Accelerated Leaf Senescence Research Articles

Mutation of Leaf Senescence 1 Encoding a C2H2 Zinc Finger Protein Induces ROS Accumulation and Accelerates Leaf Senescence in Rice.

Premature senescence of leaves causes a reduced yield and quality of rice by affecting plant growth and development. The regulatory mechanisms underlying early leaf senescence are still unclear. The Leaf senescence 1 (LS1) gene encodes a C2H2-type zinc finger protein that is localized to both the nucleus and cytoplasm. In this study, we constructed a rice mutant named leaf senescence 1 (ls1) with a premature leaf senescence phenotype using CRISPR/Cas9-mediated editing of the LS1 gene. The ls1 mutants exhibited premature leaf senescence and reduced chlorophyll content. The expression levels of LS1 were higher in mature or senescent leaves than that in young leaves. The contents of reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD) were significantly increased and catalase (CAT) activity was remarkably reduced in the ls1 plants. Furthermore, a faster decrease in pigment content was detected in mutants than that in WT upon induction of complete darkness. TUNEL and staining experiments indicated severe DNA degradation and programmed cell death in the ls1 mutants, which suggested that excessive ROS may lead to leaf senescence and cell death in ls1 plants. Additionally, an RT-qPCR analysis revealed that most senescence-associated and ROS-scavenging genes were upregulated in the ls1 mutants compared with the WT. Collectively, our findings revealed that LS1 might regulate leaf development and function, and that disruption of LS1 function promotes ROS accumulation and accelerates leaf senescence and cell death in rice.

The Relationships among "STAY-GREEN" Trait, Post-Anthesis Assimilate Remobilization, and Grain Yield in Rice (Oryza sativa L.).

The mobilization and translocation of carbohydrates and mineral nutrients from vegetative plant parts to grains are pivotal for grain filling, often involving a whole plant senescence process. Loss of greenness is a hallmark of leaf senescence. However, the relationship between crop yield and senescence has been controversial for many years. Here, in this study, the overexpression and RNA interference lines of gene of OsNYC3 (Non-Yellow Coloring 3), a chlorophyll catabolism gene, were investigated. Furthermore, exogenous phytohormones were applied, and a treatment of alternate wetting and moderate drying (AWMD) was introduced to regulate the processes of leaf senescence. The results indicated that the delayed senescence of the "STAY-GREEN" trait of rice is undesirable for the process of grain filling, and it would cause a lower ratio of grain filling and lower grain weight of inferior grains, because of unused assimilates in the stems and leaves. Through the overexpression of OsNYC3, application of exogenous chemicals of abscisic acid (ABA), and water management of AWMD, leaf photosynthesis was less influenced, a high ratio of carbohydrate assimilates was partitioned to grains other than leaves and stems as labeled by 13C, grain filling was improved, especially for inferior spikelets, and activities of starch-synthesizing enzymes were enhanced. However, application of ethephon not only accelerated leaf senescence, but also caused seed abortion and grain weight reduction. Thus, plant senescence needs to be finely adjusted in order to make a contribution to crop productivity.

The auxin response factor TaARF15-A1 negatively regulates senescence in common wheat (Triticum aestivum L.).

Auxin plays an important role in regulating leaf senescence. Auxin response factors (ARFs) are crucial components of the auxin signaling pathway; however, their roles in leaf senescence in cereal crops are unknown. In this study, we identified TaARF15-A1 as a negative regulator of senescence in wheat (Triticum aestivum L.) by analyzing TaARF15-A1 overexpression (OE) and RNA interference lines and CRISPR/Cas9-based arf15 mutants. OE of TaARF15-A1 delayed senescence, whereas knockdown lines and knockout mutants showed accelerated leaf senescence and grain ripening. RNA-seq analysis revealed that TaARF15-A1 delays leaf senescence by negatively regulating senescence-promoting processes and positively modulating senescence-delaying genes including senescence-associated phytohormone biosynthesis and metabolism genes as well as transcription factors (TFs). We also demonstrated that TaARF15-A1 physically interacts with TaMYC2, a core jasmonic acid (JA) signaling TF that positively modulates wheat senescence. Furthermore, TaARF15-A1 suppressed the expression of TaNAM-1 (TaNAM-A1 and TaNAM-D1) via protein-protein interaction and competition with TaMYC2 for binding to its promoter to regulate senescence. Finally, we identified two haplotypes of TaARF15-A1 in global wheat collections. Association analysis revealed that TaARF15-A1-HapI has undergone strong selection during wheat breeding in China, likely owing to its earlier maturity. Thus, we identify TaARF15-A1 as a negative regulator of senescence in common wheat and present another perspective on the crosstalk between auxin and JA signaling pathways in regulating plant senescence.

Research progress on the relationship between leaf senescence and quality, yield and stress resistance in horticultural plants.

Leaf senescence, the final stage of leaf development, is one of the adaptive mechanisms formed by plants over a long period of evolution. Leaf senescence is accompanied by various changes in cell structure, physiological metabolism, and gene expressions. This process is controlled by a variety of internal and external factors. Meanwhile, the genes and plant hormones involved in leaf aging affect the quality, yield and stress resistance in horticultural plants. Leaf senescence mediated by plant hormones affected plant quality at both pre-harvest and post-harvest stages. Exogenous plant growth regulators or plant hormone inhibitors has been applied to delay leaf senescence. Modification of related gene expression by over-expression or antisense inhibition could delay or accelerate leaf senescence, and thus influence quality. Environmental factors such as light, temperature and water status also trigger or delay leaf senescence. Delaying leaf senescence could increase chloroplast lifespan and photosynthesis and thus improve source strength, leading to enhanced yield. Accelerating leaf senescence promotes nutrient redistribution from old leaves into young leaves, and may raise yield under certain circumstances. Many genes and transcriptional factors involved in leaf senescence are associated with responses to abiotic and biotic stresses. WRKY transcriptional factors play a vital role in this process and they could interact with JA signalling. This review summarized how genes, plant hormones and environmental factors affect the quality, yield. Besides, the regulation of leaf senescence holds great promise to improving the resistance to plant biotic and abiotic stresses.

A NAC transcriptional factor BrNAC029 is involved in cytokinin-delayed leaf senescence in postharvest Chinese flowering cabbage

Both cytokinin and NAC transcription factors were reported to involve in leaf senescence. However, the mechanism of NAC transcription factors how to regulate cytokinin-delayed leaf senescence is still unknown. In this study, application of N-(2-chloro-4-pyridyl)-N'-phenylurea (CPPU), a cytokinin analogue, significantly delayed leaf senescence and maintained cytokinin content of Chinese flowering cabbage during storage. Meanwhile, the expression of an NAC transcriptional activator (BrNAC029) was increased but suppressed by CPPU treatment. Furthermore, BrNAC029 activated the expressions of chlorophyll catabolic genes BrPAO and BrSGR2, cytokinin oxidase gene BrCKX1 and senescence maker gene BrSAG113 by binding to their promoters. Additionally, overexpressions of BrNAC029 in tobacco and Arabidopsis accelerated leaf senescence and up-expressed the related genes. Taken together, it was suggested that BrNAC029 may serve as a transcriptional activator to activate the transcriptions of these related genes to eventually accelerate leaf senescence of Chinese flowering cabbage by promoting chlorophyll degradation and reducing endogenous cytokinin level.

Arabidopsis Ubiquitin-Conjugating Enzymes UBC4, UBC5, and UBC6 Have Major Functions in Sugar Metabolism and Leaf Senescence.

The ubiquitin-conjugating enzyme (E2) is required for protein ubiquitination. Arabidopsis has 37 E2s grouped into 14 subfamilies and the functions for many of them are unknown. We utilized genetic and biochemical methods to study the roles of Arabidopsis UBC4, UBC5, and UBC6 of the E2 subfamily IV. The Arabidopsis ubc4/5/6 triple mutant plants had higher levels of glucose, sucrose, and starch than the control plants, as well as a higher protein level of a key gluconeogenic enzyme, cytosolic fructose 1,6-bisphosphatase 1 (cyFBP). In an in vitro assay, the proteasome inhibitor MG132 inhibited the degradation of recombinant cyFBP whereas ATP promoted cyFBP degradation. In the quadruple mutant ubc4/5/6 cyfbp, the sugar levels returned to normal, suggesting that the increased sugar levels in the ubc4/5/6 mutant were due to an increased cyFBPase level. In addition, the ubc4/5/6 mutant plants showed early leaf senescence at late stages of plant development as well as accelerated leaf senescence using detached leaves. Further, the leaf senescence phenotype remained in the quadruple ubc4/5/6 cyfbp mutant. Our results suggest that UBC4/5/6 have two lines of important functions, in sugar metabolism through regulating the cyFBP protein level and in leaf senescence likely through a cyFBP-independent mechanism.

Transgenerationally Transmitted DNA Demethylation of a Spontaneous Epialleles Using CRISPR/dCas9-TET1cd Targeted Epigenetic Editing in Arabidopsis.

CRISPR/dCas9 is an important DNA modification tool in which a disarmed Cas9 protein with no nuclease activity is fused with a specific DNA modifying enzyme. A previous study reported that overexpression of the TET1 catalytic domain (TET1cd) reduces genome-wide methylation in Arabidopsis. A spontaneous naturally occurring methylation region (NMR19-4) was identified in the promoter region of the PPH (Pheophytin Pheophorbide Hydrolase) gene, which encodes an enzyme that can degrade chlorophyll and accelerate leaf senescence. The methylation status of NMR19-4 is associated with PPH expression and leaf senescence in Arabidopsis natural accessions. In this study, we show that the CRISPR/dCas9-TET1cd system can be used to target the methylation of hypermethylated NMR19-4 region to reduce the level of methylation, thereby increasing the expression of PPH and accelerating leaf senescence. Furthermore, hybridization between transgenic demethylated plants and hypermethylated ecotypes showed that the demethylation status of edited NMR19-4, along with the enhanced PPH expression and accelerated leaf senescence, showed Mendelian inheritance in F1 and F2 progeny, indicating that spontaneous epialleles are stably transmitted trans-generationally after demethylation editing. Our results provide a rational approach for future editing of spontaneously mutated epialleles and provide insights into the epigenetic mechanisms that control plant leaf senescence.

STRONG STAYGREEN inhibits DNA binding of PvNAP transcription factors during leaf senescence in switchgrass.

Fine tuning the progression of leaf senescence is important for plant fitness in nature, while the "staygreen" phenotype with delayed leaf senescence has been considered a valuable agronomic trait in crop genetic improvement. In this study, a switchgrass (Panicum virgatum L.) CCCH-type Zinc finger gene, Strong Staygreen (PvSSG), was characterized as a suppressor of leaf senescence as overexpression or suppression of the gene led to delayed or accelerated leaf senescence, respectively. Transcriptomic analysis marked that chlorophyll (Chl) catabolic pathway genes were involved in the PvSSG-regulated leaf senescence. PvSSG was identified as a nucleus-localized protein with no transcriptional activity. By yeast two-hybrid screening, we identified its interacting proteins, including a pair of paralogous transcription factors, PvNAP1/2 (NAC-LIKE, ACTIVATED BY AP3/PI). Overexpression of PvNAPs led to precocious leaf senescence at least partially by directly targeting and transactivating Chl catabolic genes to promote Chl degradation. PvSSG, through protein-protein interaction, repressed the DNA-binding efficiency of PvNAPs and alleviated its transactivating effect on downstream genes, thereby functioning as a "brake" in the progression of leaf senescence. Moreover, overexpression of PvSSG resulted in up to 47% higher biomass yield and improved biomass feedstock quality, reiterating the importance of leaf senescence regulation in the genetic improvement of switchgrass and other feedstock crops.

Morphological, physiological, and secondary metabolic responses of Taraxacum officinale to salt stress

Salt stress has a major effect on growth and secondary metabolism in medicinal plants, however, the effect of salt stress on Taraxacum officinale F. H. Wigg. is still scarce. In this study, we evaluated the effects of salt stress on the physiology, morphology, phenolic acid accumulation, and expression of genes involved in phenolic acid biosynthesis in T. officinale. We found that plants grew well at 1 g kg−1 NaCl, and the state of photosystem Ⅱ (PSⅡ) and the organization of the chloroplasts at 0.5 g kg−1 NaCl showed no significant differences compared with the control. However, 2 g kg−1 and 4 g kg−1 NaCl inhibited growth and accelerated leaf senescence. At 4 g kg−1 NaCl, the fresh and dry weights decreased to 28% and 42% of the control, while chlorosis and necrosis were observed on the leaves. Furthermore, up-regulation of the expression of ToC3′H corresponded with an increase in the levels of caffeoylquinic acids (chlorogenic acid and isochlorogenic acid A) at NaCl concentration ≤ 1 g kg−1. Expressions of four phenolic acid biosynthesis genes, ToC4H, To4CL, ToHCT, and ToHQT, were down-regulated with increasing NaCl concentrations, consistent with the observed decreases in caftaric and cichoric acids. In summary, cultivation of T. officinale under mild salt stress (NaCl ≤ 1 g kg−1) is feasible and facilitates the accumulation of caffeoylquinic acids; thus this species may be recommended for saline soils.

Responses of photosynthetic characteristics and leaf senescence in summer maize to simultaneous stresses of waterlogging and shading

A field experiment was performed to investigate the physiological mechanism of the simultaneous stresses of waterlogging and shading on leaf photosynthetic and senescence during three growth stages of summer maize. The responses of leaf gas exchange parameters and antioxidant enzyme activities of the summer maize hybrids Denghai 605 (DH605) to waterlogging (W), shading (S), and their combination (W + S) for 6 days at the third leaf stage (V3), the sixth leaf stage (V6), and the tasseling stage (VT) were recorded. Shading, waterlogging, and their combination disturbed the activities of protective enzymes and increased the contents of H2O2 and O2−, accelerating leaf senescence and disordering photosynthetic characteristics. Under waterlogging, shading and their combination, leaf Pn, the photo-assimilates and grain yield was decreased. The greatest reduction for waterlogging and the combined stresses occurred at V3 and that for shading stress occurred at VT. The individual and combined stresses reduced the activities of protective enzymes and inhibited photosynthesis, reducing the accumulation of photosynthetic compounds and thereby yield. Waterlogging and the combined stresses at the V3 stage showed the greatest effect on leaf photosynthetic and senescence, followed by the V6 and VT stages. The greatest effect for shading stress occurred at VT, followed by the V6 and V3 stages, and the combined influence of shading and waterlogging was greater than that of either single stress.

The safflower bHLH transcription factor CtbHLH41 negatively regulates SA-induced leaf senescence through interaction with CtCP1

Salicylic acid (SA) plays an important role in regulating leaf senescence. However, the molecular mechanism of leaf senescence of safflower ( Carthamus tinctorius ) is still elusive. In this study, we found that the bHLH transcription factor (TF) CtbHLH41 in Carthamus tinctorius significantly delayed leaf senescence and inhibited the expression of senescence-related genes. In order to explore how CtbHLH41 promotes leaf senescence, we carried out yeast two-hybrid screening. In this study, by exploring the mechanism of CtbHLH41 regulating CtCP1 , it was found that CtCP1 promoted the hydrolysis of CtbHLH41 protein, accelerated the transcriptional activities of salicylic acid-mediated senescence-related genes CtSAG12 and CtSAG29 , chlorophyll degradation genes CtNYC1 and CtNYE1 , and accelerated leaf senescence. We found a negative SA regulator CtANS1 , which interacted with CtbHLH41 and regulated its stability, thereby inhibiting CtCP1 -mediated leaf senescence. In short, our results provide a new insight into the mechanism of CtbHLH41 actively regulating the senescence of safflower leaves induced by SA. • In the course of our research on CtbHLH41 , we found that the overexpression of CtbHLH41 slowed down leaf senescence. • CtCP1 was included in the results of CtbHLH41 screening by yeast two-hybrid, which may promote leaf senescence of plants. • We found that CtCP1 promoted the hydrolysis of CtbHLH41 protein and accelerated leaf senescence. • We also found a gene CtANS1 , which also interacts with CtbHLH41 , thereby delaying leaf senescence caused by CtCP1 .

Fertilization Practices: Optimization in Greenhouse Vegetable Cultivation with Different Planting Years

Cucumber plant growth and the fate of N in the plant-soil system are influenced by fertilization practices, the strengths of which may vary among soils. Three soils with different years of greenhouse vegetable cultivation (0, 2, and 18 years) were fertilized differently (CK, no N fertilizer applied; CF, chemical NPK fertilizers applied; RCF, reduced chemical NPK fertilizers applied, with N, P, and K reduced by 46.5%, 68.6%, and 54.7%; RCF+CM, 75% of the total N derived from chemical fertilizer and the rest from chicken manure in the case of reduced fertilization) in a pot experiment to study the changes in cucumber (Cucumis sativus L.) growth, N uptake, residue, and losses. The original N in soil was insufficient to maintain leaf growth and chlorophyll synthesis at later growth stages, even in soil with 18 years of greenhouse vegetable cultivation, where the original N content was the highest (total N 1.73 g kg−1). However, the CF treatment with excessive N fertilization inhibited leaf growth at the early growing stage and accelerated leaf senescence later, especially in soil with longer years of greenhouse vegetable cultivation. Therefore, reduced fertilizer application (RCF and RCF+CM) is appropriate to improve cucumber growth and productivity in greenhouse cultivation with different planting years. Although the same amount of N was applied, the RCF+CM treatment performed better than the RCF treatment in terms of increasing plant N uptake (by 30.5%) and soil N pool storage (by 25.0%) while decreasing N losses (by 16.6%) in soil with 0 years of greenhouse vegetable cultivation. In soil with 2 and 18 years of greenhouse vegetable cultivation, the soil itself functions much better in exogenous N retention and supply, with the N storage and losses not significantly different between the RCF and RCF+CM treatments. We conclude that reduced fertilization with the co-application of chicken manure is optimal for plant growth promotion, output-input ratio increase, soil N fertility improvement, and environmental risk mitigation.

The transcription factor SlWRKY37 positively regulates jasmonic acid- and dark-induced leaf senescence in tomato.

Initiation and progression of leaf senescence are triggered by various environmental stressors and phytohormones. Jasmonic acid (JA) and darkness accelerate leaf senescence in plants. However, the mechanisms that integrate these two factors to initiate and regulate leaf senescence have not been identified. Here, we report a transcriptional regulatory module centred on a novel tomato WRKY transcription factor, SlWRKY37, responsible for both JA- and dark-induced leaf senescence. The expression of SlWRKY37, together with SlMYC2, encoding a master transcription factor in JA signalling, was significantly induced by both methyl jasmonate (MeJA) and dark treatments. SlMYC2 binds directly to the promoter of SlWRKY37 to activate its expression. Knock out of SlWRKY37 inhibited JA- and dark-induced leaf senescence. Transcriptome analysis and biochemical experiments revealed SlWRKY53 and SlSGR1 (S. lycopersicum senescence-inducible chloroplast stay-green protein 1) as direct transcriptional targets of SlWRKY37 to control leaf senescence. Moreover, SlWRKY37 interacted with a VQ motif-containing protein SlVQ7, and the interaction improved the stability of SlWRKY37 and the transcriptional activation of downstream target genes. Our results reveal the physiological and molecular functions of SlWRKY37 in leaf senescence, and offer a target gene to retard leaf yellowing by reducing sensitivity to external senescence signals, such as JA and darkness.

Analysis of Physiological and Transcriptomic Differences between a Premature Senescence Mutant (GSm) and Its Wild-Type in Common Wheat (Triticum aestivum L.).

Simple SummaryEarly leaf senescence is an important agronomic trait that affects crop yield and quality. To understand the molecular mechanism of early leaf senescence, a wheat (Triticum aestivum L.) premature leaf senescence mutant (GSm) and its wild type were employed in this study. We compared the physiological characteristics and transcriptome of wheat leaves between the wild type (WT) and the mutant at two-time points. Physiological characteristics and differentially expressed gene (DEG) analysis revealed many genes and metabolic pathways that were closely related to senescence. These results will not only support further gene cloning and functional analysis of GSm, but also facilitate the study of leaf senescence in wheat.Premature leaf senescence has a profound influence on crop yield and quality. Here, a stable premature senescence mutant (GSm) was obtained from the common wheat (Triticum aestivum L.) cultivar Chang 6878 by mutagenesis with ethyl methanesulfonate. The differences between the GSm mutant and its wild-type (WT) were analyzed in terms of yield characteristics, photosynthetic fluorescence indices, and senescence-related physiological parameters. RNA sequencing was used to reveal gene expression differences between GSm and WT. The results showed that the yield of GSm was considerably lower than that of WT. The net photosynthetic rate, transpiration rate, maximum quantum yield, non-photochemical quenching coefficient, photosynthetic electron transport rate, soluble protein, peroxidase activity, and catalase activity all remarkably decreased in flag leaves of GSm, whereas malondialdehyde content distinctively increased compared with those of WT. The analysis of differentially expressed genes indicated blockade of chlorophyll and carotenoid biosynthesis, accelerated degradation of chlorophyll, and diminished photosynthetic capacity in mutant leaves; brassinolide might facilitate chlorophyll breakdown and consequently accelerate leaf senescence. NAC genes positively regulated the senescence process. Compared with NAC genes, expression of WRKY and MYB genes was induced earlier in the mutant possibly due to increased levels of reactive oxygen species and plant hormones (e.g., brassinolide, salicylic acid, and jasmonic acid), thereby accelerating leaf senescence. Furthermore, the antioxidant system played a role in minimizing oxidative damage in the mutant. These results provides novel insight into the molecular mechanisms of premature leaf senescence in crops.

Understorey light quality affects leaf pigments and leaf phenology in different plant functional types.

Forest understorey plants receive most sunlight in springtime before canopy closure, and in autumn following leaf‐fall. We hypothesised that plant species must adjust their phenological and photoprotective strategies in response to large changes in the spectral composition of the sunlight they receive. Here, we identified how plant species growing in northern deciduous and evergreen forest understoreys differ in their response to blue light and ultraviolet (UV) radiation according to their functional strategy. We installed filters in a forest understorey in southern Finland, to create the following treatments attenuating: UV radiation below 350 nm, all UV radiation (< 400 nm), all blue light and UV radiation (< 500 nm), and a transparent control. In eight species, representing different functional strategies, we assessed leaf optical properties, phenology, and epidermal flavonoid contents over two years. Blue light accelerated leaf senescence in all species measured in the understorey, apart from Quercus robur seedlings, whereas UV radiation only accelerated leaf senescence in Acer platanoides seedlings. More light‐demanding species accumulated flavonols in response to seasonal changes in light quality compared to shade‐tolerant and wintergreen species and were particularly responsive to blue light. Reduction of blue and UV radiation under shade reveals an important role for microclimatic effects on autumn phenology and leaf photoprotection. An extension of canopy cover under climate change, and its associated suppression of understorey blue light and UV radiation, may delay leaf senescence for understorey species with an autumn niche.

RING-box proteins regulate leaf senescence and stomatal closure via repression of ABA transporter gene ABCG40.

Plant hormone abscisic acid (ABA) plays an indispensable role in the control of leaf senescence, during which ABA signaling depends on its biosynthesis. Nevertheless, the role of ABA transport in leaf senescence remains unknown. Here, we identified two novel RING-box protein-encoding genes UBIQUITIN LIGASE of SENESCENCE 1 and 2 (ULS1 and ULS2) involved in leaf senescence. Lack of ULS1 and ULS2 accelerates leaf senescence, which is specifically promoted by ABA treatment. Furthermore, the expression of senescence-related genes is significantly affected in mature leaves of uls1/uls2 double mutant (versus wild type (WT)) in an ABA-dependent manner, and the ABA content is substantially increased. ULS1 and ULS2 are mainly expressed in the guard cells and aging leaves, and the expression is induced by ABA. Further RNA-seq and quantitative proteomics of ubiquitination reveal that ABA transporter ABCG40 is highly expressed in uls1/uls2 mutant versus WT, though it is not the direct target of ULS1/2. Finally, we show that the acceleration of leaf senescence, the increase of leaf ABA content, and the promotion of stomatal closure in uls1/usl2 mutant are suppressed by abcg40 loss-of-function mutation. These results indicate that ULS1 and ULS2 function in feedback inhibition of ABCG40-dependent ABA transport during ABA-induced leaf senescence and stomatal closure.

Ethylene responsive factor34 mediates stress-induced leaf senescence by regulating salt stress-responsive genes.

Leaf senescence proceeds with age but is modulated by various environmental stresses and hormones. Salt stress is one of the most well-known environmental stresses that accelerate leaf senescence. However, the molecular mechanisms that integrate salt stress signalling with leaf senescence programmes remain elusive. In this study, we characterised the role of ETHYLENE RESPONSIVE FACTOR34 (ERF34), an Arabidopsis APETALA2 (AP2)/ERF family transcription factor, in leaf senescence. ERF34 was differentially expressed under various leaf senescence-inducing conditions, and negatively regulated leaf senescence induced by age, dark, and salt stress. ERF34 also promoted salt stress tolerance at different stages of the plant life cycle such as seed germination and vegetative growth. Transcriptome analysis revealed that the overexpression of ERF34 increased the transcript levels of salt stress-responsive genes including COLD-REGULATED15A (COR15A), EARLY RESPONSIVE TO DEHYDRATION10 (ERD10), and RESPONSIVE TO DESICCATION29A (RD29A). Moreover, ERF34 directly bound to ERD10 and RD29A promoters and activated their expression. Our findings indicate that ERF34 plays a key role in the convergence of the salt stress response with the leaf senescence programmes, and is a potential candidate for crop improvement, particularly by enhancing salt stress tolerance.

STAYGREEN-mediated chlorophyll a catabolism is critical for photosystem stability during heat-induced leaf senescence in perennial ryegrass.

Suppression of the chlorophyll a (Chl a) Mg-dechelatase gene, SGR/NYE1, blocks the degradation of Chl a, resulting in a 'stay-green' trait. In this study, we investigated the effect of Chl a catabolism on plant heat-induced leaf senescence in perennial ryegrass (Lolium perenne L.). Under heat stress, the LpSGR-RNAi lines not only lost the stay-green phenotype but also showed accelerated leaf senescence with increased chloroplast disruption, more loss of photosystem (PS) proteins, lower PSⅡquantum yields, higher levels of energy dissipation, increased accumulation of reactive oxygen species (ROS)and lower ROS-scavenging enzyme activities. Transcriptome analysis revealed that the suppression of LpSGR downregulated genes encoding PS proteins and ROS-scavenging enzymes and upregulated those encoding ROS-generation enzymes under heat stress. To account for the possible side-effects resulting from constitutive suppression of LpSGR on plant growth and heat tolerance, we constructed an ethanol-inducible RNAi vector to suppress LpSGR functions. In the absence of ethanol induction, these lines exhibited the same growth and heat tolerance as the wildtype(WT). Upon ethanol induction, the transgenic lines showed compromised heat tolerance and a postharvest stay-green phenotype. Taken together, SGR-mediated Chl a catabolism is required for plant heat tolerance.

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.