A protein for reducing corn plant height and application thereof

Abstract

The application discloses a protein for reducing corn plant height and application thereof, and belongs to the technical field of molecular breeding. Specifically, the protein GA2ox6 is derived from gibberellin oxidase protein, three protein variants of GA2ox6 are designed, and the protein sequences are as shown in SEQ ID NO: 2 (GA2ox6-1), SEQ ID NO: 4 (GA2ox6-2) and SEQ ID NO: 6 (GA2ox6-3). The application provides three gene sequences for reducing corn plant height and ear height, and corresponding methods, and the reduction ranges are different. According to the plant height of the corn variety to be improved, different methods can be selected to reduce the plant height to a suitable plant height, which is beneficial to breeders to quickly breed an improved line with a suitable plant height.

Description

Technical Field

[0001] This invention relates to the fields of plant genetic engineering and plant genetic breeding, and more specifically, to a protein that reduces maize plant height and its applications. Background Technology

[0002] Corn is the most widely planted crop in the world, with an annual global planting area exceeding 2 billion mu (approximately 133 million hectares) and a total output reaching 1 billion tons. Its nutritional, industrial, and feed value makes it crucial to my country's food security system. Currently, the increase in corn yield is attributed to increased planting density. Under high-density conditions, plants exhibit a shade-avoidance response, manifested as increased plant height and ear position, thinner stems, a smaller stem-leaf angle, elongated leaves, abnormal male and female development, and an increased interval between male and female reproductive stages. Ultimately, this results in reduced lodging resistance, lower photosynthetic efficiency, lower seed setting rate, and lower yield per plant. Compact plant types and low plant height generally exhibit stronger tolerance to high density. Reducing plant height often leads to thicker stems, and lowering the ear position is particularly beneficial for increasing lodging resistance. Furthermore, appropriately reducing plant height and ear position can also improve the corn harvest index. Therefore, dwarfing breeding while maintaining yield is an important direction in current corn breeding.

[0003] Currently, gene editing technology has been used to create various types of dwarfing mutants in maize inbred lines by knocking out single genes, altering gene amino acid sequences, and changing gene expression levels, achieving varying degrees of plant height reduction. Furthermore, some mutant forms show a greater reduction in ear height than in overall plant height; these mutations, which primarily reduce the length of internodes below the ear, are more beneficial for breeding lodging-resistant and densely planted maize.

[0004] Although much research has been done on plant height, and plant hormones such as auxin, gibberellin, brassinolide, and cytokinin can regulate plant height, the germplasm resources in maize that can be used for dwarfing improvement are limited. Using these limited dwarfing germplasm resources for plant height improvement through traditional breeding methods will further narrow the already scarce maize breeding germplasm resources, hindering the development of more superior maize varieties. Summary of the Invention

[0005] The technical problem to be solved by this invention is to provide a novel protein-reducing method to achieve significantly reduced plant height and ear height in transgenic plants. Corn is a monocotyledonous model plant. This invention takes corn as an example to improve corn lodging resistance, increase corn yield, and improve corn yield under high-density planting conditions.

[0006] Currently, gene editing technology has been used to create various types of dwarfing mutants in maize inbred lines by knocking out single genes, altering gene amino acid sequences, and changing gene expression levels. Alternatively, transgenic methods can achieve varying degrees of plant height reduction. However, most gene editing methods require simultaneous editing of both parents to achieve a suitable reduction in plant height in existing maize varieties; while transgenic methods may not reduce plant height sufficiently or have a significant impact on yield.

[0007] Gibberellin oxidase is a key regulatory site in the later stages of GA biosynthesis and catabolism, regulating gibberellin homeostasis in plants. Gibberellin oxidases mainly include different types such as GA20ox, GA3ox, and GA2ox.

[0008] The expression of GA2ox-like genes can regulate gibberellin levels. This invention has shown that rice overexpressing wild-type OsGA2ox6 exhibits a completely dwarfing trait.

[0009] Specifically, this invention provides three mutated genes, GA2ox6, which, when introduced into plants, have different effects on maize plant height and ear height, resulting in different reductions, so that improved lines can choose appropriate methods to reduce plant height.

[0010] The beneficial effects of this invention are:

[0011] This invention provides three gene sequences for reducing maize plant height and ear height, along with corresponding methods, each with varying degrees of reduction. Depending on the desired improvement in maize variety plant height, different methods can be chosen to reduce the plant height to a suitable level, facilitating breeders to quickly develop suitable height-adjusted lines for their specific varieties. Attached Figure Description

[0012] Figure 1 These are the spectra of carriers LP457, LP458, and LP459.

[0013] Figure 2 This is the result of routine PCR testing to verify maize plants transformed with the GA2ox6 gene.

[0014] Figure 3 This is a CDS difference diagram of GA20x6 contained in LP457, LP458, and LP459 vectors.

[0015] Figure 4 It is the amino acid sequence of GA20x6 contained in the LP457, LP458, and LP459 vectors.

[0016] Figure 5 This represents the actual plant height as observed in the field. Detailed Implementation

[0017] Example 1

[0018] Construction, preservation and detection of recombinant expression vectors

[0019] The protein GA2ox6 is derived from gibberellin oxidase protein. This invention designs three protein variants of GA2ox6 based on the wild-type GA2ox6 sequence shown in SEQ ID NO:20. The protein sequences are GA2ox6-1 as shown in SEQ ID NO:2, involving the mutation site E144A; GA2ox6-2 as shown in SEQ ID NO:4, involving the mutation site A145E; and GA2ox6-3 as shown in SEQ ID NO:6, involving the mutation site H147A.

[0020] Nanjing Genscript Biotechnology Co., Ltd. synthesized the gene encoding the gibberellin oxidase variant protein GA2ox6-1 by attaching an Nco I restriction site to its 5' end and an EcoRI restriction site to its 3' end. The nucleotide sequence of the gene GA2ox6-1 is shown in SEQ ID NO:1.

[0021] The nucleotide sequence of the synthesized GA2ox6-1 gene (SEQ ID NO:1) was ligated into the cloning vector pEASY-T5 (TransGen Biotech, Beijing, Cat. No: CT501-01), following the instructions for use of the pEASY-T5 vector. This yielded the LP-T recombinant cloning vector, which was then transformed into *E. coli* T1 competent cells (TransGen Biotech, Beijing, Cat. No: CD501) using a heat shock method. The heat shock conditions were as follows:

[0022] (1) 50 μL of Escherichia coli T1 competent cells and 10 μL of plasmid DNA (recombinant cloning vector LP-T) were incubated in a water bath at 42°C for 30 s and then incubated in a water bath at 37°C for 45 min (shaking at 200 rpm). The mixture was then spread on LB agar plates containing ampicillin (100 mg / L) (tryptone 10 g / L, yeast extract 5 g / L, NaCl 10 g / L, agar 15 g / L, pH adjusted to 7.5 with NaOH) and grown overnight.

[0023] (2) Pick white colonies and incubate them overnight at 37°C in LB liquid medium (tryptone 10g / L, yeast extract 5g / L, NaCl 10g / L, ampicillin 100mg / L, pH adjusted to 7.5 with NaOH).

[0024] (3) Alkaline extraction of plasmids: Centrifuge the bacterial culture at 12000 rpm for 1 min, remove the supernatant, and suspend the precipitated bacterial cells in 100 μL of ice-cold solution I (25 mM Tris-HCl, 10 mM EDTA (ethylenediaminetetraacetic acid), 50 mM glucose, pH 8.0); add 150 μL of freshly prepared solution II (0.2 M NaOH, 1% SDS (sodium dodecyl sulfate)), invert the tube 4 times to mix, and place on ice for 3-5 min; add 150 μL of ice-cold solution III (4 M potassium acetate, 2 M... Add acetic acid, mix thoroughly immediately, and place on ice for 5-10 minutes; centrifuge at 4°C and 12000 rpm for 5 minutes, add 2 volumes of anhydrous ethanol to the supernatant, mix well, and place at room temperature for 5 minutes; centrifuge at 4°C and 12000 rpm for 5 minutes, discard the supernatant, wash the precipitate with 70% ethanol and air dry; add 30 μL of TE (10 mM Tris-HCl, 1 mM EDTA, pH 8.0) containing RNase (20 μg / mL) to dissolve the precipitate; digest RNA in a water bath at 37°C for 30 minutes; store at -20°C for later use.

[0025] After the extracted plasmids were identified by Nco I and EcoRI digestion, the positive clones were sequenced for verification. The results showed that the nucleotide sequence inserted into the recombinant cloning vector LP-T was the nucleotide sequence of LP457 shown in SEQ ID NO:1 in the sequence listing.

[0026] The recombinant cloning vector LP-T and expression vector LP-BB1 (vector backbone: pCAMBIA3301 (available from the CAMBIA organization)) were digested with restriction endonucleases Nco I and EcoRI, respectively. The excised nucleotide sequence fragment of the target gene was then inserted between the Nco I and EcoRI sites of the expression vector LP-BB1 to construct the recombinant expression vector LP457. The vector map of LP457, which incorporates the GA2ox6-1 gene, is shown below. Figure 1 As shown.

[0027] The nucleotide sequences of the GA2ox6-2 and GA2ox6-3 genes were constructed into the recombinant expression vectors LP458 and LP459, respectively, using the same method. The nucleotide sequences of the GA2ox6-2 and GA2ox6-3 genes are shown in SEQ ID NO:3 and SEQ ID NO:5, respectively. The maps of vectors LP458 and LP459 are shown below. Figure 1 As shown;

[0028] Wherein, KanR: kanamycin gene; Ori: pUCorigin represents the replication region sequence of plasmid pUC, which can guide the double-stranded DNA replication process; RB: right boundary; pZmUbi1: from the promoter of the maize ubiquitin gene (SEQ ID NO:7); GA2ox6-1: a variant nucleotide sequence of the gene GA2ox6 (SEQ ID NO:1), similarly, GA2ox6-2 represents the second variant nucleotide sequence of the gene GA2ox6 (SEQ ID NO:3), and GA2ox6-3 represents the third variant nucleotide sequence of the gene GA2ox6 (SEQ ID NO:5); Nos: terminator of carmine synthase (SEQ ID NO:8); PAT: phosphinicotinamide acetyltransferase, used for transformation screening and conferring glufosinate herbicide resistance to transgenic plants (SEQ ID NO:9); 35S: terminator from cauliflower mosaic virus (CaMV) (SEQ ID NO:10); LB: left boundary.

[0029] Example 2:

[0030] 2.1 Plant Transformation and Detection

[0031] Transformation was performed using the conventional Agrobacterium infection method. The aseptically cultured transgenic maize embryos were co-cultured with the Agrobacterium in Example 1 to transfer the T-DNA from the constructed recombinant expression vectors LP457, LP458, and LP459 into the maize chromosome to generate transgenic maize events.

[0032] (1) Infection

[0033] For Agrobacterium-mediated maize transformation, briefly, immature embryos are isolated from maize and contacted with an Agrobacterium suspension, wherein Agrobacterium is capable of delivering the nucleic acid sequences of the gibberellin oxidase gene GA2ox6 and the PAT gene to at least one cell of one of the embryos. In this step, the embryos are specifically immersed in an Agrobacterium suspension (OD660 = 0.4-0.6, infection medium (MS salt 4.3 g / L, MS vitamins, casein 300 mg / L, sucrose 68.5 g / L, glucose 36 g / L, acetylsuccinone (AS) 40 mg / L, 2,4-dichlorophenoxyacetic acid (2,4-D) 1 mg / L, pH 5.3) to initiate inoculation.

[0034] (2) Co-cultivation

[0035] The embryos were co-cultured with Agrobacterium for a period of time (3 days). Specifically, after the infection step, the embryos were cultured on solid medium (MS salt 4.3 g / L, MS vitamins, casein 300 mg / L, sucrose 20 g / L, glucose 10 g / L, acetylsylgenone (AS) 100 mg / L, 2,4-dichlorophenoxyacetic acid (2,4-D) 1 mg / L, agar 8 g / L, pH 5.8).

[0036] (3) Recovery

[0037] Following this co-culture phase, a selective "recovery" step may be performed. In the "recovery" step, the recovery medium (MS salt 4.3 g / L, MS vitamins, casein 300 mg / L, sucrose 30 g / L, 2,4-dichlorophenoxyacetic acid (2,4-D) 1 mg / L, plant gel 3 g / L, pH 5.8) contains at least one known antibiotic that inhibits Agrobacterium growth (such as cephalosporin), without the addition of a selector for plant transformants.

[0038] (4) Choose

[0039] Specifically, the embryos were cultured on a solid medium containing antibiotics but without a selector to eliminate Agrobacterium and provide a recovery period for infected cells. Next, the inoculated embryos were cultured on a medium containing a selector (N-(phosphonocarboxymethyl)glycine) and the growing transformed callus was selected. Specifically, the embryos were cultured on a selection solid medium containing a selector (MS salt 4.3 g / L, MS vitamins, casein 300 mg / L, sucrose 30 g / L, N-(phosphonocarboxymethyl)glycine 0.25 mol / L, 2,4-dichlorophenoxyacetic acid (2,4-D) 1 mg / L, plant gel 3 g / L, pH 5.8), resulting in selective growth of transformed cells.

[0040] (5) Regeneration

[0041] Then, the callus tissue regenerates into a plant. Specifically, callus tissue grown on a medium containing a selector is cultured on solid media (MS differentiation medium and MS rooting medium) to regenerate a plant.

[0042] The selected resistant callus tissues were transferred to MS differentiation medium (MS salt 4.3 g / L, MS vitamins, casein 300 mg / L, sucrose 30 g / L, 6-benzyladenine 2 mg / L, N-(phosphonocarboxymethyl)glycine 0.125 mol / L, plant gel 3 g / L, pH=5.8) and cultured at 25℃ for differentiation. The differentiated seedlings were transferred to MS rooting medium (MS salt 2.15 g / L, MS vitamins, casein 300 mg / L, sucrose 30 g / L, indole-3-acetic acid 1 mg / L, agar 8 g / L, pH=5.8) and cultured at 25℃ until approximately 10 cm tall, then transferred to a greenhouse for further cultivation until fruit set. In the greenhouse, the seedlings were cultured at 28℃ for 16 hours daily, followed by 8 hours at 20℃.

[0043] 2.2 Detection of transgenic maize plants

[0044] (1) The maize plants transformed with the GA2ox6 gene were verified by conventional PCR using 2×EasyTaq PCRSuperMix (Cat. No: AS111-11) from Beijing TransGen Biotech Co., Ltd. The results are as follows: Figure 2 As shown.

[0045] PCR primers span the promoter and gene region:

[0046] GA2ox6-1 (LP457)

[0047] pZmUbi1-GA2ox6-1-F: 5'- GATGCTCACCCTGTTGTTTGGT -3' (SEQ ID NO: 11);

[0048] pZmUbi1-GA2ox6-1-R: 5'- GGACGTGGAACGCGGCC-3' (SEQ ID NO: 12).

[0049] PCR detection fragment size: 486bp.

[0050] GA2ox6-2 (LP458)

[0051] pZmUbi1-GA2ox6-2-F: 5'- GATGCTCACCCTGTTGTTTGGT -3' (SEQ ID NO: 13);

[0052] pZmUbi1-GA2ox6-2-R: 5'-GGGACGTGGAACTCCTCC-3' (SEQ ID NO: 14).

[0053] PCR detection fragment size: 487bp.

[0054] GA2ox6-3 (LP458)

[0055] pZmUbi1-GA2ox6-3-F: 5'- GATGCTCACCCTGTTGTTTGGT -3' (SEQ ID NO: 15);

[0056] pZmUbi1-GA2ox6-3-R: 5'- CGGGACGGCGAACGCCT-3' (SEQ ID NO: 16).

[0057] PCR detection fragment size: 488bp.

[0058] The PCR reaction conditions were: 30 cycles, each cycle being 95℃ for 30 min; 58℃ for 30 min; 72℃ for 40 min.

[0059] (2) Use TaqMan to verify maize plants transformed with the gibberellin oxidase gene GA2ox6

[0060] Approximately 100 mg of leaf samples were taken from maize plants transformed with the gibberellin oxidase gene GA2ox6 nucleotide sequence (SEQ ID NO:2). Genomic DNA was extracted using Qiagen's DNeasy Plant Maxi Kit, and the copy number of GA2ox6 was detected by Taqman probe-based quantitative real-time PCR. Wild-type maize plants (non-transgenic, recipient plants) were used as controls, and the same analysis was performed. The experiment was conducted in triplicate, and the average value was used.

[0061] The specific method is as follows:

[0062] Step 1: Take 100 mg of leaves with the transgenic gibberellin oxidase gene GA2ox6, grind them into a homogenate in a mortar with liquid nitrogen, and take 3 replicates for each sample.

[0063] Step 2: Use Qiagen's DNeasy Plant Mini Kit to extract genomic DNA from the above samples. Refer to the product instructions for specific methods.

[0064] Step 3: Determine the genomic DNA concentration of the above samples using NanoDrop 2000 (Thermo Scientific);

[0065] Step 4: Adjust the genomic DNA concentration of the above samples to the same concentration value, wherein the concentration value ranges from 80-100 ng / μl;

[0066] Step 5: The copy number of the samples was identified using TaqMan probe-based quantitative real-time PCR. Samples with known copy numbers were used as standards, and wild-type maize plants (non-transgenic, transformation recipients) were used as controls. Each sample was tested in triplicate, and the average value was taken. The primer and probe sequences for quantitative real-time PCR were as follows:

[0067] The following primers and probes are used to detect the Ubi-GA2ox6 gene sequence:

[0068] pZmUbi-F: GCCCTGCCTTCATACGCTAT, as shown in SEQ ID NO:17 in the sequence listing;

[0069] pZmUbi-R:TGTCGATGCTCACCCTGTTG, as shown in SEQ ID NO:18 in the sequence listing;

[0070] ZmGA2ox6:CGGAGTGGCAGCTACGTAAC, as shown in SEQ ID NO:19 in the sequence listing;

[0071] The PCR reaction system is as follows:

[0072] JumpStartTMTaq ReadyMixTM (Sigma): 10μL;

[0073] 50× primer / probe mixture: 1 μL;

[0074] Genomic DNA: 3 μL;

[0075] Water (ddH2O): 6 μL;

[0076] The 50× primer / probe mixture contains 45 μL of each primer at a concentration of 1 mM, 50 μL of the probe at a concentration of 100 μM, and 860 μL of 1×TE buffer, and is stored in amber tubes at 4°C.

[0077] The PCR reaction conditions are as follows:

[0078]

[0079] Data were analyzed using SDS2.3 software (Applied Biosystems).

[0080] Example 3: Ear Height Test

[0081] The CDS differences of GA20x6 contained in LP457, LP458, and LP459 vectors are as follows: Figure 3 As shown, the amino acid sequence is as follows Figure 4As shown in the table. When the three transgenic plants and the control were planted at a plant spacing of 25 cm and a row spacing of 50 cm, three plants were selected from each vector transformant (LP457, LP458, and LP459) for measurement. The control sample was a conventional variety (without exogenous vector insertion). The results showed that the plant height and ear height of the transgenic materials were significantly lower than those of the conventional control material. The reduction in plant height and ear height of LP457 and LP459 was less than that of LP458. However, in actual production applications, after large-scale yield testing, it was found that the transgenic maize plants containing the LP459 vector showed good agronomic traits in the field, with increased yield per unit area. The measurement results are shown in Tables 1-4, and the actual plant height in the field is shown in Tables 1-4. Figure 5 .

[0082] Table 1

[0083]

[0084] Table 2

[0085]

[0086] Table 3

[0087]

[0088] Table 4

[0089]

[0090] The improvement in plant height of maize plants containing the LP459 vector fell between that of LP457 and LP458, but at the same planting density, plants containing the LP459 vector exhibited higher yields. High-density planting yield experiments showed that LP459 yielded significantly higher than the control at high-density planting (6000 / 7000 / 8000 plants per mu). Specifically, at the same planting density of 8000 plants per mu, the yield increase reached 23.1%. The optimal planting density (highest yield) of LP459 at 8000 plants per mu increased the yield by 7.9% compared to the control at 6000 plants per mu, and the number of kernels increased by 25.1%. This indicates that, at the same planting area, high-density planting of maize plants containing the LP459 vector can effectively increase maize yield, demonstrating significant application value in high-density planting for yield increase and maize seed production.

[0091] The sequences involved in this invention are as follows:

[0092] Seq ID no:1 is the ga2ox6 CDS sequence of the lp457 vector:

[0093]

[0094] SEQ ID NO: 2. Amino acid sequence of the GA2OX6 protein of the LP457 vector.

[0095] MRYVAATPTPMSLVAEAAEPPlVDsYLELLRRGGGGGGIAAATEGCVQERELPLIDLTCLQGSAEAAARTTCADAMARAASewgffqvtghgvsrALLERLRAEQARLFRLPFETKAKAGLLNGSYRWGAPTAtsLRHLsWSAAFHVPLASISGTACDfGELSSLrDVVQEVADAMSrVAKTVAVALAGsLLGHDEAAAFPAGCGETTCYLRLNRYPACPFAANTFGLVPHTDSDFLTvLsQDqvGGGLQLMTdagwvavkprpdALIVNIGDLFQAWsNNLYKsVEHKVVANAaaERFSAaYFLCPSYDsLVGTcGEPSPYRdFTFGEYRRKVQEDVKRTGRKIGLPNFLKHRPPPQSRPA*

[0096]

[0097] SEQ ID NO:4 is the amino acid sequence of the GA2OX6 protein of the LP458 vector.

[0098] MRYVAATPTPMSLVAEAAEPPLVDSYLELLRRGGGGGGIAAATEGCVQERELPLIDLTCLQGSAGEAARTTCADAMARAASWGFFQVTGHGVSRALLERLRAEQARLFRLPFETKAKAGLLNGSYRWGAPTA*

[0099] SEQ ID NO:5 is the GA2OX6 CDS sequence of the LP459 vector

[0100]

[0101] seq id no:6 is the amino acid sequence of the ga2ox6 protein in the lp459 vector.

[0102] mryvaatptmpslvaesaaepplvdsylellrrggggggiaaategcvqerelplidltclqgsageaarttcadamaraasewgffqvtghgvsrallerlraeqarlfrlpfetkakagllngsyrwgaptatslrhlswseafavplasisgtacdfgelsslrdvvqevadamsrva ktvavalagsllghdeaaafpagcgettcyrlrlnrypacpfaantfglvphtdsdfltvlsqdqvgglqlmtdagwvavkprpdalivnigdlfqawsnnlyksvehkvvanaaaerfsaayflcpsydslvgtcgepspyrdftfgeyrrkvqedvkrtgrkiglpnflkhrpppqsrpa*

[0103] seq id no:7 Maize ubiquitin gene promoter

[0104]

[0105] SEQ ID NO:8 Tnos terminator

[0106] gatcgttcaaacatttggcaataaagtttcttaagattgaatcctgttgccggtcttgcgatgattatcatataatttctgttgaattacgttaagcatgtaataattaacatgtaatgcatgacgttatttatgagatgggtttttatgattagagtcccgcaattatacatttaatacgcgatagaaaacaaaatatagcgcgcaaactaggataaattatcgcgcgcggtgtcatctatgttactagatc

[0107] SEQ ID NO:9 pat

[0108] atgtctccggagaggagaccagttgagattaggccagctacagcagctgatatggccgcggtttgtgatatcgttaaccattacattgagacgtctacagtgaactttaggacagagccacaaacaccacaagagtggattgatgatctagagaggttgcaagatagatacccttggttggttgctgaggttgagggtgttgtggctggtattgcttacgctgggccctggaaggctaggaacgcttacgattggacagttgagagtactgtttacgtgtcacataggcatcaaaggttgggcctaggatccacattgtacacacatttgcttaagtctatggaggcgcaaggttttaagtctgtggttgctgttataggccttccaaacgatccatctgttaggttgcatgaggctttgggatacacagcccggggtacattgcgcgcagctggatacaagcatggtggatggcatgatgttggtttttggcaaagggattttgagttgccagctcctccaaggccagttaggccagttacccagatctga

[0109] SEQ ID NO:10 35S terminator

[0110] ctgaaatcaccagtctctctctacaaatctatctctctctataataatgtgtgagtagttcccagataagggaattagggttcttataggttcgctcatgtgttgagcatataagaaacccttagtatgtatttgtatttgtaaaatacttcttatcaataaaatttctatctaaaccaaaatccagtgg

[0111] seq id no:11 pzmubi1-ga2xo6-1-f:

[0112] gatgctcaccctgttgtttggt

[0113] seq id no:12 pzmubi1-ga2xo6-1-r:

[0114] ggacgtggaacgcggcc

[0115] seq id no:13 pzmubi1-ga2xo6-2-f:

[0116] gatgctcaccctgttgtttggt

[0117] seq id no:14 pzmubi1-ga2xo6-2-r

[0118] gggacgtggaactcctcc

[0119] seq id no:15 pzmubi1-ga2xo6-3-f:

[0120] gatgctcaccctgttgtttggt

[0121] seq id no:16 pzmubi1-ga2xo6-3-r:

[0122] cgggacggcgaacgcct

[0123] seq id no:17 pzmubi1-f:

[0124] gccctgccttcatacgctat

[0125] seq id no:18 pzmubi1-r:

[0126] tgtcgatgctcaccctgttg

[0127] seq id no:19 zmga2ox6 probe:

[0128] cggagtggcagctacgtaac

[0129] seq id no:20 Wild-type ga2ox6 sequence

[0130] mryvaatptmpslvaesaaepplvdsylellrrggggggiaaategcvqerelplidltclqgsageaarttcadamaraasewgffqvtghgvsrallerlraeqarlfrlpfetkakagllngsyrwgaptatslrhlswseafhvplasisgtacdfgelsslrdvvqevadamsrva ktvavalagsllghdeaaafpagcgettcyrlrlnrypacpfaantfglvphtdsdfltvlsqdqvgglqlmtdagwvavkprpdalivnigdlfqawsnnlyksvehkvvanaaaerfsaayflcpsydslvgtcgepspyrdftfgeyrrkvqedvkrtgrkiglpnflkhrpppqsrpa*

[0131] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A protein that reduces maize plant height, characterized in that, The protein amino acid sequence is obtained by mutating H at position 147 of the amino acid sequence shown in SEQ ID NO:20 to A.

2. A protein for reducing maize plant height as described in claim 1, characterized in that, The amino acid sequence of the protein is the amino acid sequence shown in SEQ ID NO:

6.

3. The gene sequence encoding the protein that reduces maize plant height as described in claim 1, characterized in that, The nucleotide sequence of the gene is the nucleotide sequence shown in SEQ ID NO:

5.

4. A biomaterial comprising the gene sequence of claim 3, characterized in that, The biomaterial is an expression cassette, vector, or host cell.

5. The application of the biomaterial described in claim 4 in reducing the height of maize plants.

6. The application of the biomaterials described in claim 4 in the improvement of maize germplasm resources.

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