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Plant stress resistance-related protein OsIAA18, encoding gene and application thereof

A plant stress resistance, protein technology, applied in the direction of plant genetic improvement, application, plant peptides, etc., can solve problems affecting food production, restrictions, etc., achieve broad application space and market prospects, improve stress resistance, and increase crop yields Effect

Active Publication Date: 2018-02-06
HUAIYIN INSTITUTE OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existence of large areas of salinized land has seriously affected food production and has become the main factor restricting agricultural production

Method used

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  • Plant stress resistance-related protein OsIAA18, encoding gene and application thereof
  • Plant stress resistance-related protein OsIAA18, encoding gene and application thereof
  • Plant stress resistance-related protein OsIAA18, encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Acquisition of Rice Stress Resistance-Related Protein OsIAA18 and Its Encoding Gene

[0050] 1. Experimental materials

[0051] Refer to Jan et al. (2013) [Asad Jan, Kyonoshin Maruyama, Daisuke Todaka, Satoshi Kidokoro, Mitsuru Abo, Etsuro Yoshimura, Kazuo Shinozaki, Kazuo Nakashima and Kazuko Yamaguchi-Shinozaki. OsTZF1, a CCCH-Tandem Zinc Finger Protein, Confers Delayed Tolayed Sess Rice by Regulating Stress-Related Genes.Plant Physiology, 2013, 161:1202-1216], the plant leaf material of the rice variety 'Huaidao No. 5' was removed, quick-frozen in liquid nitrogen, and stored at -80°C.

[0052] 2. Extraction and purification of total RNA from leaves

[0053] Take about 2.0 g of the leaves of Huaidao No. 5, grind them into powder in liquid nitrogen, put them into a 10 mL centrifuge tube, and use the Applygen Plant RNA Extraction Kit (Applygen Technologies Inc, Beijing) to extract the total RNA of Huaidao No. 5 leaves. Including: Plant RNA Reagent, plant tis...

Embodiment 2

[0061] Example 2 Construction of OsIAA18 Gene Overexpression Vector

[0062] The DNA fragment identified correctly by sequencing in Example 1 containing the nucleotide shown in SEQ ID NO 1 in the sequence table was double-digested with BamH I and Sac I, and the DNA fragment was reclaimed with 1% agarose gel, passed through T 4 The recovered OsIAA18 gene fragment was connected with the recombinant vector pCBGUS containing double 35S promoters by DNA ligase, and the recombinant plant expression vector pCAMBIA1301-OsIAA18 containing the rice OsIAA18 gene was obtained through enzyme digestion identification and sequence analysis. The expression vector also contains a gusA reporter gene and a kanamycin resistance marker gene with an intron, such as figure 1 shown.

Embodiment 3

[0063] Example 3 Transformation of Arabidopsis thaliana with OsIAA18 gene

[0064] The plant expression vector pCAMBIA1301-OsIAA18 of the rice OsIAA18 gene constructed in Example 2 was transformed into Arabidopsis thaliana by the method of dipping flowers, and the specific method is as follows:

[0065] 1. Preparation of Agrobacterium

[0066] (1) Transform pCAMBIA1301-OsIAA18 into Agrobacterium tumefaciens LBA4404 strain (BiovectorCo., LTD) by electric shock method to obtain a recombinant Agrobacterium containing pCAMBIA1301-OsIAA18, and spread it on a plate containing kanamycin resistance to select transformants .

[0067] (2) A single Agrobacterium was picked and inoculated in 5 mL of LB liquid medium (rifampicin 50 μg / mL, chloramphenicol 100 μg / mL), and cultured at 28° C. and 250 rpm for 20 h.

[0068] (3) Transfer 1 mL of bacterial liquid into 20-30 mL of LB liquid medium (rifampicin 50 μg / mL, chloramphenicol 100 μg / mL), culture at 28°C, 250 rpm for about 12 hours, and ...

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Abstract

The invention discloses a plant stress resistance-related protein OsIAA18, an encoding gene and an application thereof. The protein provided by the invention is (a) or (b) as follows: (a) the proteincomposed of the amino acid sequence as shown in sequence SEQ ID NO:2 in a sequence table and (b) the protein with one or more substituted and / or deleted and / or added amino acid residue of the amino acid residue sequence as shown in sequence SEQ ID NO:2 in the sequence table and derived from the plant stress resistance-related sequence SEQ ID NO:2. An experiment of the invention proves that a transgenic plant with obviously enhanced salt tolerance and drought resistance can be acquired in the manner of introducing the encoding gene of the protein into the plant cells. The protein and the encoding gene thereof provided by the invention have an important application value in culturing the plant variety with enhanced stress resistance, so that the protein and the encoding gene thereof have significance in increasing the crop yield. The plant stress resistance-related protein OsIAA18, the encoding gene and the application thereof have wide application space and market prospect in the agricultural field.

Description

technical field [0001] The invention relates to a plant stress resistance-related protein OsIAA18 and its encoding gene and application, in particular to the stress resistance-related protein OsIAA18 derived from rice and its encoding gene and application. Background technique [0002] Rice (Oryza sativa) is one of the world's major food crops, and rice cultivation requires a large amount of freshwater resources. The shortage of water resources is a serious ecological problem restricting the development of global agricultural production. Drought has long been the main limiting factor affecting food security around the world. With the increase of global temperature, the area of ​​arid and semi-arid land is increasing year by year. The area of ​​arid and semi-arid cultivated land in China accounts for about 51% of the total cultivated land area, nearly 2.5×106hm per year 2 Cultivated land is affected by drought to varying degrees. At present, with the global warming and the ...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
Inventor 王飞兵陈新红叶玉秀周青任高磊李烽生王博文杨雨霖
Owner HUAIYIN INSTITUTE OF TECHNOLOGY
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