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OsQR1 protein, encoding gene thereof, and application of protein and gene

A technology that encodes genes and proteins, applied in applications, genetic engineering, plant genetic improvement, etc., can solve the problems of slow progress, long cycle of drought-resistant rice varieties, unstable varieties, etc., and achieve the effect of improving drought resistance

Active Publication Date: 2016-06-01
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Using traditional breeding methods to cultivate drought-resistant rice varieties has a long cycle and high chance, the varieties cultivated are unstable, and the progress is relatively slow. However, the use of genetic engineering technology can directly modify the genetic background of plants at the genetic level, and the genetic traits of plants can be directional modified

Method used

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  • OsQR1 protein, encoding gene thereof, and application of protein and gene
  • OsQR1 protein, encoding gene thereof, and application of protein and gene
  • OsQR1 protein, encoding gene thereof, and application of protein and gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Acquisition of the gene encoding the drought resistance-related protein OsQR1 in upland rice

[0035] 1. Take the leaves of upland rice IRAT109 seedlings cultivated under normal conditions, extract total RNA by Trizol method, and reverse transcribe with M-MLV reverse transcriptase to obtain cDNA.

[0036] 2. Using the cDNA obtained in step 1 as a template, perform PCR amplification with primer F1 and primer R1 to obtain a PCR amplification product.

[0037] The primer sequences are as follows:

[0038] F1: 5'-G TTAATTAA CAATGGCGGTGAAGGTCT-3' (SEQ ID No. 1)

[0039] (The underlined sequence is the restriction endonuclease PacI restriction endonuclease recognition site)

[0040] R1: 5'-C GAGCTC TCAAGCGGACCCCTTGA-3' (SEQ ID No. 2)

[0041] (The underlined sequence is the restriction endonuclease SacI restriction endonuclease recognition site)

[0042] 3. Perform agarose gel electrophoresis on the PCR amplification product, recover and purify the DNA frag...

Embodiment 2

[0043] Embodiment 2, real-time fluorescent quantitative PCR analysis endogenous OsQR1 gene expression

[0044] 1. Coercion treatment

[0045] The rice variety Nipponbare and the upland rice variety IRAT109 seedlings that are 4 weeks old in hydroponics are subjected to any of the following (1) to (4) treatments respectively:

[0046] (1) ABA treatment: soak the roots of the seedlings in an aqueous solution of 100 μM ABA, and cultivate them under light for 1 hour, 4 hours, 6 hours, 8 hours, 9 hours, 16 hours, and 24 hours, then take the leaves and freeze them with liquid nitrogen, Store at 80°C for later use.

[0047] (two) H 2 o 2 Treatment: Soak the roots of the seedlings in 1mMH 2 o 2 After being placed in the aqueous solution for 1 hour, 3 hours, 6 hours, 8 hours, 9 hours, 16 hours, and 24 hours, the leaves were taken out, quickly frozen with liquid nitrogen, and stored at -80°C for later use.

[0048] (3) PEG (drought) treatment: Soak the roots of the seedlings in 200...

Embodiment 3

[0064] Embodiment 3, the construction of recombinant expression vector

[0065] PacI and SacI double-digest the PCR amplification product obtained in step 2 of Example 1 to obtain the gene fragment; PacI and SacI double-enzyme-digest the vector pMDC83 to obtain the large vector fragment; connect the gene fragment to the large vector fragment to obtain the recombinant vector 35S ::OsQR1, the recombinant vector 35S::OsQR1 is inserted between the PacI and SacI sites of the vector pMDC83 (ie downstream of the double tobacco mosaic virus promoter 35S) from the 79th to the 5' end of SEQ ID No.3 The DNA fragment indicated by 690 nucleotides was sequenced by 35S::OsQR1, and the result was correct.

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Abstract

The invention discloses an OsQR1 protein, an encoding gene thereof, and an application of the protein and the gene. The protein can be a protein (1) with the amino acid sequence represented by SEQ ID No.4, or a protein (2) obtained by substituting and / or deleting and / or adding one or more amino acid residues to the amino acid sequence represented by SEQ ID No.4 and having same functions with the protein (1). The OsQR1 protein and the encoding gene thereof provide a new gene and a new material for rice drought-resisting molecule breeding, and are important to improve the drought resistance of rice and other crops.

Description

technical field [0001] The invention relates to an OsQR1 protein and its coding gene and application, belonging to the field of biotechnology. Background technique [0002] Abiotic stress is the main stress factor that affects the normal growth and development of plants and limits the increase of crop yield. At present, the impact of drought on crop production in the world ranks first among all stress factors, and its damage to crop growth is equivalent to the sum of all natural disasters, and has become a bottleneck in agricultural development in many places. Rice is one of the main food crops in the world and is the staple food for more than 60% of the population in my country. According to statistics, rice water accounts for about 70% of agricultural water use. Since 1997, my country's rice cultivation area has continued to shrink, and more than 20 million mu of rice suffers from drought every year, and even tens of millions of mu of rice cannot be planted. Therefore, o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
Inventor 李自超刘鹏丽段俊枝熊海燕李金杰张洪亮
Owner CHINA AGRI UNIV
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