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Application of TT1 gene in improving cold tolerance of plant

A cold resistance, plant technology, applied in plant regeneration, plant products, genetic engineering and other directions, can solve the problem of few cold resistance genes, and achieve the improvement of plant cold resistance, good application prospects, simple and effective methods. Effect

Active Publication Date: 2014-03-26
SICHUAN BIODESIGN GENE ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few available cold resistance genes in this field

Method used

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  • Application of TT1 gene in improving cold tolerance of plant
  • Application of TT1 gene in improving cold tolerance of plant
  • Application of TT1 gene in improving cold tolerance of plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1. Using TT1 gene to improve cold resistance of rice

[0050] 1. Cloning and acquisition of TT1 gene

[0051] Using the atp6 (genebank gi:89279377) gene in rapeseed as the bait protein, according to the yeast two-hybrid method (see the information disclosed by Clontech), an EST sequence (shown in SEQ ID NO: 2) in rapeseed was screened, and then According to the sequence screened in this section, the gene for improving plant cold resistance of the present invention is obtained by the method of 5' RACE (see the information disclosed by Takara Company), and its nucleotide sequence is as SEQ ID NO.1 in the sequence listing shown. Design primers according to the nucleotide sequence shown in SEQ ID NO.1,

[0052] Upstream primer (SEQ ID NO.3): 5'-ATGTCGGATCATTTGAGTTTATG-3',

[0053] Downstream primer (SEQ ID NO.4): 5'-TCAGACTGGTGTTGGGTTGGATAT-3'.

[0054] Then the nucleotide sequence shown in SEQ ID NO.1 is amplified from the rapeseed cDNA by PCR. The PCR program...

Embodiment 2

[0170] Embodiment 2, using TT1 gene to improve the cold resistance of rapeseed

[0171] 1. Preparation of Rapeseed Rapeseed Plant Overexpressing TT1 Gene and Obtaining of Seeds

[0172] 1. Construction of recombinant plasmids for overexpression of target genes

[0173] Design primers according to the nucleotide sequence shown in SEQ ID NO.1,

[0174] Upstream primer (SEQ ID NO.11): 5'-CGC GGATCCATGTCGGATCATTTGAGTTTATG-3',

[0175] Downstream primer (SEQ ID NO.12): 5'-CCGGAGC TCTCAGACTGGTGTTGGGTTGGATAT-3'.

[0176] Through PCR, the nucleotide sequence shown in the complete SEQ ID NO.1 is amplified from rapeseed cDNA, and the PCR program is as follows:

[0177] 1. 95℃ for 4min (pre-denaturation)

[0178] 2. 95℃ 30s (denaturation)

[0179] 3. 53℃ 30s (refolding)

[0180] 4. 72℃ 50s (extension)

[0181] 5. 2~4 steps cycle 30 times

[0182] 6. 72℃ for 5min (final extension)

[0183] 7. Store at 4°C.

[0184] Purify the PCR product (see the information disclosed by Qiagen)...

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Abstract

The invention relates to the field of biotechnology, in particular to the application of TT1 gene in improving the cold tolerance of plant, and aims at providing a new effective alternative for the field of transgenic technology for improving the cold tolerance of plant. Proved by experiments, the plant genetically modified and overexpressed by the TT1 gene has favorable resistance to cold adverse situations. The method of cultivating the cold resistant plant is simple and effective, and provides a new effective alternative for the field.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the use of TT1 gene in improving cold resistance of plants. technical background [0002] Plants have sufficient adaptability and resistance to environmental changes and adverse environments. This stress resistance is not only controlled by phylogenetic genes, but also restricted by physiological ecology during individual development. As one of the important environmental factors, temperature plays a decisive role in certain growth and development processes of plants under the limitation of plant genetic background. The low temperature causes the plants to be damaged to varying degrees, and even cause death. The ability of plants to resist low temperature damage is the plant's cold resistance. [0003] There are many kinds of plants, widely distributed in a wide area, and are often harmed by low temperature. However, plants of different species or different varieties o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29A01H4/00A01H5/00A01H5/10
Inventor 杨毅
Owner SICHUAN BIODESIGN GENE ENG
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