ZmTCP19 gene mutant and application of ZmTCP19 gene mutant in regulating and controlling nitrogen utilization rate, growth amount and plant height of corn

Abstract

The invention relates to the technical field of biology, in particular to a ZmTCP19 gene mutant and application of the ZmTCP19 gene mutant to regulation and control of the nitrogen utilization rate, the growth amount and the plant height of corn. ZmTCP19 gene mutants (a mutant zmtcp19-1 and a mutant zmtcp19-2) are obtained through a molecular genetics means, the functions of the ZmTCP19 gene mutants are deeply analyzed and identified, and results show that under the condition of low nitrogen, the total fresh weight of the zmtcp19-1 is reduced by 30.33% compared with that of a control group, and under the condition of normal nitrogen, the total fresh weight of the zmtcp19-1 is reduced by 25.54% compared with that of the control group; under the condition of low nitrogen, the total fresh weight of zmtcp19-2 is reduced by 25.19% compared with that of a control group, and under the condition of normal nitrogen, the total fresh weight is reduced by 17.30% compared with that of the control group. Meanwhile, the ZmTCP19 gene mutant can reduce the plant height of the corn and improve the lodging resistance of the corn.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biotechnology, in particular to a ZmTCP19 gene mutant and application thereof in regulating nitrogen utilization rate, growth and plant height of corn. BACKGROUND

[0002] As a core element indispensable for plant growth and development, nitrogen is the basis for constructing key life substances such as proteins, nucleic acids and chlorophyll. Except for leguminous plants that can fix nitrogen through symbiotic rhizobia, corn and other major crops highly depend on roots to absorb inorganic nitrogen from soil. Limited arable land resources make it the main way to increase yield of corn, and insufficient supply of soil available nitrogen and low efficiency of chemical nitrogen fertilizer application have become the key bottleneck restricting the sustained increase of corn yield. Excessive nitrogen application can increase yield in the short term, but its nitrogen utilization efficiency is generally low, leading to rising production costs, resource waste, and environmental problems such as water eutrophication and greenhouse gas emissions. Therefore, it is very important to improve the absorption, assimilation and utilization efficiency of corn itself for nitrogen, and to implement the strategy of "reducing nitrogen and increasing efficiency". Developing new germplasm of corn with high nitrogen utilization efficiency is also a key path to ensure national food security and sustainable development of agriculture.

[0003] The perception, absorption, assimilation and reuse of nitrogen by plants is a signal network that is closely regulated, and transcription factors play a core regulatory role. Since the nitrogen utilization efficiency of corn and other plants is too low, it is important to improve the nitrogen utilization efficiency of these plants. At the same time, lodging can lead to reduced yield, moldy grains and poor grain quality, and is an important factor limiting the mechanized harvesting of corn. The problem of corn lodging seriously restricts its yield, quality and mechanized harvesting. With the transfer of labor force and the development of mechanization, higher requirements are put forward for the lodging resistance of corn. However, few lodging resistance genes have been found in corn, which cannot meet the needs of corn lodging resistance breeding.

[0004] In summary, breeding new corn varieties that can efficiently utilize nitrogen and breeding lodging-resistant corn varieties are key to solving the contradiction between environment and resources in agricultural production in China. SUMMARY

[0005] The purpose of the present application is to provide a ZmTCP19 gene mutant and its application in regulating nitrogen utilization rate, growth and plant height of corn, so as to solve the problems existing in the prior art.

[0006] To achieve the above purpose, the present application provides the following solutions.

[0007] The application provides a ZmTCP19 gene mutant, which comprises mutant zmtcp19-1 and mutant zmtcp19-2; the nucleotide sequence of the mutant zmtcp19-1 is shown as SEQ ID NO:1; and the nucleotide sequence of the mutant zmtcp19-2 is shown as SEQ ID NO:2.

[0008] The application provides application of the ZmTCP19 gene mutant or the biological material containing the ZmTCP19 gene mutant in regulating nitrogen utilization rate of corn.

[0009] Preferably, the biological material comprises a recombinant carrier or a recombinant bacterium.

[0010] The application provides application of the ZmTCP19 gene mutant or the biological material containing the ZmTCP19 gene mutant in regulating biomass of corn.

[0011] Preferably, the biological material comprises a recombinant carrier or a recombinant bacterium; and the biomass is fresh weight.

[0012] The application provides application of the ZmTCP19 gene mutant or the biological material containing the ZmTCP19 gene mutant in regulating plant height of corn.

[0013] Preferably, the biological material comprises a recombinant carrier or a recombinant bacterium.

[0014] The application provides application of the ZmTCP19 gene mutant or the biological material containing the ZmTCP19 gene mutant in cultivating lodging-resistant corn.

[0015] Preferably, the biological material comprises a recombinant carrier or a recombinant bacterium.

[0016] The application provides application of the ZmTCP19 gene mutant or the biological material containing the ZmTCP19 gene mutant in corn molecular breeding.

[0017] The application discloses the following technical effects:

[0018] The application first excavates a key gene that can regulate plant biomass and nitrogen utilization of plants-ZmTCP19. The application obtains ZmTCP19 gene mutants (mutants zmtcp19-1 and zmtcp19-2) through molecular genetic means, and deeply analyzes and identifies the function thereof. The results show that compared with the control, the plant height and biomass of the mutant plant of the gene are significantly reduced, and the nitrogen-dependent biological growth amount is lower than that of the control; under the condition of low nitrogen, the total fresh weight of zmtcp19-1 is reduced by 30.33% compared with the control, and under the condition of normal nitrogen, the total fresh weight is reduced by 25.54% compared with the control; under the condition of low nitrogen, the total fresh weight of zmtcp19-2 is reduced by 25.19% compared with the control, and under the condition of normal nitrogen, the total fresh weight is reduced by 17.30% compared with the control. At the same time, the ZmTCP19 gene mutant can reduce the plant height of corn and increase the lodging resistance of corn. The application further determines the role of ZmTCP8 gene and its mutant in affecting the biomass of corn plants and the nitrogen utilization efficiency. The application provides gene resources and germplasm resources for excavating key elements of nitrogen utilization and lodging resistance. BRIEF DESCRIPTION OF DRAWINGS

[0019] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed in the embodiments will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative labor on the basis of these drawings.

[0020] Figure 1 Figure 1 is a plant phenotype diagram of zmtcp19-1 mutant material (tcp19-1), zmtcp19-2 mutant material (tcp19-2) and control (B73);

[0021] Figure 2 Figure 2 is a biomass statistical diagram of zmtcp19-1 mutant material (tcp19-1), zmtcp19-2 mutant material (tcp19-2) and control (B73);

[0022] Figure 3 Figure 3 is a plant height statistical diagram of zmtcp19-1 mutant material (tcp19-1), zmtcp19-2 mutant material (tcp19-2) and control (B73);

[0023] Figure 4Biomass plot for nitrogen gradient test for zmtcp19-1 mutant material (tcp19-1), zmtcp19-2 mutant material (tcp19-2) and control (B73); wherein KCI is no nitrogen nutrient solution, LN is low nitrogen nutrient solution, and NN is normal nitrogen nutrient solution. DETAILED DESCRIPTION

[0024] Various exemplary embodiments of the present application will now be described in detail, with reference to the figures. The detailed description is not to be regarded as limiting the application, but rather as an exemplification of certain aspects, features and embodiments of the application.

[0025] It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. Additionally, the use of "including," "comprising," "having," "containing," and variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. Unless otherwise indicated, the use of the terms "or" and "and" herein is intended to mean "and / or," where such terms are used in the conjunctive sense. Further, unless otherwise indicated, the use of relational terms, such as first, second, and the like, are used solely to distinguish one from another, without necessarily implying a chronology or an order of succession.

[0026] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, the preferred methods and materials are described. All patents, patent applications, and publications mentioned herein are incorporated by reference for the disclosure and description thereof to the extent that such incorporation is permitted under copyright laws.

[0027] Many modifications and variations of this application can be made without departing from its spirit or scope, which will be apparent to those skilled in the art. Other embodiments of the application will be apparent to those skilled in the art from consideration of the specification and practice of the application disclosed herein. The specification and examples given are exemplary only.

[0028] As used herein, the terms "comprise", "comprising", "include", "including", "have", "having" and the like are open-ended and do not exclude additional elements or steps.

[0029] Unless otherwise required by context, as used herein the term "about" means approximately or nearly as understood by one of ordinary skill in the art. When used in a numerical sense with respect to a particular value, the term "about" generally means a range of values plus or minus 10% of that value.

[0030] Example 1 Source of ZmTCP19 gene mutants zmtcp19-1 and zmtcp19-2

[0031] Zmtcp19-1 mutant material (abbreviated as tcp19-1) and zmtcp19-2 mutant material (abbreviated as tcp19-2) (http: / / maizeems.qilnu.edu.cn / ) have original mutant numbers of EMS4-03446c and EMS4-034469 respectively, and before the present embodiment is studied, there is no report on the function of ZmTCP19 gene mutants. The function of ZmTCP19 gene mutant zmtcp19-1 and ZmTCP19 gene mutant zmtcp19-2 is studied for the first time in the present embodiment. The nucleotide sequences of ZmTCP19 gene mutant zmtcp19-1 and ZmTCP19 gene zmtcp19-2 are shown in SEQ ID NO: 1 and SEQ ID NO: 2.

[0032] The nucleotide sequence of ZmTCP19 gene mutant zmtcp19-1 is shown in SEQ ID NO: 1, and is specifically as follows:

[0033]

[0034] The nucleotide sequence of the ZmTCP19 gene mutant zmtcp19-2 is shown as SEQ ID NO: 2, in particular:

[0035]

[0036] Example 2 Seedling phenotypic investigation of zmtcp19-1 mutant material and zmtcp19-2 mutant material

[0037] 1. Method

[0038] In 2024, the control (maize B73 inbred line, referred to as B73), zmtcp19-1 mutant material and zmtcp19-2 mutant material (purchased from the maize EMS mutant website (http: / / maizeems.qlnu.edu.cn / )) were planted in the Damao base of the Cotton Research Institute in Jiyang District, Sanya City. Phenotype shooting and physiological index determination were performed 10 days after pollination. The index detection included plant phenotype, physiological index (total biomass, plant height), wherein the determination method of total biomass was to dig out the whole plant, wash the roots, remove excess vermiculite or soil, and weigh and measure the plant height of the whole plant.

[0039] 2. Plant phenotype and physiological index detection results

[0040] The plant phenotypes of zmtcp19-1 mutant material, zmtcp19-2 mutant material and the control are shown in Figure 1 , and the results of fresh weight (total biomass) detection and plant height detection are shown in Figure 2 and Figure 3 . The results show that the total fresh weight of the control plant is 823 g; the total fresh weight of the zmtcp19-1 mutant material is 600 g, and the biomass is reduced by 27.1% compared with the control; the total fresh weight of the zmtcp19-2 mutant material is 710 g, and the biomass is reduced by 12.7% compared with the control. The plant height of the control plant is 180 cm, while the plant height of the zmtcp19-1 mutant material is 159 cm, which is reduced by 4.3% compared with the control, and the plant height of the zmtcp19-2 mutant material is 173 cm, which is reduced by 3.9% compared with the control. It can be seen that compared with the control plant, the plant height of the mutant material is reduced, which can be used for breeding new varieties of maize resistant to lodging.

[0041] Example 3 Phenotype and physiological index detection results of zmtcp19-1 mutant material and zmtcp19-2 mutant material under nitrogen gradient treatment

[0042] 1. Method

[0043] The nitrogen screening gradient test was carried out in Chengdu, Sichuan Agricultural University in 2025. The Hoagland nutrient solution was modified to prepare a nitrogen-free nutrient solution (KCL, the rest of the components are the same as the Hoagland nutrient solution) containing 4 mM KCl, a low-nitrogen nutrient solution (LN, the rest of the components are the same as the Hoagland nutrient solution) containing 0.04 mM KNO3, and a normal-nitrogen nutrient solution (NN, the rest of the components are the same as the Hoagland nutrient solution) containing 4 mM KNO3. Then, the zmtcp19-1 mutant material, the zmtcp19-2 mutant material, and B73 were subjected to soil culture test using the aforementioned nitrogen nutrient solution. During the soil culture, the photoperiod was set to 16 h light / 8 h dark. Before the nitrogen nutrient solution treatment, the plants were cultured with deionized water to two leaves, and then transferred to the environment treated with the nitrogen nutrient solution of different concentrations for culture, and the phenotypic characteristics were observed and the physiological indexes were detected.

[0044] 2. Results

[0045] The results are shown in Table 1. Figure 4 As shown in Table 1, the results show that there is no significant difference in the biomass of the zmtcp19-1 mutant material, the zmtcp19-2 mutant material, and B73 under the condition of no nitrogen (KCl), indicating that the other functions of the mutant are not impaired. Under the condition of low nitrogen, the total fresh weight of the zmtcp19-1 mutant material is reduced by 30.33% compared with the control, and under the condition of normal nitrogen, the total fresh weight is reduced by 25.54% compared with the control. Under the condition of low nitrogen, the total fresh weight of the zmtcp19-2 mutant material is reduced by 25.19% compared with the control, and under the condition of normal nitrogen, the total fresh weight is reduced by 17.30% compared with the control. After applying a small amount of nitrogen (LN), the biomass of B73 increases, and the biomass of the zmtcp19-1 mutant material and the zmtcp19-2 mutant material does not change significantly. With the increase of the nitrogen concentration, the biomass of B73 gradually increases, and the biomass under the condition of normal nitrogen (NN) is 2.4 times that under the condition of no nitrogen (KCl), while the biomass of the zmtcp19-1 mutant material under the condition of normal nitrogen (NN) is 1.6 times that under the condition of no nitrogen (KCL), and the biomass of the zmtcp19-2 mutant material under the condition of normal nitrogen (NN) is 1.9 times that under the condition of no nitrogen (KCL).

[0046] The above-described embodiments are only to describe the preferred modes of the present application, and do not limit the scope of the present application. Without departing from the design spirit of the present application, various modifications and improvements to the technical solutions of the present application made by those skilled in the art shall fall within the protection scope determined by the claims of the present application.

Claims

1. A ZmTCP19 gene mutant, characterized in that, The ZmTCP19 gene mutant includes mutant zmtcp19-1 and mutant zmtcp19-2; the nucleotide sequence of the mutant zmtcp19-1 is shown as SEQ ID NO: 1; and the nucleotide sequence of the mutant zmtcp19-2 is shown as SEQ ID NO:

2.

2. Application of the ZmTCP19 gene mutant or the biological material containing the ZmTCP19 gene mutant of claim 1 in regulating nitrogen utilization rate of corn.

3. Use according to claim 2, characterized in that, The biological material includes a recombinant vector or a recombinant bacterium.

4. Application of the ZmTCP19 gene mutant or the biological material containing the ZmTCP19 gene mutant of claim 1 in regulating biomass of corn.

5. Use according to claim 4, characterized in that, The biological material includes a recombinant vector or a recombinant bacterium; and the biomass is fresh weight.

6. Application of the ZmTCP19 gene mutant or the biological material containing the ZmTCP19 gene mutant of claim 1 in regulating plant height of corn.

7. Use according to claim 6, characterized in that, The biological material includes a recombinant vector or a recombinant bacterium.

8. Application of the ZmTCP19 gene mutant or the biological material containing the ZmTCP19 gene mutant of claim 1 in cultivating lodging-resistant corn.

9. Use according to claim 8, characterized in that, The biological material includes a recombinant vector or a recombinant bacterium.

10. Application of the ZmTCP19 gene mutant or the biological material containing the ZmTCP19 gene mutant of claim 1 in corn molecular breeding.

Images