Leaf senescence is an orderly and highly coordinated process, and finely regulated by ethylene and nitrogen (N), ultimately affecting grain yield and nitrogen-use efficiency (NUE). However, the underlying regulatory mechanisms on the crosstalk between ethylene- and N-regulated leaf senescence remain a mystery in maize. In this study, ethylene biosynthesis gene ZmACS7 overexpressing (OE-ZmACS7) plants were used to study the role of ethylene regulating leaf senescence in response to N deficiency, and they exhibited the premature leaf senescence accompanied by increased ethylene release, decreased chlorophyll content and Fv/Fm ratio, and accelerated chloroplast degradation. Then, we investigated the dynamics changes of transcriptome reprogramming underlying ethylene-accelerated leaf senescence in response to N deficiency. The differentially expressed genes (DEGs) involved in chlorophyll biosynthesis were significantly down-regulated, while DEGs involved in chlorophyll degradation and autophagy processes were significantly up-regulated, especially in OE-ZmACS7 plants in response to N deficiency. A gene regulatory network (GRN) was predicted during ethylene-accelerated leaf senescence in response to N deficiency. Three transcription factors (TFs) ZmHSF4, ZmbHLH106, and ZmEREB147 were identified as the key regulatory genes, which targeted chlorophyll biosynthesis gene ZmLES22, chlorophyll degradation gene ZmNYC1, and autophagy-related gene ZmATG5, respectively. Furthermore, ethylene signaling key genes might be located upstream of these TFs, generating the signaling cascade networks during ethylene-accelerated leaf senescence in response to N deficiency. Collectively, these findings improve our molecular knowledge of ethylene-accelerated maize leaf senescence in response to N deficiency, which is promising to improve NUE by manipulating the progress of leaf senescence in maize.

Nitrogen (N) is an essential factor for limiting crop yields, and cultivation of crops with low nitrogen-use efficiency (NUE) exhibits increasing environmental and ecological risks. Hence, it is crucial to mine valuable NUE improvement genes, which is very important to develop and breed new crop varieties with high NUE in sustainable agriculture system. Quantitative trait locus (QTL) and genome-wide association study (GWAS) analysis are the most common methods for dissecting genetic variations underlying complex traits. In addition, with the advancement of biotechnology, multi-omics technologies can be used to accelerate the process of exploring genetic variations. In this study, we integrate the substantial data of QTLs, quantitative trait nucleotides (QTNs) from GWAS, and multi-omics data including transcriptome, proteome, and metabolome and further analyze their interactions to predict some NUE-related candidate genes. We also provide the genic resources for NUE improvement among maize, rice, wheat, and sorghum by homologous alignment and collinearity analysis. Furthermore, we propose to utilize the knowledge gained from classical cases to provide the frameworks for improving NUE and breeding N-efficient varieties through integrated genomics, systems biology, and modern breeding technologies.

Nitrogen (N), one of the most important nutrients, limits plant growth and crop yields in sustainable agriculture system, in which phytohormones are known to play essential roles in N availability. Hence, it is not surprising that massive studies about the crosstalk between N and phytohormones have been constantly emerging. In this review, with the intellectual landscape of N and phytohormones crosstalk provided by the bibliometric analysis, we trace the research story of best-known crosstalk between N and various phytohormones over the last 20 years. Then, we discuss how N regulates various phytohormones biosynthesis and transport in plants. In reverse, we also summarize how phytohormones signallings modulate root system architecture (RSA) in response to N availability. Besides, we expand to outline how phytohormones signallings regulate uptake, transport, and assimilation of N in plants. Further, we conclude advanced biotechnology strategies, explain their application, and provide potential phytohormones-regulated N use efficiency (NUE) targets in crops. Collectively, this review provides not only a better understanding on the recent progress of crosstalk between N and phytohormones, but also targeted strategies for improvement of NUE to increase crop yields in future biotechnology breeding of crops.

BackgroundAuxin plays a crucial role in nitrate (NO3-)-mediated root architecture, and it is still unclear that if NO3- supply modulates auxin reallocation for regulating root formation in maize (Zea mays L.). This study was conducted to investigate the role of auxin efflux carrier ZmPIN1a in the root formation in response to NO3- supply.ResultsLow NO3- (LN) promoted primary root (PR) elongation, while repressed the development of lateral root primordia (LRP) and total root length. LN modulated auxin levels and polar transport and regulated the expression of auxin-responsive and -signaling genes in roots. Moreover, LN up-regulated the expression level of ZmPIN1a, and overexpression of ZmPIN1a enhanced IAA efflux and accumulation in PR tip, while repressed IAA accumulation in LRP initiation zone, which consequently induced LN-mediated PR elongation and LR inhibition. The inhibition rate of PR length, LRP density and number of ZmPIN1a-OE plants was higher than that of wild-type plants after auxin transport inhibitor NPA treatment under NN and LN conditions, and the degree of inhibition of root growth in ZmPIN1a-OE plants was more obvious under LN condition.ConclusionThese findings suggest that ZmPIN1a was involved in modulating auxin levels and transport to alter NO3--mediated root formation in maize.

Brassinosteroids (BRs) are steroid hormones that function in plant growth and development and response to environmental stresses and nutrient supplies. However, few studies have investigated the effect of BRs in modulating the physiological response to nitrogen (N) supply in maize. In the present study, BR signaling-deficient mutant zmbri1-RNAi lines and exogenous application of 2,4-epibrassinolide (eBL) were used to study the role of BRs in the regulation of physiological response in maize seedlings supplied with N. Exogenous application of eBL increased primary root length and plant biomass, but zmbri1 plants showed shorter primary roots and less plant biomass than wild-type plants under low N (LN) and normal N (NN) conditions. LN induced the expression of the BR signaling-associated genes ZmDWF4, ZmCPD, ZmDET2, and ZmBZR1 and the production of longer primary roots than NN. Knockdown of ZmBRI1 weakened the biological effects of LN-induced primary root elongation. eBL treatment increased N accumulation in shoots and roots of maize seedlings exposed to LN or NN treatment. Correspondingly, zmbri1 plants showed lower N accumulation in shoots and roots than wild-type plants. Along with reduced N accumulation, zmbri1 plants showed lower NO3− fluxes and 15NO3− uptake. The expression of nitrate transporter (NRT) genes (ZmNPF6.4, ZmNPF6.6, ZmNRT2.1, ZmNRT2.2) was lower in zmbri1 than in wild-type roots, but eBL treatments up-regulated the transcript expression of NRT genes. Thus, BRs modulated N physiological response and regulated the transcript expression of NRT genes to promote N uptake in maize.

Summary Nitrogen (N), one of the most important nutrients, limits plant growth and crop yields in sustainable agriculture system, in which phytohormones are known to play essential roles in N availability. Hence, it is not surprising that massive studies about the crosstalk between N and phytohormones have been constantly emerging. In this review, with the intellectual landscape of N and phytohormones crosstalk provided by the bibliometric analysis, we trace the research story of best‐known crosstalk between N and various phytohormones over the last 20 years. Then, we discuss how N regulates various phytohormones biosynthesis and transport in plants. In reverse, we also summarize how phytohormones signallings modulate root system architecture (RSA) in response to N availability. Besides, we expand to outline how phytohormones signallings regulate uptake, transport, and assimilation of N in plants. Further, we conclude advanced biotechnology strategies, explain their application, and provide potential phytohormones‐regulated N use efficiency (NUE) targets in crops. Collectively, this review provides not only a better understanding on the recent progress of crosstalk between N and phytohormones, but also targeted strategies for improvement of NUE to increase crop yields in future biotechnology breeding of crops.

Dodder is a holoparasitic flowering plant that re-establishes parasitism with the host when broken off from the host. However, how in vitro dodder shoots recycle stored nutrients to maintain growth for reparasitizing hosts is not well characterized. Here, the spatial and temporal distribution characteristics of carbohydrates and reactive oxygen species (ROS) were analysed to explore the mechanism of recycling stored nutrients in dodder shoots in vitro. Our results showed that in vitro dodder shoots grew actively for more than 10 d, while dry mass decreased continuously. During this process, the transcript levels and activities of amylases gradually increased until 2 d and then declined in basal stems, which induced starch degradation at the tissue, cellular and subcellular levels. Additionally, the distribution characteristics of H2O2 and the activities and transcript levels of antioxidant enzymes indicated that shoot tips exhibited more robust ROS-scavenging capacity, and basal stems maintained higher ROS accumulation. Comparative proteomics analysis revealed that starch in basal stems acted as an energy source, and the glycolysis, TCA cycle and pentose phosphate pathway represented the energy supply for shoot tip elongation with time. These results indicated that efficient nutrient recycling and ROS modulation facilitated the parasitism of dodder grown in vitro by promoting shoot elongation growth to reach the host.

Coronatine (COR), a structural and functional mimic of jasmonates, is involved in a wide array of effects on plant development and defense response. The present study aims to investigate the role of COR, in counteracting drought stress by modulating reactive oxygen species (ROS) homeostasis, water balance, and antioxidant regulation in detached maize plants. Our results showed that COR can markedly decrease the water loss rate, but the antioxidants diphenyleneiodonium chloride (DPI) and dimethylthiourea (DMTU) eliminate the effect of water loss induced by COR. Using the dye 2′,7′-dichlorofluorescein diacetate (H2DCF-DA) loaded in the maize epidermis guard cells, it is observed that COR could increase ROS production, and then antioxidants DPI and DMTU decreased ROS production induced by COR. In addition, the expression of ZmRBOHs genes, which were associated with ROS generation was increased by COR in levels and ZmRBOHC was highly expressed in the epidermis guard cells. Moreover, COR-treated plants increased H2O2 and O2−· accumulation, antioxidant enzyme activities in control plants, while COR relieved the ROS accumulation and antioxidant enzyme activities under PEG treatment. These results indicated that COR could improve maize performance under drought stress by modulating ROS homeostasis to maintain water loss rate and antioxidant enzyme activities.

Modified gibberellin (GA) signaling leads to semi-dwarfism with low nitrogen (N) use efficiency (NUE) in crops. An understanding of GA-mediated N uptake is essential for the development of crops with improved NUE. The function of GA in modulating N uptake capacity and nitrate (NO3−) transporters (NRTs) was analyzed in the GA synthesis-deficient mutant zmga3ox grown under low (LN) and sufficient (SN) N conditions. LN significantly suppressed the production of GA1, GA3, and GA4, and the zmga3ox plants showed more sensitivity in shoots as well as LN stress. Moreover, the higher anthocyanin accumulation and the decrease of chlorophyll content were also recorded. The net NO3− fluxes and 15N content were decreased in zmga3ox plants under both LN and SN conditions. Exogenous GA3 could restore the NO3− uptake in zmga3ox plants, but uniconazole repressed NO3− uptake. Moreover, the transcript levels of ZmNRT2.1/2.2 were downregulated in zmga3ox plants, while the GA3 application enhanced the expression level. Furthermore, the RNA-seq analyses identified several transcription factors that are involved in the GA-mediated transcriptional operation of NRTs related genes. These findings revealed that GAs influenced N uptake involved in the transcriptional regulation of NRTs and physiological responses in maize responding to nitrogen supply.

Use of growth retardants enables post-planting optimization of vegetative growth, which is particularly important given ongoing climate change. Mepiquat chloride is an economical and safe retardant widely applied in cotton farming, but it is not uniformly effective. Here, identification of its molecular target as the ent -copalyl diphosphate synthase that initiates gibberellin biosynthesis enabled the introduction of selective agrochemical inhibition, leaving intact more specialized metabolism important for resistance to biotic and abiotic stresses.

Salinity stress impairs plant growth and causes crops to yield losses worldwide. Reduction of in vivo gibberellin acid (GA) level is known to repress plant size but is beneficial to plant salt tolerance. However, the mechanisms of in vivo GA deficiency-enhanced salt tolerance in maize are still ambiguous. In this study, we generated two independent maize knockout mutant lines of ent-copalyl diphosphate synthase (one of the key enzymes for early steps of GA biosynthesis), zmcps-1 and zmcps-7, to explore the role of GA in maize salt tolerance. The typical dwarf phenotype with lower GA content and delayed leaf senescence under salinity was observed in the mutant plants. The leaf water potential and cell turgor potential were significantly higher in zmcps-1 and zmcps-7 than in the wild type (WT) under salt stress. The mutant plants exhibited a lower superoxide anion production rate in leaves and also a downregulated relative expression level of NAPDH oxidase ZmRbohA-C than the WT maize under salt stress. Also, the mutant plants had higher enzymatic activities of superoxide dismutase (SOD) and catalase (CAT) and higher content of soluble sugars and proline under salt stress. The Na+/K+ ratio was not significantly different between the mutant maize plants and WT plants under salt stress conditions, but the Na+ and K+ content was increased in zmcps-1 and zmcps-7 leaves and shoots. Na+ fluorescent dye staining showed that the mutant leaves have significantly higher vacuolar Na+ intensity than the WT maize. The expression level of vacuolar Na+/H+ exchanger gene ZmNHX1 and vacuolar proton pump genes ZmVP1-1 and ZmVP2 were upregulated in the zmcps-1 and zmcps-7 plants under salinity, further proving that in vivo GA deficiency enhanced vacuolar Na+ sequestration in zmcps-1 and zmcps-7 leaves cells to avoid Na+ cytotoxicity. Together, our results suggested that maintaining ROS homeostasis and enhancing vacuolar Na+ sequestration could be involved in GA deficiency-improved maize salt tolerance.

Ethephon efficiently regulates plant growth to modulate the maize (Zea mays L.) stalk strength and yield potential, yet there is little information on how ethylene governs a specific cellular response for altering internode elongation. Here, the internode elongation kinetics, cell morphological and physiological properties and transcript expression patterns were investigated in the ethephon-treated elongating internode. Ethephon decreased the internode elongation rate, shortened the effective elongation duration, and advanced the growth process. Ethephon regulated the expression patterns of expansin and secondary cell wall-associated cellulose synthase genes to alter cell size. Moreover, ethephon increased the activities and transcripts level of phenylalanine ammonia-lyase and peroxidase, which contributed to lignin accumulation. Otherwise, ethephon-boosted ethylene evolution activated ethylene signal and increased ZmGA2ox3 and ZmGA2ox10 transcript levels while down-regulating ZmPIN1a, ZmPIN4 and ZmGA3ox1 transcript levels, which led to lower accumulation of gibberellins and auxin. In addition, transcriptome profiles confirmed previous results and identified several transcription factors that are involved in the ethephon-modulated transcriptional regulation of cell wall biosynthesis and modification and responses to ethylene, gibberellins and auxin. These results indicated that ethylene-modulated auxin and gibberellins signaling mediated the transcriptional operation of cell wall modification to regulate cell elongation in the ethephon-treated maize internode.

Propiconazole (Pcz) has been widely used as fungistat in crop production. Meanwhile, Pcz can be used as plant growth regulator to influence plant growth. There is less study on the application of Pcz in maize. The present experiment using maize variety Zhengdan 958 was conducted to study the effect of Pcz on maize seedling growth, cell morphology and hormone signal. Pcz treatment inhibited the growth of mesocotyl and coleoptile, reduced plant height, shortened the length of sheath and leaf, and decreased leaf angle significantly. At the same time, longitudinal growth of sheath and leaf was inhibited and the shape of pulvinus cells changed from loosened rectangle type or oval type to closely spaced spindle cell. Furthermore, the GA (GA1and GA3)content was reduced significantly and the expression of GA biosynthesis gene GA3ox1 was down-regulated,the expression of GA2ox5 and GA2ox8, genes involved in GA inactivation, was up-regulated, and the expression of GA biosynthesis gene GA20ox1 was up-regulated at the beginning and down-regulated later;the BR content was reduced significantly and the expres-sion of BR biosynthesis genes CPD and DWF4 was up-regulated,which might be feedback regulated by endogenous BR levels. In addition, the expression of expansin genes EXPA4, EXPA5 and xyloglucan endotransglucosylase/hydrolase genes XTH1 and XET1 was down-regulated.In conclusion,Pcz treatment can adjust the signal transduction of GA and BR,inhibit the accumulation of GA and BR,regulate the expression of EXPs and XTHs,control the cell growth,and adjust plant type of maize effectively.