 knockout soybeans that show delayed leaf senescence and improved yield under natural field conditions, indicating potential applications in enhancing soybean production by manipulating the leaf senescence trait.

The NAC transcription factor MdNAC4 positively regulates nitrogen deficiency-induced leaf senescence by enhancing ABA biosynthesis in apple

Although it is well established that nitrogen (N) deficiency induces leaf senescence, the molecular mechanism of N deficiency-induced leaf senescence remains largely unknown. Here, we show that an abscisic acid (ABA)-responsive NAC transcription factor (TF) is involved in N deficiency-induced leaf senescence. The overexpression of MdNAC4 led to increased ABA levels in apple calli by directly activating the transcription of the ABA biosynthesis gene MdNCED2. In addition, MdNAC4 overexpression promoted N deficiency-induced leaf senescence. Further investigation showed that MdNAC4 directly bound the promoter of the senescence-associated gene (SAG) MdSAG39 and upregulated its expression. Interestingly, the function of MdNAC4 in promoting N deficiency-induced leaf senescence was enhanced in the presence of ABA. Furthermore, we identified an interaction between the ABA receptor protein MdPYL4 and the MdNAC4 protein. Moreover, MdPYL4 showed a function similar to that of MdNAC4 in ABA-mediated N deficiency-induced leaf senescence. These findings suggest that ABA plays a central role in N deficiency-induced leaf senescence and that MdPYL4 interacts with MdNAC4 to enhance the response of the latter to N deficiency, thus promoting N deficiency-induced leaf senescence. In conclusion, our results provide new insight into how MdNAC4 regulates N deficiency-induced leaf senescence.Graphical

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 invivo 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.

Comparative Gene Enrichment Analysis for Drought Tolerance in Contrasting Maize Genotype.

Drought stress is a significant abiotic factor influencing maize growth and development. Understanding the molecular mechanism of drought tolerance is critical to develop the drought tolerant genotype. The identification of the stress responsive gene is the first step to developing a drought tolerant genotype. The aim of the current research was to pinpoint the genes that are essential for conserved samples in maize drought tolerance. In the current study, inbred lines of maize, 478 and H21, a drought-tolerant and susceptible line, were cultivated in the field and various treatments were applied. The circumstances during the vegetative stage (severe drought, moderate drought and well-watered environments) and RNA sequencing were used to look into their origins. In 478, 68%, 48% and 32% of drought-responsive genes (DRGs) were found, with 63% of DRGs in moderate drought and severe drought conditions in H21, respectively. Gene ontology (GO) keywords were explicitly enriched in the DRGs of H21, which were considerably over-represented in the two lines. According to the results of the GSEA, "phenylpropanoid biosynthesis" was exclusively enriched in H21, but "starch and sucrose metabolism" and "plant hormone signal transduction" were enhanced in both of the two lines. Further investigation found that the various expression patterns of genes linked to the trehalose biosynthesis pathway, reactive oxygen scavenging, and transcription factors, may have a role in maize's ability to withstand drought. Our findings illuminate the molecular ways that respond to lack and offer gene resources for maize drought resistance. Similarly, SNP and correlation analysis gave us noticeable results that urged us to do the same kind of analysis on other crops. Additionally, we isolated particular transcription factors that could control the expression of genes associated to photosynthesis and leaf senescence. According to our findings, a key factor in tolerance is the equilibrium between the induction of leaf senescence and the preservation of photosynthesis under drought.

Ectopic expression of rice malate synthase in Arabidopsis revealed its roles in salt stress responses

Expression of rice malate synthase (OsMS), one of the two key genes in the glyoxylate cycle, is highly upregulated under salt stress. In this study, we aimed to investigate the role of OsMS in salt stress responses using the Arabidopsis T-DNA insertional mutant line of malate synthase (AtMS), an OsMS orthologous gene, for ectopic expression. Germination of the Atms mutant under salt stress was lower than that of Arabidopsis Col-0 wildtype (WT); furthermore, the two Atms mutant lines ectopically expressing OsMS reversed the salt-sensitive phenotype. Atms mutants salt-treated for 3 days exhibited higher electrolyte leakage, higher Na+/K+ ratio, lower expression of stress-responsive genes, and lower soluble sugar content than WT and the two OsMS-expressing Atms mutant lines. Consistently, Atms mutants salt-treated for 3 days followed by a 5-day recovery period displayed greater fresh-weight reduction. Notably, leaf greenness and chlorophyll and total carotenoid contents were higher in the Atms mutant lines than in the WT under stress. OsMS-expressing Atms mutants exhibited a change in pigment content closer to that of WT. During dark-induced senescence, an Atms mutant, Aticl, mutant (the other key gene in the glyoxylate cycle), and three double mutant lines of Atms and Aticl exhibited lower decreases in leaf greenness than the WT and OsMS-expressing Atms mutant lines. Furthermore, SAG12 expression levels in the Atms mutant, Aticl mutant, and three double mutant lines were lower than those in OsMS-expressing Atms mutant lines. Altogether, our data indicate that OsMS likely plays a key role in salt stress responses, possibly through the induction of leaf senescence.

Oilseed rape MPK1 mediates reactive oxygen species-dependent cell death and jasmonic acid-induced leaf senescence

Leaf senescence is the terminal phase of leaf development, which involves remobilization of nutrients from leaves to other organs and is affected by many different endogenous (e.g. hormonal) and external stimuli. A proper control of leaf senescence is essential for fitness of annual plants. Although great progress has been made in identifying key genes or proteins controlling leaf senescence in Arabidopsis and several other plants, the relevant genes controlling leaf senescence in oilseed rape ( Brassica napus ) remain largely elusive. In this study, we found that a Mitogen-activated Protein Kinase, BnaMPK1 from oilseed rape positively modulated reactive oxygen species (ROS) -dependent cell death and jasmonic acid (JA)-induced leaf senescence. The expression of BnaMPK1 dramatically increased in senescing leaves of oilseed rape. Transient expression of BnaMPK1 in Nicotiana benthamiana and rapeseed protoplasts resulted in accumulation of ROS and cell death. Moreover, ectopic expression of BnaMPK1 in Arabidopsis exhibited accelerated JA-induced leaf senescence while atmpk1 mutants showed delayed leaf senescence. Furthermore, the expression of chlorophyll degradation and senescence marker genes Non-Yellowing 1 ( NYE1 , SGR1 ), Non-Yellow Coloring 1 ( NYC1 ), Senescence-Associated Gene 29 ( SAG29 ) and two well-known gene mediating JA-induced senescence ORESARA ( ORE ) 1 and ORE9 were induced in BnaMPK1 overexpression lines, whereas most of them were repressed in the atmpk1 mutant. A bimolecular fluorescence complementation (BiFC) and co-immunoprecipitation (Co-IP) assay revealed that BnaMKK3 interacted with BnaMPK1 and, constitutively active BnaMKK3 phosphorylated BnaMPK1 whereas inactive BnaMKK3 can not. Taken together, these results indicate that BnaMPK1 acts as a positive regulator of ROS- and JA-dependent leaf senescence. • BnaMPK1 expression level is up-regulated in senescing leaves. • BnaMPK1 expression induces ROS-dependent cell death in leaves and protoplasts of oilseed rape. • BnaMPK1 modulates jasmonic acid (JA)-induced leaf senescence. • BnaMPK1 expression induces the transcription of a few senescence-associated marker genes. • BnaMPK1 interacts with upstream BnaMKK3 and is phosphorylated by BnaMKK3.

Unraveling the genetics of polyamine metabolism in barley for senescence-related crop improvement

We explored the polyamine (PA) metabolic pathway genes in barley (Hv) to understand plant development and stress adaptation in Gramineae crops with emphasis on leaf senescence. Bioinformatics and functional genomics tools were utilized for genome-wide identification, comprehensive gene features, evolution, development and stress effects on the expression of the polyamine metabolic pathway gene families (PMGs). Three S-adenosylmethionine decarboxylases (HvSAMDCs), two ornithine decarboxylases (HvODCs), one arginine decarboxylase (HvADC), one spermidine synthase (HvSPDS), two spermine synthases (HvSPMSs), five copper amine oxidases (HvCuAOs) and seven polyamine oxidases (HvPAOs) members of PMGs were identified and characterized in barley. All the HvPMG genes were found to be distributed on all chromosomes of barley. The phylogenetic and comparative assessment revealed that PA metabolic pathway is highly conserved in plants and the prediction of nine H. vulgare miRNAs (hvu-miR) target sites, 18 protein-protein interactions and 961 putative CREs in the promoter region were discerned. Gene expression of HvSAMDC3, HvCuAO7, HvPAO4 and HvSPMS1 was apparent at every developmental stage. SPDS/SPMS gene family was found to be the most responsive to induced leaf senescence. This study provides a reference for the functional investigation of the molecular mechanism(s) that regulate polyamine metabolism in plants as a tool for future breeding decision management systems.

The apple GARP family gene MdHHO3 regulates the nitrate response and leaf senescence.

The regulation of plant gene expression by nitrate is a complex regulatory process. Here, we identified 90 GARP family genes in apples by genome-wide analysis. As a member of the GARP gene family, the expression of MdHHO3 (Malus domestica HYPERSENSITIVITY TO LOW PHOSPHATE-ELICITED PRIMARY ROOT SHORTENING1 HOMOLOG 3) is upregulated under N (nitrogen) supply. The results of DNA-binding site analysis and electrophoretic mobility shift assays (EMSA) showed that MdHHO3 binds to the motif-containing GAATC. Furthermore, MdHHO3 binds to its promoter sequence and inhibits its activity. In addition, the overexpression of MdHHO3 in apple calli resulted in less accumulation of nitrate in 35S:MdHHO3-GFP calli and downregulated the expression of the nitrate transport-related genes but upregulated the expression of the nitrate assimilation-related genes. Similarly, the expression of the nitrate transport-related genes was downregulated and the expression of the nitrate assimilation-related genes was upregulated in MdHHO3 overexpression Arabidopsis and tobacco plants. Interaction experiments showed that MdHHO3 could bind to the promoter MdNRT2.1 (NITRATE TRANSPORTER 2.1) and negatively regulate its expression. Moreover, the exposure of MdHHO3-overexpressing Arabidopsis and tobacco to nitrate deficiency resulted in an early senescence phenotype as compared to the WT plants. These results show that MdHHO3 can not only negatively regulate nitrate accumulation in response to nitrate but also promote early leaf senescence under nitrate deficiency. This information may be useful to further reveal the mechanism of the nitrate response and demonstrates that nitrate deficiency induces leaf senescence in apples.

MdNAC4 Interacts With MdAPRR2 to Regulate Nitrogen Deficiency-Induced Leaf Senescence in Apple (Malus domestica).

Nitrogen (N) is one of the important macronutrients in plants, and N deficiency induces leaf senescence. However, the molecular mechanism underlying how N deficiency affects leaf senescence is unclear. Here, we report an apple NAC TF, MdNAC4, that participates in N deficiency-induced leaf senescence. The senescence phenotype of apple leaves overexpressing MdNAC4 was enhanced after N deficiency. Consistently, the chlorophyll content of transgenic leaves was significantly lower than that in the WT control leaves, the expression of chlorophyll catabolism-related genes (MdNYC1, MdPAO, and MdSGR1) was significantly higher than that in the WT controls, and the expression of chlorophyll synthesis-related genes (MdHEMA, MdCHLI, and MdCHLM) was significantly lower than that in the WT control leaves. Furthermore, MdNAC4 was found to directly activate the transcription of the chlorophyll catabolism-related genes MdNYC1 and MdPAO. Additionally, MdNAC4 was proven to interact with MdAPRR2 proteins both in vitro and in vivo, and overexpression of MdAPRR2 seemed to delay N deficiency-induced leaf senescence. Correspondingly, the chlorophyll loss of MdAPRR2-overexpressing (MdAPRR2-OE) lines was significantly lower than in WT control plants. Although downregulated, the expression of the chlorophyll synthesis-related genes MdHEMA, MdCHLI, and MdCHLM in the transgenic plants was more than twice that in the WT control plants. Taken together, our results enrich the regulatory network of leaf senescence induced by N deficiency through the interaction between MdNAC4 and MdAPRR2.

Low temperature and short daylength interact to affect the leaf senescence of two temperate tree species.

Temperature and photoperiod are two major environmental cues shaping the leaf senescence of temperate tree species. However, how the control of leaf senescence is split between photoperiod and temperature is unknown for many ecologically important species. Here, we conducted a growth chamber experiment to test the effects of temperature (6, 9, 18 and 21°C) and photoperiod (8 and 16h daylength) on leaf senescence of two temperate tree species (Quercus mongolica Fisch. and Larix principis-rupprechtii Mayr.) distributed in montane forest of China. The results showed that low temperature (LT) alone could induce leaf senescence of both species under long daylength (LD) conditions, but the leaf senescence of L. principis-rupprechtii was more sensitive to the decrease in temperature than that of Q. mongolica under the LD condition. Short daylength (SD) alone could only induce the leaf senescence of L. principis-rupprechtii, suggesting that the photoperiod sensitivity varies between species. SD could accelerate the LT-induced senescence, but the effect of SD reduced with the decrease in temperature. Based on these findings, we developed a new autumn phenology model by incorporating interspecific differences in the photoperiod sensitivity of leaf senescence. Compared with the three existing process-based autumn phenology models, the new model was more robust in simulating the experimental data. When employing these models to available long-term phenological data, our new model also performed best in reproducing the observed leaf senescence date of two closely related species (Quercus robur L. and Larix decidua Mill.). These results enhance our understanding of how LT and SD control leaf senescence. The prediction of the climate change impacts on forest carbon uptake could be improved by incorporating this new autumn phenological model into the terrestrial biosphere models.

Transcriptome analysis uncovering regulatory networks and hub genes of Populus photosynthesis and chlorophyll content

Photosynthesis plays vital role in plant growth and development throughout its life, and it is influenced by environmental signals and circadian rhythms. We analyzed the transcriptome landscape of the two poplars progeny with contrasting photosynthesis rates at three times point (ZT4, ZT16, ZT22), constructed gene regulatory network that related to circadian rhythm and photosynthesis. We suggest that the differences in photosynthetic rate between the progenies may originate from plant endogenous circadian oscillators prepare poplar plants for photosynthesis by regulating photosynthesis-associated nuclear genes and carotenoid metabolism genes before dawn, genes associated with plant hormone signal transduction and transcription factor increase leaf size and stomatal movement, the influence of other core regulatory factors on chlorophyll accumulation. Furthermore, overexpression of candidate regulatory gene, AP3 (Potri.007G017000), induced leaf senescence and reduced the content of chlorophyll. These results demonstrated that many potential key regulators are integrated closely with chlorophyll content and photosynthesis.

ZmbHLH121, a maize bHLH transcription factor inducing leaf senescence by accelerating ROS accumulation in maize (Zea mays L.)

Leaf senescence is the final stage of leaf development which is induced by many genetic and environmental factors. To reveal the role of transcription factors in leaf senescence, we studied the function of a bHLH transcription factor ZmbHLH121 in maize. The overexpression of ZmbHLH121 exhibited phenotypes with leaf senescence. Higher expression pattern of ZmbHLH121 was observed in the immature leaves through expression analysis at early vegetative developmental stages. Results of 3,3-diaminobenzidine (DAB), nitroblue tetrazolium chloride (NBT) histochemical analysis, quantitative measurement of reactive oxygen species (ROS), chlorophyll quantification and transcriptome analysis revealed that, ZmbHLH121 positively regulated leaf senescence by accelerating ROS production and chlorophyll breakdown in maize leaves. It is suggested that, ZmbHLH121 regulated leaf senescence might be due to the excess amount of ROS production and chlorophyll loss in leaves. This study provided useful clue about the mechanism of ZmbHLH121 mediated leaf senescence.

Transcriptome alterations of radish shoots exposed to cadmium can be interpreted in the context of leaf senescence.

Till now few transcriptome studies have described shoot responses of heavy metal (HM)-sensitive plants to excess Cd and still a unifying model of Cd action is lacking. Using RNA-seq technique, the transcriptome responses of radish (Raphanus sativus L.) leaves to Cd stress were investigated in plants raised hydroponically under control and 5.0mg L-1 Cd. The element was mainly accumulated in roots and led to declined biomass and photosynthetic pigments, increased H2O2 and lipid peroxidation, and the accumulation of sugars, protein thiols, and phytochelatins. Out of 524 differentially expressed genes (DEGs), 244 and 280 upregulated and downregulated ones were assigned to 82 and 115 GO terms, respectively. The upregulated DEGs were involved in osmotic regulation, protein metabolism, chelators, and carbohydrate metabolisms, whereas downregulated DEGs were related to photosynthesis, response to oxidative stress, glucosinolate, and secondary metabolite biosynthesis. Our transcriptome data suggest that Cd triggers ROS production and photosynthesis decline associated with increased proteolysis through ubiquitin-proteasome system (UPS)- and chloroplast-proteases and in this way brings about re-mobilization of N and C stores into amino acids and sugars. Meanwhile, declined glucosinolate metabolism in favor of chelator synthesis and upregulation of dehydrins as inferred from transcriptome analysis confers shoots some tolerance to the HM-derived ionic/osmotic imbalances. Thus, the induction of leaf senescence might be a major long-term response of HM-sensitive plants to Cd toxicity.

Regulation of Arabidopsis Matrix Metalloproteinases by Mitogen-Activated Protein Kinases and Their Function in Leaf Senescence

Leaf senescence is a developmentally programmed cell death process that is influenced by a variety of endogenous signals and environmental factors. Here, we report that MPK3 and MPK6, two Arabidopsis mitogen-activated protein kinases (MAPKs or MPKs), and their two upstream MAPK kinases (MAPKKs or MKKs), MKK4 and MKK5, are key regulators of leaf senescence. Weak induction of constitutively active MAPKKs driven by steroid-inducible promoter, which activates endogenous MPK3 and MPK6, induces leaf senescence. This gain-of-function phenotype requires functional endogenous MPK3 and MPK6. Furthermore, loss of function of both MKK4 and MKK5 delays leaf senescence. Expression profiling leads to the identification of matrix metalloproteinases (MMPs), a family of zinc- and calcium-dependent endopeptidases, as the downstream target genes of MPK3/MPK6 cascade. MPK3/MPK6 activation-triggered leaf senescence is associated with rapid and strong induction of At3-MMP and At2-MMP. Expression of Arabidopsis MMP genes is strongly induced during leaf senescence, qualifying them as senescence-associated genes (SAGs). In addition, either constitutive or inducible overexpression of At3-MMP is sufficient to trigger leaf senescence. Based on these findings, we conclude that MPK3/MPK6 MAPK cascade and MMP target genes further downstream are involved in regulating leaf senescence in Arabidopsis.