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Method for improving salt stress resistance capacity of plants

A salt stress, plant technology, applied in the field of improving the ability of plants to resist salt stress, can solve the problems of reducing the performance of salt stress resistance of plants, and achieve the effect of improving the salt resistance of plants and improving the ability of plants to resist salt stress

Inactive Publication Date: 2014-01-22
EAST CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is generally believed that auxin is not directly related to plant salt stress resistance, and it has also been reported that auxin reduces plant salt stress resistance

Method used

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  • Method for improving salt stress resistance capacity of plants
  • Method for improving salt stress resistance capacity of plants
  • Method for improving salt stress resistance capacity of plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1, Extracting Arabidopsis Genomic DNA

[0037] Reagent:

[0038] Lysis Buffer: Tris-HCl, pH8.0 (0.2M); Urea (7M); Sodium Lauryl Sarcosine (2%); EDTA (0.05M) Steps:

[0039] (1) Put an appropriate amount of green leaves or flowers of the plant in a 1.5ml centrifuge tube, add 200μl of lysate, grind it into a slurry with a small rod, and then wash the small rod with 400μl of lysate.

[0040] (2) Add 600 μl of phenol-chloroform and shake vigorously for 20 seconds to denature the protein.

[0041] (3) After centrifuging at 12000rpm for 5 minutes, take the supernatant.

[0042] (4) Add 50 μl of sodium acetate and 600 μl of isopropanol, mix well, and centrifuge at 12,000 rpm for 5 minutes.

[0043] (5) Remove the supernatant, and dissolve the pellet in 400 μl TE (Tris-EDTA).

[0044] (6) Add 40μl 3mol / L NaAC (PH5.2), 1ml absolute ethanol, mix well, and centrifuge at 12000rpm for 5min.

[0045] (7) Remove the supernatant, wash the precipitate once with 70% ethanol,...

Embodiment 2

[0047] Example 2 (a), PCR method to obtain the target gene fragment

[0048] (1) Using the Arabidopsis DNA extracted in Example 1 as a template

[0049] The polynucleotide sequence of URO gene is shown as sequence 1 in the sequence listing.

[0050] The polynucleotide sequence of the promoter is shown as sequence 2 in the sequence listing.

[0051] (2) Primer design

[0052] According to the URO gene and its promoter sequence, and the convenience of purchasing and cloning, design a pair of primers, URO-F and URO-R, wherein the former primer has a restriction endonuclease site for Xhol, and the primer sequence is :

[0053] Upstream primer URO-F: 5' ctc gag atg aac cac cgg gac aaa c, as shown in SEQ ID NO: 3.

[0054] The downstream primer URO-R: 5'tta atg atg acg atg acc g is shown in sequence 4.

[0055] (3) PCR amplification of the target fragment

[0056] PCR reagent: KOD-Plus enzyme was purchased from TOYOBO company

[0057] PCR reaction system:

[0058]

[0059...

Embodiment 3

[0075] Embodiment 3, the cloning of PCT product

[0076] Reagent: The T-Vector used is pMD18-T Simple Vector, purchased from Takara

[0077] Connection system:

[0078]

[0079] Place the ligation system at 4°C or 16°C overnight, or at 25°C for two hours to obtain a recombinant plasmid.

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Abstract

The invention provides a method for improving salt stress resistance capacity of plants. The plants are transformed by expression vectors containing URO (Urobilinogen) genes and promoter sequences, and thus the salt stress resistance capacity of the plants is improved. Nucleotide sequences of the URO genes are shown as a sequence 1 in a sequence table; nucleotide sequences of promoters are shown as a sequence 2 in the sequence table; the expression vectors containing the URO genes and the promoter sequences are binary vectors containing T-DNA (Transferred-Deoxyribose Nucleic Acid) sequences. According to the method, through changing the expression of a transcriptional regulation factor, plant cell differentiation states are changed, and thus the purpose of increasing the salt stress resistance capacity of the plants is realized.

Description

technical field [0001] The invention relates to the technical field of improving the performance of agricultural crops, in particular to a method for improving the ability of plants to resist salt stress, so as to improve the ability of plants to resist salt stress. Background technique [0002] With the increase of global temperature, the evaporation of water on the earth's surface accelerates, the sea level rises, and the long-term irrigation of agricultural production leads to the accumulation of salt in the soil year by year, which has become one of the reasons for the reduction of crop production. On the other hand, the pressure of the sharply increasing population on the limited arable land area is also increasing year by year, and the effective use of a large number of abandoned saline-alkali lands is also an important direction for agricultural development. Therefore, improving the salt stress tolerance of commercial crops is one of the important directions of commer...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/29C12N15/66A01H5/00
Inventor 孙越刘水李玲
Owner EAST CHINA NORMAL UNIVERSITY
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