Genome-wide identification and expression analysis of NIN-like Protein(NLP)genes reveals their potential roles in the response to nitrate signaling in watermelon

To balance the relationship between high yield and low nitrogen supply,the nitrogen utilization efficiency of watermelon needs to be improved urgently.Nodule inception-like Protein(NLP)transcription factors play a key node role in nitrate response and growth and development of plant,however,comprehensive analysis of the NLP gene family in watermelon is unclear.This study explored the functional classification,evolutionary characteristics,and expression profile of the ClNLP gene family.Three ClNLPs were categorized into three groups according to their gene structure and phylogeny.All of them contained the conserved RWP-RK and PB1 domains.Evolutionary analysis of ClNLPs revealed that ClNLP1 and ClNLP3 underwent strong purified selection.In addition,cis-acting elements related to plant hormones and abiotic stresses were present in the ClNLP promoter.According to tissue-specific analysis ClNLP was widely expressed in roots,stems,leaves,flowers and fruits,and ClNLP1 was significantly induced in the roots of different nitrogen utilization varieties under different nitrate nitrogen supply.The SRTING functional protein association network suggested that ClNLP1 is associated with most genes,such as NRT1.1,NRT2.1,NIA1,and NIR1,and the dual-luciferase reporter assay found that ClNLP1 positively regulates the expression of ClNRT2.1.We speculated that ClNLP1 might play a central role in regulating the response of watermelon to nitrate nitrogen.

Nitrate signaling and use efficiency in crops

Nitrate(NO3–)is not only an essential nutrient but also an important signaling molecule for plant growth.Low nitrogen use efficiency(NUE)of crops is causing increasingly serious environmental and ecological problems.Understanding the molecular mechanisms of NO3–regulation in crops is crucial for NUE improvement in agriculture.During the last several years,significant progress has been made in understanding the regulation of NO3–signaling in crops,and some key NO3–signaling factors have been shown to play important roles in NO3–utilization.However,no detailed reviews have yet summarized these advances.Here,we focus mainly on recent advances in crop NO3–signaling,including short-term signaling,long-term signaling,and the impact of environmental factors.We also review the regulation of crop NUE by crucial genes involved in NO3–signaling.This review provides useful information for further research on NO3–signaling in crops and a theoretical basis for breeding new crop varieties with high NUE,which has great significance for sustainable agriculture.

Nuclear translocation of OsMADS25 facilitated by OsNAR2.1 in reponse to nitrate signals promotes rice root growth by targeting OsMADS27 and OsARF7

Nitrate is an important nitrogen source and signaling molecule that regulates plant growth and development.Although several components of the nitrate signaling pathway have been identified,the detailed mechanisms are still unclear.Our previous results showed that OsMADS25 can regulate root development in response to nitrate signals,but the mechanism is still unknown.Here,we try to answer two key questions:how does OsMADS25 move from the cytoplasm to the nucleus,and what are the direct target genes activated by OsMADS25 to regulate root growth after it moves to the nucleus in response to nitrate?Our results demonstrated that OsMADS25 moves from the cytoplasm to the nucleus in the presence of nitrate in an OsNAR2.1-dependentmanner.Chromatin immunoprecipitation sequencing,chromatin immunoprecipitation qPCR,yeast one-hybrid,and luciferase experiments showed that OsMADS25 directly activates the expression of OsMADS27 and OsARF7,which are reported to be associated with root growth.Finally,OsMADS25-RNAi lines,the Osnar2.1 mutant,and OsMADS25-RNAi Osnar2.1 lines exhibited significantly reduced root growth compared with the wild type in response to nitrate supply,and expression of OsMADS27 and OsARF7 was significantly suppressed in these lines.Collectively,these results reveal a new mechanismby which OsMADS25 interacts with OsNAR2.1.This interaction is required for nuclear accumulation of OsMADS25,which promotes OsMADS27 and OsARF7 expression and root growth in a nitratedependent manner.

HBI1-TCP20 interaction positively regulates the CEPs-mediated systemic nitrate acquisition

Nitrate is the main source of nitrogen for plants but often distributed heterogeneously in soil.Plants have evolved sophisticated strategies to achieve adequate nitrate by modulating the root system architecture. The nitrate acquisition system is triggered by the short mobile peptides C-TERMINALLY ENCODED PEPTIDES(CEPs)that are synthesized on the nitrate-starved roots,but induce the expression of nitrate transporters on the other nitrate-rich roots through an unclear signal transduction pathway. Here,we demonstrate that the transcription factors HBI1 and TCP20 play important roles in plant growth and development in response to fluctuating nitrate supply. HBI1 physically interacts with TCP20, and this interaction was enhanced by the nitrate starvation. HBI1 and TCP20 directly bind to the promoters of CEPs and cooperatively induce their expression. Mutation in HBIs and/or TCP20 resulted in impaired systemic nitrate acquisition response. Our solid genetic and molecular evidence strongly indicate that the HBI1-TCP20 module positively regulates the CEPs-mediated systemic nitrate acquisition.

Say “NO” to ABA signaling in guard cells by S-nitrosylation of OST1

Nitric oxide（NO）,a gaseous compound,plays important roles in plant immunity,abiotic stress response and plant development[1].In plants,NO is synthesized through either oxidative or reductive route that is dependent on the nitrate reductases（NADH）1（NIA1）and NIA2.NO bioactivity is realized through redox-based post-translational protein

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.