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CN-121992000-A - Application of gene ASN3 in improving corn nitrogen utilization efficiency

CN121992000ACN 121992000 ACN121992000 ACN 121992000ACN-121992000-A

Abstract

The invention discloses application of asparagine synthase 3, namely ASN3, in improving nitrogen utilization efficiency of plants. The corn source asparagine synthase 3 gene Zm00001d028750 can effectively improve the utilization efficiency of corn nitrogen, can improve the biomass of corn plants, and can be used for cultivating new varieties of high-efficiency corn.

Inventors

  • WU YONGRUI
  • HUANG YONGCAI
  • WANG HAIHAI
  • ZHU YIDONG
  • CUI YAHUI
  • WANG QIONG

Assignees

  • 中国科学院分子植物科学卓越创新中心

Dates

Publication Date
20260508
Application Date
20241105

Claims (10)

  1. 1. Use of asparagine synthase 3, ASN3, or an expressed gene thereof for increasing nitrogen utilization efficiency in plants.
  2. 2. The use according to claim 1, wherein the plant is a gramineous crop selected from the group consisting of maize, rice, wheat, soybean, barley, oat, rye and sorghum.
  3. 3. The use according to claim 1, wherein the asparagine synthase 3 is a maize-derived protein ZmASN (NCBI accession number Zm00001d 028750) having the amino acid sequence shown in SEQ ID No. 1 or a conservatively variant polypeptide thereof, said conservatively variant polypeptide having more than 90% homology to ZmASN3 and having the function of asparagine synthase 3.
  4. 4. The use according to claim 3, wherein the nucleotide sequence of the CDS sequence, which is the coding region of the expressed gene ZmASN3 of the protein ZmASN, is SEQ ID NO. 2 or a nucleotide sequence having a homology of 90% or more with SEQ ID NO. 2.
  5. 5. The use according to claim 2, wherein the gramineous crop is maize.
  6. 6. The method of claim 4, wherein the maize is selected from the group consisting of existing wild species, inbred lines, farmed species and hybrid species.
  7. 7. The use according to claim 1 for creating new varieties of nitrogen-efficient maize.
  8. 8. The use according to claim 1, wherein the plant nitrogen utilization efficiency is increased by over-expressing the protein ASN3 or a gene encoding the same.
  9. 9. Use according to claim 8, wherein the over-expressed protein ASN3 or a gene encoding it, e.g. ZmASN3, is achieved by: A. Cloning a gene ZmASN encoding a protein ASN3, e.g., zmASN3, e.g., a gene ZmASN having a nucleotide sequence of SEQ ID NO:2, on a plasmid vector suitable for expression in Agrobacterium to form a recombinant plasmid, i.e., an ASN3 overexpression vector, transforming the plant by Agrobacterium-mediated transformation to obtain a transgenic plant overexpressing the protein ASN3, or B. the gene ZmASN3 encoding the protein ASN3 such as ZmASN, for example, the gene whose nucleotide sequence is SEQ ID NO. 2 is cloned on a plant chromosome by a gene editing technology to obtain a transgenic plant over-expressing the protein ASN 3.
  10. 10. Use according to claim 8, wherein the plant is maize and the ubi promoter is used to regulate expression of a gene encoding protein ZmASN, such as gene ZmASN having the nucleotide sequence of SEQ ID No. 2.

Description

Application of gene ASN3 in improving corn nitrogen utilization efficiency Technical Field The invention belongs to the technical field of agricultural biology, and particularly relates to application of an asparagine synthase 3 (ASN 3) gene in improving nitrogen utilization efficiency of plants. Background Corn (Zea mays L.) is one of three major food crops, and is also the crop with the widest planting area worldwide (FAO (2020). FAOSTAT data.food and Agricultural Organization, italy, rome. Http:// faostat.fao.org /). Corn is mainly fed or used as an industrial raw material in China, and with the rapid development of economy in China, the living standard of people is continuously improved, and people need to be fully eaten and better eaten, so that the demands on the yield and quality of corn are continuously increased. Nitrogen is one of essential elements for plant growth and development, and proper application of Nitrogen fertilizer can promote yield increase and stable yield of crops, but excessive application of Nitrogen fertilizer not only causes waste of shortage resources for producing Nitrogen fertilizer, but also causes a series of environmental pollution (Wani,S.H.et al.Nitrogen use efficiency(NUE):elucidated mechanisms,mapped genes and gene networks in maize(Zea mays L.).Physiol.Mol.Biol.Plants 27,2875-2891(2021).). such as soil nitrosation, water eutrophication and the like, and the NUE is one of important measures for cultivating and excavating corn varieties with high Nitrogen utilization rate by measuring important indexes (Ciampitti,I.A.&Lemaire,G.From use efficiency to effective use of nitrogen:a dilemma for maize breeding improvement.Sci.Total Environ.826,154125(2022).), for Nitrogen absorption and utilization. However, few reports on new nitrogen efficient materials in corn are currently available. The efficient property of corn nitrogen is complex agronomic property influenced by multiple genes and multiple paths, due to the abuse of chemical fertilizers, related genes such as absorption and transportation related nitrogen utilization in many inbred lines are selected from wild corn to domestication of cultivated corn, and in the early stage, the gene is cloned from the wild corn to a key gene THP9 for controlling the content of corn protein, can obviously improve the content of protein in corn kernels, roots, stems and leaves, can obviously improve the nitrogen utilization efficiency of corn under the condition of reducing nitrogen, and is predicted to have important application potential in cultivating novel ecological friendly corn (Huang et al.THP9 enhances seed protein content and nitrogen-use efficiency in maize.Nature.612(7939):292-300(2022).). Disclosure of Invention In the early stage, we clone a corn asparagine synthase 3 gene ASN3 (ASPARAGINE SYNTHETASE, NCBI number Zm00001d 028750), see patent document CN202410783823X, and found that over-expression of the ASN3 gene in corn can remarkably improve the protein content of corn kernels and plants and increase the biomass of corn. Recently, we performed field tests of Nitrogen Utilization Efficiency (NUE) on these overexpressing materials, and found that ASN3 overexpressing lines also have nitrogen-efficient function. Tests were carried out in the nitrogen pool of the Shanghai Songjiang base under both normal nitrogen application (40 g/plant; normal nitrogen conditions) and no nitrogen application (low nitrogen conditions), and ASN3 overexpressing transgenic maize grew better than B73 plants. Under normal nitrogen application conditions, ASN3 over-expressed plants increased significantly from plant height, aerial and subsurface compared to B73. Under the condition of no nitrogen application, the ASN3 over-expression plant and the B73 plant are dwarf compared with the normal nitrogen treatment plant, the fresh weight of the overground part and the underground part is reduced, but the ASN3 over-expression plant grows better than the B73 plant, the leaf color is dark green, and the plant height, the overground part and the underground part are obviously increased compared with the B73. Therefore, based on the nitrogen high-efficiency function of ASN3 and the over-expression transgene material, the gene material is introduced into a main variety such as Zhengdan 958, so that important genetic material can be provided for cultivating novel corn with high nitrogen efficiency. Specifically, the invention comprises the following technical scheme. The invention provides application of asparagine synthase 3, namely ASN3 (ASPARAGINE SYNTHETASE 3, ASN 3) or an expression gene thereof in improving plant nitrogen utilization efficiency, and provides a method for improving plant nitrogen utilization efficiency by using asparagine synthase 3, namely ASN3 (ASPARAGINE SYNTHETASE, ASN 3) or an expression gene thereof. The plant is preferably a monocotyledonous plant, preferably a gramineous crop selected from the group consisting of maize, rice,