Application of proteins or their encoding genes in regulating plant lipid biosynthesis

Abstract

The present invention discloses the use of proteins or their encoding genes in regulating plant lipid biosynthesis, relating to the field of biotechnology. The amino acid sequence of the ZmNAC128 protein is shown in SEQ ID NO.1; the amino acid sequence of the ZmNAC130 protein is shown in SEQ ID NO.2. The present invention demonstrates that the ZmNAC128 and ZmNAC130 proteins regulate lipid biosynthesis by activating the expression of lipid biosynthesis-related genes LPP1, DGATII1, and DGATII2. Mutations in the ZmNAC128 and ZmNAC130 genes can lead to reduced triacylglycerol content in grains. Therefore, the ZmNAC128 and ZmNAC130 proteins and related biomaterials provided by the present invention can be used to regulate lipid biosynthesis and have significant application and promotion value in increasing the oil content of plants.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, in particular to the application of proteins or their encoding genes in regulating plant lipid biosynthesis. Background Art

[0002] In modern agricultural production, corn (Zea mays) is the world's most important food and feed crop. Lipids are the fundamental building blocks of all biological cell membranes, playing a vital role in maintaining cell structural integrity and regulating cell function. In plants, lipids are not only the main components of cell membranes but also key substances for energy storage and plant metabolism.

[0003] The lipid biosynthesis pathway primarily involves fatty acid synthesis and lipid assembly. In this complex series of biochemical reactions, fatty acid synthase catalyzes the production of long-chain fatty acids, which are then converted through a series of enzymatic reactions into various lipids, such as triacylglycerols, phospholipids, and sterols. These processes are precisely regulated by a variety of enzymes and regulatory factors to ensure efficient lipid synthesis and product quality.

[0004] The Illinois Long-Term Selection Experiment, which began in 1896, is the longest continuous genetic experiment in higher plants. A striking observation from the Illinois Long-Term Selection Experiment was that corn kernel yield and starch content were negatively correlated with lipid content, likely because starch synthesis is more energy-efficient than lipid synthesis. In both the Illinois high-oil and Alexho synthetic populations, increasing kernel oil content resulted in an increased proportion of the embryo to the kernel, leading to a smaller kernel. A study of corn kernel texture revealed that non-starch lipids, primarily composed of triacylglycerols, accumulated much more in the starchy endosperm than in the vitreous endosperm. Therefore, rationally reducing lipid content could help improve corn kernel yield and firmness, offering broad application prospects.

[0005] However, there is still a lack of research on the regulatory factors of lipid biosynthesis in corn. This invention aims to provide a theoretical basis and practical guidance for improving corn kernel yield and hardness by studying the regulatory factors and regulatory mechanisms of lipid synthesis in plant kernels. Summary of the Invention

[0006] The present invention aims to address the aforementioned problems of the prior art by providing applications of proteins or their encoding genes for regulating plant lipid biosynthesis. The present invention has discovered that the ZmNAC128 and ZmNAC130 proteins and their encoding genes can be used to regulate lipid biosynthesis, demonstrating significant application and promotion value in increasing the oil content of plants such as corn.

[0007] To achieve the above object, the present invention provides the following solutions:

[0008] The present invention provides the use of a protein or a gene encoding the protein in regulating plant lipid biosynthesis, wherein the protein is a protein shown in any one of the following (a1) to (a4):

[0009] (a1) ZmNAC128 protein and / or ZmNAC130 protein; the amino acid sequence of the ZmNAC128 protein is shown in SEQ ID NO.1; the amino acid sequence of the ZmNAC130 protein is shown in SEQ ID NO.2;

[0010] (a2) a protein having the same function as the protein shown in SEQ ID NO. 1 and / or SEQ ID NO. 2, wherein one or more amino acid residues are substituted and / or deleted and / or added;

[0011] (a3) a protein having an amino acid sequence identity of 80% or more to that of the protein shown in (a1) or (a2) and having the same function;

[0012] (a4) A fusion protein obtained by ligating a tag to the end of any of the proteins shown in (a1) to (a3).

[0013] Furthermore, the plant lipid is plant seed triglyceride.

[0014] Furthermore, the plant is corn.

[0015] Furthermore, the nucleotide sequence of the gene encoding the ZmNAC128 protein is shown in SEQ ID NO.3; the nucleotide sequence of the gene encoding the ZmNAC130 protein is shown in SEQ ID NO.4.

[0016] The present invention also provides applications of zmnac128 protein and zmnac130 protein, wherein the applications are applications in regulating plant lipid biosynthesis;

[0017] The zmnac128 protein is a protein obtained by mutating the phenylalanine residue at position 83 of the protein shown in SEQ ID NO.1 to a leucine residue, deleting the tyrosine residue at position 84, and causing all subsequent amino acids to undergo frameshift mutations; the zmnac130 protein is a protein obtained by deleting the tyrosine residue at position 83 and the glutamine residue at position 84 of the protein shown in SEQ ID NO.2, mutating the lysine residue at position 85 to an arginine residue, and causing all subsequent amino acids to undergo frameshift mutations.

[0018] The present invention also provides an application of the biomaterial, wherein the application is an application in regulating plant lipid biosynthesis;

[0019] The relevant biological material is any one of the following (b1)-(b3):

[0020] (b1) a gene encoding the zmnac128 protein and a gene encoding the zmnac130 protein;

[0021] (b2) vectors for knocking out the ZmNAC128 gene and vectors for knocking out the ZmNAC130 gene;

[0022] (b3) a microbial strain containing the ZmNAC128 knockout gene and a vector containing the ZmNAC130 knockout gene;

[0023] The nucleotide sequence of the ZmNAC128 gene is shown in SEQ ID NO.3; the nucleotide sequence of the ZmNAC130 gene is shown in SEQ ID NO.4.

[0024] Furthermore, the plant lipid is plant seed triglyceride.

[0025] Furthermore, the plant is corn.

[0026] The present invention also provides a method for reducing the triacylglycerol content in corn kernels, comprising the following steps:

[0027] The ZmNAC128 gene knockout strain of maize was constructed to obtain ZmNAC128 gene knockout plants;

[0028] The ZmNAC130 gene knockout strain of maize was constructed to obtain ZmNAC130 gene knockout plants;

[0029] Crossing the ZmNAC128 gene knockout plant with the ZmNAC130 gene knockout plant to obtain a double mutant material, namely a corn plant with low triacylglycerol content in the grain;

[0030] The nucleotide sequence of the ZmNAC128 gene is shown in SEQ ID NO.3; the nucleotide sequence of the ZmNAC130 gene is shown in SEQ ID NO.4.

[0031] The present invention discloses the following technical effects:

[0032] The present invention demonstrates that the ZmNAC128 and ZmNAC130 proteins regulate lipid biosynthesis by activating the expression of lipid biosynthesis-related genes LPP1, DGATII1, and DGATII2. Mutations in the ZmNAC128 and ZmNAC130 genes can lead to reduced triacylglycerol content in grains. Therefore, the ZmNAC128 and ZmNAC130 proteins, Zmnac128 and Zmnac130 proteins, and related biomaterials provided by the present invention can be used to regulate lipid biosynthesis and have significant application and promotion value in increasing oil content, yield, and grain hardness in plants (such as corn). BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0034] Figure 1 Statistical graph of LPP1 gene expression in WT and zmnac128 / 130 mutant grains;

[0035] Figure 2 Statistical graph of the expression level of DGATII1 gene in WT and zmnac128 / 130 mutant grains;

[0036] Figure 3 Statistical graph of the expression level of DGATII2 gene in WT and zmnac128 / 130 mutant grains;

[0037] Figure 4 The figure shows the results of a dual luciferase assay of ZmNAC128 protein and ZmNAC130 protein on the LPP1 promoter in maize leaf protoplasts;

[0038] Figure 5 The figure shows the results of a dual luciferase assay of ZmNAC128 protein and ZmNAC130 protein on the DGATII1 promoter in maize leaf protoplasts;

[0039] Figure 6 This is a graph showing the results of a dual luciferase assay of ZmNAC128 and ZmNAC130 proteins on the DGATII2 promoter in maize leaf protoplasts;

[0040] Figure 7 Statistical graph of triacylglycerol content in grains of WT and zmnac128 / 130 mutant;

[0041] Figure 8Transmission electron micrographs of the aleurone layer of WT and zmnac128 / 130 mutant grains; arrows indicate oil bodies, Nu indicates nuclei, and scale bars are 10 μm.

[0042] Figure 9 Statistical graph of the number of oil bodies in the aleurone layer of WT and zmnac128 / 130 mutant grains. DETAILED DESCRIPTION

[0043] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0044] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. The intermediate value within any stated value or stated range, and each smaller range between any other stated value or intermediate value within the stated range, is also encompassed within the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0045] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0046] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be illustrative only.

[0047] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0048] Unless otherwise specified, the experimental methods in the following examples are conventional methods and were performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials and reagents used in the following examples, unless otherwise specified, were all commercially available.

[0049] Unless otherwise specified, GraphPad Prism 8 statistical software was used to process the data. The experimental results were expressed as mean values ​​and analyzed using Student's t-test.

[0050] The overexpression vector pHB and the fluorescent reporter vector pGreen 0800II have been disclosed in the following literature: Li, CB, et al. (2018). The ZmbZIP22 transcription factor regulates 27-kDγ-zein gene transcription during maize endosperm development. The Plant Cell, 10: 2402.

[0051] The CRISPR / Cas9 gene knockout vector has been disclosed in the following literature: Qi, WW, et al. (2016). High-efficiency CRISPR / Cas9 multiplex gene editing using the glycine tRNA-processing system-based strategy in maize. BMC biotechnology, 16: 58.

[0052] The maize Hi-II hybrid line (the hybrid line Hi II) used in the following examples has been disclosed in the following literature: Frame, BR, et al. (2002). Agrobacterium tumefaciens-mediated transformation of maize embryos using a standard binary vector system. Plant Physiology, 129: 13-22.

[0053] The amino acid sequence of the ZmNAC128 protein in the following examples is shown in SEQ ID NO. 1, the CDS sequence of the protein is shown in SEQ ID NO. 5, and the nucleotide sequence of the gene encoding the protein (ZmNAC128) is shown in SEQ ID NO. 3. The amino acid sequence of the ZmNAC130 protein is shown in SEQ ID NO. 2, the CDS sequence of the protein is shown in SEQ ID NO. 6, and the nucleotide sequence of the gene encoding the protein (ZmNAC130) is shown in SEQ ID NO. 4.

[0054] The nucleotide sequence of the zmnac128 gene in the following embodiments is a continuous nucleotide sequence after the deletion of positions 682 to 686 in the ZmNAC128 gene sequence. The nucleotide sequence of the zmnac130 gene is a continuous nucleotide sequence after the deletion of positions 855 to 861 in the ZmNAC130 gene sequence. The amino acid sequence of the protein encoded by the zmnac128 gene (zmnac128 protein) is a mutation of the phenylalanine residue at position 83 of the ZmNAC128 protein amino acid sequence to a leucine residue, a deletion of the tyrosine at position 84, and a frameshift mutation of all subsequent amino acids. The amino acid sequence of the protein encoded by the zmnac130 gene (zmnac130 protein) is a deletion of the tyrosine residue at position 83 and the glutamine residue at position 84 in the ZmNAC130 protein amino acid sequence, a mutation of the lysine residue at position 85 to an arginine residue, and a frameshift mutation of all subsequent amino acids. The CDS of the zmnac128 protein is the continuous nucleotide sequence after the deletion of positions 249 to 253 of the CDS of the ZmNAC128 protein. The CDS of the zmnac130 protein is the continuous nucleotide sequence after the deletion of positions 247 to 253 of the CDS of the ZmNAC130 protein.

[0055] RNA Detection Method: Immature corn kernels were harvested and their endosperms were used for total RNA extraction, which was then reverse-transcribed into cDNA. Real-time fluorescence quantitative analysis of relevant genes was performed, using the maize ACTIN gene (GenBank accession number NM_001155179.2) as an internal control. Expression levels of relevant genes were calculated relative to the expression level of the ACTIN gene.

[0056] Triglyceride content assay: Mature corn kernels were harvested and their endosperms were used for triglyceride extraction. A kit (Solarbio, Cat. No. BC0625) was used to assay triglyceride content.

[0057] Example 1 CRISPR / Cas9 knockout of ZmNAC128 and ZmNAC130 genes

[0058] (1) Construction of ZmNAC128 gene knockout maize transgenic plants

[0059] ① Construction of CRISPR / Cas9 knockout vector for ZmNAC128 gene

[0060] A synthetic fragment containing the 20-bp target nucleotide sequence (SEQ ID NO. 7) in the ZmNAC128 gene was digested with Xba I and ligated between the maize U6 promoter and terminator. Subsequently, a fragment containing the U6 promoter-synthetic fragment-U6 terminator was digested with Pst I and ligated into the CRISPR / Cas9 knockout vector to complete the construction of the ZmNAC128 gene knockout vector.

[0061] ②CRISPR / Cas9 knockout vector transformation into maize embryos

[0062] The ZmNAC128 gene knockout vector was transformed into Agrobacterium tumefaciens EHA105, and then the vector was transformed into immature embryos of maize Hi-II hybrid using the Agrobacterium-mediated method to obtain transgenic T0 generation plants.

[0063] ③ Editing and identification of transgenic plants

[0064] PCR amplified the fragment containing the target site, followed by TA cloning and sequencing, to obtain the editing information of the ZmNAC128 gene. Plants with ZmNAC128 knockout were named zmnac128-cas9 mutants (zmnac128-cas9). In this mutant, #1, a C was deleted at position 17 of the ZmNAC128 gene target site, while #14, a CTACC was deleted between positions 13 and 17 of the ZmNAC128 gene target site. Both events introduced a premature stop codon, resulting in premature termination of protein translation.

[0065] Since the ZmNAC130 gene also contains a 20bp target nucleotide with the same sequence (SEQ ID NO.7), PCR was used to amplify the fragment containing the target, and the fragment was sequenced after TA cloning to obtain the edited plant of the ZmNAC130 gene. The plant with the ZmNAC130 gene knocked out was named zmnac130 mutant (zmnac130), among which 1# deleted a large fragment of nucleotides starting from the 17th position of the target site of the ZmNAC130 gene, and there were a large number of single nucleotide mutations near the target. 14# deleted TACCAGA between the 14th and 20th positions of the first target site of the ZmNAC130 gene. Both of the above events introduced the stop codon prematurely, causing the protein translation to terminate prematurely.

[0066] (2) Hybridization of ZmNAC128 knockout plants and ZmNAC130 knockout plants

[0067] The zmnac128 and zmnac130 plants from event 14# were planted in the field and hybridized. The heterozygous plants were then planted in the field and self-pollinated to obtain double mutants of the ZmNAC128 and ZmNAC130 genes, named zmnac128 / 130.

[0068] Example 2: Reduced expression of lipid biosynthesis-related genes in the zmnac128 / 130 mutant

[0069] After reverse transcription of the extracted total RNA into cDNA, the expression levels of lipid biosynthesis-related genes LPP1, DGATII1, and DGATII2 in wild-type (WT) and mutant (zmnac128 / 130) materials were detected. The detection primers are as follows:

[0070] LPP1-F: GGTCTTACAGTTGCTTCGTTTT (SEQ ID NO. 8);

[0071] LPP1-R: CTCCCATGGTATCTGAGACATC (SEQ ID NO.9);

[0072] DGATII1-F: GACCACACTAGAACTAGAACCC (SEQ ID NO. 10);

[0073] DGATII1-R: TTGCTCAGGCCATTATTGGTTC (SEQ ID NO. 11);

[0074] DGATII2-F: TACATGCAGCTGTATCATCAGT (SEQ ID NO. 12);

[0075] DGATII2-R:GTTCTTCAGACAATGACACAGC (SEQ ID NO. 13);

[0076] ACTIN-F: GCTACGAGATGCCTGATGGTC (SEQ ID NO. 14);

[0077] ACTIN-R: CCCCCACTGAGGACAACG (SEQ ID NO. 15).

[0078] The reaction system was: ddH2O 8.2 μL, qPCR SYBR Green Master Mix (Yeasen) 10 μL, primer F 0.4 μL, primer R 0.4 μL, and cDNA 1 μL.

[0079] The reaction procedure (two-step method) is as follows:

[0080] Pre-denaturation: 95°C for 5 min, cycle number 1; 95°C for 10 sec, 60°C for 30 sec, cycle number 40.

[0081] RT-qPCR results showed that the expression levels of LPP1, DGATII1 and DGATII2 genes were decreased in zmnac128 / 130 compared with WT ( Figure 1-3 ).

[0082] Example 3 ZmNAC128 and ZmNAC130 regulate lipid biosynthesis

[0083] 1. Construction of overexpression vector and fluorescent reporter vector

[0084] (1) The full-length coding sequences of genes ZmNAC128 (SEQ ID NO. 5) and ZmNAC130 (SEQ ID NO. 6) were amplified using specific primers. The primers are as follows:

[0085] pHB-ZmNAC128-F: ATCACCAGTCTCTCTCTCCAAGCTTATGGCGGACCAGCAGCAGC C (SEQ ID NO. 16);

[0086] pHB-ZmNAC128-R: CGATGATACGAACGAAAGCTCTAGATCAGTACTTGTACTTCCA TA (SEQ ID NO. 17);

[0087] pHB-ZmNAC130-F: ATCACCAGTCTCTCTCTCCAAGCTTATGGCGGCGGACCAGCAGC C (SEQ ID NO. 18);

[0088] pHB-ZmNAC130-R: CGATGATACGAACGAAAGCTCTAGATCAGTACTTCCACACGCC AT (SEQ ID NO. 19).

[0089] (2) The fragment containing the full-length coding sequence was ligated to the pHB vector that had been double-digested with Hind III and Xba I, thereby constructing the overexpression vector for genes ZmNAC128 and ZmNAC130.

[0090] (3) Specific primers were used to amplify the promoter sequences of lipid biosynthesis-related genes LPP1, DGATII1, and DGATII2 containing the conserved binding motifs of ZmNAC128 and ZmNAC130, respectively. The primers are as follows:

[0091] pLPP1-F:TCGACGGTATCGATAAGCTTAACCCACGGTTTTACGGGTT (SEQ ID NO. 20);

[0092] pLPP1-R: GTGGATCCCCCGGGCTGCAGAGGTGGAGGAAGGTGGGTC (SEQ ID NO. 21);

[0093] pDGATII1-F: TCGACGGTATCGATAAGCTTGGGAGAGCGGAAGACAAGAG (SEQ ID NO. 22);

[0094] pDGATII1-R: GTGGATCCCCCGGGCTGCAGCGCCACGACTCTGCATCAAT (SEQ ID NO. 23);

[0095] pDGATII2-F:TCGACGGTATCGATAAGCTTGAGAACCACTCCTAGCCAGC (SEQ ID NO. 24);

[0096] pDGATII2-R: GTGGATCCCCCGGGCTGCAGTACAGCAAGGCCTGATGGAC (SEQ ID NO. 25).

[0097] (4) The fragment containing the promoter sequence was connected to the pGreen 0800II vector that had been double-digested with Hind III and Pst I, thus completing the construction of the promoter fluorescent reporter vector.

[0098] 2. Dual luciferase assay test

[0099] The cauliflower mosaic virus (CaMV) 35S promoter was used to drive the expression of genes ZmNAC128 and ZmNAC130. The reporter vector used the p35S promoter to drive the expression of the Renilla luciferase gene (REN) as an internal reference, and the lipid biosynthesis-related gene promoter was used to drive the expression of the firefly luciferase gene (LUC). The overexpression vectors of genes ZmNAC128 and ZmNAC130 and the reporter vector were co-transformed into corn leaf protoplasts, and luciferase substrate was added for signal detection. The results showed that ZmNAC128 and ZmNAC130 significantly activated the expression of LPP1, DGATII1 and DGATII2 genes ( Figure 4-6 The triglyceride content of WT and zmnac128 / 130 mature grains was detected. The results showed that the triglyceride content in zmnac128 / 130 mature grains was significantly reduced ( Figure 7 ).

[0100] Example 4: Reduced Oil Body Number in Immature Kernels of WT and zmnac128 / 130 Mutants

[0101] Transmission electron microscopy was used to observe the aleurone layer in immature grains of WT and zmnac128 / 130 mutants. It was found that aleurone cells were filled with a large number of oil bodies ( Figure 8 ).like Figure 9 As shown, by counting 10 areas of 100 μm 2 The number of oil bodies in the region of α-glucose phosphate-buffered saline (ΔGTP-glucose) indicated that the number of oil bodies was reduced in zmnac128 / 130.

[0102] SEQ ID NO.1:

[0103] MADQQQPQQQPQEMDVDRTGGLELPPGFRFHPSDFEIINDYLTKKVHDRDYSCIAIADADLNKTEPWDLPKVAKMGEKEWYFFYQKDRKYPTGLRANRATEAGYWKATGKDKEVYNPFAAEGLLLVGMKKTLVFYKGRAPRGDKTNWVMHEYRLEGSGRLPASPASASGSATNIAAAMMKASASACKDEWVVVCRVF NKTTGIKKTAAPAYQVAMAGPEMDQNQNNIPAIPIPMPLQLPLPVPMQMQFPILPDFAMDPVAPYYPNPNAGAGMMPPMALAGMGGAGGLQINGALFGNPVPAPLPMNFYHHQMGMGAAAGQVDMGAAAGQMDMGAAGAGAGGFDVAAPESRPSSMVSQKDEQANAAEISSMMSVTGPGSATTTIEMDGIWKYKY.

[0104] SEQ ID NO.2:

[0105] MAADQQPQLQEEMNDAAGGGLRLPGFRFHPSDFEIVSFYLTNKVLNTRFTCTAITEADLNKIEPWDLPSKAKMGEKEWYFFYQKDRKYPTGLRANRATEAGYWKATGKDKEVYNAAEGVAVLVGMKKTLVFYRGRAPRGDKTNWVMHEYRLEGSGRLPAGLASATGSAAANAAAALKAS AYKQDEWVVCRVFHKTTGIKKTTAAPAYQVAMAGAEMDQNQNNFPGIPFPMPMQFPMLPDFSLDPVPPYYPNAAGAGMSMLPMAAGIGGGAGGFQLNGAALFGNPMAAPQPMSFYHQMGAAGTACAGGFDVSAPESRPSSMVSQKDDQANGAEISSMMSVAGPGPATTTIEMDGVWKY。

[0106] SEQ ID NO.3:

[0107]

[0108] SEQ ID NO.4:

[0109]

[0110] SEQ ID NO.5:

[0111]

[0112] SEQ ID NO.6:

[0113] ATGGCGGCGGACCAGCAGCCGCAGCTGCAGGAGGAGATGAACGACGCTGCCGGCGGCGGCCTCAGGCTGCCTCCAGGGTTCCGCTTCCACCCGAGCGACTTCGAGATTGTCAGCTTCTACCTCACCAACAAGGTGCTCAACACGCGCTTCACCTGCACCGCCATCACGGAGGCCGACCTAAACAAGATTGAGCCATGGGACCTCCCTAGCAAGGCGAAGATGGGCGAGAAAGAGTGGTACTTCTTCTACCAGAAGGACCGCAAGTACCCGACGGGGCTGAGGGCGAACCGGGCCACCGAGGCCGGTTACTGGAAGGCGACGGGCAAGGACAAGGAGGTCTACAACGCCGCGGAAGGGGTGGCGGTACTGGTCGGCATGAAGAAGACGCTCGTCTTCTACAGGGGCAGGGCTCCCAGGGGTGACAAGACAAACTGGGTCATGCACGAGTACAGGCTCGAAGGCAGCGGCAGGCTCCCCGCCGGCCTCGCGTCCGCAACCGGCTCAGCCGCCGCCAACGCCGCGGCGGCCTTGAAAGCTTCTGCTTATAAGCAGGATGAGTGGGTAGTGTGTCGTGTGTTCCACAAGACCACTGGGATCAAGAAGACCACTGCTGCACCGGCGTACCAGGTGGCCATGGCCGGCGCTGAGATGGATCAGAATCAGAACAACTTCCCGGGCATCCCCTTCCCCATGCCGATGCAATTTCCCATGCTGCCAGACTTCTCCTTGGACCCGGTGCCCCCCTACTACCCCAACGCCGCTGGCGCGGGGATGTCGATGCTTCCTATGGCAGCAGGTATAGGTGGTGGCGCCGGTGGGTTCCAGCTCAACGGCGCCGCCCTGTTCGGCAATCCGATGGCCGCGCCGCAGCCCATGAGCTTCTACCACCAGATGGGCGCGGCGGGGACAGCTTGCGCTGGCGGCTTCGATGTTTCTGCGCCGGAGAGTAGGCCGTCCTCGATGGTGTCGCAGAAGGACGACCAGGCTAATGGCGCTGAGATCTCGTCGATGATGTCCGTGGCCGGCCCAGGGCCTGCGACCACCACCACCATAGAGATGGATGGCGTGTGGAAGTACTGA。

[0114] SEQ ID NO.7:GTGGTACTTCTTCTACCAGA。

[0115] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.

Claims

1. The application of biomaterials in reducing lipid biosynthesis in corn, characterized in that: The biomaterial is any one of the following (1)-(2): (1) Vectors for knocking out the ZmNAC128 gene and vectors for knocking out the ZmNAC130 gene; (2) a microbial strain containing the vector for knocking out the ZmNAC128 gene and a microbial strain containing the vector for knocking out the ZmNAC130 gene; The nucleotide sequence of the ZmNAC128 gene is shown in SEQ ID NO.3; the nucleotide sequence of the ZmNAC130 gene is shown in SEQ ID NO.

4.

2. The use according to claim 1, characterized in that The corn lipid is plant seed triacylglycerol.

3. A method for reducing the triglyceride content in corn kernels, characterized in that: The following steps are involved: The ZmNAC128 gene knockout strain of maize was constructed to obtain ZmNAC128 gene knockout plants; The ZmNAC130 gene knockout strain of maize was constructed to obtain ZmNAC130 gene knockout plants; Crossing the ZmNAC128 gene knockout plant with the ZmNAC130 gene knockout plant to obtain a double mutant material, namely a corn plant with low triacylglycerol content in the grain; The nucleotide sequence of the ZmNAC128 gene is shown in SEQ ID NO.3; the nucleotide sequence of the ZmNAC130 gene is shown in SEQ ID NO.4.

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