CN-122012609-A - Rice endogenous space-time specific promoter OsMOT, 1pro and application thereof

Abstract

The invention belongs to the technical field of agricultural biology, and particularly relates to a rice endogenous space-time specific promoter OsMOT < 1 >, 1pro and application thereof. The nucleotide sequence of the promoter OsMOT1 and 1pro is shown as SEQ ID NO. 1. The OsMOT & lt 1 & gt, 1pro & gt is a space-time special expression regulatory element derived from rice, can efficiently and stably drive a target gene to express in plant callus, and can realize targeted efficient and stable expression in root and stem tissues in plant vegetative growth period. The promoter can effectively replace a non-plant source promoter commonly used in the existing plant genetic engineering, has important industrialized application value in the field of plant genetic engineering, can not only improve the targeting property and stability of target gene expression in transgenic plants, but also remarkably reduce the potential biosafety risk of the transgenic plants caused by introducing exogenous DNA into plant genome.

Inventors

• LI YINGJIANG
• LIU XIAOLIN
• SHENG XIABING
• ZHANG TAO
• TANG JIE
• CHEN YIYAN
• LIU YUHAO
• HU YUANYI

Assignees

• 三亚市国家耐盐碱水稻技术创新中心
• 湖南杂交水稻研究中心

Dates

Publication Date

20260512

Application Date

20260416

Claims (4)

1. 1. The application of plant space-time specific promoter OsMOT and 1pro is characterized in that the application is any one of the following: 1) Application in preparing transgenic rice; 2) The application of driving the expression of exogenous genes in the callus, root and stem of rice, wherein the exogenous genes are GUS genes; the nucleotide sequence of the promoter OsMOT1 and 1pro is shown as SEQ ID NO. 1.
2. 2. The use according to claim 1, wherein in the preparation of transgenic rice, the GUS gene expression cassette is introduced into rice to obtain transgenic rice, wherein the GUS gene expression cassette comprises a promoter OsMOT1 and 1pro drives transcription of the GUS gene.
3. 3. The use according to claim 1, wherein the promoter OsMOT 1pro is amplified from the primer pair set forth in SEQ ID NO. 2-3.
4. 4. Use of a biomaterial related to the promoter OsMOT, 1pro as claimed in claim 1, characterized in that the use is any of the following: 1) Application in preparing transgenic rice; 2) The application of driving the exogenous gene to express in the callus, root and stem in vegetative growth period of rice, wherein the exogenous gene is GUS gene; The biological material is any one of the following A1) to A3): a1 1pro, an expression cassette comprising said promoter OsMOT; a2 1pro, or a recombinant vector comprising the expression cassette of A1); A3 A recombinant microorganism containing said promoter OsMOT1;1pro, or a recombinant microorganism containing said expression cassette of A1), or a recombinant microorganism containing said recombinant vector of A2).

Description

Rice endogenous space-time specific promoter OsMOT, 1pro and application thereof Technical Field The invention belongs to the technical field of agricultural biology, and particularly relates to a rice endogenous space-time specific promoter OsMOT < 1 >, 1pro and application thereof. Background The promoter is a key region for specific recognition, binding and transcription initiation of RNA polymerase as a core DNA cis-acting element for regulating and controlling gene transcription initiation, contains a conserved motif necessary for transcription initiation, is mostly positioned upstream of a transcription initiation site of a structural gene, and does not participate in transcription process. According to the gene expression regulation mode, the promoters can be divided into three types of constitutive, inducible and space-time specific types, wherein the space-time specific type promoters can mediate the accurate expression of genes in specific tissues, organs or development stages, are core elements for realizing the accurate regulation and control of gene expression and avoiding the side effects of ectopic expression, have irreplaceable unique values in directional property improvement and metabolic pathway regulation, and are vital to the accurate development of plant genetic engineering. In the current plant genetic engineering, although the application of the space-time specific promoter is very valuable, available high-quality resources are relatively deficient, and the existing identified space-time specific promoter has the problems of insufficient expression activity, low tissue specificity and the like, so that the actual requirement of accurately regulating and controlling the target gene expression is difficult to meet. Part of the space-time specific promoters derived from heterologous species also have the defects of poor suitability and unstable regulation efficiency, and further limit the wide application of the promoters in plant transgenic operation. Therefore, the novel efficient, high-specificity and wide-adaptability plant origin space-time specific promoter is excavated, and the novel efficient, high-specificity and wide-adaptability plant origin space-time specific promoter has become a core requirement for breaking through the bottleneck of accurate regulation of plant genes and pushing genetic engineering to develop to high efficiency and accuracy. Disclosure of Invention The invention aims to dig and verify a safe and efficient space-time specific promoter from rice so as to overcome the limitation of the existing transgenic promoter in safety and application effect. In order to achieve the above purpose, the present invention provides the following technical solutions: the first aspect of the invention provides a plant space-time specific promoter OsMOT1, 1pro, the nucleotide sequence of which is shown as SEQ ID NO.1, and the promoter can be obtained by high-efficiency amplification of primer pairs shown as SEQ ID NO.2 and SEQ ID NO. 3. In a second aspect, the invention provides plant space-time specific promoters OsMOT1, 1 pro-related biomaterials: 1) An expression cassette containing said promoter OsMOT, 1 pro. In an example, the present inventors constructed an expression cassette OsMOT1 comprising the promoter, 1pro-GUS-nosT, which also comprises a functional gene and a terminator, the nucleotide sequence of which is shown in SEQ ID NO. 4. The structures in the expression cassette are functionally linked to each other in the direction of transcription. 2) Recombinant vectors containing said promoter OsMOT, 1pro or the expression cassette described above. In the examples, the present inventors inserted the above promoter or expression cassette between HindIII and NcoI double cleavage sites of the 1305gusplus vector to construct the recombinant vector 1305gusplus-OsMOT1, 1pro-2020. 3) Recombinant microorganisms containing said promoter OsMOT, 1pro or the above-mentioned expression cassette or the above-mentioned recombinant vector. 4) A transgenic plant cell line containing said promoter OsMOT1, 1pro or the expression cassette described above or the recombinant vector described above. In a third aspect, the invention provides a plant space-time specific promoter OsMOT1, 1pro and any one of the following applications of the relevant biological materials described above: 1) Use in the preparation of transgenic plants; 2) The application of driving the expression of exogenous genes in plant callus and/or vegetative growth phase tissue; The exogenous gene is GUS gene, and the plant comprises rice, corn and wheat; When the plant is rice, driving GUS gene to express in callus, root and stem; when the plant is corn, driving GUS gene to express in callus, root and stem; When the plant is wheat, GUS gene is driven to express in callus, roots and stems. Further, the application is that the OsMOT1, 1pro promoter is constructed into a vector and then is introduced into a plant to prepa