Rice melatonin deficiency causes premature leaf senescence via DNA methylation regulation

In a study of DNA methylation changes in melatonin-deficient rice mutants,mutant plants showed premature leaf senescence during grain-filling and reduced grain yield.Melatonin deficiency led to transcriptional reprogramming,especially of genes involved in chlorophyll and carbon metabolism,redox regulation,and transcriptional regulation,during dark-induced leaf senescence.Hypomethylation of mCG and mCHG in the melatonin-deficient rice mutants was associated with the expression change of both protein-coding genes and transposable element-related genes.Changes in gene expression and DNA methylation in the melatonin-deficient mutants were compensated by exogenous application of melatonin.A decreased S-adenosyl-L-methionine level may have contributed to the DNA methylation variations in rice mutants of melatonin deficiency under dark conditions.

Synergistic effects of planting density and nitrogen fertilization on chlorophyll degradation and leaf senescence after silking in maize

Regulating planting density and nitrogen(N)fertilization could delay chlorophyll(Chl)degradation and leaf senescence in maize cultivars.This study measured changes in ear leaf green area(GLA_(ear)),Chl content,the activities of Chl a-degrading enzymes after silking,and the post-silking dry matter accumulation and grain yield under multiple planting densities and N fertilization rates.The dynamic change of GLA_(ear)after silking fitted to the logistic model,and the GLA_(ear) duration and the GLAearat 42 d after silking were affected mainly by the duration of the initial senescence period(T_(1))which was a key factor of the leaf senescence.The average chlorophyllase(CLH)activity was 8.3 times higher than pheophytinase activity and contributed most to the Chl content,indicating that CLH is a key enzyme for degrading Chl a in maize.Increasing density increased the CLH activity and decreased the Chl content,T1,GLAear,and GLA_(ear) duration.Under high density,appropriate N application reduced CLH activity,increased Chl content,prolonged T1,alleviated high-density-induced leaf senescence,and increased post-silking dry matter accumulation and grain yield.

Rice ONAC016 promotes leaf senescence through abscisic acid signaling pathway involving OsNAP

Senescence-induced NAC(senNAC)TFs play a crucial role in senescence during the final stage of leaf development.In this study,we identified a rice senNAC,ONAC016,which functions as a positive regulator of leaf senescence.The expression of ONAC016 increased rapidly in rice leaves during the progression of dark-induced and natural senescence.The onac016-1 knockout mutant showed a delayed leaf yellowing phenotype,whereas the overexpression of ONAC016 accelerated leaf senescence.Notably,ONAC016 expression was upregulated by abscisic acid(ABA),and thus detached leaves of the onac016-1 mutant remained green much longer under ABA treatment.Quantitative RT-PCR analysis showed that ONAC016 upregulates the genes associated with chlorophyll degradation,senescence,and ABA signaling.Yeast one-hybrid and dual-luciferase assays revealed that ONAC016 binds directly to the promoter regions of OsNAP,a key gene involved in chlorophyll degradation and ABA-induced senescence.Taken together,these results suggest that ONAC016 plays an important role in promoting leaf senescence through the ABA signaling pathway involving OsNAP.

Leaf Senescence Characteristics and Responses to Water Shortage of Different Hybrid Rice Cultivars during Grain Filling Stage

[Objective] The pattern of leaves senescence was studied to provide reference for the planting technology and breeding rice cultivars resistant to premature senescence. [Method] Six hybrid rice cultivars were cultivated under limited and conventional irrigation managements; after the heading of rice, the root activities, nitrogen content in leaves, chlorophyll content （SPAD value）, net photosynthetic rate and the activities of superoxide dismutase （SOD）, peroxidase （POD）, catalase （CAT） were determined. [Result] The rhythms in attenuation of xylem sap flow rate, leaf nitrogen content, chlorophyll content （SPAD value） and net photosynthetic rate of different rice cultivars were significantly different in attenuation starting time, attenuation frequency and attenuation range. Physiological indexes of different cultivars responded inconsistently to water shortage. Extremely significantly positive correlations were found between the decline rates of chlorophyll content and leaf nitrogen content and xylem sap flow rate of roots, the photosynthetic rate and chlorophyll content. The changes in activities of SOD, CAT and POD were different, and responded to water shortage differently in different cultivars. The change dynamic of leaf nitrogen content and chlorophyll content was positively related with the activity dynamic of SOD and CAT. The change dynamic of leaf nitrogen content and chlorophyll content was negatively related with the malondialdehyde （MDA） content. The difference in the attenuation rhythm of physiological indexes of leaves and response to water shortage of different hybrid rice cultivars were related to genotype difference. Increasing roots activities and protecting enzyme activities of leaves benefit to defer attenuation of chlorophyll content and nitrogen content of leaves and to keep stronger photosynthetic function. [Conclusion] The senescence resistance of rice is a combined result of genotype differences and physiological response to environment.

Slight shading after anthesis increases photosynthetic productivity and grain yield of winter wheat (Triticum aestivum L.) due to the delaying of leaf senescence

The solar radiation intensity and duration are continuously decreasing in the major wheat planting area of China. As a con- sequence, leaf senescence, photosynthesis, grain filling and thus wheat yield shall be affected by light deficiency. Therefore, two winter wheat （Triticum aestivum L.） cultivars, Tainong 18 （a large-spike cultivar） and Ji＇nan 17 （a multiple-spike cultivar）, were subjected to shading during anthesis and maturity under field condition in 2010-2011 and 2011-2012. Under the slight shading treatment （$1,88% of full sunshine）, leaf senescence was delayed, net photosynthesis rate （Po） and canopy apparent photosynthesis rate （CAP） were improved, and thus thousand-kernel weight （TKW） and grain yield were higher as compared with the control. However, mid and severe shading （S2 andS3, 67 and 35% of full sunshine, respectively） led to negative effects on these traits substantially. Moreover, superoxide dismutase （SOD）, peroxidase （POD） and cat- alase （CAT） activities in flag leaf were significantly greater under slight shading than those in other treatments, while the malondialdehyde （MDA） content was less than that under other treatments. In addition, the multiple-spike cultivar is more tolerant to shading than large-spike cultivar. In conclusion, slight shading after anthesis delayed leaf senescence, enhanced photosynthesis and grain filling, and thus resulted in higher grain yield.

Effects of Aerated Irrigation on Leaf Senescence at Late Growth Stage and Grain Yield of Rice

With the japonica inbred cultivar Xiushui 09, indica hybrid combinations Guodao 6 and Liangyoupeijiu as materials, field experiments were conducted in 2007 and 2008 to study the effects of aerated irrigation on leaf senescence at late growth stage and grain yield of rice. The dissolved oxygen concentration of aerated water evidently increased and decreased at a slow rate. The soil oxidation-reduction potential under aerated irrigation treatment was significantly higher than that of the CK, contributing to significant increases in effective panicles, seed setting rate and grain yield. In addition, the aerated irrigation improved root function, increased superoxide dismutase activity and decreased malondialdehyde content in flag leaves at post-flowering, which delayed leaf senescence process, prolonged leaf functional activity and led to enhanced grain filling.

Exogenous abscisic acid coordinating leaf senescence and transport of assimilates into wheat grains under drought stress by regulating hormones homeostasis

Drought at the grain filling stage of wheat will cause premature leaf senescence, thus leading to considerable loss of wheat yield. Therefore, this paper aims to establish a cultivation technology for strong drought resistance, delayed senescence, and yield improvement based on the analysis of hormones homeostasis obtained by applying chemical control substances. Experiments were conducted with two genotypes of wheat. Four water irrigation treatments were applied to impose the water deficit, including well-watered control treatment(WW), mild water deficit(MiWD), moderate water deficit(MoWD), and severe water deficit(SWD). Exogenous abscisic acid(ABA) was sprayed on the plants at the anthesis stage of the wheat. As a result, exogenous ABA reduced initial senescence rate(r0), total duration of chlorophyll(Chltotal), rapid senescence phase(Chlloss), and the accumulated temperature at an inflection point(M) but improved the persistence phase(Chlper) of flag leaves under all of the four treatments. However, exogenous ABA produced inconsistent effects on photoassimilate relocation and grain weight under different treatments. It produced positive regulatory effects on grain weight under WW, MiWD, and MoWD treatments. On the one hand, spraying ABA during the persistence phase of flag leaves reduced the ratios of zeatin to gibberellin(Z/GA3), spermine to spermidine(Spm/Spd), and salicylic acid to ABA(SA/ABA),which prolonged active photosynthesis by stimulating high level of proline(Pro) and increased the activities of antioxidant enzymes, such as superoxide dismutase(SOD), peroxidase(POD), catalase(CAT), and ascorbate peroxidase(APX). Therefore, drought tolerance was enhanced, and more photosynthetic assimilates were accumulated. On the other hand, the rapid senescence phase and the transport rate of assimilates into grains were accelerated, resulting in higher grain weight, yield, and water use efficiency(WUE). However, under SWD treatment, exogenous ABA improved the ratio of SA/ABA, leading to low Pro content and low antioxidant enzyme activity of flag leaves in the rapid loss phase. Meanwhile,drought resistance declined and the transport duration of assimilates into grains was shortened, thus making photosynthetic assimilates redundant. Therefore, exogenous ABA can lead to the reduction in grain weight, yield, and WUE of wheat under SWD treatment.

Mapping a leaf senescence gene els1 by BSR-Seq in common wheat

Leaf senescence is normally the last stage of plant development. Early senescence of functional leaves significantly reduces the photosynthetic time and efficiency, seriously affecting grain yield and quality in wheat. Discovering genes responsible for early leaf senescence(els) are necessary for developing novel germplasms and cultivars with delayed leaf-senescence through molecular manipulation and marker assisted selection. In this study, we identified an early leaf senescence line M114 in a derivative of a wheat breeding population. Genetic analysis indicated that early leaf senescence in M114 is controlled by a single recessive gene, provisionally designated els1. By applying bulked segregant analysis and RNA-Seq(BSR-Seq), seven polymorphic markers linked to els1 were developed and the gene was located on chromosome arm 2 BS in a 1.5 c M genetic interval between markers WGGB303 and WGGB305. A co-segregating marker, WGGB302, provide a starting point for fine mapping and map-based cloning of els1.

Interactions between cytokinin and nitrogen contribute to grain mass in wheat cultivars by regulating the flag leaf senescence process

Premature senescence after anthesis reduces crop yields.Delaying leaf senescence could maintain photosynthetic activity for a longer period and lead to a higher photosynthetic rate.Recent studies have provided some insights into the interaction between cytokinin and nitrogen(N)in the regulation of plant development.In the present study,foliar application of exogenous 6-benzylaminopurine(6-BA)and lovastatin,an inhibitor of cytokinin synthesis,was combined with three N rates[0 kg ha^(-1)(low nitrogen,LN),240 kg ha^(-1)(normal nitrogen,NN),and 360 kg ha^(-1)(high nitrogen,HN)]in two wheat cultivars,Wennong 6(with a staygreen phenotype)and Jimai 20(with a non-staygreen phenotype).Flag leaf senescence was assessed using a Gompertz growth curve.Grain mass,dry matter accumulation and distribution,total N of flag leaf,and concentrations of zeatin riboside(ZR)and abscisic acid(ABA)were also used to evaluate the functional characteristics of flag leaves.Grain mass was negatively correlated with initial senescence rate(r_0)and duration of rapid chlorophyll loss(Chl_(loss)),whereas it was positively correlated with maximum senescence rate(r_(max)),average senescence rate(r_(aver)),persistence phase(Chl_(per)),total duration of flag leaf(Chl_(total))and inflection point cumulative temperature(M).Compared to Jimai 20,Wennong 6 had larger r_(aver),Chl_(per),and Chl_(total).The concentration of ZR was highest under the 6-BA×NN treatment,followed by the 6-BA×HN and 6-BA×LN treatments.However,the concentration of ABA showed the opposite trend.It was concluded that the staygreen phenotype Wennong 6 was associated with greater grain mass and altered cytokinin metabolism and could be classified as a functional staygreen type.Foliar application of 6-BA interacting with N at the NN level(240 kg ha^(-1))may be a beneficial strategy for improving grain yield of wheat by regulating endogenous hormones and the flag leaf senescence process.Increasing endogenous cytokinin promoted the transport of dry matter to grain.

A rice XANTHINE DEHYDROGENASE gene regulates leaf senescence and response to abiotic stresses

Xanthine dehydrogenase, a member of the molybdenum enzyme family, participates in purine metabolism and catalyzes the generation of ureides from xanthine and hypoxanthine. However, the mechanisms by which xanthine dehydrogenase affects rice growth and development are poorly understood. In the present study, we identified a mutant with early leaf senescence and reduced tillering that we named early senescence and less-tillering 1(esl1). Map-based cloning revealed that ESL1 encodes a xanthine dehydrogenase, and it was expressed in all tissues. Chlorophyll content was reduced and chloroplast maldevelopment was severe in the esl1 mutant. Mutation of ESL1 led to decreases in allantoin, allantoate, and ABA contents. Further analysis revealed that the accumulation of reactive oxygen species in esl1 resulted in decreased photosynthesis and impaired chloroplast development, along with increased sensitivity to abscisic acid and abiotic stresses. Ttranscriptome analysis showed that the ESL1 mutation altered the expression of genes involved in the photosynthesis process and reactive oxygen species metabolism.Our results suggest that ESL1 is involved in purine metabolism and the induction of leaf senescence.These findings reveal novel molecular mechanisms of ESL1 gene-mediated plant growth and leaf senescence.

Yield, Leaf Senescence, and Cry1Ac Expression in Response to Removal of Early Fruiting Branches in Transgenic Bt Cotton

Two-year field experiments were conducted at Linqing, Yellow River valley of China, to study the plant response to the removal of early fruiting branches in transgenic Bt （Bacillus thuringiensis） cotton （Gossypium hirsutum L.） from 2003 to 2004. Plants were undamaged and treated by removing two basal fruiting branches （FB） at squaring to form the control and the removal treatment, respectively. The plant height, leaf area （LA）, dry weight of fruiting forms （DWFF）, the number of fruiting nodes （NFN）, photosynthetic （Pn） rate, and levels of leaf chlorophyll （Chl）, N, P, K, and Cry lAc protein in main- stem leaves were measured at a 10- or 20-d interval after FB removal, and the sink/source ratio as indicated by NFN/LA and DWFF/LA was determined. FB removal significantly increased the plant height, LA, and plant biomass in both years. Lint yields were increased 7.5 and 5.2% by removal compared with their controls in 2003 and 2004, respectively. Significant increases in boll size （5.7 and 5.1%） were also observed in removal than in control for both years. Either NFN/LA or DWFF/LA was significantly reduced by removal before 40 d after removal; however, both NFN/LA and DWFF/LA were significantly enhanced by FB removal at 80 d after removal compared to the untreated control. There was no significant difference in fiber quality in the first two harvests between removal and control, but fiber strength and micronarie in the third harvest were significantly improved by FB removal. In terms of leaf Chl, Pn rate, levels of total N, P, and K in late season, leaf senescence was considerably delayed by FB removal. Levels of CrylAc protein in the fully expanded young leaves were considerably higher in FB-excised plants than in control, indicating FB removal enhanced CrylAc expression. It is suggested that the yield and quality improvement with FB removal may be attributed to the increased NFN/LA or DWFF/LA in late season and delayed leaf senescence, respectively. FB removal can be a potential practice incorporated into the intensive cultivation system for enhancing transgenic Bt cotton production.

The Changes of Photosynthetic Properties and Cell Microstructure in Peanut Leaves during Leaf Senescence

The changes of photosynthetic properties and cell microstructure in peanut leaves during leaf senescence were studied with two high-yielding peanut cultivars (cv. Luhuall and Fu8707). The main results showed that during the whole process of leaf growth and senescence, changes in the photosynthesis rate (Pn) and contents of chlorophyll in leaves, could be described with a parabolic function, y = A + Bx + Cx2 (where y refers to the values of the above parameters and x to the days after leaf unfolding). During peanut leaf senescence , the shape of chloroplast changed gradually from long ellipses to circles.The starch globule in chloroplast altered gradually from more and larger sizes to fewer and smaller, but the oil globule from fewer and smaller to more and larger. The grana lamellae varied progressively: from thinness and length to thickness and shortness; from ranking along the long axle direction of chloroplast to disorderly arrangment and finally blurring. At last, the membrane envelope of chloroplast broke, so the inclusion seeped out to the cell and the chloroplast broke up.

AtWRKY75 positively regulates age-triggered leaf senescence through gibberellin pathway

WRKY transcription factors play essential roles during leaf senescence.However,the mechanisms by which they regulate this process remains largely unknown.Here,we identified the transcription factor WRKY75 as a positive regulator during leaf senescence.Mutations of WRKY75 caused a delay in agetriggered leaf senescence,whereas overexpression of WRKY75 markedly accelerated this process.Expression of senescence-associated genes(SAGs)was suppressed in WRKY75 mutants but increased in WRKY75-overexpressing plants.Further analysis demonstrated that WRKY75 directly associates with the promoters of SAG12 and SAG29,to activate their expression.Conversely,GAI and RGL1,two DELLA proteins,can suppress the WRKY75-mediated activation,thereby attenuating SAG expression during leaf senescence.Genetic analyses showed that GAI gain-of-function or RGL1 overexpression can partially rescue the accelerated senescence phenotype caused by WRKY75 overexpression.Furthermore,WRKY75 can positively regulate WRKY45 expression during leaf senescence.Our data thus imply that WRKY75 may positively modulate age-triggered leaf senescence through the gibberellin-mediated signaling pathway。

Effects of 4PU-30 on leaf senescence and degration of protein and nucleic acid in rice

4PU—30[N—phenyl—’N—(2—chloro—4—pyridyl) urea] is a new type of plant growth regulator with cytokinin properties. It has been confirmed to delay rice leaf senescence effectively. In order to elucidate the physiological role of 4PU—30 in delaying senescence, the changes of protein, nucleic acid contents, and the related activities of degradative enzymes were studied. Shanyou 63, an indica hybrid rice was used for this experiment. In the in vitro experiment, two full—developed leaves from the top during heading stage were collected and cut into 5.0cm segments, They were floated on the surface of distilled water containing 0.1mg/14PU—30 and incubated in darkness at 30 C. The leaves floated on distilled water were used as control.It was observed that chlorophyll content in controlled leaves declined rapidly started from the second day and dropped by 93.4% on the 6th day while that in leaves treated with 4PU—30 declined by 41.4% only. During senescence, specific activities of hemoglobin—digesting

Effects of a GA3/4PU-30 compound (90-09) on leaf senescence of hybrid rice

90-09 is a cmpound made with GA3,4PU-30and some trace elements(B ete.).To studythe effects of 90-09 on the leaf senescence ofhybrid rice leaf,the changes of activities ofsome degradation enzyme and the contents ofendogenous hormones(ABA,ZRs,GAs,andIAA),protein,and nucleic acids were mea-sured during leaf senescence. Shanyou 63,an indica hybrid rice,wasused in this experiment.Rice plants were firstsprayed with 90-09(115 ml/hm~2) solution 10d after heading,followed by two more sprayswith once a week.The controlled plants weresprayed with water only.

Functional divergences of natural variations of TaNAM-A1 in controlling leaf senescence during wheat grain filling

Leaf senescence is an essential physiological process related to grain yield potential and nutritional quality.Green leaf duration(GLD)after anthesis directly reflects the leaf senescence process and exhibits large genotypic differences in common wheat;however,the underlying gene regulatory mechanism is still lacking.Here,we identified TaNAM-A1 as the causal gene of the major loci q GLD-6A for GLD during grain filling by map-based cloning.Transgenic assays and TILLING mutant analyses demonstrated that TaNAM-A1 played a critical role in regulating leaf senescence,and also affected spike length and grain size.Furthermore,the functional divergences among the three haplotypes of TaNAM-A1 were systematically evaluated.Wheat varieties with TaNAM-A1d(containing two mutations in the coding DNA sequence of TaNAM-A1)exhibited a longer GLD and superior yield-related traits compared to those with the wild type TaNAM-A1a.All three haplotypes were functional in activating the expression of genes involved in macromolecule degradation and mineral nutrient remobilization,with TaNAM-A1a showing the strongest activity and TaNAM-A1d the weakest.TaNAM-A1 also modulated the expression of the senescencerelated transcription factors TaNAC-S-7A and TaNAC016-3A.TaNAC016-3A enhanced the transcriptional activation ability of TaNAM-A1a by protein–protein interaction,thereby promoting the senescence process.Our study offers new insights into the fine-tuning of the leaf functional period and grain yield formation for wheat breeding under various geographical climatic conditions.

Arabidopsis WRKY1 promotes monocarpic senescence by integrative regulation of flowering,leaf senescence,and nitrogen remobilization

Monocarpic senescence,characterized by whole-plant senescence following a single flowering phase,is widespread in seed plants,particularly in crops,determining seed harvest time and quality.However,how external and internal signals are systemically integrated into monocarpic senescence remains largely unknown.Here,we report that the Arabidopsis thaliana transcription factor WRKY1 plays essential roles in multiple key steps of monocarpic senescence.WRKY1 expression is induced by age,salicylic acid(SA),and nitrogen(N)deficiency.Flowering and leaf senescence are accelerated in the WRKY1 overexpression lines but are delayed in the wrky1 mutants.The combined DNA affinity purification sequencing and RNA sequencing analyses uncover the direct target genes of WRKY1.Further studies show that WRKY1 coordinately regulates three processes in monocarpic senescence:(1)suppressing FLOWERING LOCUS C gene expression to initiate flowering,(2)inducing SA biosynthesis genes to promote leaf senescence,and(3)activating the N assimilation and transport genes to trigger N remobilization.In summary,our study reveals how one stress-responsive transcription factor,WRKY1,integrates flowering,leaf senescence,and N remobilization processes into monocarpic senescence,providing important insights into plant lifetime regulation.

AtVQ25 promotes salicylic acid-related leaf senescence by fine-tuning the self-repression of AtWRKY53

Most mechanistic details of chronologically ordered regulation of leaf senescence are unknown.Regulatory networks centered on AtWRKY53 are crucial for orchestrating and integrating various senescence-related signals.Notably,AtWRKY53binds to its own promoter and represses transcription of AtWRKY53,but the biological significance and mechanism underlying this selfrepression remain unclear.In this study,we identified the VQ motif-containing protein AtVQ25as a cooperator of AtWRKY53.The expression level of AtVQ25 peaked at mature stage and was specifically repressed after the onset of leaf senescence.AtVQ25-overexpressing plants and atvq25 mutants displayed precocious and delayed leaf senescence,respectively.Importantly,we identified AtWRKY53 as an interacting partner of AtVQ25.We determined that interaction between AtVQ25 and AtWRKY53 prevented AtWRKY53from binding to W-box elements on the AtWRKY53promoter and thus counteracted the selfrepression of AtWRKY53.In addition,our RNA-sequencing data revealed that the AtVQ25-AtWRKY53 module is related to the salicylic acid(SA)pathway.Precocious leaf senescence and SA-induced leaf senescence in AtVQ25-overexpressing lines were inhibited by an SA pathway mutant,atsid2,and Nah G transgenic plants;AtVQ25-overexpressing/atwrky53 plants were also insensitive to SA-induced leaf senescence.Collectively,we demonstrated that AtVQ25 directly attenuates the self-repression of AtWRKY53 during the onset of leaf senescence,which is substantially helpful for understanding the timing of leaf senescence onset modulated by AtWRKY53.

Defect in an immune regulator gene BrSRFR1 leads to premature leaf senescence in Chinese cabbage

Leaf senescence is the final stage of leaf development, where the nutrients and energy of senescent leaves are redistributed to developing tissues or organs for plant growth, reproduction, and defense. Outer leaves are photosynthetic organs that usually senesce at the late heading stage in Chinese cabbage, and premature leaf senescence often reduces leafy head yield and quality. In this study, 11 premature leaf senescence mutants were screened from an ethyl methanesulfonate-mutagenized population of the double haploid line ‘FT' in Chinese cabbage. At the early heading stage, the mutants exhibited edge yellowing within its outer leaves, and at the mature stage, its leafy head weight decreased significantly. Genetic analysis revealed that the mutated trait of all 11 mutants corresponds to single gene recessive inheritance. Semi-diallel cross tests showed that 5 of the 11 were allelic mutants. MutMap and Kompetitive Allele Specific PCR genotyping revealed that BraA01g001400.3C was the candidate gene, which is orthologous of Arabidopsis SUPPRESSOR OF rps4-RLD 1, encoding an immune regulator, so we named it as BrSRFR1. All the BrSRFR1 in the five allelic mutants exhibited single nucleotide polymorphisms at different positions on their exons and led to premature translation termination, which confirmed that defect in BrSRFR1 led to premature leaf senescence. These results verify the role of Br SRFR1 on leaf senescence and provide a new insight into the mechanisms of leaf senescence in Chinese cabbage, which reveals a novel function of SRFR1 in plant development.

A molecular framework underlying low-nitrogeninduced early leaf senescence in Arabidopsis thaliana

Nitrogen(N)deficiency causes early leaf senescence,resulting in accelerated whole-plant maturation and severely reduced crop yield.However,the molecular mechanisms underlying N-deficiency-induced early leaf senescence remain unclear,even in the model species Arabidopsis thaliana.In this study,we identified Growth,Development and Splicing 1(GDS1),a previously reported transcription factor,as a new regulator of nitrate(NO3)signaling by a yeast-one-hybrid screen using a NO3enhancer fragment from the promoter of NRT2.1.We showed that GDS1 promotes NO3 signaling,absorption and assimilation by affecting the expression of multiple NO3 regulatory genes,including Nitrate Regulatory Gene2(NRG2).Interestingly,we observedthat gds1mutants show early leaf senescence as well as reduced NO3-contentand Nuptake under N-deficient conditions.Further analyses indicated that GDS1 binds to the promoters of several senescence-related genes,including Phytochrome-lnteracting Transcription Factors 4 and 5(PIF4 and PIF5)and represses their expression.Interestingly,we found that N deficiency decreases GDS1 protein accumulation,and GDS1 could interact with Anaphase Promoting Complex Subunit 10(APC10).Genetic and biochemical experiments demonstrated that Anaphase Promoting Complex or Cyclosome(APC/C)promotes the ubiquitination and degradation of GDS1 under N deficiency,resulting in loss of PIF4 and PiF5 repression and consequent early leaf senescence.Furthermore,we discovered that overexpression of GDS1 could delay leaf senescence and improve seed yield and N-use efficiency(NUE)in Arabidopsis.In summary,our study uncovers a molecular framework illustrating a new mechanism underlying low-N-induced early leaf senescence and provides potential targets for genetic improvement of crop varieties with increased yield and NUE.