CN-122012507-A - Adversity stress inducible promoter proLc and application thereof

Abstract

The invention provides an adversity stress inducible promoter proLc and application thereof, 15 cis-elements possibly induced by drought in a classical NCED, LEA, DHN gene promoter regulated by drought are integrated in leymus chinensis, and a drought-induced promoter proLc is synthesized, and the nucleotide sequence of the drought-induced promoter proLc is shown as SEQ ID NO. 1. The proLc318,318 promoter has transcription activity induced by drought stress, can drive the target gene at the downstream of the promoter to be efficiently expressed in plants, can be used for activating the expression of the target gene under the drought stress condition, and further provides an efficient tool for breeding and researching transgenic plants.

Inventors

• Li Gawen
• LIU QIKUN
• TIAN CHUNYU
• WU ZINIAN

Assignees

• 北京大学
• 中国农业科学院草原研究所

Dates

Publication Date

20260512

Application Date

20260409

Claims (9)

1. 1. A DNA molecule which is any one of the following 1) to 2): 1) A nucleotide sequence shown in SEQ ID NO. 1; 2) A DNA molecule which hybridizes to the nucleotide sequence set forth in 1) under stringent conditions and has promoter activity.
2. 2. A biomaterial comprising the DNA molecule of claim 1, wherein the biomaterial is any one of the following: A1 A DNA expression cassette; a2 A nucleic acid construct; a3 A recombinant vector; a4 A recombinant microorganism; a5 A recombinant cell line.
3. 3. Use of a DNA molecule according to claim 1, comprising: B1 Use as promoter; b2 As a promoter to promote expression of a gene of interest.
4. 4. A method for improving the expression level of a target gene under drought stress of a plant is characterized by comprising the steps of introducing the DNA molecule of claim 1 into a target plant to obtain a transgenic plant, enabling the target gene in the transgenic plant to start transcription of the target gene by taking the DNA molecule of claim 1 as a promoter, and improving the expression level of the target gene after transcription, wherein the plant is tobacco or alfalfa.
5. 5. The method of claim 4, wherein the gene of interest is an endogenous gene or an exogenous gene of the plant of interest.
6. 6. The method according to claim 4, wherein the gene of interest is a gene capable of combating abiotic stress.
7. 7. The method of claim 4, wherein the abiotic stress is drought stress, high salt stress, abscisic acid stress, low temperature stress, and/or alkali stress.
8. 8. Use of a DNA molecule according to claim 1, or a biomaterial according to claim 2, or a method of preparation according to any one of claims 4 to 7, for the initiation of gene expression of interest in plants, such as tobacco or alfalfa.
9. 9. Use of a DNA molecule according to claim 1, or a biomaterial according to claim 2, or a method of preparation according to any one of claims 4 to 7, for the cultivation of a transgenic plant, said transgenic plant being tobacco or alfalfa.

Description

Adversity stress inducible promoter proLc and application thereof Technical Field The invention relates to the technical field of genetic engineering, in particular to an adversity stress inducible promoter proLc318,318 and application thereof. Background The leymus chinensis (Leymus chinensis (Trin.) Tzvel.) is a typical drought-tolerant rural gramineous plant in northern grasslands in China, and forms a multi-level drought-tolerant regulation network in a long-term evolution process, so that a key gene resource and a regulation mechanism basis are provided for drought-tolerant genetic improvement of crops. In the regulation network, NAC, bZIP and other transcription factors can specifically identify and combine with the conserved cis-elements in the gene promoter region, so as to activate NCED, LEA, DHN and other core drought-resistant gene expression, and form a drought response core regulation module. The regulation mechanism is highly conserved in the plant kingdom, and provides a blueprint of a molecular level for the design of an artificial promoter. Furthermore, by modularly assembling the above-described critical cis-elements, a synthetic promoter having a small length can be constructed. The promoter has high specificity and strong inducibility when facing drought stress, can greatly improve the expression efficiency of downstream stress resistance genes in time and space, and opens up a new way for realizing the design of the accurate synthetic promoter with low background and high inducibility. Disclosure of Invention The invention integrates various cis-elements possibly induced by drought in classical promoters of genes subjected to drought regulation in the leymus chinensis, synthesizes an adversity stress induction type promoter, improves the expression efficiency of downstream adversity resistance genes in time and space, and opens up a new way for the design of a low-background high-induction precise synthetic promoter. The technical scheme of the invention is realized as follows: The first aspect of the present invention proposes a DNA molecule which is any one of the following 1) to 3): 1) A nucleotide sequence shown in SEQ ID NO. 1; 2) DNA molecules having 75% or more identity with the nucleotide sequence defined in 1) and having promoter function, or 3) A DNA molecule which hybridizes to the nucleotide sequence set forth in 1) under stringent conditions and has promoter activity. In a second aspect of the invention, a biomaterial comprising a DNA molecule according to the first aspect is provided, said biomaterial being any one of the following: A1 A DNA expression cassette; a2 A nucleic acid construct; a3 A recombinant vector; a4 A recombinant microorganism; a5 A recombinant cell line. In a third aspect, the invention provides the use of a DNA molecule according to the first aspect, comprising: B1 Use as promoter; b2 As a promoter to promote expression of a gene of interest. In a fourth aspect, the present invention provides a method for increasing the expression level of a target gene under drought stress in a plant, comprising introducing the DNA molecule of the first aspect into a target plant to obtain a transgenic plant, and allowing the target gene in the transgenic plant to start transcription of the target gene with the DNA molecule of the first aspect as a promoter, thereby increasing the expression level of the target gene after transcription. Further, the target gene is an endogenous gene or an exogenous gene of the target plant. Further, the target gene is a gene capable of resisting abiotic stress. Further, the abiotic stress is drought stress, high salt stress, abscisic acid stress, low temperature stress and/or alkali stress. In a fifth aspect, the invention provides the use of a DNA molecule according to the first aspect, or a biological material according to the second aspect, or a method of preparation according to the third aspect, for initiating expression of a gene of interest. In a sixth aspect, the invention provides the use of a DNA molecule according to the first aspect, or a biological material according to the second aspect, or a method of preparation according to the third aspect, for the cultivation of transgenic plants. Further, the plant is tobacco or alfalfa. Compared with the prior art, the invention has the beneficial effects that: According to the invention, 15 cis-elements possibly induced by drought in a classical NCED, LEA, DHN gene promoter regulated by drought are integrated in leymus chinensis for the first time, and an artificial promoter induced by drought is synthesized. Through detection, the promoter has transcription activity induced by drought stress, can drive the target gene at the downstream of the promoter to be efficiently expressed in plants, and opens up a new way for the design of a low-background high-induction accurate synthetic promoter. The DNA molecule provided by the invention is used as an drought stress inducible promoter