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Protein related with plant tolerance and coding gene and applications thereof

A technology of plant stress tolerance and related proteins, which is applied in the field of plant stress tolerance related proteins and their coding genes and applications, can solve problems such as the complexity of plant stress resistance mechanisms, achieve broad application and market prospects, and improve drought tolerance Effect

Active Publication Date: 2013-02-06
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The mechanism of plant resistance to stress is very complex. Although traditional breeding methods have some effect on the improvement of crop stress resistance, they are still far from the desired goal.

Method used

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  • Protein related with plant tolerance and coding gene and applications thereof
  • Protein related with plant tolerance and coding gene and applications thereof
  • Protein related with plant tolerance and coding gene and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1, the discovery of ZmbZIP72 protein and its coding gene

[0041] A new protein was discovered from maize variety CN165 through observation of a large number of traits and molecular detection.

[0042] The protein shown in Sequence 1 of the sequence listing is named ZmbZIP72 protein. The gene encoding the ZmbZIP72 protein is named ZmbZIP72 gene, its cDNA is shown in sequence 2 of the sequence listing, and the open reading frame is the 181st to 1074th nucleotides from the 5' end of the sequence 2 of the sequence listing.

Embodiment 2

[0043] Embodiment 2, the acquisition of transgenic plants

[0044] 1. Construction of recombinant expression vector

[0045] The construction flowchart of recombinant plasmid pCAMBIA3301-35S::ZmbZIP72 is shown in figure 1 .

[0046] 1. Extract the total RNA of corn variety CN165 and reverse transcribe it into cDNA.

[0047] 2. Using the cDNA extracted in step 1 as a template, perform PCR amplification with a primer pair composed of F1 and R1 to obtain a PCR amplification product.

[0048] F1: 5'-TTA AGATCT ATGGACGAGCTGCTCCAG-3';

[0049] R1: 5'-TAT GGTCACC TCACCAGGGGGCCGTCAACGT-3'.

[0050] 3. The PCR amplified product of step 2 was double digested with restriction endonucleases Bgl II and BstE II, and the digested product was recovered.

[0051] 4. Digest the plasmid pCAMBIA3301 with restriction endonucleases Bgl II and BstE II to recover the vector backbone (about 9265 bp).

[0052] 5. Ligate the digested product of step 3 with the vector backbone of step 4 to obta...

Embodiment 3

[0067] Embodiment 3, identification of transgenic plants

[0068] 1. RT-PCR identification

[0069] T 3 The total RNA of generation plants was used to identify the expression of ZmbZIP72 gene by RT-PCR. The primer pair identified by RT-PCR consisted of F3 and R3, and the target sequence was about 100bp.

[0070] F3: 5'-GCTTATACAATGGAGTTAGAAGCCG-3';

[0071] R3: 5'-CTCGTTGTTCTGCATTTCCATAATTTC-3'.

[0072] The results of four transgenic lines (TL5-2, TL6-1, TL11-3, TL12-3) are shown in Figure 4 . The results showed that the Colombian ecotype Arabidopsis did not get the expected band, which was identified as a negative result. The plants of the 9 transgenic lines all obtained the expected bands, which were identified as positive results.

[0073] 2. Abscisic acid stress experiment

[0074] The seeds of the transgenic line TL6-1 (T 3 Generation plants), seeds of transgenic line TL12-3 (T 3 Generation plants), seeds of control plants transformed with empty vector (T 3 G...

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Abstract

The invention discloses a protein related with plant tolerance and a coding gene and applications thereof. The protein is (a) or (b) as follows: (a) a protein prepared from an amino acid sequence as shown in a sequence 1 in a sequence table; and (b) a protein derived by the sequence 1, namely the amino acid sequence of the sequence 1 is subjected to the substitution and / or deficiency and / or addition of one or several amino acid residue and is related with the plant tolerance. The coding gene of the protein is introduced into the plants, so that the drought resistance of the plants can be enhanced. The method has an important theoretical and practical significance to the research in a drought-tolerance molecule mechanism of the plants, the seed selection of drought-tolerance varieties of the plants and the breeding of the drought-tolerance molecules of the plants, and the invention provides an economic, fast and effective way for increasing the drought tolerance of the plants. The protein related with plant tolerance and the coding gene and the applications thereof have wide application and market prospects in the agricultural field.

Description

technical field [0001] The invention relates to a plant stress tolerance-related protein, its coding gene and application. Background technique [0002] Water scarcity and soil salinization are serious facts facing global agricultural production. Due to frequent droughts, the average annual drought-affected area in my country is 327 million mu. With global warming, the drought will become more and more serious, and the area of ​​farmland threatened by drought may exceed 700 million mu. According to estimates, abiotic stresses such as drought, low temperature, and salinity have an impact on crop yields of up to 40%. Water shortages in major grain-producing areas in northern my country reduce yields by 70-80 billion kilograms each year; Frequent occurrence, for example, the drought-affected area of ​​Guangdong every year exceeds 7.5 million mu; the drought-affected area of ​​crops in Sichuan Province in 2006 reached 21 million mu, accounting for 35% of the sown area, and 3.97 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/21C12N15/11C12N15/82A01H5/00C12R1/91
Inventor 黎裕英生张登峰石云素宋燕春王天宇
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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