ZmPAR1 gene and its application

Abstract

This application relates to the field of maize technology, specifically to the ZmPAR1 gene and its applications. A gene encoding a nucleoside kinase was discovered in the maize genome. This nucleoside kinase was obtained through in vitro recombinant expression of the encoded gene. Testing revealed that this nucleoside kinase exhibits binding activity to various nucleoside substrates and can catalyze the synthesis of monophosphate nucleotides from nucleosides. This gene influences maize plant height, leaf angle, ear height, leaf length, and leaf width. Knocking down or overexpressing this gene can yield new maize materials, providing technical support for the breeding of new maize materials and the construction of germplasm resources.

Description

Technical Field

[0001] This application relates to the field of maize technology, specifically to the ZmPAR1 gene and its applications. Background Technology

[0002] Corn is an important dual-purpose crop for both food and feed, and also a crucial industrial raw material. With increasing planting density, the leaves of corn plants often shade each other, easily triggering shade avoidance syndrome, which leads to increased plant height, thinner stems, narrower leaves, and premature aging. Improving the corn plant architecture, including plant height, leaf angle, leaf length, and leaf width, can reduce the occurrence of shade avoidance syndrome to some extent. Improving the corn plant architecture relies on cloning plant architecture genes. However, the number of known plant architecture genes is currently limited, and more new genetic resources are needed to breed varieties with ideal plant architectures. Summary of the Invention

[0003] The inventors of this application discovered a gene encoding a nucleoside kinase in the maize genome. This gene is located at position 189856343–189862099 bp on chromosome 1, with the reference genome number Zm-B73-REFERENCE-GRAMENE-4.0 (database (https: / / maizegdb.org / )). The nucleoside kinase was obtained through in vitro recombinant expression of this gene. Testing revealed that this nucleoside kinase exhibits binding activity to various nucleoside substrates and can catalyze the synthesis of monophosphate nucleotides from nucleosides.

[0004] Furthermore, the inventors of this application have discovered that this gene affects maize plant height, leaf angle, ear height, leaf length, and leaf width. Knocking down or overexpressing this gene can yield new maize materials, providing technical support for the breeding of new maize materials and the construction of germplasm resources.

[0005] Therefore, the embodiments of this application disclose at least the following technical solutions:

[0006] In one aspect, the embodiments disclose an enzyme preparation using a protein as shown in SEQ ID NO:1 as the active ingredient.

[0007] Secondly, the embodiments disclose a method for preparing ZmPAR1 protein. The method includes providing an expression vector containing a nucleotide sequence encoding the ZmPAR1 protein, transforming the expression vector into *E. coli* to obtain an expression strain, and obtaining the ZmPAR1 protein from the culture medium of the expression strain. The nucleotide sequence encoding the ZmPAR1 protein is shown in SEQ ID NO:2.

[0008] Thirdly, the embodiments disclose a method for in vitro synthesis of monophosphate nucleotides, including the step of incubating a protein as shown in SEQ ID NO:1 with a nucleoside in vitro.

[0009] Fourthly, the embodiments disclose a CRISPR vector, including a guide sequence targeting the maize ZmPAR1 gene and an expression sequence for Cas9. The ZmPAR1 gene is located at position 189856343–189862099 bp on chromosome 1 of the maize genome Zm-B73-REFERENCE-GRAMENE-4.0. The guide sequence is shown in SEQ ID NO:6. The expression sequence for Cas9 is shown in SEQ ID NO:5.

[0010] Fifthly, the embodiments disclose a method for deleting the maize ZmPAR1 gene, including the step of introducing the CRISPR vector described in the fourth aspect into wild maize plants. The ZmPAR1 gene is located at position 189856343–189862099 bp on chromosome 1 of the maize genome Zm-B73-REFERENCE-GRAMENE-4.0.

[0011] Sixthly, the embodiments disclose a method for improving maize phenotype, including the step of introducing the CRISPR vector described in the fourth aspect into wild maize plants. The improvement of maize phenotype includes reducing plant height, increasing leaf angle, reducing ear height, shortening leaf length, and reducing leaf width.

[0012] In a seventh aspect, the embodiments disclose an overexpression vector comprising a nucleic acid molecule as shown in SEQ ID NO:2.

[0013] Eighthly, the embodiments disclose a method for overexpressing the ZmPAR1 gene, including the step of introducing the overexpression vector described in the seventh aspect into wild maize plants.

[0014] Ninthly, the embodiments disclose a method for improving maize phenotypes, including the step of introducing the overexpression vector described in the first aspect into wild maize plants. The modified maize phenotype includes reduced leaf angle, increased plant height and ear height, and increased leaf length and width. The ZmPAR1 gene is located at position 189856343–189862099 bp on chromosome 1 of the maize genome Zm-B73-REFERENCE-GRAMENE-4.0.

[0015] Tenthly, the embodiments disclose the application of the protein as shown in SEQ ID NO:1 or the nucleic acid molecule as described in SEQ ID NO:2. The application is selected from:

[0016] 1) Preparation of nucleoside kinase:

[0017] 2) Preparation of catalysts for the in vitro synthesis of monophosphate nucleotides;

[0018] 3) Prepare a vector to knock down the ZmPAR1 gene, wherein the ZmPAR1 gene is located at position 189856343~189862099bp on chromosome 1 of the maize genome Zm-B73-REFERENCE-GRAMENE-4.0;

[0019] 4) Improve the first maize phenotype, which includes reducing plant height, increasing leaf angle, reducing ear height, shortening leaf length, and reducing leaf width.

[0020] 5) Prepare a vector that overexpresses the ZmPAR1 gene, wherein the ZmPAR1 gene is located at position 189856343~189862099bp on chromosome 1 of the maize genome Zm-B73-REFERENCE-GRAMENE-4.0;

[0021] 6) Improve the second maize phenotype, which includes reducing the leaf angle, increasing plant height and ear height, and increasing leaf length and width;

[0022] 7) Preparation of new corn materials. Attached Figure Description

[0023] Figure 1 The prokaryotic expression results of the ZmPAR1 protein provided in the examples. Figure 1 A is a Coomassie staining diagram. Crude represents the crude extract, Unbound represents the flow buffer, 5thwash represents the fifth wash buffer, E1 represents the first elution buffer, E2 represents the second elution buffer, and the arrows indicate the location of the fusion protein. Figure 1 B is the Western blot development diagram. After development, the target protein was found in the crude extract, the flow buffer, and the second elution buffer. No target protein was found in the fifth wash buffer and the first elution buffer. The arrow indicates the location of the fusion protein after development.

[0024] Figure 2 The LC-MS spectrum of the ZmPAR1 protein as a nucleoside affinity enzyme, provided for the example. Figure 2 A represents the AMP, ADP, and ATP signals detected by LC-MS before and after incubation at 30°C for 50 min with 2 μg PAR1 protein in the presence of 100 μM ATP, using 2 μg PAR1 protein and 50 μM adenosine. The horizontal axis represents the peak time, and the vertical axis represents the signal response value. Figure 2 B represents the incubation of PAR1 with inosine, with LC-MS used to detect IMP, ADP, and ATP signals before the reaction and 50 min after the reaction. Figure 2C represents PAR1 incubated with uridine. UMP, ADP, and ATP signals were detected by LC-MS before the reaction and 50 min after the reaction. Figure 2 D represents PAR1 incubated with cytidine. CMP, ADP, and ATP signals were detected by LC-MS before the reaction and 50 min after the reaction. Figure 2 E represents PAR1 incubated with guanosine. GMP, ADP, and ATP signals were detected by LC-MS before the reaction and 50 min after the reaction.

[0025] Figure 3 The kinetic constants of inosine (A) and uridine (B) provided in the examples are shown. The horizontal axis represents the substrate concentration, the left vertical axis represents the reaction rate, and the right vertical axis represents the ratio of substrate to reaction rate. Kinetic curves were plotted using the software Graph, with an error of SD. Each substrate concentration was represented by three biological replicates.

[0026] Figure 4 The gene sequences (A) and protein sequences (B) of the cr-1 mutant, cr-2 mutant, and wild-type (WT) strains provided in the examples are shown in the figure.

[0027] Figure 5 The mature phenotypic diagrams of the cr-1 mutant, cr-2 mutant, and wild-type (WT) strains provided in the examples are shown in the figure. Scale bar: 10 cm.

[0028] Figure 6 Phenotypic statistics of the Cr-1 mutant, Cr-2 mutant, and wild-type (WT) strains provided in the examples. Phenotypes include: plant height, ear height, leaf length, leaf width, and leaf angle. Asterisks indicate statistical differences: ***, p < 0.001; *, p < 0.05; ns, p > 0.05.

[0029] Figure 7 Images and phenotypic statistics of fruits from the Cr-1 mutant, Cr-2 mutant, and wild-type (WT) strains provided for the examples. Phenotypic data includes ear length and ear diameter. Scale bar: 2 cm. Asterisks indicate statistical differences, ***, p < 0.001.

[0030] Figure 8 Western blot images of the inbred line B104 provided for the examples, and the overexpression transgenic families PAR1-Flag-24, PAR1-Flag-21, PAR1-Flag-20, PAR1-Flag-8 and PAR1-Flag-5.

[0031] Figure 9 OE provided for the embodiments 21 Strain, OE 24 Images and phenotypic statistics of fruits from both wild-type and wild-type plants (WT). Phenotypic data include: hundred-grain weight, grain length, grain width, ear length, ear diameter, and kernel row numbers. Scale bar: 2 cm. Asterisks indicate statistical differences: **, p < 0.01; ns, p > 0.05. Error is SD. Detailed Implementation

[0032] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the scope of this application. Reagents not specifically described in detail herein are all conventional reagents and are commercially available; methods not specifically described in detail are all conventional experimental methods and can be learned from the prior art.

[0033] Preparation of ZmPAR1 protein

[0034] Some embodiments provide a method for preparing the ZmPAR1 protein. The method includes providing an expression vector containing a nucleotide sequence encoding the ZmPAR1 protein, transforming the expression vector into *E. coli* to obtain an expression strain, and obtaining the ZmPAR1 protein from the culture medium of the expression strain. The nucleotide sequence encoding the ZmPAR1 protein is shown in SEQ ID NO:1.

[0035] In some embodiments, the expression vector is a pET28a-MBP-T7-6×HIS plasmid (CB113269532, Beijing Solarbio) that clones the coding sequence of ZmPAR1 (shown in SEQ ID NO:2).

[0036] In some embodiments, the preparation method of ZmPAR1 protein includes:

[0037] 1) The DNA molecule shown in SEQ ID NO:2 was amplified by PCR. The DNA molecule was ligated with the pET28a-MBP-T7-6×HIS restriction fragment using a one-step cloning method (ClonExpressII OneStepCloningKit, C112-01). The resulting ligated fragment was transformed into E. coli, positive clones were screened, and expression plasmids were extracted from the culture of positive clones.

[0038] 2) The expression plasmid was transfected into RIL cells (BL21-CodonPlus(DE3)-RIL, ZY1025, Shanghai Zeye Biotechnology Co., Ltd.), positive clones were screened, and the cells were seeded at a volume ratio of 1:100 into LB liquid medium containing Kan antibiotic. The cells were cultured at 37℃ and 180r / min for 4h with shaking. The OD600 value was approximately 0.8 to 1.0.

[0039] 3) Add IPTG inducer to the mixture from step 2) to a final concentration of 0.2 mM. Pre-cool the mixture to 16°C on a shaker and incubate at 180 rpm for 16-18 hours. Centrifuge the fermentation broth at 5000 rpm for 10 minutes to collect the cells. Add 20 mL of lysis buffer (QC25-05102, Qincheng Biotechnology), and vigorously shake to suspend the cells. Simultaneously, add PMSF protease inhibitor to a final concentration of 0.1 mM to prevent protein degradation.

[0040] 4) Use a high-pressure homogenizer to break down the protein. Perform high-pressure homogenization 2-3 times at 1000 mPa and 4℃. Transfer the bacterial culture to a 50 mL centrifuge tube and centrifuge at 14000 r / min and 4℃ for 1 h. Collect the supernatant after centrifugation for later use.

[0041] 5) The supernatant obtained in step 4) was purified using an affinity chromatography column with an MBP tag (Amylose Resin HighFlow, NEB, E8022S packing material) and the sugars were removed by chromatography to obtain the ZmPAR1 fusion protein with a C-terminal MBP tag, with an expected size of 86.4 kDa.

[0042] like Figure 1 As shown in Figure A, a protein of the expected size was found in the second eluent tube (E2).

[0043] like Figure 1 As shown in Figure B, after incubation and color development of ZmPAR1 protein with MBP antibody (ABclonal, catalog number AE016), the target protein was detected in crude extract, unbound fluid, and lane E2.

[0044] ZmPAR1 protein acts as a nucleoside affinity enzyme.

[0045] This test case utilizes nucleosides as substrates to characterize the enzymatic activity of the ZmPAR1 protein. In this test case, five different nucleosides (adenosine, inosine, uridine, cytidine, and guanosine) were used as substrates. ZmPAR1 protein was incubated with each of these five nucleosides in vitro for 50 min, and changes in the reaction products were detected. Following the ZmPAR1 protease activity reaction, adenosine monophosphate (AMP), inosine monophosphate (IMP), uridine monophosphate (UMP), cytidine monophosphate (CMP), and guanosine monophosphate (GMP) were generated accordingly. This enzyme activity assay uses ATP as energy; one molecule of ATP is consumed to produce one molecule of ADP.

[0046] Preparation of buffer stock solution: 1M NH4HCO3 (pH=7.5), 0.1M MgCl2, 0.5M KCl, 1M DTT (dithiothreitol), 10mM ATP.

[0047] Preparation of substrate stock solution: 500mM inosine, 500mM uridine, 500mM cytidine, 500mM adenosine, 50mM guanosine.

[0048] After prokaryotic expression and purification, the protein concentration was adjusted to 20 ng / μL using the BSA standard curve detection method.

[0049] The enzyme activity test procedure is as follows:

[0050] Prepare the following components in a 250 μL PCR tube: buffer of 40 mM NH4HCO3, 2 mM MgCl2, 10 mM KCl, 1 mM DTT, and 1 mM ATP; 2 μg protein; 100 μM substrate. The total reaction volume is 100 μL. Incubate at 30 °C for 50 min. Add an equal volume of chloroform / isoamyl alcohol (1:1), invert or vortex to mix, and centrifuge at low speed for 10 min. Collect the supernatant and extract again. Centrifuge at high speed for 10 min, and collect the supernatant. Dilute by two-fold and aliquot 100 μL into a mass spectrometry vial for LC-MS detection.

[0051] like Figure 2 As shown, monophosphate product AMP can be detected in the reaction solution. Figure 2 A) IMP Figure 2 B), UMP Figure 2 C), CMP Figure 2 D) GMP Figure 2 The production of E) and the product of ATP consumption, ADP ( Figure 2 (AE), which indicates that ZmPAR1 has phosphorylation activity on all five substrates, thus demonstrating that ZmPAR1 has broad nucleoside substrate activity.

[0052] ZmPAR1 protein exhibits stronger catalytic activity towards inosine and uridine.

[0053] Test examples, by measuring the kinetic constants of inosine and uridine, revealed that ZmPAR1 has high catalytic activity for the substrates inosine and uridine.

[0054] For the determination of the kinetic constants of inosine and uridine, the amount of monophosphate nucleotide product was used to determine the enzyme rate. The enzyme reaction buffer concentration and volume remained constant, and six different substrate concentration gradients were set. Each gradient was repeated in triplicate. The substrate concentrations for inosine were set as follows: 1 mM, 2 mM, 5 mM, 10 mM, 15 mM, and 20 mM. The substrate concentrations for uridine were set as follows: 5 mM, 20 mM, 40 mM, 60 mM, 80 mM, and 100 mM. The reaction buffer consisted of 40 mM NH4HCO3, 2 mM MgCl2, 10 mM KCl, 1 mM MTT, and 1 mM ATP. First, a reaction solution containing 2 μg of protease was prepared and pre-chilled on ice. Then, the substrate sample was added to an RNase-free eight-tube (the eight-tube was placed on an ice box), followed by the pre-chilled protease reaction solution. The total reaction volume was 100 μL. The mixture was centrifuged for 1 min. Temperature control was performed using a PCR instrument, with reaction times of 3 min and 6 min. After the reaction, the sample was immediately placed on ice to maintain a low temperature. Two min later, phenol-chloroform was added for the first extraction. The mixture was inverted several times to mix, centrifuged at low speed for 10 min, and 80 μL of the supernatant was collected. An equal volume of phenol-chloroform was then added for the second extraction, centrifuged at high speed for 10 min, and 60 μL of the supernatant was collected. Finally, an equal volume of double-distilled water was added for dilution. 50–80 μL of the diluted sample was analyzed by LC-MS.

[0055] The product was quantified using a standard curve method. First, the substance to be detected was diluted to seven concentrations: 10 ppb, 20 ppb, 50 ppb, 100 ppb, 200 ppb, 500 ppb, 1000 ppb, and 2000 ppb. The peak areas at the corresponding concentrations were detected by LC-MS, and a standard curve was generated using the formula with peak area on the x-axis and concentration on the y-axis. R0 was used. 2 A standard curve with a peak area greater than 0.99 was generated. The concentration of the substance was then calculated based on the measured peak area.

[0056] like Figure 3 As shown in A, the K of inosine m The value was 1.39 mM, while the turnover rate of inosine, K... cat It is 0.054s -1 The catalytic efficiency K of inosine cat / K m 38.84M -1 s -1 .like Figure 3 As shown in B, the K of uridine m The value was 11.33 mM, while the turnover rate of uridine K was...cat It is 0.171s -1 catalytic efficiency K of uridine cat / K m 15.09M -1 s -1 This indicates that ZmPAR1 preferentially binds to inosine and has higher catalytic efficiency for inosine, but the substrate turnover rate of uridine is higher than that of inosine.

[0057] Application of ZmPAR1 (knock-down)

[0058] 1. Preparation of ZmPAR1 knockdown editing plasmid

[0059] Referring to "High-Throughput CRISPR / Cas9 Mutagenesis Streamlines Trait GeneIdentification in Maize, The Plant Cell, Vol.32:1397~1413, May 2020", the sgRNA shown in SEQ ID NO:6 was cloned into plasmid pCXB053 (Li,C.,Liu,C.,Qi,X.,Wu,Y.,Fei,X.,Mao,L.,Cheng,B.,Li,X.,and Xie,C.(2017).RNA-guided Cas9 as an in vivo desired-target mutator in maize.Plant Biotechnol.J.15:1566–1576.) by enzyme digestion and ligation, thus obtaining the ZmPAR1 knockdown editing plasmid.

[0060] 2. Constructing ZmPAR1 knockdown maize plants

[0061] The obtained ZmPAR1 knockdown editing plasmid was transformed into Agrobacterium tumefaciens (Beijing Tianenze Gene Technology Co., Ltd., 140383). Positive clones were screened from the transformants. The positive clones were then used to infect immature embryo callus tissue of maize KN5585 (Weimi Biotechnology). Regenerated plants were cultured and screened with herbicides to obtain transgenic regenerated plants, resulting in cr-1 mutant and cr-2 mutant strains.

[0062] 3. Sequencing of Cr-1 and Cr-2 mutant strains

[0063] like Figure 4As shown in A and 4B, the cr-1 mutant lacks two bases, leading to premature transcription termination and ultimately encoding 193 amino acids. The cr-2 mutant has a one-base insertion, which also causes premature transcription termination, ultimately encoding 194 amino acids. Although both the cr-1 and cr-2 families result in early transcription termination, the first domain of the ZmPAR1 protein is partially preserved, and both the cr-1 and cr-2 mutants lack the D2 domain.

[0064] 4. Phenotypic observation of cr-1 mutant and cr-2 mutant strain

[0065] like Figure 5 and 6 As shown, the plant height and ear height of the cr-1 mutant were significantly lower than those of the wild type (“WT”, KN5585, 24.4%, 25.3% in the figure), while the leaf length and leaf width were significantly shorter than those of the wild type (7.4%, 22.2%). The plant height and ear height of the cr-2 mutant were significantly lower than those of the wild type (49.5%, 59.7%), while the leaf length and leaf width were significantly shorter than those of the wild type (37.9%, 38.8%). Furthermore, the leaf angle of the cr-1 mutant was significantly larger than that of the wild type (12.6%), while the leaf angle of the cr-2 mutant showed no significant change compared to the wild type.

[0066] like Figure 7 As shown, the spike length of the cr-1 mutant was significantly shorter than that of the wild type (“WT” in the figure, 48.4%), and the spike diameter was significantly shorter than that of the wild type (16.1%). The spike length of the cr-2 mutant was significantly shorter than that of the wild type (28.2%), and the spike diameter was significantly shorter than that of the wild type (12.1%). Since the cr-1 and cr-2 mutants still contain some functional domains, it is speculated that the cr-1 and cr-2 mutants may belong to weakly mutant alleles.

[0067] Therefore, knocking down ZmPAR1 can reduce maize plant height, increase the angle between maize leaves, reduce the height of maize ears, shorten maize leaf length, and reduce maize leaf width. Applications of ZmPAR1 (overexpression)

[0068] 1. Construction of ZmPAR1 overexpression vector

[0069] The lac promoter in plasmid pCAMBIA3300 (SnapGene) was replaced with the Ubi promoter (SEQ ID NO:3) to obtain plasmid WMV014.

[0070] A fusion DNA molecule containing a maize ubiquitin promoter, a ZmPAR1 coding sequence, and a Flag tag (pUBI::ZmPar1-Flag, SEQ ID NO:4) was synthesized.

[0071] The fusion DNA molecule was ligated with the linearized WMV014 fragment, the ligation product was transformed into E. coli, positive clones were screened from the transformants, and the ZmPAR1 overexpression vector was extracted from the culture of the positive clones.

[0072] 2. Construction of ZmPAR1 overexpression maize strain

[0073] The obtained ZmPAR1 overexpression vector was transformed into Agrobacterium tumefaciens (Beijing Tianenze Gene Technology Co., Ltd., 140383). Positive clones were screened from the transformants. The positive clones were then used to infect the immature embryo callus tissue of the maize B104 inbred line (Weimi Biotechnology). Regenerated plants were cultured and screened with herbicides to obtain transgenic regenerated plants. Five overexpression transgenic families were obtained: PAR1-Flag-24, PAR1-Flag-21, PAR1-Flag-20, PAR1-Flag-8, and PAR1-Flag-5.

[0074] 3. Western Blot Detection

[0075] The ZmPAR1 protein level in five overexpressing maize plants was detected by Western blotting.

[0076] like Figure 8 As shown, among the five overexpressing transgenic families of the maize B104 inbred line, PAR1-Flag-21 (abbreviated as OE) is the most common transgenic line. 21 ) and PAR1-Flag-24 (abbreviated as OE) 24 The ZmPAR1 protein level was significantly increased.

[0077] 4. Field experiments

[0078] like Figure 9 As shown, relative to wild maize plants (“WT”, B104 in the figure), OE 21 Plant height increased by 8.9%, OE 21 Ear height increased by 22.9%, leaf length of OE21 increased by 7.9%, and OE 21 Leaf width increased by 10.3%, OE 21 The leaf angle decreased by 7.2%, OE 21 The diameter of the ear increased by 5.0%.

[0079] like Figure 9 As shown, relative to wild maize plants (“WT” in the figure), OE 24 Plant height increased by 7.1%, ear height of OE24 increased by 10.9%, OE 24 Leaf length increased by 3.9%, OE 24 The diameter of the spike increased by 3.1%.

[0080] This demonstrates that overexpressing the ZmPAR1 gene can reduce the leaf angle, increase plant height and ear height, and increase leaf length and width, providing technical support for breeding maize varieties.

[0081] The above description is merely a preferred embodiment of this application, but the scope of protection of this application is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in this application should be included within the scope of protection of this application.

Claims

1. A method for synthesizing monophosphate nucleotides in vitro, comprising the step of incubating ZmPAR1 protein as shown in SEQ ID NO: 1 with nucleosides in vitro. 2.A method for improving a corn phenotype, comprising the step of introducing a CRISPR vector knocking down ZmPAR1 gene into a wild corn plant, wherein the corn phenotype is improved by reducing plant height, increasing leaf angle, reducing ear height, shortening leaf length, and reducing leaf width. wherein The CRISPR vector comprises a guide sequence targeting ZmPAR1 gene of corn and an expression sequence of Cas9; the nucleic acid sequence of ZmPAR1 gene is shown in SEQ ID NO: 2; the guide sequence is shown in SEQ ID NO: 6; and the expression sequence of Cas9 is shown in SEQ ID NO:

5. 3.A method for improving a corn phenotype, comprising the step of introducing an overexpression vector overexpressing ZmPAR1 gene into a wild corn plant, wherein the corn phenotype is improved by reducing leaf angle, increasing plant height and ear height, increasing leaf length and leaf width. wherein The overexpression vector comprises ZmPAR1 gene; and the nucleic acid sequence of ZmPAR1 gene is shown in SEQ ID NO:

2. 4.Use of ZmPAR1 protein as shown in SEQ ID NO: 1 or ZmPAR1 gene as shown in SEQ ID NO: 2, wherein the use is selected from: 1) preparing nucleoside kinase; 2) preparing a catalyst for synthesizing monophosphate nucleotides in vitro; 3) knocking down ZmPAR1 gene to improve a first corn phenotype, wherein the first corn phenotype is improved by reducing plant height, increasing leaf angle, reducing ear height, shortening leaf length, and reducing leaf width; 4) overexpressing ZmPAR1 gene to improve a second corn phenotype, wherein the second corn phenotype is improved by reducing leaf angle, increasing plant height and ear height, increasing leaf length and leaf width.

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