Application of PLPB1 gene in regulation and control of plant nitrogen efficiency

Abstract

The invention provides an application of a PLPB1 gene or a protein coded by the PLPB1 gene in regulation and control of plant nitrogen efficiency. The application proves that the PLPB1 gene participates in the nitrogen metabolism regulation process of the corn, and the nitrogen utilization efficiency of the corn can be remarkably improved by inhibiting the expression and / or activity of the PLPB1 gene. The discovery not only enriches the understanding of a corn nitrogen metabolism mechanism, but also provides a gene resource for cultivating a new variety with high nitrogen efficiency, and is beneficial to high efficiency and environmental protection of agricultural production. In addition, the analysis of the PLPB1 gene function also lays a foundation for nitrogen efficient improvement of other crops.

Description

Technical Field

[0001] The present application relates to the field of biotechnology, and in particular, to the application of the PLPB1 gene in regulating plant nitrogen efficiency. Background Art

[0002] Maize (Zea mays L.), the world's most widely cultivated cereal crop, accounts for over 37% of global grain production, making it a key crop for ensuring international food security (FAO, http: / / faostat.fao.org / ). Nitrogen fertilizer plays an indispensable role in ensuring global food security, and large-scale application of chemical nitrogen fertilizers is one of the most effective ways to increase crop yields. However, excessive nitrogen fertilizer application poses a dual challenge to agricultural ecosystems: while significantly increasing crop yields, it also leads to soil nutrient imbalances and environmental pollution.

[0003] As global food demand continues to grow, nitrogen fertilizer application is on a rising trend, posing a dilemma for agricultural production: reducing nitrogen use and increasing efficiency. Against this backdrop, improving maize nitrogen use efficiency (NUE) has become a sustainable solution for balancing high crop yields with environmental protection. Therefore, studying the molecular mechanisms of efficient nitrogen use in maize, identifying superior nitrogen-efficient genetic resources, and developing new maize varieties with high nitrogen efficiency are crucial for global food security and sustainable agricultural development.

[0004] Nitrogen use efficiency is a complex agronomic trait that involves nitrogen absorption, transport, assimilation and utilization, as well as nitrogen recycling. Nitrogen in the soil includes inorganic nitrogen (nitrate nitrogen and ammonium nitrogen) and organic nitrogen (urea, amino acids). The nitrogen absorption process of corn is that the nitrogen receptors in the roots sense the nitrogen in the soil, and then, under the action of various nitrogen transporters (NPF family, AMT family, urease, cell membrane ion channels), the absorbed inorganic nitrogen and organic nitrogen are transferred to the leaves. Corn converts the absorbed nitrogen in different forms into amino acids that can be directly absorbed through a series of complex physiological and biochemical processes in the leaves. This process is called assimilation. After flowering, corn enters the reproductive growth stage. At this time, the leaves will act as source organs to store nitrogen and metabolites involved in the nitrogen cycle (NO3 - NH4 + , Glu, Gln, Asn, Asp) are gradually transferred into corn kernels, and this process is called the recycling process.

[0005] Although several genes have been identified that may improve nitrogen use efficiency in various nitrogen metabolic processes, including nitrogen uptake and transport, assimilation and utilization, and recycling, significant gaps remain in the understanding of the entire nitrogen metabolism process in maize. Genetic resources regulating efficient nitrogen use are relatively scarce, and the molecular basis is weak, making it difficult to effectively breed new varieties with high nitrogen utilization. Therefore, further in-depth research on these processes and the discovery of relevant genetic resources are crucial. Summary of the Invention

[0006] In response to the problems existing in the prior art, the purpose of this application is to provide an application of the PLPB1 gene in regulating plant nitrogen efficiency.

[0007] Specifically, this application involves the following aspects:

[0008] 1. Application of the PLPB1 gene or the protein encoded by the PLPB1 gene in regulating plant nitrogen efficiency;

[0009] Preferably, the plant is corn.

[0010] 2. The use according to item 1, wherein the nucleotide sequence of the PLPB1 gene is shown as SEQ ID NO: 1, or the amino acid sequence of the protein encoded by the PLPB1 gene is shown as SEQ ID NO: 2.

[0011] 3. A PLPB1 gene mutant, wherein the PLPB1 gene mutant comprises a substitution, deletion or insertion of one or more nucleotides based on a reference sequence, the nucleotide sequence of the reference sequence being shown in SEQ ID NO: 1.

[0012] 4. Use of the PLPB1 gene mutant according to item 3 in regulating plant nitrogen efficiency;

[0013] Preferably, the plant is corn.

[0014] 5. A biomaterial, wherein the biomaterial comprises:

[0015] An expression cassette for inhibiting the action of the PLPB1 gene;

[0016] A recombinant vector capable of inhibiting the PLPB1 gene;

[0017] A recombinant microorganism capable of inhibiting the PLPB1 gene; or

[0018] A plant cell line comprising the PLPB1 gene mutant according to item 3.

[0019] 6. The biomaterial according to item 5, wherein the nucleotide sequence of the PLPB1 gene is shown as SEQ ID NO: 1.

[0020] 7. Use of the biomaterial according to item 6 in regulating plant nitrogen efficiency;

[0021] Preferably, the plant is corn.

[0022] 8. Application of PLPB1 gene inhibitors in regulating nitrogen efficiency;

[0023] Preferably, the plant is corn.

[0024] 9. The use according to item 8, wherein the nucleotide sequence of the PLPB1 gene is as shown in SEQ ID NO: 1, and the inhibitor inhibits the expression and / or activity of the PLPB1 gene;

[0025] Preferably, the inhibitor comprises a small molecule compound, a polypeptide, an inhibitory nucleic acid molecule that hybridizes with a PLPB1 nucleic acid molecule, or a gene editor.

[0026] 10. A method for regulating plant nitrogen efficiency, wherein the method comprises:

[0027] By inhibiting the expression and / or activity of the PLPB1 gene in plants, the nitrogen efficiency of plants is regulated;

[0028] Preferably, the plant is corn.

[0029] 11. The method according to item 10, wherein the nucleotide sequence of the PLPB1 gene is shown as SEQ ID NO: 1.

[0030] 12. Application of the PLPB1 gene or the protein encoded by the PLPB1 gene in the breeding of nitrogen-efficient plants;

[0031] Preferably, the plant is corn.

[0032] 13. The use according to item 12, wherein the nucleotide sequence of the PLPB1 gene is shown as SEQ ID NO: 1, or the amino acid sequence of the protein encoded by the PLPB1 gene is shown as SEQ ID NO: 2.

[0033] 14. Use of the PLPB1 gene mutant according to item 3, or the biomaterial according to item 5 or 6, in nitrogen-efficient plant breeding;

[0034] Preferably, the plant is corn.

[0035] 15. Application of PLPB1 gene inhibitors in nitrogen-efficient plant breeding.

[0036] Preferably, the plant is corn.

[0037] 16. The use according to item 15, wherein the nucleotide sequence of the PLPB1 gene is as shown in SEQ ID NO: 1, and the inhibitor inhibits the expression and / or activity of the PLPB1 gene;

[0038] Preferably, the inhibitor comprises a small molecule compound, a polypeptide, an inhibitory nucleic acid molecule that hybridizes with a PLPB1 nucleic acid molecule, or a gene editor.

[0039] Beneficial effects:

[0040] This application demonstrates that the PLPB1 gene is involved in regulating nitrogen metabolism in maize and that inhibiting its expression and / or activity can significantly improve the nitrogen utilization efficiency of maize. This discovery not only enriches our understanding of the nitrogen metabolism mechanism of maize but also provides a new genetic engineering target, which is of great significance for future crop improvement. In addition, by combining traditional breeding with modern biotechnology, based on this application's research on the PLPB1 gene, new nitrogen-efficient varieties can be cultivated. This provides a new path for achieving efficient and environmentally friendly agricultural production and helps address global food security and environmental pollution issues. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1A-Figure 1C The transcription level of PLPB1 gene is affected by different nitrogen conditions. Figure 1A The results of the detection of PLPB1 gene expression in maize seedlings after treatment with different concentrations of NH4Cl or KNO3; Figure 1B The results of the detection of PLPB1 gene expression in maize seedlings at different time periods after nitrogen source supply was removed; Figure 1C Figure 3 shows the expression of PLPB1 gene in maize seedlings at different time periods after resupply of NH4Cl or KNO3. Different lowercase letters indicate significant differences (P < 0.05).

[0042] Figure 2A-2C To obtain PLPB1 gene editing materials. Figure 2A Schematic diagram of the CRISPR / Cas9 gene knockout vector structure of the PLPB1 gene; Figure 2B This is a target sequence analysis diagram for gene editing; Figure 2C Schematic diagram of the mutation locations of plpb1-1 and plpb1-2.

[0043] Figures 3A-3D The growth phenotypes and phenotypic analysis results of wild-type control B104 corn material (WT) and PLPB1 gene-edited materials under low nitrogen and normal nitrogen treatments at the seedling stage. Figure 3A This is a graph showing the phenotypes of PLPB1 gene-edited materials and WT under normal nitrogen treatment at the seedling stage; Figure 3BThis is a graph showing the phenotypes of PLPB1 gene-edited materials and WT under low nitrogen treatment at the seedling stage; Figure 3C The results of plant height, leaf nitrogen content, and chlorophyll content testing of PLPB1 gene-edited materials and WT under normal nitrogen treatment are shown; Figure 3D Figure 3. Plant height, leaf nitrogen content, and chlorophyll content of PLPB1 gene-edited materials and WT under low nitrogen treatment. Scale bar = 5 cm. Different lowercase letters indicate significant differences (P < 0.05).

[0044] Figures 4A-4C The results of yield and yield trait analysis of wild-type control B104 corn material (WT) and PLPB1 gene-edited material under normal nitrogen field and low nitrogen field conditions are shown. Figure 4A The results of ear phenotypes of PLPB1 gene-edited materials and WT under normal nitrogen field and low nitrogen field conditions;

[0045] Figure 4B The results of ear length detection of PLPB1 gene-edited materials and WT under normal nitrogen field and low nitrogen field conditions; Figure 4C The results of ear weight detection of PLPB1 gene-edited materials and WT under normal nitrogen field and low nitrogen field conditions. Scale bar = 5 cm. DETAILED DESCRIPTION

[0046] The present application is further described below with reference to examples. It should be understood that the examples are only used to further illustrate and explain the present application and are not intended to limit the present application.

[0047] Unless otherwise defined, technical and scientific terms used in this specification have the same meaning as those commonly understood by those skilled in the art. Although methods and materials similar or identical to those described herein may be used in experiments or practical applications, the materials and methods are described herein below. In the event of a conflict, the present specification, including definitions, will prevail. In addition, the materials, methods, and examples are provided for illustrative purposes only and are not intended to be limiting. The present application is further described below with reference to specific examples, which are not intended to limit the scope of this application.

[0048] definition

[0049] As used herein, the term "amino acid" or "amino acid sequence" refers to an oligopeptide, peptide, polypeptide, or protein sequence, or a fragment of any of these, and refers to a naturally occurring or synthetic molecule. When "amino acid sequence" is described herein as referring to the amino acid sequence of a naturally occurring protein molecule, "amino acid sequence" and similar terms are not intended to limit the amino acid sequence to the complete native amino acid sequence associated with the described protein molecule.

[0050]

[00146] "Amino acids" may be referred to herein by their name, by their commonly known three letter symbols, or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission.

[0051] As used herein, the terms "nucleic acid," "nucleic acid sequence," "nucleotide sequence," "polynucleotide," "polynucleotide sequence," "RNA sequence," or "DNA sequence" refer to oligonucleotides, nucleotides, or polynucleotides, and fragments and portions thereof, and to DNA or RNA of genomic or synthetic origin, which may be single-stranded or double-stranded and represent the sense or antisense strand. The sequence may be a non-coding sequence, a coding sequence, or a mixture of the two. The nucleic acid sequences of the present application can be prepared using standard techniques well known to those skilled in the art.

[0052] As used herein, the term "mutation" generally refers to any type of change or modification to a sequence (nucleic acid or amino acid sequence), including deletion, truncation, inactivation, disruption, substitution, translocation, or insertion of amino acids or nucleotides.

[0053] As used herein, the term "mutant" or "variant" refers to a molecule having certain differences in its base or amino acid sequence compared to a reference polynucleotide or polypeptide (naturally occurring), respectively, and these differences are artificially synthesized or naturally occurring. These differences include substitutions, insertions, deletions, or any desired combination of such changes in the natural polynucleotide encoding the amino acid sequence.

[0054] As used herein, the term "expression cassette" refers to a DNA sequence capable of expressing a protein or nucleic acid of interest in a host cell. The expression cassette may also include a single-stranded or double-stranded nucleic acid molecule containing all regulatory sequences necessary to express the nucleic acid molecule for any of the proteins described above. The regulatory sequences are capable of directing the expression of any of the proteins described above from the coding sequence in a suitable host cell under compatible conditions. The regulatory sequences include, but are not limited to, a leader sequence, a polyadenylation sequence, a propeptide sequence, a promoter, a signal sequence, and a transcription terminator. At a minimum, the regulatory sequences include a promoter and termination signals for transcription and translation. The regulatory sequences may be provided with linkers to introduce specific restriction enzyme sites into the vector for ligation to the coding region of the protein-encoding nucleic acid sequence. The regulatory sequence may be a suitable promoter sequence, i.e., a nucleic acid sequence recognized by the host cell in which the nucleic acid sequence is to be expressed. The promoter sequence contains transcriptional regulatory sequences that mediate protein expression. The promoter may be any nucleic acid sequence that is transcriptionally active in the selected host cell, including mutant, truncated, and hybrid promoters, and may be derived from genes encoding extracellular or intracellular proteins that are homologous or heterologous to the host cell. A regulatory sequence may also be a suitable transcriptional terminator sequence, i.e., a sequence recognized by the host cell to terminate transcription. The terminator sequence may be operably linked to the 3' end of the nucleic acid sequence encoding the protein. Any terminator that is functional in the selected host cell may be used in this application. A regulatory sequence may also be a suitable leader sequence, i.e., an untranslated region of an mRNA that is important for translation in the host cell. The leader sequence may be operably linked to the 5' end of the nucleic acid sequence encoding the protein. Any leader sequence that is functional in the selected host cell may be used in this application. A regulatory sequence may also be a signal peptide coding region, which encodes an amino acid sequence attached to the amino terminus of the protein that directs the encoded protein into the cell's secretory pathway. Any signal peptide coding region that directs the expressed protein into the secretory pathway of the selected host cell may be used in this application. It may also be desirable to add regulatory sequences that can regulate protein expression based on the growth conditions of the host cells. Examples of regulatory systems are those that can turn gene expression on or off in response to chemical or physical stimuli (including in the presence of regulatory compounds). Other examples of regulatory sequences are those that enable gene amplification. In these instances, the protein-encoding nucleic acid sequence should be operably linked to the regulatory sequences.

[0055] As used herein, the term "vector" generally refers to a vehicle capable of transporting exogenous DNA or a gene of interest into host cells for amplification and / or expression. The vector can be a cloning vector or an expression vector. A vector can be introduced into a host cell via transformation, transduction, or transfection, allowing the genetic material it carries to be amplified and / or expressed in the host cell. Those skilled in the art can select an appropriate vector based on the purpose of the genetic engineering project and the properties of the recipient cell. The vector includes, but is not limited to, a plasmid, a phage (e.g., lambda phage or M13 phage), a cosmid (i.e., cosmid), a phagemid, a shuttle vector (e.g., a yeast expression vector), a Ti plasmid, an artificial chromosome (e.g., a yeast artificial chromosome (YAC), a bacterial artificial chromosome (BAC), a P1 artificial chromosome (PAC), or a Ti plasmid artificial chromosome (TAC)), a viral vector (e.g., a baculovirus vector, a retrovirus (including a lentivirus), adenovirus, adeno-associated virus, poxvirus, papillomavirus, papillomavirus (e.g., SV40), or a herpes virus (e.g., herpes simplex virus)). A vector may contain a variety of elements for controlling expression, including, but not limited to, a promoter sequence, a transcription initiation sequence, an enhancer sequence, a selection element, and a reporter gene. In addition, the vector may also contain a replication initiation site.

[0056] As used herein, the term "recombinant vector" generally refers to a recombinant DNA molecule constructed by connecting an exogenous target gene to a vector in vitro. It can be constructed in any suitable manner, as long as the constructed recombinant vector can carry the exogenous target gene into the recipient cell and provide the exogenous target gene with the ability to replicate, integrate, amplify and / or express in the recipient cell.

[0057] As used herein, the term "microorganism" generally includes bacteria, viruses, fungi, actinomycetes, rickettsia, mycoplasma, chlamydia, spirochetes, algae, etc. For example, the bacteria may be from the genus Escherichia sp. (such as Escherichia coli), Erwinia sp., Agrobacterium sp. (such as Agrobacterium tumefaciens), Flavobacterium sp., Alcaligenes sp., Pseudomonas sp., and Bacillus sp. (such as Bacillus), etc. The virus may include rotavirus, baculovirus, retrovirus (such as lentivirus), adenovirus, adeno-associated virus, poxvirus, papillomavirus, influenza virus, papillomavirus (such as SV40), and herpes virus (such as herpes simplex virus), etc. The fungi may be from the genus Saccharomyces (e.g., Saccharomyces cerevisiae, Methanol yeast, Pichia pastoris), Fusarium sp., Rhizoctonia sp., Verticillium sp., Penicillium sp., Aspergillus sp., and Cephalosporium sp., etc. The actinomycetes may be from the genus Streptomyces sp. (e.g., Streptomyces). The algae may be from the phylum Cyanophyta (e.g., Cyanobacteria), Fucus sp., Achnanthes sp., Amphiprora sp., Amphora sp., Ankistrodesmus sp., Asteromonas sp., and Boekelovia sp., etc.

[0058] As used herein, the term "recombinant microorganism" generally refers to a recombinant microorganism whose genes have been manipulated and modified to produce functionally altered recombinant microorganisms. This can include introducing an exogenous gene of interest or a recombinant vector into the microorganism, or directly editing the endogenous genes of the microorganism.

[0059] As used herein, the term "percent identity," for example, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, or 99.5% identity, refers to a degree of similarity between amino acid sequences or nucleotide sequences determined by sequence alignment of 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 98.5%, 99%, or 99.5%. For example, the percentage of positions with identical bases or amino acid residues is determined as a ratio of the total number of positions after two sequences have been aligned to have identical residues at as many positions as possible, such as by introducing gaps. Percent identity can be determined using software programs known in the art. Preferably, the alignment is performed using default parameters. A preferred alignment program is BLAST. Preferred programs are BLASTN and BLASTP. Details of these programs can be found on the Internet at the following address: blast.ncbi.nlm.nih.gov / Blast.cgi.

[0060] As used herein, the term "nitrogen efficiency" refers to the efficiency with which plants utilize nitrogen resources, that is, the ability to obtain higher yields or economic benefits per unit nitrogen input while reducing the negative impact of nitrogen loss on the environment.

[0061] As used herein, the term "plant" refers to any plant at any stage of development, particularly seed plants. The term "plant" includes the entire plant or any part or derivative thereof, such as a plant cell, seed, plant protoplast, plant callus or callus, meristematic cells, microspores, embryos, immature embryos, pollen, ovules, anthers, fruits, flowers, leaves, cotyledons, pistils, seeds, seed coats, roots, root tips, and the like.

[0062] PLPB1 gene and its encoded protein

[0063] This application identifies a highly efficient nitrogen utilization gene, PLPB1. Through testing under varying nitrogen conditions, the inventors found that the expression of the PLPB1 gene in maize seedlings is affected by the concentration of the nitrogen source. Furthermore, in the presence of a nitrogen source, the expression of the PLPB1 gene significantly increases over time, indicating that the PLPB1 gene is involved in nitrogen metabolism in maize.

[0064] The nucleotide sequence of the PLPB1 gene is shown in SEQ ID NO: 1, and the amino acid sequence of the protein encoded by the PLPB1 gene is shown in SEQ ID NO: 2.

[0065] Among them, the amino acid sequence shown in SEQ ID NO:2 is the protein encoded by the gene PLPB1 (SEQ ID NO:1, gene ID is Zm00001d039530) derived from maize.

[0066] The sequence of SEQ ID NO:1 is as follows:

[0067] CACGCCGCCTCACCTCGCCGGGCGGCGTGATGGCGACTTCCGGGGCCGGCGACCACG

[0068] GGCAGGGCCAGGGCCAGGGGCACGGCCGCTTCGCGTCGTCCCTGACGGCGCGCTACTCGG [[ID=ll]]

[0069] ACTGGGTGCTGGAGACGCTGGACGAGCTCCCAGGGAGCTTCCTGCTCACGGACCCCGCCCT

[0070] GCCGGGCCACCCCATCGTGTACGCTTCCCGGGGCCTCGCCGCGCTCACCGGCTACGCGCCG

[0071] CGCGACGTGCTGGGCCGCAACGCGCGCCTCTTCCAGGGCGCCGCCACCGACCGCGCCACC

[0072] GTCGCCGGCGTCCGCGAGGCCGTCCGCGCGCACCGCCCGCACCAGGCCGCCATCCTCAACT

[0073] ACCGCCGCGACGGCGCCCCGCACTGGGTCCTCCTCCACCTCGCCCCCCTATTCCACGCCCG

[0074] CGACGGCACCCTGCTCCACTTCCTCGCCGTCCAGGTGCCCATCGCCCCCGCCGCCGTCGCG ]>

[0075] CGCGGGGCGACGTGCCACACGACGGGGCATCTGCTAGCCGCGTGCCGTGACGAGGCCAGC

[0076] AGGGTCGCGGAGGATTTCCCCTGCGCCACCCACGCGGGCAAGGTGTTTGTCGATATGGACA

[0077] AGAGAGGTAATACTGCTAGTATGCTGCACTAGATAATAACTTACTCATGCTCGATTTTCCCTA

[0078] GAGTTTCAAGCATTAATTGCATCTGAGTTCAATTAAATCCCTTCCTCGTTCCTAAAGCTGAAC

[0079] GTAGTGCGTTCCAAGCTTGACATTCCTTGGTTGGTTGAATTACCAGTTCAGCTATCGTATATT

[0080] TATTATAGTTGCACTAATCTCATTTGTGGGCATAAACAGACAAACCGCTGAATAAGTAGGTGT

[0081] AGACTGTAGTGGCAGATTACTAAGAGAGTCAGATTACTCCCTTGTTCCATCTGGTCTTGGTT

[0082] CAAAGTCTGTTAATCGTGGCTATACGTTTATATTGCCCTTTTACCCAATGCAAACTATGCATGG

[0083] TTGCACGCACAAACACATATATGTGATAACTCTTTCCTTGCGCGGTGCTTACATAATGCGTTC

[0084] AACCCAAAAGAAAAAAAAAACTAATACGTTTTGGGATCAGGACAAACCACATTATGTGATT

[0085] TCCAGGATGAAGATTCAACAGCGTGCATGTTAGTTACTTGTTCTTAAATGCTGAAGATCAAG

[0086] AACAAGACAACACAATTATTAATTATTATGGACATTCGTCCTCCAAAGCATTATCTCCATATAG

[0087] GAATAATGCTTATCAGACTAAGGTCACGAAAGACTTATCCCCATTTAATATATGAATGAGAAT

[0088] GTAAAAGAATGAAGACAATAACTTCATTTTATTAATTCATATATATTTGCCTTTCATAAAACAT

[0089] ATATGAATGTTGACAATATTAATGCTACATTGATACTTTCGGCTTGCTCGAAGGTGAAGACGC

[0090] GAAAGAGAGATTATAATTCAACGTAAATAGTATGGTGTTACTGTTCATCTATTTATAGGCACG

[0091] AGACACGGTCCGAGTAAAAGTACATTTATACCCTTTATATTCATCTATAAACATAACACGAAT

[0092] CATTAAGGACTGAGTAGTCTTTGTCCCTTTTCGGTCATCATCATAATTGGTCTTCATCATCACA

[0093] TCAAGCCGAAGCTCATCGGCTATAGCTTCATCGTCTATTCTTATCACCTTCGGGCTACATCTTT

[0094] ACCCTTTCATGAGAAAAGGCCTTCATCCCGAAACCGAAGCCCCCTGTAATAATTCATGTCAT

[0095] ACTAAAAACATATGGTTAGCCAACACTGCAGTAGTTTACGAGTCTTCATTACTGGCAGTTTG

[0096] AGCTGGGGTTTATAGTTGACTTAACACTTCAATGTTTTCACTCTTACACACAAGTATCGAACA

[0097] AAACATAAGTGAACCAAGTTGTGAATATCTTGTTGCTAGTTTGCTGTTGCAGGGCTGGAGAC

[0098] CGAGGAACCACGTGTACCTAGCGATAGTGAGAAAGATATGGCAATAAGTACGGCTAACAGC

[0099] ATTGTCTCTGCGCTGAACAGCTACAGCAAGCTAACTGGTCTAGTGGTCAGCAGGAAGAGAT

[0100] GTGACTCGATTGGCATCCCAGCACTCAGTTCGTCGTTAAAACTCTCTCTCGGTAGAATCAAA

[0101] CAGAGCTTTGTATTGTGAGTTATTTGCATATTTTGTCCATTATTTTGTTGTCTACTTGTCTCGTC

[0102] TTATTTTTGTGTTGAAAACATTGTATGTCCAATTCATGCGTGCTTCATTATTAAACTGATCATT

[0103] TTGAACTGATCACAGGACTGACTCCCGCTTGCCTAACATGCCAATTATTTATGCTAGTGATGC

[0104] TTTTACATCATTAACAGGTAATTGTTTTTGACTGTTAGTACCAGAGTACGCATTCTAGCAGTAT

[0105] GTAATATGTGCAACTTCCAATATATTACTGTTATATTTTCAATATATTGTTTTCACTGTCTCATGT

[0106] TAATCAAACTGATTGTATATACTTGGTGCATGCATACATAAAACTCATGTGTTTTGGGATTGTC

[0107] ATGTGAAAAAAGAGAGCAATTTTTCACCCACTGCAACAGACAGCATCCTGGCTTTTTTTATT

[0108] TAGGACAGGGGTATGCGCTACATTTTCAATTTTTTCACAAAAATAAGACAACCTCGAGCATT

[0109] TTGCTAAGAACAAGACCTTCTCAAGCAGGTTGAGAAAACCTCCAAACCCCTGTCTGCCCAA

[0110] TATAAAGCGACACCATAGTACATGCGAGAACGACCATGACTAGGACTGAACCTTAAATATGT

[0111] GCTTTGACGTAGGATAGATGAGAGGACCTTTTAAACCTCAGACTGAAATTTGCTCTCATGGA

[0112] AAGTCGAACACAAGAACCTAAGGAGTGCTACTAGAGCCATCTAACCAACTTGAGGCCCTTT

[0113] CAGAAATGATGTAAAAATAGAAGTTAAGACCTGTTTTTGGTGATGCTATTGAGGTGACATAC

[0114] TTATTAATGCACTTTTCTTGTACAAGTGAACACAGTTTTAATATAAGTTGTTTAGAGACAATA

[0115] CATTCTTTCTTTGGTAGTTGTGCATGCTAGTAGTTGTGGTTCTTTTGGGTGATGTTTATTTTG

[0116] GTTAAAGCTTGCCTATGTTGCCAACTTGACATGGGGTCCATTACTCGGTGGCTGTGCATTATA

[0117] TTTCACTAGGAAATCATTTTTTTTCTTACAGGCAGAACTTATATTGCTCTTAAACTTTAACTTC

[0118] TTCAGGTTACTCAAGAGAAGAAATATTGGGCTGTAACTGCAAAGTCTTAAATGGGGCCAGGC

[0119] ACTAGCTTGGAGGTTTTAGAGGAGGTGCGGGCCAGGATTATATAGCTTGTATGACCACATTC

[0120] TCCATAATATAGCTTACCTTTTAAAATTTATATTCAAAATTGGACGTACAGATAAATCAGCATA

[0121] TTTGTTCTGAGCAGGCATGCACGGTGGATCTACTGAGTTACAGGTGCGTTGCTGCGTGTTTG

[0122] TACTTGTAGCTACCAGCTTTATGAGGTTATGCAAGGAACTTGACAGAAGTTTTGTTGCCTCTA

[0123] CTTTGTATCAGGAAAGATGGAAGTTCATTCTGTGATCATCTACATGTATCTCCTATCCGAGAT

[0124] GCTTCGGGCAAGGTTCGCTTACATCTTTCTCAGGTCTTCCCTGAAAGAAACTGTCTCTCAAA

[0125] TGCATTAAACAATCACTACACAATTGCTATGTTTGTGTGCTATGAAGCTCTAAACTACGGAAC

[0126] AGAATAGATTTGCAGTTTGCTATCAGTCAGATATTCCGCTCTTGTACAACCATCATGGGGATT

[0127] TTTTTTTTCCCGACTGACTTCATACCTACGCAGGTGGCCTTTCATATCTGGGTTCACCTTGAC

[0128] ATGGGTGCAAAGCACGATTTTAGTGGGCTGACCCCCGAGAAATGGCTGCTTGGCGCTGTTG

[0129] GTGCGGTGAGGGTTGCTGTGAGAGGCTTGTCGGCGTCAGGTAGCCTGTTGAGACCATCCCA

[0130] ATAGCAGTTTTAGTCTCCTCGCCATTTCATGACCACGGCGACCGGCCTTCCACGATGTACATT

[0131] GCTGCTTGCTATTCTGTGTAAAGCAACTGCTTAGCATGACAAAGTAAATGAGTTCGTTTTGA

[0132] AGGAAAATCCATGGTAAATACAAGATGTTGCTGTGACTAT;

[0133] The sequence of SEQ ID NO:2 is as follows:

[0134] MATSGAGDHGQGQGQGHGRFASSLTARYSDWVLETLDELPGSFLLTDPALPGHPIVYASRG

[0135] LAALTGYAPRDVLGRNARLFQGAATDRATVAGVREAVRAHRPHQAAILNYRRDGAPHWVLLH

[0136] LAPLFHARDGTLLHFLAVQVPIAPAAVARGATCHTTGHLLAACRDEASRVAEDFPCATHAGKVF

[0137] VDMDKRGLETEEPRVPSDSEKDMAISTANSIVSALNSYSKLTGLVVSRKRCDSIGIPALSSSLKLS

[0138] LGRIKQSFVLTDSRLPNMPIIYASDAFTSLTGYSREEILGCNCKVLNGPGTSLEVLEEINQHICSEQ

[0139] ACTVDLLSYRKDGSSFCDHLHVSPIRDASGKVAFHIWVHLDMGAKHDFSGLTPEKWLLGAVG

[0140] AVRVAVRGLSASGSLLRPSQ。

[0141] The present application is not intended to limit the source of the PLPB1 gene and its encoded protein. Specifically, as long as the homologous protein in the selected plant has similar functions to corn PLPB1, it meets the requirements of the present application. For example, the inventors of the present application used the MaizeGDB website to perform sequence analysis and found that sorghum (Sorghum bicolor L.) has a homologous protein (gene number Sb03g003350) with approximately 90% identity to the maize PLPB1 gene; Setaria viridis (L.) P.Beauv. has a homologous protein (gene number Si001684m.g) with approximately 80% identity to the maize PLPB1 gene; Rice (Oryza sativa L.) has a homologous protein (gene number LOC_Os01g09120) with approximately 75% identity to the maize PLPB1 gene; and Brachypodium distachyon (L.) P.Beauv. has a homologous protein (gene number Bradi2g05520) with approximately 75% identity to the maize PLPB1 gene. Therefore, it should be understood that although the PLPB1 gene provided in the examples of this application is derived from corn, sequences derived from other similar plants and having a certain identity with the sequence shown in SEQ ID NO: 1 of this application (for example, 70%, 75%, 80%, 85%, 90%, 95% or 100% identity) are also included in this application, as long as those skilled in the art can easily isolate and obtain the sequence from other plants based on the information provided in this application after reading this application.

[0142] Furthermore, those skilled in the art will appreciate that the nucleotide sequence of the PLPB1 gene is not limited to the specific sequences listed above, and should include sequences that contain one, two, or three or more nucleotide mutations compared to the sequence shown in SEQ ID NO: 1 but are still substantially functionally identical to the sequence shown in SEQ ID NO: 1, sequences that have 95%, 96%, 97%, 98%, or 99% sequence identity to the sequence shown in SEQ ID NO: 1, and sequences that have one or more nucleotides deleted, one or more nucleotides added, or one or more nucleotides substituted based on the sequence shown in SEQ ID NO: 1 but are 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence shown in SEQ ID NO: 1. Similarly, those skilled in the art will appreciate that the amino acid sequence of the protein encoded by the PLPB1 gene is not limited to the specific sequences listed above.

[0143] Based on this, the present application provides the use of the above-mentioned PLPB1 gene or the protein encoded by the PLPB1 gene in regulating plant nitrogen efficiency.

[0144] The plant is, for example, sorghum, foxtail grass, millet, Brachypodium distichum, rice or corn. In some embodiments, the plant is corn.

[0145] PLPB1 gene mutants

[0146] The present application provides a PLPB1 gene mutant, wherein the PLPB1 gene mutant comprises a substitution, deletion or insertion of one or more nucleotides based on a reference sequence, and the nucleotide sequence of the reference sequence is shown in SEQ ID NO: 1.

[0147] The nucleotide sequence of the PLPB1 gene mutant is shown in SEQ ID NO: 5 or SEQ ID NO: 6.

[0148] The sequence of SEQ ID NO:5 is as follows:

[0149] CACGCCGCCTCACCTCGCCGGGCGGCGTGATGGCGACTTCCGGGGCCGGCGACCACG

[0150] GGCAGGGCCAGGGCCAGGGGCACGGCCGCTTCGCGTCGTCCCTGACGGCGCGCTAACTCG

[0151] GACTGGGTGCTGGAGACGCCACCGACCGCGCGCACCGCCCGCACCAGGCCGCCATCCTCA

[0152] ACTACCGCCGCGACGGCGCCCCGCACTGGGTCCTCCTCCACCTCGCCCCCCTATTCCACGCC

[0153] CGCGACGGCACCCTGCTCCACTTCCTCGCCGTCCAGGTGCCATCGCCCCCGCCGCCGTCG

[0154] CGCGCGGGGCGACGTGCCACACGACGGGGCATCTGCTAGCCGCGTGCCGTGACGAGGCCA

[0155] GCAGGGTCGCGGAGGATTTCCCCTGCGCCACCCACGCGGGCAAGGTGTTTGTCGATATGGA

[0156] CAAGAGAGGTAATAGCTGCTAGTATGCTGCACTAGATAATAACTTACTCATGCTCGATTTTCCCC

[0157] TAGAGTTTCAAGCATTAATTGCATCTGAGTTCAATTAAATCCCTTCCTCGTTCCTAAAGCTGA

[0158] ACGTAGTGCGTTCCAAGCTTGACATTCCTTGGTTGGTTGAATTACCAGTTCAGCTATCGTATA

[0159] TTTATTATAGTTGCACTAATCTCATTTGTGGGCATAAACAGACAAACCGCTGAATAAGTAGGT

[0160] GTAGACTGTAGTGGCAGATTACTAAGAGAGTCAGATTACTCCCTTGTTCCATCTGGTCTTGG

[0161] TTCAAAGTCTGTTAATCGTGGCTATACGTTTATATTGCCCTTTACCCAATGCAAACTATGCAT

[0162] GGTTGCACGCACAAACACATATATGTGATAACTCTTTCCTTGCGCGGTGCTTACATAATGCGT

[0163] TCAACCCAAAAAAAAAAAAAAACTAATACGTTTTGGGATCAGGACAAACCACATTAGTGA

[0164] TTTCCAGGATGAAGATTCAACAGCGTGCATGTTAGTTACTTGTTCTTAAATGCTGAAGATCA

[0165] AGAACAAGACAACACAATTATTAATTATTATGGACATTCGTCCTCCCAAAGCATTATCTCCATAT

[0166] AGGAATAATGCTTATCAGACTAAGGTCACGAAAGACTTATCCCCATTTAAATATATGAAGAGA

[0167] ATGTAAAAGAATGAAGACAATAACTTCATTTTATTAATTCATATATATTTGCCTTTCATAAAAC

[0168] ATATATGAATGTTGACAATATTAATGCTACATTGATACTTTCGGCTTGCTCGAAGGTGAAGAC

[0169] GCGAAAGAGAGATTATAATTCAACGTAAATAGTATGGTGTTACTGTTCATCTATTTATAGGCA

[0170] CGAGACACGGTCCGAGTAAGTACATTTATACCCTTTATATTCATCTATAAACATAACACGA

[0171] ATCATTAAGGACTGAGTAGTCTTTGTCCCTTTTCGGTCATCATCATAATTGGTCTTCATCATCA

[0172] CATCAAGCCGAAGCTCATCGGCTATAGCTTCATCGTCTATTCTTATCACCTTCGGGCTACATCT

[0173] TTACCCTTTCATGAGAAAAGGCCTTCATCCCGAAACCGAAGCCCCCTGTAATAATTCATGTC

[0174] ATACTAAAAACATATGGTTAGCCAACACTGCAGTAGTTTACGAGTCTTCATTACTGGCAGTTT

[0175] GAGCTGGGGTTTATAGTTGACTTAACACTTCAATGTTTTCACTCTTACACACAAGTATCGAAC

[0176] AAAACATAAGTGAACCAAGTTGTGAATATCTTGTTGCTAGTTTGCTGTTGCAGGGCTGGAGA

[0177] CCGAGGAACCACGTGTACCTAGCGATAGTGAGAAAGATATGGCAATAAGTACGGCTAACAG

[0178] CATTGTCTCTGCGCTGAACAGCTACAGCAAGCTAACTGGTCTAGTGGTCAGCAGGAAGAGA

[0179] TGTGACTCGATTGGCATCCCAGCACTCAGTTCGTCGTTAAAACTCTCTCTCGGTAGAATCAA

[0180] ACAGAGCTTTGTATTGTGAGTTATTTGCATATTTTGTCCATTATTTTGTTGTCTACTTGTCTCGT

[0181] CTTATTTTTGTGTTGAAAACATTGTATGTCCAATTCATGCGTGCTTCATTATTAAACTGATCAT

[0182] TTTGAACTGATCACAGGACTGACTCCCGCTTGCCTAACATGCCAATTATTTATGCTAGTGATG

[0183] CTTTTACATCATTAACAGGTAATTGTTTTTGACTGTTAGTACCAGAGTACGCATTCTAGCAGT

[0184] ATGTAATATGTGCAACTTCCAATATATTACTGTTATATTTTCAATATATTGTTTTCACTGTCTCAT

[0185] GTTAATCAAACTGATTGTATATACTTGGTGCATGCATACATAAAACTCATGTGTTTTGGGATTG

[0186] TCATGTGAAAAAAGAGAGCAATTTTTCACCCACTGCAACAGACAGCATCCTGGCTTTTTTTA

[0187] TTTAGGACAGGGGTATGCGCTACATTTTCAATTTTTTCACAAAAATAAGACAACCTCGAGCA

[0188] TTTTGCTAAGAACAAGACCTTCTCAAGCAGGTTGAGAAAACCTCCAAACCCCTGTCTGCCC

[0189] AATATAAAGCGACACCATAGTACATGCGAGAACGACCATGACTAGGACTGAACCTTAAATAT

[0190] GTGCTTTGACGTAGGATAGATGAGAGGACCTTTTAAACCTCAGACTGAAATTTGCTCTCATG

[0191] GAAAGTCGAACACAAGAACCTAAGGAGTGCTACTAGAGCCATCTAACCAACTTGAGGCCCT

[0192] TTCAGAAATGATGTAAAAATAGAAGTTAAGACCTGTTTTTGGTGATGCTATTGAGGTGACAT

[0193] ACTTATTAATGCACTTTTCTTGTACAAGTGAACACAGTTTTAATATAAGTTGTTTAGAGACAA

[0194] TACATTTCTTTCTTTGGTAGTTGTGCATGCTAGTAGTTGTGGTTCTTTTGGGTGATGTTTATTT

[0195] TGGTTAAAGCTTGCCTATGTTGCCAACTTGACATGGGGTCCATTACTCGGTGGCTGTGCATTA

[0196] TATTTCACTAGGAAATCATTTTTTTTCTTACAGGCAGAACTTATATTGCTCTTAAACTTTAACT

[0197] TCTTCAGGTTACTCAAGAGAAGAAATATTGGGCTGTAACTGCAAAGTCTTAAATGGGGCCAG

[0198] GCACTAGCTTGGAGGTTTTAGAGGAGGTGCGGGCCAGGATTATATAGCTTGTATGACCACAT

[0199] TCTCCATAATATAGCTTACCTTTTAAAATTTATATTCAAAATTGGACGTACAGATAAATCAGCA

[0200] TATTTGTTCTGAGCAGGCATGCACGGTGGATCTACTGAGTTACAGGTGCGTTGCTGCGTGTT

[0201] TGTACTTGTAGCTACCAGCTTTATGAGGTTATGCAAGGAACTTGACAGAAGTTTTGTTGCCT

[0202] CTACTTTGTATCAGGAAAGATGGAAGTTCATTCTGTGATCATCTACATGTATCTCCTATCCGA

[0203] GATGCTTCGGGCAAGGTTCGCTTACATCTTTCTCAGGTCTTCCCTGAAAGAAACTGTCTCTC

[0204] AAATGCATTAAACAATCACTACACAATTGCTATGTTTGTGTGCTATGAAGCTCTAAACTACGG

[0205] AACAGAATAGATTTGCAGTTTGCTATCAGTCAGATATTCCGCTCTTGTACAACCATCATGGGG

[0206] ATTTTTTTTTTCCCGACTGACTTCATACCTACGCAGGTGGCCTTTCATATCTGGGTTCACCTTG

[0207] ACATGGGTGCAAAGCACGATTTTAGTGGGCTGACCCCCGAGAAATGGCTGCTTGGCGCTGT

[0208] TGGTGCGGTGAGGGTTGCTGTGAGAGGCTTGTCGGCGTCAGGTAGCCTGTTGAGACCATCC

[0209] CAATAGCAGTTTTAGTCTCCTCGCCATTTCATGACCACGGCGACCGGCCTTCCACGATGTAC

[0210] ATTGCTGCTTGCTATTCTGTGTAAAGCAACTGCTTAGCATGACAAAGTAAATGAGTTCGTTTT

[0211] GAAGGAAAATCCATGGTAAATACAAGATGTTGCTGTGACTAT;

[0212] The sequence of SEQ ID NO:6 is as follows:

[0213] CACGCCGCCTCACCTCGCCGGGCGGCGTGATGGCGACTTCCGGGGCCGGCGACCACG

[0214] GGCAGGGCCAGGGCCAGGGGCACGGCCGCTTCGCGTCGTCCCTGACGGCGCGCTTACTCG

[0215] GACTGGGTGCTGGAGACGCTGGACGAGCTCCCAGGGAGCTTCCTGCTCACGGACCCCGCC

[0216] CTGCCGGGCCACCCCATCGTACGCTTCCCGGGGCCTCGCCGCGCTCACCGGCTACGCGCCG

[0217] CGCGACGTGCTGGGCCGCAACGCGCGCCTCTTCCAGGGCGCCGCCACCGACCGCGCCACC

[0218] GTCGCCGGCGTCCGCGAGGCCGTCCGCGCGCACCGCCCGCACCAGGCCGCCATCCTCAACT

[0219] ACCGCCGCGACGGCGCCCCGCACTGGGTCCTCCTCCACCTCGCCCCCCTATTCCACGCCCG

[0220] CGACGGCACCCTGCTCCACTTCCTCGCCGTCCAGGTGCCCATCGCCCCCGCCGCCGTCGCG

[0221] CGCGGGGCGACGTGCCACACGACGGGGCATCTGCTAGCCGCGTGCCGTGACGAGGCCAGC

[0222] AGGGTCGCGGAGGATTTCCCCTGCGCCACCCACGCGGGCAAGGTGTTTGTCGATATGGACA

[0223] AGAGAGGTAATACTGCTAGTATGCTGCACTAGATAATAACTTACTCATGCTCGATTTTCCCTA

[0224] GAGTTTCAAGCATTAATTGCATCTGAGTTCAATTAAATCCCTTCCTCGTTCCTAAAGCTGAAC

[0225] GTAGTGCGTTCCAAGCTTGACATTCCTTGGTTGGTTGAATTACCAGTTCAGCTATCGTATATT

[0226] TATTATAGTTGCACTAATCTCATTTGTGGGCATAAACAGACAAACCGCTGAATAAGTAGGTGT

[0227] AGACTGTAGTGGCAGATTACTAAGAGAGTCAGATTACTCCCTTGTTCCATCTGGTCTTGGTT

[0228] CAAAGTCTGTTAATCGTGGCTATACGTTTATATTGCCCTTTTACCCAATGCAAACTATGCATGG

[0229] TTGCACGCACAAACACATATATGTGATAACTCTTTCCTTGCGCGGTGCTTACATAATGCGTTC

[0230] AACCCAAAAGAAAAAAAAAACTAATACGTTTTGGGATCAGGACAAACCACATTATGTGATT

[0231] TCCAGGATGAAGATTCAACAGCGTGCATGTTAGTTACTTGTTCTTAAATGCTGAAGATCAAG

[0232] AACAAGACAACACAATTATTAATTATTATGGACATTCGTCCTCCAAAGCATTATCTCCATATAG

[0233] GAATAATGCTTATCAGACTAAGGTCACGAAAGACTTATCCCCATTTAATATATGAATGAGAAT

[0234] GTAAAAGAATGAAGACAATAACTTCATTTTATTAATTCATATATATTTGCCTTTCATAAAACAT

[0235] ATATGAATGTTGACAATATTAATGCTACATTGATACTTTCGGCTTGCTCGAAGGTGAAGACGC

[0236] GAAAGAGAGATTATAATTCAACGTAAATAGTATGGTGTTACTGTTCATCTATTTATAGGCACG

[0237] AGACACGGTCCGAGTAAAAGTACATTTATACCCTTTATATTCATCTATAAACATAACACGAAT

[0238] CATTAAGGACTGAGTAGTCTTTGTCCCTTTTCGGTCATCATCATAATTGGTCTTCATCATCACA

[0239] TCAAGCCGAAGCTCATCGGCTATAGCTTCATCGTCTATTCTTATCACCTTCGGGCTACATCTTT

[0240] ACCCTTTCATGAGAAAAGGCCTTCATCCCGAAACCGAAGCCCCCTGTAATAATTCATGTCAT

[0241] ACTAAAAACATATGGTTAGCCAACACTGCAGTAGTTTACGAGTCTTCATTACTGGCAGTTTG

[0242] AGCTGGGGTTTATAGTTGACTTAACACTTCAATGTTTTCACTCTTACACACAAGTATCGAACA

[0243] AAACATAAGTGAACCAAGTTGTGAATATCTTGTTGCTAGTTTGCTGTTGCAGGGCTGGAGAC

[0244] CGAGGAACCACGTGTACCTAGCGATAGTGAGAAAGATATGGCAATAAGTACGGCTAACAGC

[0245] ATTGTCTCTGCGCTGAACAGCTACAGCAAGCTAACTGGTCTAGTGGTCAGCAGGAAGAGAT

[0246] GTGACTCGATTGGCATCCCAGCACTCAGTTCGTCGTTAAAACTCTCTCTCGGTAGAATCAAA

[0247] CAGAGCTTTGTATTGTGAGTTATTTGCATATTTTGTCCATTATTTTGTTGTCTACTTGTCTCGTC

[0248] TTATTTTTGTGTTGAAAACATTGTATGTCCAATTCATGCGTGCTTCATTATTAAACTGATCATT

[0249] TTGAACTGATCACAGGACTGACTCCCGCTTGCCTAACATGCCAATTATTTATGCTAGTGATGC

[0250] TTTTACATCATTAACAGGTAATTGTTTTTGACTGTTAGTACCAGAGTACGCATTCTAGCAGTAT

[0251] GTAATATGTGCAACTTCCAATATATTACTGTTATATTTTCAATATATTGTTTTCACTGTCTCATGT

[0252] TAATCAAACTGATTGTATATACTTGGTGCATGCATACATAAAACTCATGTGTTTTGGGATTGTC

[0253] ATGTGAAAAAAGAGAGCAATTTTTCACCCACTGCAACAGACAGCATCCTGGCTTTTTTTATT

[0254] TAGGACAGGGGTATGCGCTACATTTTCAATTTTTTCACAAAAATAAGACAACCTCGAGCATT

[0255] TTGCTAAGAACAAGACCTTCTCAAGCAGGTTGAGAAAACCTCCAAACCCCTGTCTGCCCAA

[0256] TATAAAGCGACACCATAGTACATGCGAGAACGACCATGACTAGGACTGAACCTTAAATATGT

[0257] GCTTTGACGTAGGATAGATGAGAGGACCTTTTAAACCTCAGACTGAAATTTGCTCTCATGGA

[0258] AAGTCGAACACAAGAACCTAAGGAGTGCTACTAGAGCCATCTAACCAACTTGAGGCCCTTT

[0259] CAGAAATGATGTAAAAATAGAAGTTAAGACCTGTTTTTGGTGATGCTATTGAGGTGACATAC

[0260] TTATTAATGCACTTTTCTTGTACAAGTGAACACAGTTTTAATATAAGTTGTTTAGAGACAATA

[0261] CATTCTTTCTTTGGTAGTTGTGCATGCTAGTAGTTGTGGTTCTTTTGGGTGATGTTTATTTTG

[0262] GTTAAAGCTTGCCTATGTTGCCAACTTGACATGGGGTCCATTACTCGGTGGCTGTGCATTATA

[0263] TTTCACTAGGAAATCATTTTTTTTCTTACAGGCAGAACTTATATTGCTCTTAAACTTTAACTTC

[0264] TTCAGGTTACTCAAGAGAAGAAATATTGGGCTGTAACTGCAAAGTCTTAAATGGGGCCAGGC

[0265] ACTAGCTTGGAGGTTTTAGAGGAGGTGCGGGCCAGGATTATATAGCTTGTATGACCACATTC

[0266] TCCATAATATAGCTTACCTTTTAAAATTTATATTCAAAATTGGACGTACAGATAAATCAGCATA

[0267] TTTGTTCTGAGCAGGCATGCACGGTGGATCTACTGAGTTACAGGTGCGTTGCTGCGTGTTTG

[0268] TACTTGTAGCTACCAGCTTTATGAGGTTATGCAAGGAACTTGACAGAAGTTTTGTTGCCTCTA

[0269] CTTTGTATCAGGAAAGATGGAAGTTCATTCTGTGATCATCTACATGTATCTCCTATCCGAGAT

[0270] GCTTCGGGCAAGGTTCGCTTACATCTTTCTCAGGTCTTCCCTGAAAGAAACTGTCTCTCAAA

[0271] TGCATTAAACAATCACTACACAATTGCTATGTTTGTGTGCTATGAAGCTCTAAACTACGGAAC

[0272] AGAATAGATTTGCAGTTTGCTATCAGTCAGATATTCCGCTCTTGTACAACCATCATGGGGATT

[0273] TTTTTTTTCCCGACTGACTTCATACCTACGCAGGTGGCCTTTCATATCTGGGTTCACCTTGAC

[0274] ATGGGTGCAAAGCACGATTTTAGTGGGCTGACCCCCGAGAAATGGCTGCTTGGCGCTGTTG

[0275] GTGCGGTGAGGGTTGCTGTGAGAGGCTTGTCGGCGTCAGGTAGCCTGTTGAGACCATCCCA

[0276] ATAGCAGTTTTAGTCTCCTCGCCATTTCATGACCACGGCGACCGGCCTTCCACGATGTACATT

[0277] GCTGCTTGCTATTCTGTGTAAAGCAACTGCTTAGCATGACAAAGTAAATGAGTTCGTTTTGA

[0278] AGGAAAATCCATGGTAAATACAAGATGTTGCTGTGACTAT.

[0279] Those skilled in the art will understand that the PLPB1 gene mutants are not limited to the specific sequences listed above. The PLPB1 gene mutants should include sequences that contain one, two, or three or more nucleotide mutations compared to the sequence shown in SEQ ID NO: 5 or SEQ ID NO: 6, but still have substantially the same functional identity as the sequence shown in SEQ ID NO: 5 or SEQ ID NO: 6, and sequences that have 95%, 96%, 97%, 98%, or 99% sequence identity to the sequence shown in SEQ ID NO: 5 or SEQ ID NO: 6, and sequences that have one or more nucleotides deleted, one or more nucleotides added, or one or more nucleotides substituted based on the sequence shown in SEQ ID NO: 5 or SEQ ID NO: 6 but have 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the sequence shown in SEQ ID NO: 5 or SEQ ID NO: 6.

[0280] Those skilled in the art can use any known method, such as random mutagenesis, directed mutagenesis, and directed evolution, to mutate the nucleotide sequence of the PLPB1 gene of the present application.

[0281] The present application also provides the use of any of the above-mentioned PLPB1 gene mutants in regulating plant nitrogen efficiency.

[0282] The plant is, for example, corn.

[0283] Biomaterials

[0284] The present application provides a biomaterial, comprising:

[0285] An expression cassette for inhibiting the action of the above-mentioned PLPB1 gene;

[0286] A recombinant vector capable of inhibiting the PLPB1 gene;

[0287] A recombinant microorganism capable of inhibiting the above-mentioned PLPB1 gene; or

[0288] A plant cell line comprising any one of the above-mentioned PLPB1 gene mutants.

[0289] The present application also provides the use of the above-mentioned biological materials in regulating plant nitrogen efficiency.

[0290] The plant is, for example, sorghum, foxtail grass, millet, Brachypodium distichum, rice or corn. In some embodiments, the plant is corn.

[0291] PLPB1 gene inhibitor

[0292] The inventors of the present application have discovered that inhibiting the function of the PLPB1 gene can effectively improve the nitrogen utilization efficiency of corn and increase corn yield under both low-nitrogen and normal-nitrogen conditions.

[0293] Based on this, the present application provides a PLPB1 gene inhibitor and provides the use of the PLPB1 gene inhibitor in regulating plant nitrogen efficiency.

[0294] The PLPB1 gene inhibitor regulates plant nitrogen efficiency by inhibiting the expression and / or activity of the PLPB1 gene.

[0295] The nucleotide sequence of the PLPB1 gene is shown in SEQ ID NO: 1.

[0296] The present application is not intended to limit the source of the PLPB1 gene. Specifically, as long as the homologous protein in the selected plant has a function similar to that of corn PLPB1, it meets the requirements of the present application.

[0297] Furthermore, those skilled in the art will appreciate that the nucleotide sequence of the PLPB1 gene is not limited to the specific sequences listed above, but should include sequences that contain one, two, or three or more nucleotide mutations compared to the sequence shown in SEQ ID NO: 1, but are substantially functionally identical thereto, sequences that have 95%, 96%, 97%, 98%, or 99% sequence identity to the sequence shown in SEQ ID NO: 1, and sequences that have one or more nucleotides deleted, one or more nucleotides added, or one or more nucleotides substituted based on the sequence shown in SEQ ID NO: 1 but are 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence shown in SEQ ID NO: 1.

[0298] The plant is, for example, sorghum, foxtail grass, millet, Brachypodium distichum, rice or corn. In some embodiments, the plant is corn.

[0299] In some embodiments, the PLPB1 gene inhibitor comprises a small molecule compound, a polypeptide, an inhibitory nucleic acid molecule that hybridizes with a PLPB1 nucleic acid molecule, or a gene editor.

[0300] Examples of inhibitory nucleic acid molecules include, but are not limited to, antisense nucleic acid molecules, small interfering RNA (siRNA), and short hairpin RNA (shRNA). Such inhibitory nucleic acid molecules can be designed to target any region of a PLPB1 nucleic acid molecule. In some embodiments, the antisense RNA, siRNA, or shRNA hybridizes to a sequence within a PLPB1 nucleic acid molecule or mRNA molecule and reduces expression of a PLPB1 polypeptide. In some embodiments, a PLPB1 gene inhibitor comprises an antisense molecule that hybridizes to a PLPB1 nucleic acid molecule or mRNA molecule and reduces expression of a PLPB1 polypeptide. In some embodiments, a PLPB1 gene inhibitor comprises an siRNA that hybridizes to a PLPB1 nucleic acid molecule or mRNA molecule and reduces expression of a PLPB1 polypeptide. In some embodiments, a PLPB1 gene inhibitor comprises an shRNA that hybridizes to a PLPB1 nucleic acid molecule or mRNA molecule and reduces expression of a PLPB1 polypeptide.

[0301] In the present application, the inhibitory nucleic acid molecule may comprise RNA or DNA, or both RNA and DNA. The inhibitory nucleic acid molecule may also be connected or fused to a heterologous nucleic acid sequence (such as a heterologous nucleic acid sequence in a vector) or a heterologous marker. For example, the inhibitory nucleic acid molecule may be in a vector comprising the inhibitory nucleic acid molecule and the heterologous nucleic acid sequence or as an exogenous donor sequence comprising the inhibitory nucleic acid molecule and the heterologous nucleic acid sequence. The inhibitory nucleic acid molecule may also be connected or fused to a heterologous marker. The marker may be directly detectable (such as a fluorophore) or indirectly detectable (such as a hapten, enzyme, or fluorophore quencher). Such markers may be detected by spectroscopy, photochemistry, biochemistry, immunochemistry, or chemical means. Such markers include, for example, radioactive labels, pigments, dyes, chromogens, spin labels, and fluorescent markers. The marker may also be, for example, a chemiluminescent substance; a metal-containing substance; or an enzyme. The term "marker" may also refer to a "tag" or a hapten that selectively binds to a conjugated molecule so that the conjugated molecule is used to generate a detectable signal when subsequently added with a substrate. Exemplary tags that can be used as tags to facilitate purification include, but are not limited to, MYC, HA, FLAG or 3XFLAG, 6XHis or polyhistidine, glutathione-S-transferase (GST), maltose binding protein, epitope tags, or the Fc portion of an immunoglobulin. Many tags include, for example, particles, fluorophores, haptens, enzymes and their calorimetric, fluorescent and chemiluminescent substrates, and other labels.

[0302] In the present application, the gene editor includes a DNA gene editor and an RNA gene editor. The gene editor includes a gene editing protein and an optional sgRNA.

[0303] Preferably, the gene editing protein is a Cas protein. In the present application, suitable Cas proteins include, for example, wild-type Cas9 protein and wild-type Cpf1 protein (such as FnCpf1). The Cas protein can have complete cutting activity to produce double-strand breaks in the PLPB1 nucleic acid molecule, or it can be a nickase that produces single-strand breaks in the PLPB1 nucleic acid molecule. Additional examples of Cas proteins include, but are not limited to, Cas1, Cas1B, Cas2, Cas3, Cas4, Cas5, Cas5e (CasD), Cas6, Cas6e, Cas6f, Cas7, Cas8a1, Cas8a2, Cas8b, Cas8c, Cas9 (Csn1 or Csx12), Cas10, Cas10d, CasF, CasG, CasH, Csy1, Csy2, Csy3, Cse1 (CasA), Cse2 (CasB), C se3 (CasE), Cse4 (CasC), Csc1, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4, Csm5, Csm6, Cmr1, Cmr3, Cmr4, Cmr5, Cmr6, Csb1, Csb2, Csb3, Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, Csx1, Csx15, Csf1, Csf2, Csf3, Csf4 and Cu1966, and homologs or modified versions thereof.

[0304] In some embodiments, the gene editor comprises a Cas protein and an sgRNA.

[0305] In this application, the terms "sgRNA", "guide RNA" and "CRISPR guide sequence" are used interchangeably throughout and refer to nucleic acids that contain sequences that determine the specificity of the Cas protein of the CRISPR / Cas system. The sgRNA hybridizes (partially or fully complementary) to a target nucleic acid sequence in the genome of the host cell. The length of the sgRNA or portion thereof that hybridizes to the target nucleic acid may be between 15-25 nucleotides, 18-22 nucleotides, or 19-21 nucleotides. In some embodiments, the length of the sgRNA sequence that hybridizes to the target nucleic acid may be 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides. In some embodiments, the length of the sgRNA sequence that hybridizes to the target nucleic acid is between 10-30 or 15-25 nucleotides.

[0306] In this application, the term "Cas protein" refers to a CRISPR-associated protein, which is a related protein in the CRISPR / Cas system.

[0307] In the present application, the small molecule compound can be natural or chemically synthesized.

[0308] Methods for regulating plant nitrogen efficiency

[0309] The present application provides a method for regulating plant nitrogen efficiency, wherein the method comprises: regulating plant nitrogen efficiency by inhibiting the expression and / or activity of the PLPB1 gene in the plant.

[0310] The nucleotide sequence of the PLPB1 gene is shown in SEQ ID NO: 1.

[0311] The present application is not intended to limit the source of the PLPB1 gene. Specifically, as long as the homologous protein in the selected plant has a function similar to that of corn PLPB1, it meets the requirements of the present application.

[0312] Furthermore, those skilled in the art will appreciate that the nucleotide sequence of the PLPB1 gene is not limited to the specific sequences listed above, but should include sequences that contain one, two, or three or more nucleotide mutations compared to the sequence shown in SEQ ID NO: 1, but are substantially functionally identical thereto, sequences that have 95%, 96%, 97%, 98%, or 99% sequence identity to the sequence shown in SEQ ID NO: 1, and sequences that have one or more nucleotides deleted, one or more nucleotides added, or one or more nucleotides substituted based on the sequence shown in SEQ ID NO: 1 but are 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence shown in SEQ ID NO: 1.

[0313] The plant is, for example, sorghum, foxtail grass, millet, Brachypodium distichum, rice or corn. In some embodiments, the plant is corn.

[0314] In some embodiments, the method for regulating plant nitrogen efficiency comprises: inhibiting the expression and / or activity of the PLPB1 gene by using any of the above-mentioned PLPB1 gene inhibitors, thereby regulating the plant nitrogen efficiency.

[0315] In some embodiments, methods for regulating plant nitrogen efficiency include: regulating plant nitrogen efficiency by using any of the aforementioned biological materials. For example, an expression cassette or recombinant vector capable of inhibiting the PLPB1 gene can be introduced into the plant. Suitable introduction methods include, but are not limited to, biolistic methods, PEG-mediated protoplast transformation, Agrobacterium-mediated transformation, plant virus-mediated transformation, pollen tube passage, and ovary injection.

[0316] In some embodiments, the method for regulating plant nitrogen efficiency includes: screening and identifying superior haplotypes in natural conditions to regulate plant nitrogen efficiency. The haplotypes can inhibit the expression and / or activity of the PLPB1 gene.

[0317] In the above, the regulation of plant nitrogen efficiency is reflected in that: when the expression level and / or activity of the PLPB1 gene in the plant decreases, the nitrogen utilization efficiency of the plant increases; conversely, when the expression level and / or activity of the PLPB1 gene in the plant increases, the nitrogen utilization efficiency of the plant decreases.

[0318] Application in plant breeding

[0319] The present application provides the use of the above-mentioned PLPB1 gene or the protein encoded by the PLPB1 gene in nitrogen-efficient plant breeding.

[0320] The present application also provides the use of any of the aforementioned PLPB1 gene mutants, any of the aforementioned biological materials, or any of the aforementioned PLPB1 gene inhibitors in nitrogen-efficient plant breeding.

[0321] The present application also provides a method for cultivating nitrogen-efficient plants, the method comprising: obtaining the nitrogen-efficient plants by inhibiting the expression and / or activity of the PLPB1 gene in the plants.

[0322] The plant is, for example, sorghum, foxtail grass, millet, Brachypodium distichum, rice or corn. In some embodiments, the plant is corn.

[0323] This application confirms that the PLPB1 gene is involved in the regulation of nitrogen metabolism in corn. By inhibiting the expression and / or activity of the PLPB1 gene in corn, this application successfully constructed nitrogen-efficient PLPB1 gene-edited materials. Compared with the wild-type B104 corn material, the PLPB1 gene-edited material exhibited higher nitrogen utilization efficiency and achieved increased yield under both low-nitrogen and normal-nitrogen conditions. This discovery not only enriches our understanding of the nitrogen metabolism mechanism of corn, but also provides genetic resources for the cultivation of new nitrogen-efficient varieties, contributing to the efficiency and environmental protection of agricultural production. In addition, the analysis of the function of the PLPB1 gene also lays the foundation for the improvement of nitrogen efficiency in other crops.

[0324] Example

[0325] The present application will be described below in conjunction with specific examples, but the scope of the present application is not limited thereto. Unless otherwise specified, the reagents and instruments used in the following examples are conventional reagents and instruments in this area and can be obtained commercially. Quantitative experiments are all performed with more than 5 replicates. The methods used are all conventional experimental methods, and those skilled in the art can undoubtedly implement the scheme and obtain corresponding results based on the embodiments.

[0326] Example 1 Response of PLPB1 to different nitrogen conditions

[0327] To investigate whether the expression of the PLPB1 gene (Zm00001d039530, the nucleotide sequence of which is shown in SEQ ID NO: 1) is affected by different nitrogen sources, B73 maize materials were cultured to the two-leaf and one-heart stage using the paper roll method, and then the endosperm was removed and transferred to NH 4 Hydroponics treatment was carried out in a nutrient solution containing Cl or KNO3 nitrogen sources (two nitrogen sources, four concentration gradients of 0.01, 0.4, 4, and 10 mM); after 3 days of hydroponic treatment, corn seedlings cultured in different concentrations of NH4Cl or KNO3 were used for fluorescence quantitative PCR experiments, where the primer sequences are shown in SEQ ID NO: 3 (CAGGCACTAGCTTGGAGGTT) and SEQ ID NO: 4 (CCCATGTCAAGGTGAACCCA).

[0328] The results are as follows Figure 1A As shown, with the addition of KNO3 and NH 4 As the concentration of Cl increased, the transcription level of the PLPB1 gene gradually increased, indicating that the expression of the PLPB1 gene in maize seedlings was affected by different concentrations of nitrogen sources.

[0329] Furthermore, in order to verify whether the expression of the PLPB1 gene is involved in the response of corn to nitrogen, the B73 corn material was cultured using the roll paper method until it reached the two-leaf and one-heart stage. After the endosperm was removed, the corn seedlings were transferred to a nutrient solution containing 2 mM NH4NO3 nitrogen source for hydroponic treatment. After 7 days of hydroponic treatment, the corn seedlings were transferred to a nitrogen-free culture solution and cultured for another 4 days. At 0 h, 24 h, 48 h, and 96 h, corn seedlings with consistent growth were taken for fluorescence quantitative PCR experiments. After the nitrogen starvation treatment, 4 mM NH4Cl or KNO3 nitrogen source was added again, and samples were taken at 0 h, 1 h, 3 h, 6 h, 18 h, 24 h, 48 h, and 72 h after the nitrogen supply was restored for fluorescence quantitative PCR experiments.

[0330] The results are as follows Figure 1B-1C As shown in Figure 2, after nitrogen source removal, the expression level of PLPB1 gene was significantly downregulated over time ( Figure 1B ); After nitrogen supply was restored, the expression of PLPB1 gene was significantly upregulated over time ( Figure 1C ). In summary, the PLPB1 gene is involved in the nitrogen metabolism process of maize.

[0331] Example 2 Obtaining PLPB1 gene editing material

[0332] In order to study the function of the PLPB1 gene, a CRISPR / Cas9 gene knockout vector for the PLPB1 gene was constructed based on the wild-type B104 corn material, and genetic transformation of corn plants was carried out using the Agrobacterium-mediated method. Figure 2A As shown, homozygous gene-edited materials (plpb1-1 and plpb1-2) were finally obtained through PCR identification and first-generation sequencing (Sanger). The target sequences of gene editing are shown in Figure 2B shown.

[0333] Among them, the sgRNA sequences used in the CRISPR / Cas9 gene knockout vector are as follows:

[0334] GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTAT CAACTTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTT (SEQ ID NO: 7).

[0335] Among them, the mutation of plpb1-1 is the insertion of 1bp at the first target site, and the insertion of 2bp and deletion of 184bp at the second target site, resulting in a frameshift mutation; the mutation of plpb1-2 is the insertion of 1bp at the first target site and the deletion of 2bp at the second target site, which also leads to a frameshift mutation ( Figure 2C The nucleotide sequence of the PLPB1 gene after mutation in plpb1-1 is shown in SEQ ID NO: 5, and the nucleotide sequence of the PLPB1 gene after mutation in plpb1-2 is shown in SEQ ID NO: 6.

[0336] Example 3 Phenotypes of PLPB1 gene-edited materials after low nitrogen and normal nitrogen treatments at the seedling stage

[0337] To verify whether PLPB1 is associated with efficient nitrogen use, the homozygous gene-edited materials plpb1-1 and plpb1-2, as well as the corresponding wild-type control B104 maize material (WT), were subjected to low nitrogen and normal nitrogen treatments at the seedling stage. The specific steps are as follows:

[0338] After the maize seedlings were cultured to the two-leaf and one-heart stage, they were cultured with Hoagland nutrient solution containing 0.04mM KNO3 and 4mM KNO3, respectively, and the nutrient solution was watered every three days; after 7 days of low nitrogen and normal nitrogen treatment, the plant height, leaf nitrogen content (determined using an N-Pen N110 plant nitrogen content meter) and leaf chlorophyll content (determined using a SPAD-502 chlorophyll meter) of plpb1-1, plpb1-2 and WT were measured.

[0339] The results are as follows Figures 3A-3D As shown, the growth of plpb1-1 and plpb1-2 was significantly better than that of WT ( Figure 3A and Figure 3B ); and after collecting phenotypic data, it was further found that the plant height, leaf nitrogen content, and chlorophyll content of plpb1-1 and plpb1-2 were significantly higher than those of WT under low nitrogen and normal nitrogen treatments ( Figure 3C and Figure 3D The above results indicate that editing the PLPB1 gene can effectively improve the nitrogen use efficiency of maize at the seedling stage.

[0340] Example 4 Yield Traits of PLPB1 Gene Edited Materials in Low Nitrogen Fields and Normal Nitrogen Fields

[0341] To further verify that PLPB1 can improve nitrogen use efficiency in maize, the homozygous gene-edited materials plpb1-1 and plpb1-2, as well as the corresponding wild-type control B104 maize material (WT), were subjected to normal nitrogen and low nitrogen treatments in Langfang, Hebei Province. The specific steps are as follows:

[0342] The normal nitrogen treatment included 191 kg / ha of urea (approximately 88 kg of pure nitrogen / ha), 750 kg / ha of superphosphate, and 135 kg / ha of potassium chloride before sowing; an additional 200 kg / ha of urea (approximately 92 kg of pure nitrogen / ha) was applied during the jointing stage. The low nitrogen treatment followed the same fertilization pattern as the normal nitrogen treatment, except that no nitrogen fertilizer was applied.

[0343] After harvesting corn ears, it was found that the yield per plant of plpb1-1 and plpb1-2 was significantly higher than that of WT ( Figure 4A ); After yield measurement, it was found that the ear length and ear weight of corn were significantly higher than those of WT ( Figure 4B and Figure 4C These results indicate that editing the PLPB1 gene can improve nitrogen use efficiency and yield of maize under low-nitrogen and normal-nitrogen conditions in the field.

[0344] The above description is merely a preferred embodiment of the present application and is not intended to limit the present application in any other manner. Any person skilled in the art may utilize the above disclosed technical content to modify or modify the present application into equivalent embodiments with equivalent variations. However, any simple modifications, equivalent variations, and modifications to the above embodiments that do not depart from the technical content of the present application and are based on the technical essence of the present application shall still fall within the scope of protection of the present application.

Claims

1. Application of the PLPB1 gene or the protein encoded by the PLPB1 gene in regulating plant nitrogen efficiency; Preferably, the plant is corn.

2. The use according to claim 1, wherein the nucleotide sequence of the PLPB1 gene is shown as SEQ ID NO: 1, or the amino acid sequence of the protein encoded by the PLPB1 gene is shown as SEQ ID NO:

2.

3. A PLPB1 gene mutant, wherein the PLPB1 gene mutant comprises a substitution, deletion or insertion of one or more nucleotides based on a reference sequence, the nucleotide sequence of the reference sequence being shown in SEQ ID NO:

1.

4. Use of the PLPB1 gene mutant according to claim 3 in regulating plant nitrogen efficiency; Preferably, the plant is corn.

5. A biomaterial, wherein the biomaterial comprises: An expression cassette for inhibiting the action of the PLPB1 gene; A recombinant vector capable of inhibiting the PLPB1 gene; A recombinant microorganism capable of inhibiting the PLPB1 gene; or A plant cell line comprising the PLPB1 gene mutant according to claim 3. The biomaterial according to claim 5 , wherein the nucleotide sequence of the PLPB1 gene is as shown in SEQ ID NO:

1.

7. Use of the biomaterial according to claim 5 or 6 in regulating plant nitrogen efficiency; Preferably, the plant is corn.

8. Application of PLPB1 gene inhibitors in regulating plant nitrogen efficiency; Preferably, the plant is corn.

9. The use according to claim 8, wherein the nucleotide sequence of the PLPB1 gene is as shown in SEQ ID NO: 1, and the inhibitor inhibits the expression and / or activity of the PLPB1 gene; Preferably, the inhibitor comprises a small molecule compound, a polypeptide, an inhibitory nucleic acid molecule that hybridizes with a PLPB1 nucleic acid molecule, or a gene editor.

10. A method for regulating plant nitrogen efficiency, wherein the method comprises: By inhibiting the expression and / or activity of the PLPB1 gene in the plant, the nitrogen efficiency of the plant is regulated; preferably, the plant is corn.

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