Nitrogen high efficiency related gene ZmCAMTA5 and application thereof

Abstract

The present application relates to a kind of nitrogen high-efficiency related gene ZmCAMTA5 and its application.The present application can obtain the plant with higher low nitrogen stress resistance relative to wild type by overexpressing ZmCAMTA5 gene in corn, the plant shows the higher low nitrogen stress resistance character of higher height in above-ground part, leaf is greener;ZmCAMTA5 gene can be applied to the breeding of nitrogen high-efficiency corn.In addition, by laser confocal microscope observation overexpressing ZmCAMTA5-GFP material, it is found that ZmCAMTA5 protein is located in cell nucleus.

Description

Technical Field

[0001] This invention relates to the field of genetic engineering technology, specifically to a nitrogen-efficient gene ZmCAMTA5 and its applications. Background Technology

[0002] Corn is one of the world's three major food crops and occupies an important position in my country's agriculture. In recent years, it has even surpassed wheat to become the second most planted food crop in China. Besides providing food, corn also plays a role in bioenergy, medicine, and environmental protection. Nitrogen, as an essential element for plant growth, plays a crucial role in the growth and development of corn. Plants mainly obtain essential nitrogen from the soil, and the inorganic nitrogen in the soil is mainly in the form of nitrate (NO3). - Nitrate exists in the form of nitrogen. To increase crop uptake of nitrate nitrogen and thus promote yield, nitrogen fertilizers are often used. However, excessive application of nitrogen fertilizers not only wastes nitrogen but also pollutes soil and water bodies with excess nitrate, affecting industrial and agricultural production and harming human health. Breeding and promoting nitrogen-efficient crops is of great significance for reducing nitrogen fertilizer application, protecting the environment, and safeguarding human health. Researching nitrogen-efficient genes in maize, and thus breeding nitrogen-efficient maize varieties, will help promote the rational application of nitrogen fertilizer while ensuring maize yield, and drive sustainable agricultural development.

[0003] Nitrogen is crucial for plant growth and development, and a lack of nitrogen leads to a series of obvious symptoms. In nitrogen-deficient corn plants, yellowing begins with older leaves. Initially, the first leaf is pale yellowish-green, gradually spreading from the tip to the base, turning yellow until the first leaf withers and dies. This process repeats over time, affecting the second and third leaves, eventually affecting the entire plant. Nitrogen deficiency also slows plant growth, resulting in stunted growth, fewer tillers, and shorter, narrower leaves. Another prominent characteristic of nitrogen-deficient plants is increased root length and a higher root-to-shoot ratio. This is a response to low nitrogen stress, helping the plant utilize limited nitrogen more efficiently; therefore, a pronounced phenotype suggests good tolerance to low nitrogen. Studies have shown that within a certain treatment period, plants treated with low nitrogen have longer roots than those under normal nitrogen conditions, but prolonged low nitrogen treatment eventually inhibits root development. Nitrogen deficiency during the growth period can also affect maize yield. Insufficient nitrogen supply leads to less accumulation of maize kernel biomass, which in turn affects yield traits such as ear length, number of rows per ear, number of kernels per row, ear weight, and 100-kernel weight. Summary of the Invention

[0004] To address the shortage of nitrogen-efficient varieties, this invention aims to provide a nitrogen-efficient gene, ZmCAMTA5, and its applications. By overexpressing the ZmCAMTA5 gene in maize, plants exhibiting low-nitrogen stress resistance compared to wild-type maize can be obtained. These plants show taller above-ground parts and higher chlorophyll content in their leaves, demonstrating resistance to low-nitrogen stress. The ZmCAMTA5 gene can be applied to the breeding of nitrogen-efficient maize.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] A nitrogen-efficient gene, ZmCAMTA5, is characterized by its coding sequence as shown in SEQ ID No. 1.

[0007] A protein encoded by the aforementioned gene ZmCAMTA5, characterized in that its amino acid sequence is shown in SEQ ID No. 2.

[0008] The application of the nitrogen-efficient gene ZmCAMTA5 in nitrogen-efficient maize breeding is characterized by the following: the application involves overexpressing the gene ZmCAMTA5 to carry out nitrogen-efficient maize breeding.

[0009] A detection primer pair for detecting the overexpression level of the above-mentioned gene ZmCAMTA5, characterized in that the sequences of the detection primer pair are shown in SEQ ID No. 3 and SEQ ID No. 4.

[0010] The beneficial effects of the nitrogen-efficient gene ZmCAMTA5 and its application described in this invention are as follows:

[0011] By overexpressing the gene ZmCAMTA5 in maize, plants with low nitrogen stress resistance compared to the wild type can be obtained. These plants exhibit taller above-ground parts and higher chlorophyll content in their leaves, thus showing resistance to low nitrogen stress. The gene ZmCAMTA5 can be applied to the breeding of nitrogen-efficient maize. Attached Figure Description

[0012] The present invention includes the following figures:

[0013] Figure 1 Identification of ZmCAMTA5 overexpression materials.

[0014] Figure 2 For the genetic verification of the ZmCAMTA5 gene; the left figure shows the growth of wild-type and ZmCAMTA5 overexpressing lines (OE#2 and OE#4) after growing for 2 weeks under normal nitrogen or low nitrogen treatment conditions; the right figure shows the yellowing of the first leaf of the corresponding plant.

[0015] Figure 3 for Figure 2The second leaf of the corresponding plant is turning yellow.

[0016] Figure 4 Phenotypic data: A represents the SPAD value of the first leaf under normal nitrogen and low nitrogen conditions; B represents the SPAD value of the second leaf.

[0017] Figure 5 The protein encoded by ZmCAMTA5 is located in the cell nucleus. Detailed Implementation

[0018] The present invention will be further described in detail below with reference to the accompanying drawings.

[0019] The following embodiments are provided to better understand the present invention, but are not limited thereto. These embodiments are for illustrative purposes only and do not limit the scope of protection of the present invention in any way.

[0020] Unless otherwise specified, the equipment and reagents used in each embodiment are all commercially available.

[0021] Example 1

[0022] Functional analysis of the nitrogen-efficient gene ZmCAMTA5

[0023] To investigate the function of ZmCAMTA5, overexpression materials of ZmCAMTA5 were constructed. The CDS sequence of ZmCAMTA5 was constructed into the Xc2840-CeGFP vector, resulting in two overexpression materials (OE#2 and OE#4). RT-qPCR detection showed that OE#2 and OE#4 were overexpressed by 9-fold and 13-fold, respectively. Figure 1 ).

[0024] Equal volumes of sieved sand and vermiculite were mixed and placed into a red plastic basin, which was then placed on a tray; a solution containing 4 mM NO3 was prepared. - (for FN treatment) and 0.05mM NO3 - (For LN treatment) Pour 4L of nutrient solution into each tray and wait for the sand to fully absorb the nutrient solution; Sow 4 plump and uniformly sized corn seeds in each plastic box and cover with an appropriate amount of sand after sowing; The seeds will germinate in about a week. Observe the phenotype when the seedlings grow to two leaves and one heart, and replenish the nutrient solution regularly during this period.

[0025] like Figure 2 and Figure 3 As shown: After 14 days of LN treatment, the first leaf of WT turned yellow before that of OE material, but there was no significant difference between WT and OE materials during FN treatment, and the phenotype of the second leaf was also the same.

[0026] like Figure 4As shown, under low nitrogen stress, OE#2 and OE#4 exhibited stronger resistance compared to WT, as evidenced by higher chlorophyll content in their leaves, indicating that the ZmCAMTA5 gene is indeed a nitrogen-efficient gene.

[0027] Example 2

[0028] Subcellular localization of the nitrogen-efficient gene ZmCAMTA5

[0029] Ten days after the ZmCAMTA5-OE#2 overexpression line was grown, the root tip was transversely cut with a blade, and the localization of the cross-section was observed using a laser confocal microscope. Figure 5 It can be seen that ZmCAMTA5-GFP is located in the cell nucleus.

[0030] The contents not described in detail in this specification are existing technologies known to those skilled in the art.

Claims

1. A nitrogen-efficient related gene, ZmCAMTA5, characterized in that, Its encoded sequence is shown in SEQ ID No.

1.

2. A protein encoded by the gene ZmCAMTA5 according to claim 1, characterized in that, Its amino acid sequence is shown in SEQ ID No.

2.

3. The application of the gene ZmCAMTA5 as described in claim 1 in high-nitrogen-efficiency breeding of maize, characterized in that, The application involves nitrogen-efficient maize breeding through overexpression of the ZmCAMTA5 gene.

4. A detection primer pair for detecting the overexpression level of the gene ZmCAMTA5 as described in claim 3, characterized in that, The sequences of the detection primer pair are shown in SEQ ID No. 3 and SEQ ID No. 4.

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