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Rice root specific promoter and application thereof

A root-specific, promoter technology, applied in the field of genetic engineering, to achieve the effect of improving efficient absorption and enhancing resistance

Inactive Publication Date: 2012-07-11
SHANGHAI AGROBIOLOGICAL GENE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are no related reports about the rice gene OsGCW14 (Gramene ID: LOC_Os10g31680) promoter regulating the root-specific expression of exogenous genes and its application

Method used

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  • Rice root specific promoter and application thereof
  • Rice root specific promoter and application thereof
  • Rice root specific promoter and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1: Screening of root-specifically expressed genes:

[0040] The Affymetrix rice microarray data were downloaded from the CREP gene expression database platform (http: / / crep.ncpgr.cn / ), and the microarray data were normalized by RMAExpress software for gene-specific expression screening. In order to search for root-specifically expressed genes, the expression microarray data of two different varieties of rice (Zhenshan 97 and Minghui 63) were normalized to find genes highly expressed in roots but low expressed in other tissues, and the t test (one- way ANOVA t test) to screen the genes with significant differences, and found that 4 genes Os10g31680, Os10g31640, Os10g31660 and Os10g31720 were specifically highly expressed in roots, and their expression levels were extremely low in other tissues, especially shoot organs (see figure 1 ), these four genes are all glycine-rich cell wall structural protein 2 family genes. Among them, Os10g31680 (OsGCW14) has the hi...

Embodiment 2

[0041] Example 2: Quantitative polymerase chain reaction (PCR) expression analysis in rice tissue

[0042] 2.1 Material preparation

[0043] After the seeds of the rice variety Nipponbare germinated, they were transplanted into liquid medium (tap water was prepared with 1 / 5 MS macroelements). After the seedlings grow for 15 days, cut the leaves and roots and put them into liquid nitrogen for storage, and use them to extract the RNA of the leaves and roots; sow the rice in the field, and when the rice starts heading and the length of the young ears is 2 cm, cut the young ears Quickly put into liquid nitrogen for storage, used to extract the RNA of the ear; inoculate the seeds on the callus induction medium (MS+2, 4-D 2.0mg / L), and the induced callus will be Quickly put into liquid nitrogen storage, used to extract the RNA of callus.

[0044] DNA-free total RNA preparation

[0045] Extract according to the instruction manual of the plant leaf RNA mini-extraction kit provided ...

Embodiment 3

[0054] Embodiment 3: OsGCW14 gene promoter regulates the expression analysis of GUS reporter gene

[0055] 3.1 Construction of OsGCW14 gene promoter fusion GUS vector:

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Abstract

The invention belongs to the technical field of genetic engineering and specifically relates to an OsGCW14 gene promoter which is used for regulating and controlling specific expression of exogenous genes of rice in roots and an application thereof. The invention discloses a rice gene promoter, and the nucleotide sequence of the rice gene promoter is as shown in SEQ (sequence) ID (identity) No.1 (number 1). The invention further introduces the application of the OsGCW14 gene promoter for constructing a transgenic plant carrier with root specific expression and transforming a plant host. The OsGCW14 gene promoter has the advantages that the OsGCW14 gene promoter can perform biological repair on soil pollution, or provide drought resistance and salt and alkali tolerance of plants, or improve high-efficiency absorption of nutrients in the soil of the plants, or enhance the resistance to pathogenic microorganisms in the soil and the like, and simultaneously transform the architecture of root systems of the plants for performing genetic improvement on crops.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to a rice gene OsGCW14 gene promoter sequence, that is, a nucleotide sequence between -1 and 1920bp from the upstream of the gene ATG, and the promoter sequence can be used in a rice transgenic regulation system Drive the specific expression and application of target genes in roots. Background technique [0002] The normal expression of plant genes depends on the promoter sequence upstream of the gene. In plant genetic engineering, we can use promoters with different expression regulation levels to drive the expression of target genes. Among them, the most widely used promoter in plants is the CaMV35S promoter. The promoter is the promoter of the tobacco mosaic virus gene, which can promote the expression of heterologous genes in most plants, and is a constitutive promoter (Chua NH, Benfey PN. The cauliflower mosaic virus35S promoter: combinatorial regulatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/63C12N1/21C12N5/10C12N15/82C12N15/84A01H5/06
Inventor 余舜武吴金红陈晨罗利军
Owner SHANGHAI AGROBIOLOGICAL GENE CENT
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