Organ senescence is an important developmental process in plants that enables recycling of nutrients, such as nitrogen, to maximize reproductive success. Nitrogen is the mineral nutrient required in greatest amount by plants, although soil-N limits plant productivity in many natural and agricultural systems, especially systems that receive little or no fertilizer-N. Use of industrial N-fertilizers in agriculture increased crop yields several fold over the past century, although at substantial cost to fossil energy reserves and the environment. Therefore, it is important to optimize nitrogen use efficiency (NUE) in agricultural systems. Organ senescence contributes to NUE in plants and manipulation of senescence in plant breeding programs is a promising approach to improve NUE in agriculture. Much of what we know about plant senescence comes from research on annual plants, which provide most of the food for humans. Relatively little work has been done on senescence in perennial plants, especially perennial grasses, which provide much of the forage for grazing animals and promise to supply much of the biomass required by the future biofuel industry. Here, we review briefly what is known about senescence from studies of annual plants, before presenting current knowledge about senescence in perennial grasses and its relationship to yield, quality, and NUE. While higher yield is a common target, desired N-content diverges between forage and biofuel crops. We discuss how senescence programs might be altered to produce high-yielding, stress-tolerant perennial grasses with high-N (protein) for forage or low-N for biofuels in systems optimized for NUE.© The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Crop productivity relies heavily on nitrogen (N) fertilization. Production and application of N fertilizers consume huge amounts of energy, and excess is detrimental to the environment; therefore, increasing plant N use efficiency (NUE) is essential for the development of sustainable agriculture. Plant NUE is inherently complex, as each step-including N uptake, translocation, assimilation, and remobilization-is governed by multiple interacting genetic and environmental factors. The limiting factors in plant metabolism for maximizing NUE are different at high and low N supplies, indicating great potential for improving the NUE of current cultivars, which were bred in well-fertilized soil. Decreasing environmental losses and increasing the productivity of crop-acquired N requires the coordination of carbohydrate and N metabolism to give high yields. Increasing both the grain and N harvest index to drive N acquisition and utilization are important approaches for breeding future high-NUE cultivars.

AtAMT2 is an ammonium transporter that is only distantly related to the five members of the AtAMT1 family of high-affinity ammonium transporters in Arabidopsis. The short-lived radioactive ion13NH4  + was used to show that AtAMT2, expressed in yeast (Saccharomyces cerevisiae), is a high-affinity transporter with a K  m for ammonium of about 20 μm. Changes in external pH between 5.0 and 7.5 had little effect on the K  m for ammonium, indicating that NH4  +, not NH3, is the substrate for AtAMT2. TheAtAMT2 gene was expressed in all organs of Arabidopsis and was subject to nitrogen (N) regulation, at least in roots where expression was partially repressed by high concentrations of ammonium nitrate and derepressed in the absence of external N. Although expression of AtAMT2 in shoots responded little to changes in root N status, transcript levels in leaves declined under high CO2 conditions. Transient expression of an AtAMT2-green fluorescent protein fusion protein in Arabidopsis leaf epidermal cells indicated a plasma membrane location for the AtAMT2 protein. Thus, AtAMT2 is likely to play a significant role in moving ammonium between the apoplast and symplast of cells throughout the plant. However, a dramatic reduction in the level ofAtAMT2 transcript brought about by dsRNA interference with gene expression had no obvious effect on plant growth or development, under the conditions tested.

The AMMONIUM TRANSPORTER (AMT) family comprises six isoforms in Arabidopsis thaliana. Here, we describe the complete functional organization of root-expressed AMTs for high-affinity ammonium uptake. High-affinity influx of 15N-labeled ammonium in two transposon-tagged amt1;2 lines was reduced by 18 to 26% compared with wild-type plants. Enrichment of the AMT1;2 protein in the plasma membrane and localization of AMT1;2 promoter activity in the endodermis and root cortex indicated that AMT1;2 mediates the uptake of ammonium entering the root via the apoplasmic transport route. An amt1;1 amt1;2 amt1;3 amt2;1 quadruple mutant (qko) showed severe growth depression under ammonium supply and maintained only 5 to 10% of wild-type high-affinity ammonium uptake capacity. Transcriptional upregulation of AMT1;5 in nitrogen-deficient rhizodermal and root hair cells and the ability of AMT1;5 to transport ammonium in yeast suggested that AMT1;5 accounts for the remaining uptake capacity in qko. Triple and quadruple amt insertion lines revealed in vivo ammonium substrate affinities of 50, 234, 61, and 4.5 μM for AMT1;1, AMT1;2, AMT1;3, and AMT1;5, respectively, but no ammonium influx activity for AMT2;1. These data suggest that two principle means of achieving effective ammonium uptake in Arabidopsis roots are the spatial arrangement of AMT1-type ammonium transporters and the distribution of their transport capacities at different substrate affinities.

Contents Summary 35 I. Introduction 35 II. Nitrogen acquisition and assimilation 36 III. Root-to-shoot transport of nitrogen 38 IV. Nitrogen storage pools in vegetative tissues 39 V. Nitrogen transport from source leaf to sink 40 VI. Nitrogen import into sinks 42 VII. Relationship between source and sink nitrogen transport processes and metabolism 43 VIII. Regulation of nitrogen transport 43 IX. Strategies for crop improvement 44 X. Conclusions 46 Acknowledgements 47 References 47 SUMMARY: Nitrogen is an essential nutrient for plant growth. World-wide, large quantities of nitrogenous fertilizer are applied to ensure maximum crop productivity. However, nitrogen fertilizer application is expensive and negatively affects the environment, and subsequently human health. A strategy to address this problem is the development of crops that are efficient in acquiring and using nitrogen and that can achieve high seed yields with reduced nitrogen input. This review integrates the current knowledge regarding inorganic and organic nitrogen management at the whole-plant level, spanning from nitrogen uptake to remobilization and utilization in source and sink organs. Plant partitioning and transient storage of inorganic and organic nitrogen forms are evaluated, as is how they affect nitrogen availability, metabolism and mobilization. Essential functions of nitrogen transporters in source and sink organs and their importance in regulating nitrogen movement in support of metabolism, and vegetative and reproductive growth are assessed. Finally, we discuss recent advances in plant engineering, demonstrating that nitrogen transporters are effective targets to improve crop productivity and nitrogen use efficiency. While inorganic and organic nitrogen transporters were examined separately in these studies, they provide valuable clues about how to successfully combine approaches for future crop engineering.© 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

In the majority of agricultural growing regions, crop production is highly dependent on the supply of exogenous nitrogen (N) fertilizers. Traditionally, this dependency and the use of N‐fertilizers to restore N depleted soils has been rewarded with increased plant health and yields. In recent years, increased competition for non‐renewable fossil fuel reserves has directly elevated prices of N‐fertilizers and the cost of agricultural production worldwide. Furthermore, N‐fertilizer based pollution is becoming a serious issue for many regions where agriculture is highly concentrated. To help minimize the N footprint associated with agricultural production there is significant interest at the plant level to develop technologies which can allow economically viable production while using less applied N. To complement recent reviews examining N utilization efficiency in agricultural plants, this review will explore those strategies operating specifically at the root level, which may directly contribute to improved N use efficiencies in agricultural crops such as cereals, where the majority of N‐fertilizers are used and lost to the environment. Root specific phenotypes that will be addressed in the context of improvements to N acquisition and assimilation efficiencies include: root morphology; root to shoot ratios; root vigour, root length density; and root N transport and metabolism.

Amino acids are not only a nitrogen source that can be directly absorbed by plants, but also the major transport form of organic nitrogen in plants. A large number of amino acid transporters have been identified in different plant species. Despite belonging to different families, these amino acid transporters usually exhibit some general features, such as broad expression pattern and substrate selectivity. This review mainly focuses on transporters involved in amino acid uptake, phloem loading and unloading, xylem-phloem transfer, import into seed and intracellular transport in plants. We summarize the other physiological roles mediated by amino acid transporters, including development regulation, abiotic stress tolerance and defense response. Finally, we discuss the potential applications of amino acid transporters for crop genetic improvement.

Drought stress is an important environmental factor limiting plant productivity. In this study, we screened drought-resistant transgenic plants from 65 promoter-pyrabactin resistance 1-like (PYL) abscisic acid (ABA) receptor gene combinations and discovered that pRD29A::PYL9 transgenic lines showed dramatically increased drought resistance and drought-induced leaf senescence in both Arabidopsis and rice. Previous studies suggested that ABA promotes senescence by causing ethylene production. However, we found that ABA promotes leaf senescence in an ethylene-independent manner by activating sucrose nonfermenting 1-related protein kinase 2s (SnRK2s), which subsequently phosphorylate ABA-responsive element-binding factors (ABFs) and Related to ABA-Insensitive 3/VP1 (RAV1) transcription factors. The phosphorylated ABFs and RAV1 up-regulate the expression of senescence-associated genes, partly by up-regulating the expression of Oresara 1. The pyl9 and ABA-insensitive 1-1 single mutants, pyl8-1pyl9 double mutant, and snrk2.2/3/6 triple mutant showed reduced ABA-induced leaf senescence relative to the WT, whereas pRD29A::PYL9 transgenic plants showed enhanced ABA-induced leaf senescence. We found that leaf senescence may benefit drought resistance by helping to generate an osmotic potential gradient, which is increased in pRD29A::PYL9 transgenic plants and causes water to preferentially flow to developing tissues. Our results uncover the molecular mechanism of ABA-induced leaf senescence and suggest an important role of PYL9 and leaf senescence in promoting resistance to extreme drought stress.

Leaf senescence is an important developmental process involving orderly disassembly of macromolecules for relocating nutrients from leaves to other organs and is critical for plants’ fitness. Leaf senescence is the response of an intricate integration of various environmental signals and leaf age information and involves a complex and highly regulated process with the coordinated actions of multiple pathways. Impressive progress has been made in understanding how senescence signals are perceived and processed, how the orderly degeneration process is regulated, how the senescence program interacts with environmental signals, and how senescence regulatory genes contribute to plant productivity and fitness. Employment of systems approaches using omics-based technologies and characterization of key regulators have been fruitful in providing newly emerging regulatory mechanisms. This review mainly discusses recent advances in systems understanding of leaf senescence from a molecular network dynamics perspective. Genetic strategies for improving the productivity and quality of crops are also described.

It has long been established that premature leaf senescence negatively impacts the yield stability of rice, but the underlying molecular mechanism driving this relationship remains largely unknown. Here, we identified a dominant premature leaf senescence mutant, prematurely senile 1 (ps1-D). PS1 encodes a plant-specific NAC (no apical meristem, Arabidopsis ATAF1/2, and cup-shaped cotyledon2) transcriptional activator, Oryza sativa NAC-like, activated by apetala3/pistillata (OsNAP). Overexpression of OsNAP significantly promoted senescence, whereas knockdown of OsNAP produced a marked delay of senescence, confirming the role of this gene in the development of rice senescence. OsNAP expression was tightly linked with the onset of leaf senescence in an age-dependent manner. Similarly, ChIP-PCR and yeast one-hybrid assays demonstrated that OsNAP positively regulates leaf senescence by directly targeting genes related to chlorophyll degradation and nutrient transport and other genes associated with senescence, suggesting that OsNAP is an ideal marker of senescence onset in rice. Further analysis determined that OsNAP is induced specifically by abscisic acid (ABA), whereas its expression is repressed in both aba1 and aba2, two ABA biosynthetic mutants. Moreover, ABA content is reduced significantly in ps1-D mutants, indicating a feedback repression of OsNAP on ABA biosynthesis. Our data suggest that OsNAP serves as an important link between ABA and leaf senescence. Additionally, reduced OsNAP expression leads to delayed leaf senescence and an extended grain-filling period, resulting in a 6.3% and 10.3% increase in the grain yield of two independent representative RNAi lines, respectively. Thus, fine-tuning OsNAP expression should be a useful strategy for improving rice yield in the future.

Leaf senescence is a very important trait that limits yield and biomass accumulation of agronomic crops and reduces post-harvest performance and the nutritional value of horticultural crops. Significant advance in physiological and molecular understanding of leaf senescence has made it possible to devise ways of manipulating leaf senescence for agricultural improvement. There are three major strategies in this regard: (i) plant hormone biology-based leaf senescence manipulation technology, the senescence-specific gene promoter-directed IPT system in particular; (ii) leaf senescence-specific transcription factor biology-based technology; and (iii) translation initiation factor biology-based technology. Among the first strategy, the P SAG12 -IPT autoregulatory senescence inhibition system has been widely explored and successfully used in a variety of plant species for manipulating senescence. The vast majority of the related research articles (more than 2000) showed that crops harbouring the autoregulatory system displayed a significant delay in leaf senescence without any abnormalities in growth and development, a marked increase in grain yield and biomass, dramatic improvement in horticultural performance, and/or enhanced tolerance to drought stress. This technology is approaching commercialization. The transcription factor biology-based and translation initiation factor biology-based technologies have also been shown to be very promising and have great potentials for manipulating leaf senescence in crops. Finally, it is speculated that technologies based on the molecular understanding of nutrient recycling during leaf senescence are highly desirable and are expected to be developed in future translational leaf senescence research. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

为了解针太行山地区针、阔叶森林凋落物分解特征，以刺槐和侧柏人工林为对象，采用样地调查法，对其凋落物养分归还和营养元素周转进行了研究，结果表明：侧柏和刺槐林年凋落总量分别为7494.71和5372.34kg/hm²；凋落物中落叶比重最大。为减少养分流失，阔叶林树刺槐的N、P养分回流现象明显，其中N回流率约为34%~53%，P回流率约为28%~56%；而针叶树侧柏的养分回流现象不明显。森林凋落物3种元素含量以Ca最高，其次是N，含量最低是P；阔叶林凋落物中N、P含量明显高于针叶林。凋落物N、P迁移符合富集—释放模式；Ca为直接释放类型。刺槐凋落物分解速率明显快于侧柏，其完全分解时间分别为7.7和12.8a。侧柏在养分年归还总量、Ca的年释放率以及P、Ca的周转时间要优于刺槐，而刺槐在N、P年释放率以及N周转时间比侧柏有优势。

为了解针太行山地区针、阔叶森林凋落物分解特征，以刺槐和侧柏人工林为对象，采用样地调查法，对其凋落物养分归还和营养元素周转进行了研究，结果表明：侧柏和刺槐林年凋落总量分别为7494.71和5372.34kg/hm²；凋落物中落叶比重最大。为减少养分流失，阔叶林树刺槐的N、P养分回流现象明显，其中N回流率约为34%~53%，P回流率约为28%~56%；而针叶树侧柏的养分回流现象不明显。森林凋落物3种元素含量以Ca最高，其次是N，含量最低是P；阔叶林凋落物中N、P含量明显高于针叶林。凋落物N、P迁移符合富集—释放模式；Ca为直接释放类型。刺槐凋落物分解速率明显快于侧柏，其完全分解时间分别为7.7和12.8a。侧柏在养分年归还总量、Ca的年释放率以及P、Ca的周转时间要优于刺槐，而刺槐在N、P年释放率以及N周转时间比侧柏有优势。

Aim  Nutrient resorption from senescing leaves is an important mechanism of nutrient conservation in plants, but the patterns of nutrient resorption at the global scale are unknown. Because soil nutrients vary along climatic gradients, we hypothesize that nutrient resorption changes with latitude, temperature and precipitation.

Studies of small trees growing in pots have established that individual amino acids or amides are translocated in the xylem sap of a range of tree species following bud burst, as a consequence of nitrogen (N) remobilization from storage. This paper reports the first study of N translocation in the xylem of large, deciduous, field-grown trees during N remobilization in the spring. We applied 15N fertilizer to the soil around 10-year-old Prunus avium L. and Populus trichocharpa Torr. & Gray ex Hook var. Hastata (Dode) A. Henry x Populus balsamifera L. var. Michauxii (Dode) Farwell trees before bud burst to label N taken up by the roots. Recovery of unlabeled N in xylem sap and leaves was used to demonstrate that P. avium remobilizes N in both glutamine (Gln) and asparagine (Asn). Sap concentrations of both amides rose sharply after bud burst, peaking 14 days after bud burst for Gln, and remaining high some 45 days for Asn. There was no 15N enrichment of either amide until 21 days after bud burst. In the Populus trees, nearly all the N was translocated in the sap as Gln, the concentration of which peaked and then declined before the amide was enriched with 15N, 40 days after bud burst. Xylem sap of clonal P. avium trees was sampled at different positions in the crown to assess if the amino acid and amide composition of the sap varied within the crown. Sap was sampled during remobilization (when the concentration of Gln was maximal), at the end of remobilization and at the end of the experiment (68 days after bud burst). Although the date of sampling had a highly significant effect on sap composition, the effect of position of sampling was marginal. The results are discussed in relation to N translocation in adult trees and the possibility of measuring N remobilization by calculating the flux of N translocation in the xylem.

以6年生烟富3/SH6/平邑甜茶为试材,用整株破坏性解析的方法,研究了萌芽期至果实成熟期7个时期下的树体生长和氮素积累动态,并借助¹⁵N同位素示踪技术研究了树体对肥料氮的吸收利用和分配特性,以期阐明苹果树的氮积累动态和肥料氮的最大效率期,从而为科学施氮提供理论依据.结果表明: 萌芽期(3月25日)至果实成熟期(萌芽后210 d)红富士苹果幼树整株干物质净积累量为4.51 kg,其中果实占66.5%,叶梢(叶片与新梢,下同)占20.2%,多年生器官占13.3%;叶梢干物质积累量在萌芽后30～60 d增长幅度较大,占其整个处理时期的42.9%;果实干物质积累量在萌芽后120～180 d增长幅度大,占整个处理时期的70%.整株氮素净积累量为29.1 g,在萌芽后30～60 d和120～180 d增长较快,分别为7.2和12.8 g,占整个处理时期的24.7%和44%;叶梢在萌芽后0～60 d氮积累速率较快,占其整个时期的69.1%;果实的氮积累量在萌芽后120～180 d最快,占其整个时期的60.8%;多年生器官的氮积累量在处理周期内呈先下降后上升的趋势,并在萌芽后 60 d到达最低水平.树体在不同时期的¹⁵N利用率差异显著,分别在萌芽后30～60、120～150和150～180 d处于较高水平,¹⁵N利用率分别为2.3%、4.1%和4.0%;多年生器官在各个时期的¹⁵N分配率均呈现较高水平,新生器官的¹⁵N分配率均为先上升后下降的趋势,其中叶片新梢在萌芽后30～60 d达到最高水平,为38.4%;果实在萌芽后120～150 d和150～180 d到达最高水平,分别为15.0%和16.6%.因此,叶片和新梢氮素积累的关键时期为萌芽后30～60 d;果实氮素积累的关键时期为萌芽后120～180 d;树体对肥料氮的最大效率期为萌芽后30～60 d和120～180 d.

Taking 6-year-old Yanfu3/SH6/<i>Malus hupehensis</i> Rehd. as the test material, the dynamics of plant growth and nitrogen (N) accumulation under seven periods from germination stage to fruit maturity stage were examined by destructive analysis. The absorption, utilization, and distribution of fertilizer N were studied by ¹⁵N isotope tracer technique to clarify the N accumulation dynamics of apple trees and the maximum efficiency period of fertilizer N, and to provide theoretical basis for scientific application of N fertilizer. The results showed that the net accumulation of dry matter was 4.51 kg in germination stage (March 25) to fruit maturity stage (210 d after budbreak), with fruit accounting for 66.5%, the leaves and new shoots accounting for 20.2%, and the perennial organs accounting for 13.3%. The dry matter accumulation in 30-60 d after budbreak was the fastest, accounting for 42.9% of the whole treatment period. The fruit dry matter accumulation in 120-180 d after budbreak was the fastest, which accounted for 70% of the whole treatment period. The total N accumulation of the plant was 29.1 g, which increased rapidly in the 30-60 d and 120-180 d after budbreak by 7.2 g and 12.8 g, respectively accounting for 24.7% and 44% of the whole treatment period. The N accumulation of leaves and new shoots was the fastest in 0-60 d after budbreak, which accounted for 69.1% of the whole period. The N accumulation of fruit was the fastest in 120-180 d after budbreak, accounting for 60.8% of the whole period. The N accumulation of the perennial organ decreased first and then increased, and reached the lowest level at 60 d after budbreak. The ¹⁵N utilization rate of plant differed significantly in different periods which was at a high level in 30-60 d,120-150 d and 150-180 d after budbreak with 2.3%, 4.1% and 4.0% respectively. The ¹⁵N distribution rate in perennial organs in each period showed a high level, that of the new born organ increased first and then decreased which reached the highest level of 38.4% in 30-60 d after budbreak. The fruit reached the highest in 120-150 d and 150-180 d after budbreak by 15.0% and 16.6% respectively. Therefore, the key period of N accumulation in leaves and shoots was 30-60 d after budbreak, and that in fruit was 120-180 d after budbreak. The period with maximum efficiency for fertilizer N was at 30-60 d and 120-180 d after budbreak.

Plant senescence is a natural phenomenon known for the appearance of beautiful autumn colors and the ripening of cereals in the field. Senescence is a controlled process that plants utilize to remobilize nutrients from source leaves to developing tissues. While during the past decades, molecular components underlying the onset of senescence have been intensively studied, knowledge remains scarce on the age-dependent mechanisms that control the onset of senescence. Recent advances have uncovered transcriptional networks regulating the competence to senesce. Here, gene regulatory networks acting as internal timing mechanisms for the onset of senescence are highlighted, illustrating that early and late leaf developmental phases are highly connected. Copyright © 2015 Elsevier Ltd. All rights reserved.

Leaf senescence constitutes the final stage of leaf development and is critical for plants’ fitness as nutrient relocation from leaves to reproducing seeds is achieved through this process. Leaf senescence involves a coordinated action at the cellular, tissue, organ, and organism levels under the control of a highly regulated genetic program. Major breakthroughs in the molecular understanding of leaf senescence were achieved through characterization of various senescence mutants and senescence-associated genes, which revealed the nature of regulatory factors and a highly complex molecular regulatory network underlying leaf senescence. The genetically identified regulatory factors include transcription regulators, receptors and signaling components for hormones and stress responses, and regulators of metabolism. Key issues still need to be elucidated, including cellular-level analysis of senescence-associated cell death, the mechanism of coordination among cellular-, organ-, and organism-level senescence, the integration mechanism of various senescence-affecting signals, and the nature and control of leaf age.

Two cDNA libraries were prepared, one from leaves of a field-grown aspen (Populus tremula) tree, harvested just before any visible sign of leaf senescence in the autumn, and one from young but fully expanded leaves of greenhouse-grown aspen (Populus tremula x tremuloides). Expressed sequence tags (ESTs; 5,128 and 4,841, respectively) were obtained from the two libraries. A semiautomatic method of annotation and functional classification of the ESTs, according to a modified Munich Institute of Protein Sequences classification scheme, was developed, utilizing information from three different databases. The patterns of gene expression in the two libraries were strikingly different. In the autumn leaf library, ESTs encoding metallothionein, early light-inducible proteins, and cysteine proteases were most abundant. Clones encoding other proteases and proteins involved in respiration and breakdown of lipids and pigments, as well as stress-related genes, were also well represented. We identified homologs to many known senescence-associated genes, as well as seven different genes encoding cysteine proteases, two encoding aspartic proteases, five encoding metallothioneins, and 35 additional genes that were up-regulated in autumn leaves. We also indirectly estimated the rate of plastid protein synthesis in the autumn leaves to be less that 10% of that in young leaves.

We have developed genomic tools to allow the genus Populus (aspens and cottonwoods) to be exploited as a full-featured model for investigating fundamental aspects of tree biology. We have undertaken large-scale expressed sequence tag (EST) sequencing programs and created Populus microarrays with significant gene coverage. One of the important aspects of plant biology that cannot be studied in annual plants is the gene activity involved in the induction of autumn leaf senescence.On the basis of 36,354 Populus ESTs, obtained from seven cDNA libraries, we have created a DNA microarray consisting of 13,490 clones, spotted in duplicate. Of these clones, 12,376 (92%) were confirmed by resequencing and all sequences were annotated and functionally classified. Here we have used the microarray to study transcript abundance in leaves of a free-growing aspen tree (Populus tremula) in northern Sweden during natural autumn senescence. Of the 13,490 spotted clones, 3,792 represented genes with significant expression in all leaf samples from the seven studied dates.We observed a major shift in gene expression, coinciding with massive chlorophyll degradation, that reflected a shift from photosynthetic competence to energy generation by mitochondrial respiration, oxidation of fatty acids and nutrient mobilization. Autumn senescence had much in common with senescence in annual plants; for example many proteases were induced. We also found evidence for increased transcriptional activity before the appearance of visible signs of senescence, presumably preparing the leaf for degradation of its components.

During leaf senescence, the final stage of leaf development, nutrients are recycled from leaves to other organs, and therefore proper control of senescence is thus critical for plant fitness. Although substantial progress has been achieved in understanding leaf senescence in annual plants, the molecular factors that control leaf senescence in perennial woody plants are largely unknown. Using RNA sequencing, we obtained a high-resolution temporal profile of gene expression during autumn leaf senescence in poplar (Populus tomentosa). Identification of hub transcription factors (TFs) by co-expression network analysis of genes revealed that senescence-associated NAC-family TFs (Sen-NAC TFs) regulate autumn leaf senescence. Age-dependent alternative splicing (AS) caused an intron-retention (IR) event in the pre-mRNA encoding PtRD26, a NAC-TF. This produced a truncated protein PtRD26IR, which functions as a dominant-negative regulator of senescence by interacting with multiple hub Sen-NAC TFs, thereby repressing their DNA-binding activities. Functional analysis of senescence-associated splicing factors identified two U2 auxiliary factors that are involved in AS of PtRD26IR. Correspondingly, silencing of these factors decreased PtRD26IR transcript abundance and induced early senescence. We propose that an age-dependent increase of IR splice variants derived from Sen-NAC TFs is a regulatory program to fine tune the molecular mechanisms that regulate leaf senescence in trees.

Leaf senescence is a unique developmental process that is characterized by massive programmed cell death and nutrient recycling. The underlying molecular regulatory mechanisms are not well understood. Here we report the functional analysis of AtNAP, a gene encoding a NAC family transcription factor. Expression of this gene is closely associated with the senescence process of Arabidopsis rosette leaves. Leaf senescence in two T‐DNA insertion lines of this gene is significantly delayed. The T‐DNA knockout plants are otherwise normal. The mutant phenotype can be restored to wild‐type by the intact AtNAP, as well as by its homologs in rice and kidney bean plants that are also upregulated during leaf senescence. Furthermore, inducible overexpression of AtNAP causes precocious senescence. These data strongly suggest that AtNAP and its homologs play an important role in leaf senescence in Arabidopsis and possibly in other plant species.

Enhancing the nutritional value of food crops is a means of improving human nutrition and health. We report here the positional cloning of Gpc-B1, a wheat quantitative trait locus associated with increased grain protein, zinc, and iron content. The ancestral wild wheat allele encodes a NAC transcription factor (NAM-B1) that accelerates senescence and increases nutrient remobilization from leaves to developing grains, whereas modern wheat varieties carry a nonfunctional NAM-B1 allele. Reduction in RNA levels of the multiple NAM homologs by RNA interference delayed senescence by more than 3 weeks and reduced wheat grain protein, zinc, and iron content by more than 30%.

Over-application of nitrogen fertilizer in fields has had a negative impact on both environment and human health. Domesticated rice varieties with high nitrogen use efficiency (NUE) reduce fertilizer for sustainable agriculture. Here, we perform genome-wide association analysis of a diverse rice population displaying extreme nitrogen-related phenotypes over three successive years in the field, and identify an elite haplotype of nitrate transporter OsNPF6.1 that enhances nitrate uptake and confers high NUE by increasing yield under low nitrogen supply. OsNPF6.1 differs in both the protein and promoter element with natural variations, which are differentially trans-activated by OsNAC42, a NUE-related transcription factor. The rare natural allele OsNPF6.1, derived from variation in wild rice and selected for enhancing both NUE and yield, has been lost in 90.3% of rice varieties due to the increased application of fertilizer. Our discovery highlights this NAC42-NPF6.1 signaling cascade as a strategy for high NUE and yield breeding in rice.

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 over-expression 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 Y2H screening, we identified its interacting proteins, including a pair of paralogous transcription factors, PvNAP1&2 (NAC-LIKE, ACTIVATED BY AP3/PI). Over-expression 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, over-expression 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.© American Society of Plant Biologists 2022. All rights reserved. For permissions, please email: journals.permissions@oup.com.