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Promoter of rice root specific expression gene OsHyPRP06/R3L1 and application thereof

A technology for expressing genes and promoters, applied in the field of plant genetic engineering, to achieve the effect of promoting root system development and great application value

Inactive Publication Date: 2021-11-02
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research and application of root-specific promoters are not as extensive and in-depth as aboveground tissues (Potenza et al., 2004). Using root-specific promoters to study the development and morphogenesis of plant roots is of great significance for crop genetic improvement

Method used

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  • Promoter of rice root specific expression gene OsHyPRP06/R3L1 and application thereof
  • Promoter of rice root specific expression gene OsHyPRP06/R3L1 and application thereof
  • Promoter of rice root specific expression gene OsHyPRP06/R3L1 and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0024] The promoter of the rice root-specific expression gene OsHyPRP06 / R3L1 of the present invention is located in the upstream sequence of the gene coding region of the rice root-specific expression gene OsHyPRP06 / R3L1, and its nucleotide sequence is shown in SEQ ID NO:1.

[0025] Analysis of the expression pattern of the OsHyPRP06 / R3L1 coding sequence driven by this promoter:

[0026] 1. Conventional cultivation of rice

[0027]Soak plump Nipponbare rice seeds in 10% sodium hypochlorite solution, disinfect for 10 minutes, and rinse with water 5-6 times. Soak the sterilized seeds in an appropriate amount of water, and germinate at 30°C for 1-2 days in a dark environment. Afterwards, plant them in 96-well culture holes, and in the incubator, maintain the culture conditions of 28°C / 25°C, 16 hours / 8 hours, and a relative humidity of 60% for ordinary hydroponic culture for 3-4 days. Afterwards, the rice seedlings are transferred to 1 / 2 culture nutrient solution (PH value 4.8-5...

Embodiment 2

[0054] This example is the application of the promoter of the rice root-specific expression gene OsHyPRP06 / R3L1 in the construction of transgenic rice. It is used in rice to regulate the specific expression of exogenous genes in roots, and to promote the specific expression of downstream coding genes in rice roots.

[0055] By constructing a plant expression vector (gene fragment vector containing the sequence) containing the promoter, it is used in rice to regulate the specific expression of foreign genes in roots, and to construct transgenic rice.

[0056] The following is the rice root-specific promoter OsHyPRP06 / R3L1 fusion pCAMBIA1301 vector (including GUS, figure 2 shown) build.

[0057] 1. Rice Genomic DNA Extraction

[0058] Genomic DNA was extracted using the CTAB method, and the specific steps were as follows:

[0059] 1) Take about 0.2g of fresh rice tissue, put it into a 2mL EP tube, freeze it in liquid nitrogen and grind it thoroughly.

[0060] 2) Add 650 μL ...

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Abstract

The invention relates to plant genetic engineering, in particular to a promoter of a rice root specific expression gene OsHyPRP06 / R3L1 and application of the promoter. The invention relates to a promoter of a rice root specific expression gene OsHyPRP06 / R3L1. The promoter is a nucleotide sequence as shown in SEQ ID NO: 1. The promoter of the rice root specific expression gene OsHyPRP06 / R3L1 is used for promoting specific expression of a downstream coding gene in a rice root and carrying out plant transformation of rice root specific expression. The promoter is used for constructing transgenic rice, a specific gene is connected to the downstream of the promoter, so that the specific gene is specifically enhanced and expressed in roots, efficient plant transformation of root specific expression is carried out, root development is promoted, yield and income increase of rice is facilitated, and the promoter has great application value only for targeted genetic improvement of roots.

Description

technical field [0001] The invention relates to plant genetic engineering, in particular to a promoter sequence for cloning the upstream of rice OsHyPRP06 / R3L1 gene coding region and the use of the promoter. Background technique [0002] The root system is an important organ for plants to absorb water and nutrients, and the development and morphogenesis of the rice root system are crucial to high and stable rice yields (Shigenori et al., 1986). The function of plant root system is complex, such as the root system has secretion function, and the secreted organic acids, enzymes, alkaloids and other substances have a profound impact on the effectiveness of minerals and fertilizers in the soil, and also affect the types and numbers of microorganisms in the root system, etc. ( Chang Xuexiu et al., 2000). The root system also synthesizes some physiologically active substances, such as cytokinin CTK and gibberellin ABA, to regulate the development of shoots (Pan Xiaohua et al., 20...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82A01H5/06A01H6/46
CPCC07K14/415C12N15/8205C12N15/8227C12N15/8261
Inventor 赵文丽王坤安世恒张博常彦鹏
Owner HENAN AGRICULTURAL UNIVERSITY
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