Accelerated Leaf Senescence Research Articles

Overexpression of BnaAGL11, a MADS-Box Transcription Factor, Regulates Leaf Morphogenesis and Senescence in Brassica napus.

Previous studies have reported that SEEDSTICK/AGAMOUS-LIKE 11 (AtSTK/AtAGL11), a MADS-box transcription factor, plays important roles in many biological processes in Arabidopsis thaliana. However, the function of BnaAGL11, an AtAGL11 homologous gene from Brassica napus, in leaf development remains unknown. Here, we found that the ectopic expression of any copy of Bna.C09.AGL11, Bna.A03.AGL11, and Bna.A09.AGL11 in A. thaliana led to smaller and curly leaves and promoted leaf senescence. Consistently, the overexpression of Bna.C09.AGL11 in B. napus also caused smaller and curly leaves and accelerated leaf senescence. Furthermore, we demonstrated that Bna.C09.AGL11 controlled leaf morphogenesis by indirectly downregulating the genes of Bna.A01.DWF4 and Bna.C07.PGX3 and promoted leaf senescence by indirectly upregulating the genes of Bna.A04.ABI5, Bna.A05.ABI5, Bna.C04.ABI5-1, and Bna.C01.SEN4 and directly activating the transcription of Bna.C04.ABI5-2 and Bna.C03.SEN4 genes. Our results provide new insights into the underlying regulatory mechanism of BnaAGL11 during leaf development in B. napus.

A long transcript mutant of the rubisco activase gene RCA upregulated by the transcription factor Ghd2 enhances drought tolerance in rice.

The transcription factor Ghd2 increases rice yield potential under normal conditions and accelerates leaf senescence under drought stress. However, its mechanism on the regulation of leaf senescence under drought stress remains unclear. In the present study, to unveil the mechanism, one target of Ghd2, the Rubisco activase gene RCA, was identified through the combined analysis of Ghd2-CRISPR transcriptome data and Ghd2-overexpression microarray data. Ghd2 binds to the 'CACA' motif in the RCA promoter by its CCT domain and upregulates RCA expression. RCA has alternative transcripts, RCAS and RCAL, which are predominantly expressed under normal conditions and drought stress, respectively. Similar to Ghd2-overexpressing plants, RCAL-overexpressing plants were more sensitive to drought stress than the wild-type. However, the plants overexpressing RCAS showed a weak drought-sensitive phenotype. Moreover, RCAL knockdown and knockout plants did not show yield loss under normal conditions, but exhibited enhanced drought tolerance and delayed leaf senescence. The chlorophyll content, the free amino acid content and the expression of senescence-related genes in the RCAL mutant were lower than those in the wild-type plants under drought stress. In summary, Ghd2 induces leaf senescence by upregulating RCAL expression under drought stress, and the RCAL mutant has important values in breeding drought-tolerant varieties.

The NAC factor LpNAL delays leaf senescence by repressing two chlorophyll catabolic genes in perennial ryegrass.

Expression of chlorophyll (Chl) catabolic genes during leaf senescence is tightly controlled at the transcriptional level. Here, we identified a NAC family transcription factor, LpNAL, involved in regulating Chl catabolic genes via the yeast one-hybrid system based on truncated promoter analysis of STAYGREEN (LpSGR) in perennial ryegrass (Lolium perenne L.). LpNAL was found to be a transcriptional repressor, directly repressing LpSGR as well as the Chl b reductase gene, NONYELLOWING COLORING1. Perennial ryegrass plants over-expressing LpNAL exhibited delayed leaf senescence or stay-green phenotypes, whereas knocking down LpNAL using RNA interference accelerated leaf senescence. Comparative transcriptome analysis of leaves at 30 d after emergence in wild-type, LpNAL-overexpression, and knock-down transgenic plants revealed that LpNAL-regulated stay-green phenotypes possess altered light reactions of photosynthesis, antioxidant metabolism, ABA and ethylene synthesis and signaling, and Chl catabolism. Collectively, the transcriptional repressor LpNAL targets both Chl a and Chl b catabolic genes and acts as a brake to fine-tune the rate of Chl degradation during leaf senescence in perennial ryegrass.

The NAC Transcription Factor ANAC087 Induces Aerial Rosette Development and Leaf Senescence in Arabidopsis.

CmNACP1 mRNA has been shown to move long distance through the phloem in Cucurbita maxima (pumpkin) and through a graft junction. Whereas the phloem transport of several different mRNAs has been documented in other systems as well, its function remains, for most of these RNAs, largely unknown. To gain insight into the possible role of these RNAs, we searched for the closest homologs of CmNACP1 in Arabidopsis, a model plant much more amenable for analysis. A phylogenetic approach using the predicted NAC domain indicated that ANAC059, ANAC092, ANAC079, ANAC100, ANAC046, and ANAC087 form a single clade with CmNACP1. In the present work, we analyzed the possible function of the ANAC087 gene in more detail. The promoter region of this gene directed expression in the vasculature, and also in trichomes, stem, apexes, and developing flowers which supports the notion that ANAC087 and CmNACP1 are orthologs. Overexpression of the ANAC087 gene induced increased branching in inflorescence stem, and also development of ectopic or aerial rosettes in T1 and T2 plants. Furthermore, overexpression of ANAC087 leads to accelerated leaf senescence in 44 days post-germination (dpg). Interestingly, a similar phenotype was observed in plants expressing the ANAC087 gene upstream region, also showing an increase in ANAC087 transcript levels. Finally, the results shown in this work indicate a role for ANAC087 in leaf senescence and also in rosette development.

OsWRKY53 Promotes Abscisic Acid Accumulation to Accelerate Leaf Senescence and Inhibit Seed Germination by Downregulating Abscisic Acid Catabolic Genes in Rice.

Abscisic acid (ABA) largely promotes leaf senescence and inhibits seed germination in plants. Endogenous ABA content is finely tuned by many transcription factors. In this study, we showed that OsWRKY53 is a positive regulator of leaf senescence and a negative regulator of seed germination in rice. OsWRKY53 expression was induced in leaves under aging, dark, and ABA treatment. The OsWRKY53-overexpressing (OsWRKY53-oe) plants showed early yellowing leaves, while the OsWRKY53 (oswrky53) knockout mutants maintained green leaves than the wild type under natural, dark-induced, and ABA-induced senescence conditions. Transcriptional analysis revealed that ABA catabolic genes, namely, OsABA8ox1 and OsABA8ox2, two key genes participating in ABA catabolism harboring ABA 8′-hydroxylase activity, were markedly downregulated in OsWRKY53-oe leaves. Chromatin immunoprecipitation and protoplast transient assays revealed that OsWRKY53 directly bound to the promoters of OsABA8ox1 and OsABA8ox2 to repress their transcription, resulting in elevated endogenous ABA contents that promoted premature leaf senescence in the OsWRKY53-oe plants. It indicates that OsWRKY53 is a positive regulator through regulating ABA accumulation to promote leaf senescence. In addition, accumulated ABA simultaneously inhibited seed germination and post-germination growth in OsWRKY53-oe plants. Taken together, OsWRKY53 suppresses the transcript of ABA catabolic genes to promote ABA accumulation to modulate ABA-induced leaf senescence and ABA-mediated inhibition of seed germination.

MicroRNA840 (MIR840) accelerates leaf senescence by targeting the overlapping 3'UTRs of PPR and WHIRLY3 in Arabidopsis thaliana.

MicroRNAs negatively regulate gene expression by promoting target mRNA cleavage and/or impairing its translation, thereby playing a crucial role in plant development and environmental stress responses. In Arabidopsis, the MIR840 gene is located within the overlapping 3'UTR of the PPR and WHIRLY3 (WHY3) genes, both being predicted targets of miR840* and miR840, the short maturation products of MIR840. Gain- and loss-of-function of MIR840 in Arabidopsis resulted in opposite senescence phenotypes. The highest expression levels of the MIR840 precursor transcript pre-miR840 were observed at senescence initiation, and pre-miR840 expression is significantly correlated with a reduction in PPR, but not WHY3, transcript levels. Although a reduction of transcript level of PPR, but not WHY3 transcript levels were not significantly affected by MIR840 overexpression, its protein levels were strongly reduced. Mutating the cleavage sites or replacing the target sequences abolishes the miR840*/miR840-mediated degradation of PPR transcripts and accumulation of WHY3 protein. In support for this, concurrent knockdown of both PPR and WHY3 in wild-type plants resulted in a senescence phenotype resembling that of the MIR840-overexpressing plant. This indicates that both PRR and WHY3 are targets in the MIR840-mediated senescence pathway. Moreover, single knockout mutants of PPR and WHY3 show a convergent upregulated subset of senescence-associated genes, which are also found among those induced by MIR840 overexpression. Our data provide evidence for a regulatory role of MIR840 in plant senescence.

The impact of drought and heat stress at flowering on maize kernel filling: Insights from the field and laboratory

Drought and exposure to high temperature at flowering strongly affect maize production. However, previous studies mostly focused on the effects of the stressors on kernel number. The impacts of drought and heat in isolation and combination at flowering on maize kernel filling and final kernel weight have received limited attention. Here we investigated the effects of three stressors (drought, heat, and drought plus heat) at flowering phase (from tasseling to seed setting completed) on maize kernel filling in field site plots covered with a rainout shelter and under laboratory conditions with kernel in vitro culture. In two experiments, the kernel weight of plants exposed to flowering stressors was significantly decreased relative to controls. In the field experiment, exposure to stressors decreased kernel weight by 5.0–8.3% for the basal kernel and 5.6–11.0% for the apical kernel. Accelerated leaf senescence and decreased rate of photosynthesis were observed in the field. Kernel weight was also reduced by 18.0–37.6% in the laboratory with kernel in vitro culture, suggesting that early stressors at flowering led to a sink (kernel) limit. Carbohydrate metabolism and starch biosynthesis were impaired by drought and heat individually and their combination. During the early kernel filling stage, the activities of sucrose invertase, sucrose synthase, ADP-Glc pyrophosphorylase, and starch synthase were decreased at 10 and 20 days after pollination (DAP) in the stress treatment. For both basal and apical kernels, the correlation analyses indicated the mean kernel filling rate correlated positively with zeatin riboside content at DAP 10 and indole-3-acetic acid content at DAP 20. This study provides useful information to support the development of drought- and heat-tolerant hybrids and effective management strategies to improve crop yields.

LpNOL-knockdown suppression of heat-induced leaf senescence in perennial ryegrass involving regulation of amino acid and organic acid metabolism.

The nonyellow COLORING 1-like gene (NOL) is known for its roles in accelerating leaf senescence, but the underlying metabolic mechanisms for heat-induced leaf senescence remain unclear. The objectives of this study were to identify metabolites and associated metabolic pathways regulated by knockdown of NOL in perennial ryegrass (Lolium perenne) and to determine the metabolic mechanisms of NOL controlling heat-induced leaf senescence. Wild-type (WT; cv. "Pinnacle") and two lines (Noli-1 and Noli-2) of perennial ryegrass with LpNOL knockdown were exposed to heat stress at 35/33°C (day/night) or nonstress control temperatures at 25/22°C (day/night) for 30 days in growth chambers. Leaf electrolyte leakage, chlorophyll (Chl) content, photochemical efficiency (Fv /Fm ), and net photosynthetic rate (Pn) were measured as physiological indicators of leaf senescence, while gas chromatography-mass spectrometry was performed to identify metabolites regulated by LpNOL. Knockdown of LpNOL suppressed heat-induced leaf senescence and produced a stay-green phenotype in perennial ryegrass, as manifested by increased Chl content, photochemical efficiency, net photosynthetic rate, and cell membrane stability in Noli-1 and Noli-2. Five metabolites (valine, malic acid, threonic acid, shikimic acid, chlorogenic acid) were uniquely upregulated in LpNOL plants exposed to heat stress, and six metabolites (aspartic acid, glutamic acid, 5-oxoproline, phenylalanine, proline, tartaric acid) exhibited more pronounced increases in their content in LpNOL plants than the WT. LpNOL could regulate heat-induced leaf senescence in perennial ryegrass through metabolic reprogramming in the pathways of respiration, secondary metabolism, antioxidant metabolism, and protein synthesis.

Ethephon-Induced Ethylene Enhances Starch Degradation and Sucrose Transport with an Interactive Abscisic Acid-Mediated Manner in Mature Leaves of Oilseed rape (Brassica napus L.).

The leaf senescence process is characterized by the degradation of macromolecules in mature leaves and the remobilization of degradation products via phloem transport. The phytohormone ethylene mediates leaf senescence. This study aimed to investigate the ethephon-induced ethylene effects on starch degradation and sucrose remobilization through their interactive regulation with other hormones. Ethephon (2-chloroethylphosphonic acid) was used as an ethylene-generating agent. Endogenous hormonal status, carbohydrate compounds, starch degradation-related gene expression, sucrose transporter gene expression, and phloem sucrose loading were compared between the ethephon-treated plants and controls. Foliar ethephon spray enhanced the endogenous ethylene concentration and accelerated leaf senescence, as evidenced by reduced chlorophyll content and enhanced expression of the senescence-related gene SAG12. Ethephon-enhanced ethylene prominently enhanced the endogenous abscisic acid (ABA) level. accompanied with upregulation of ABA synthesis gene 9-cis-epoxycarotenoid dioxygenase (NCED3), ABA receptor gene pyrabactin resistance 1 (PYR1), and ABA signaling genes sucrose non-fermenting 1 (Snf1)-related protein kinase 2 (SnRK2), ABA-responsive element binding 2 (AREB2), and basic-helix-loop-helix (bHLH) transcription factor (MYC2).) Ethephon treatment decreased starch content by enhancing expression of the starch degradation-related genes α-amylase 3 (AMY3) and β-amylase 1 (BAM1), resulting in an increase in sucrose content in phloem exudates with enhanced expression of sucrose transporters, SUT1, SUT4, and SWEET11. These results suggest that a synergistic interaction between ethylene and ABA might account for sucrose accumulation, mainly due to starch degradation in mature leaves and sucrose phloem loading in the ethephon-induced senescent leaves.

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 invivo 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 invitro and invivo. 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.

The apple C2H2-type zinc finger transcription factor MdZAT10 positively regulates JA-induced leaf senescence by interacting with MdBT2

Jasmonic acid (JA) plays an important role in regulating leaf senescence. However, the molecular mechanisms of leaf senescence in apple (Malus domestica) remain elusive. In this study, we found that MdZAT10, a C2H2-type zinc finger transcription factor (TF) in apple, markedly accelerates leaf senescence and increases the expression of senescence-related genes. To explore how MdZAT10 promotes leaf senescence, we carried out liquid chromatography/mass spectrometry screening. We found that MdABI5 physically interacts with MdZAT10. MdABI5, an important positive regulator of leaf senescence, significantly accelerated leaf senescence in apple. MdZAT10 was found to enhance the transcriptional activity of MdABI5 for MdNYC1 and MdNYE1, thus accelerating leaf senescence. In addition, we found that MdZAT10 expression was induced by methyl jasmonate (MeJA), which accelerated JA-induced leaf senescence. We also found that the JA-responsive protein MdBT2 directly interacts with MdZAT10 and reduces its protein stability through ubiquitination and degradation, thereby delaying MdZAT10-mediated leaf senescence. Taken together, our results provide new insight into the mechanisms by which MdZAT10 positively regulates JA-induced leaf senescence in apple.

Photosynthetic acclimation during low-light-induced leaf senescence in post-anthesis maize plants.

Low light conditions not only induce leaf senescence, but also photosynthetic acclimation. This study aimed to determine whether plants exhibit photosynthetic acclimation during low-light-induced leaf senescence. The influences of shading on leaf senescence and photosynthetic acclimation were explored in post-anthesis maize plants. The results showed that whole shading (WS) of maize plants accelerated leaf senescence, whereas partial shading (PS) slowed leaf senescence. WS led to larger decreases in the photosynthetic rate (Pn) and stomatal conductance (Gs) compared to those of the PS treatment. Interestingly, chlorophyll afluorescence (ChlF) demonstrated that the absorption flux (ABS/CSo) and trapped energy flux (TRo/CSo) per cross section in leaves remained relatively stable under WS, whereas significant decreases in the active PSII reaction centers (RC/CSo) resulted in considerable increases in absorption (ABS/RC) and trapped energy flux (TRo/RC) per reaction center. ABS/CSo, TRo/CSo, ABS/RC, and TRo/RC increased markedly under PS, whereas there were slight decreases in RC/CSo and electron transport activity. These results suggest that the PS treatment resulted in obvious improvements in the absorption and capture of light energy in shaded leaves. Further analysis demonstrated that both the WS and PS treatments resulted in a greater decrease in the activity of Rubisco compared to that of phosphoenolpyruvate carboxylase (PEPC). Moreover, PEPC activity in PS was maintained at a high level. Consequently, the current study proposed that the improvement of the absorption and capture of light energy and the maintenance of PEPC activity of mesophyll cells were due to photosynthetic acclimation of low-light-induced leaf senescence in maize plants. In addition, the rate of senescence of vascular bundle cells in maize leaves exceeded that of mesophyll cells under low light, showing obvious tissue specificity.

Growth and Photosynthetic Responses of Seedlings of Japanese White Birch, a Fast-Growing Pioneer Species, to Free-Air Elevated O3 and CO2

Plant growth is not solely determined by the net photosynthetic rate (A), but also influenced by the amount of leaves as a photosynthetic apparatus. To evaluate growth responses to CO2 and O3, we investigated the effects of elevated CO2 (550–560 µmol mol−1) and O3 (52 nmol mol−1; 1.7 × ambient O3) on photosynthesis and biomass allocation in seedlings of Japanese white birch (Betula platyphylla var. japonica) grown in a free-air CO2 and O3 exposure system without any limitation of root growth. Total biomass was enhanced by elevated CO2 but decreased by elevated O3. The ratio of root to shoot (R:S ratio) showed no difference among the treatment combinations, suggesting that neither elevated CO2 nor elevated O3 affected biomass allocation in the leaf. Accordingly, photosynthetic responses to CO2 and O3 might be more important for the growth response of Japanese white birch. Based on A measured under respective growth CO2 conditions, light-saturated A at a light intensity of 1500 µmol m−2 s−1 (A1500) in young leaves (ca. 30 days old) exhibited no enhancement by elevated CO2 in August, suggesting photosynthetic acclimation to elevated CO2. However, lower A1500 was observed in old leaves (ca. 60 days old) of plants grown under elevated O3 (regulated to be twice ambient O3). Conversely, light-limited A measured under a light intensity of 200 µmol m−2 s−1 (A200) was significantly enhanced by elevated CO2 in young leaves, but suppressed by elevated O3 in old leaves. Decreases in total biomass under elevated O3 might be attributed to accelerated leaf senescence by O3, indicated by the reduced A1500 and A200 in old leaves. Increases in total biomass under elevated CO2 might be attributed to enhanced A under high light intensities, which possibly occurred before the photosynthetic acclimation observed in August, and/or enhanced A under limiting light intensities.

NOL-mediated functional stay-green traits in perennial ryegrass (Lolium perenne L.) involving multifaceted molecular factors and metabolic pathways regulating leaf senescence.

Loss of chlorophyll (Chl) is a hallmark of leaf senescence, which may be regulated by Chl catabolic genes, including NON-YELLOW COLORING 1 (NYC1)-like (NOL). The objective of this study was to determine molecular factors and metabolic pathways underlying NOL regulation of leaf senescence in perennial grass species. LpNOL was cloned from perennial ryegrass (Lolium perenne L.) and found to be highly expressed in senescent leaves. Transient overexpression of LpNOL accelerated leaf senescence and Chl b degradation in Nicotiana benthamiana. LpNOL RNA interference (NOLi) in perennial ryegrass not only significantly blocked Chl degradation in senescent leaves, but also delayed initiation and progression of leaf senescence. This study found that NOL, in addition to functioning as a Chl b reductase, could enact the functional stay-green phenotype in perennial grass species, as manifested by increased photosynthetic activities in NOLi plants. Comparative transcriptomic analysis revealed that NOL-mediated functional stay-green in perennial ryegrass was mainly achieved through the modulation of Chl catabolism, light harvesting for photosynthesis, photorespiration, cytochrome respiration, carbohydrate catabolism, oxidative detoxification, and abscisic acid biosynthesis and signaling pathways.

AtWAKL10, a Cell Wall Associated Receptor-Like Kinase, Negatively Regulates Leaf Senescence in Arabidopsis thaliana.

Receptor-like kinases (RLKs) constitute a large group of cell surface receptors that play crucial roles in multiple biological processes. However, the function of most RLKs in plants has not been extensively explored, and much less for the class of cell wall associated kinases (WAKs) and WAK-like kinases (WAKLs). In this study, analyses of developmental expression patterns uncovered a putative role of AtWAKL10 in modulating leaf senescence, which was further investigated at physiological and molecular levels. The expression level of AtWAKL10 increased with the developmental progression and was rapidly upregulated in senescing leaf tissues. The promoter of AtWAKL10 contains various defense and hormone responsive elements, and its expression could be significantly induced by exogenous ABA, JA and SA. Moreover, the loss-of-function atwakl10 mutant showed earlier senescence along the course of natural development and accelerated leaf senescence under darkness and hormonal stresses, while plants overexpressing AtWAKL10 showed an opposite trend. Additionally, some defense and senescence related WRKY transcription factors could bind to the promoter of AtWAKL10. In addition, deletion and overexpression of AtWAKL10 caused several specific transcriptional alterations, including genes involved in cell extension, cell wall modification, defense response and senescence related WRKYs, which may be implicated in regulatory mechanisms adopted by AtWAKL10 in controlling leaf senescence. Taken together, these results revealed that AtWAKL10 negatively regulated leaf senescence.

Ethylene and salicylic acid synergistically accelerate leaf senescence in Arabidopsis.

The phytohormones ethylene and salicylic acid (SA) have long been known to promote senescence, but their interplay during this process remains elusive. Here we report the synergistic effects of ethylene and SA on promoting leaf senescence in Arabidopsis. EIN3, a key transcription factor of ethylene signaling, physically interacted with the core SA signaling regulator NPR1 in senescing leaves. EIN3 and NPR1 synergistically promoted the expression of the senescence-associated genes ORE1 and SAG29. The senescence phenotype was more delayed for the ein3eil1npr1 triple mutant than ein3eil1 or npr1 with ethylene or/and SA treatment. NPR1-promoted leaf senescence may depend on functional EIN3/EIL1.

Senescence-Specific Expression of RAmy1A Accelerates Non-structural Carbohydrate Remobilization and Grain Filling in Rice (Oryza sativa L.).

Remobilization of pre-anthesis NSCs (non-structural carbohydrates) is significant for effective grain filling in rice (Oryza sativa L.). However, abundant starch particles as an important component of NSCs are still present in the leaf sheath and stem at the late stage of grain filling. There are no studies on how bioengineering techniques can be used to improve the efficiency of NSC remobilization. In this study, RAmy1A was expressed under the senescence-specific promoter of SAG12, which was designed to degrade starch in the leaf sheath and stem during grain filling. RAmy1A mRNA successfully accumulated in the leaf, stem, and sheath of transgenic plants after anthesis. At the same time, the starch and total soluble sugar content in the leaf, stem, and leaf sheath were obviously decreased during the grain-filling period. The photosynthetic rate of transgenic lines was higher than that of the wild types by an average of 4.0 and 9.9%, at 5 and 10 days after flowering, respectively. In addition, the grain-filling rate of transgenic lines was faster than that of the wild types by an average of 26.09%. These results indicate an enhanced transport efficiency of NSCs from source tissues in transgenic rice. Transgenic rice also displayed accelerated leaf senescence, which was hypothesized to contribute to decreased grain weight.

The NPR1-WRKY46-WRKY6 signaling cascade mediates probenazole/salicylic acid-elicited leaf senescence in Arabidopsis thaliana.

Endogenous salicylic acid (SA) regulates leaf senescence, but the underlying mechanism remains largely unexplored. The exogenous application of SA to living plants is not efficient for inducing leaf senescence. By taking advantage of probenazole (PBZ)-induced biosynthesis of endogenous SA, we previously established a chemical inducible leaf senescence system that depends on SA biosynthesis and its core signaling receptor NPR1 in Arabidopsis thaliana. Here, using this system, we identified WRKY46 and WRKY6 as key components of the transcriptional machinery downstream of NPR1 signaling. Upon PBZ treatment, the wrky46 mutant exhibited significantly delayed leaf senescence. We demonstrate that NPR1 is essential for PBZ/SA-induced WRKY46 activation, whereas WRKY46 in turn enhances NPR1 expression. WRKY46 interacts with NPR1 in the nucleus, binding to the W-box of the WRKY6 promoter to induce its expression in response to SA signaling. Dysfunction of WRKY6 abolished PBZ-induced leaf senescence, while overexpression of WRKY6 was sufficient to accelerate leaf senescence even under normal growth conditions, suggesting that WRKY6 may serve as an integration node of multiple leaf senescence signaling pathways. Taken together, these findings reveal that the NPR1-WRKY46-WRKY6 signaling cascade plays a critical role in PBZ/SA-mediated leaf senescence in Arabidopsis.

Salicylic acid is a key player of Arabidopsis autophagy mutant susceptibility to the necrotrophic bacterium Dickeya dadantii

Autophagy is a ubiquitous vesicular process for protein and organelle recycling in eukaryotes. In plant, autophagy is reported to play pivotal roles in nutrient recycling, adaptation to biotic and abiotic stresses. The role of autophagy in plant immunity remains poorly understood. Several reports showed enhanced susceptibility of different Arabidopsis autophagy mutants (atg) to necrotrophic fungal pathogens. Interaction of necrotrophic bacterial pathogens with autophagy is overlooked. We then investigated such interaction by inoculating the necrotrophic enterobacterium Dickeya dadantii in leaves of the atg2 and atg5 mutants and an ATG8a overexpressing line. Overexpressing ATG8a enhances plant tolerance to D. dadantii. While atg5 mutant displayed similar susceptibility to the WT, the atg2 mutant exhibited accelerated leaf senescence and enhanced susceptibility upon infection. Both phenotypes were reversed when the sid2 mutation, abolishing SA signaling, was introduced in the atg2 mutant. High levels of SA signaling in atg2 mutant resulted in repression of the jasmonic acid (JA) defense pathway known to limit D. dadantii progression in A. thaliana. We provide evidence that in atg2 mutant, the disturbed hormonal balance leading to higher SA signaling is the main factor causing increased susceptibility to the D. dadantii necrotroph by repressing the JA pathway and accelerating developmental senescence.

Effects of thidiazuron and ethephon on the grain filling and dehydration characteristics of maize in Northeast China

ABSTRACT A mixture of ethephon (ETH) and thidiazuron (TDZ) (called ETTD) has been proven to effectively reduce maize grain moisture content and improve yield and quality of mechanical grain harvesting, but the underlying mechanism is unclear. Field trials were conducted in 852 Farm, Heilongjiang Province, China, in 2017 and 2018, to study the effects of ETH, TDZ and ETTD sprayed on Xianyu335 at 25 days after flowering (appropriate time in the pre-experiment) on grain filling and dehydration characteristics. ETH, TDZ and ETTD significantly accelerated leaf senescence, increased grain, bract, and cob dehydration rates, and decreased grain filling duration and moisture content. TDZ and ETTD significantly increased grain filling rate and auxin (IAA), zeatin riboside (ZR), and abscisic acid (ABA) and decreased gibberellic acid (GA). ETH had no significant effect on ABA and GA, but significantly reduced IAA and ZR in the later period. The grain dehydration rate was significantly negatively correlated with grain filling duration and positively correlated with rates of grain filling and bract and cob dehydration. ETTD significantly decreased the rates of breakage, impurity, and increased mechanical grain harvesting yield. In conclusion, ETTD can be an effective measure to improve yield and quality of maize mechanical grain harvesting.