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Plant resistance related protein ATSAR 25, and encoding genes and application thereof

A gene and encoding technology, applied in the field of plant resistance-related protein ATSAR25 and its encoding gene and application, can solve the problems of polluted environment and high cost, and achieve the effect of enhancing disease resistance

Active Publication Date: 2012-03-07
FUJIAN SANAN SINO SCI PHOTOBIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the effect of this measure is ideal, it is costly and pollutes the environment

Method used

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  • Plant resistance related protein ATSAR 25, and encoding genes and application thereof
  • Plant resistance related protein ATSAR 25, and encoding genes and application thereof
  • Plant resistance related protein ATSAR 25, and encoding genes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Embodiment 1, the acquisition of ATSAR25 protein and its coding gene

[0048] 1. According to the existing NCBI database and literature, set a pair of primers as follows:

[0049] F1 (forward primer): 5'-GC TCTAGA GCAAAATAAGAAATTCTATAAGTCCT-3' (XbaI digestion recognition sequence is underlined);

[0050] R1 (reverse primer): 5'-CG GGATCC gatgtctctcaagatctgcca-3' (underlined BamHI restriction recognition sequence).

[0051] 2. Genomic DNA was extracted from leaves of Arabidopsis thaliana ecotype Columbia.

[0052] 3. Using genomic DNA as a template, use the primer pair designed in step 1, and perform PCR amplification with PrimeSTAR HS high-fidelity enzyme from TaKaRa Company.

[0053] 4. The PCR amplification product is sequenced, as shown in sequence 2 of the sequence listing.

[0054] The protein shown in Sequence 1 of the sequence listing is named ATSAR25 protein. The gene encoding the ATSAR25 protein is named as the ATSAR25 gene, and its genomic DNA is shown...

Embodiment 2

[0055] Embodiment 2, the cloning of each gene and the construction of its recombinant expression vector

[0056] 1. Acquisition of ATSAR25 gene and construction of recombinant plasmid 326-ATSAR25-FLAG

[0057] 1. Genomic DNA was extracted from leaves of Arabidopsis thaliana ecotype Columbia.

[0058] 2. Using the genomic DNA in step 1 as a template, use the primer pair composed of F1 and R1 to carry out PCR amplification under the action of TaKaRa's PrimeSTAR HS high-fidelity enzyme to obtain PCR amplification products. The agarose gel electrophoresis of the PCR amplification product is shown in figure 1 (M represents a nucleotide marker, which is DL2000 from TaKaRa Company).

[0059] 3. The PCR amplification product of step 2 was double-digested with restriction enzymes XbaI and BamHI, and the digested product was recovered.

[0060] 4. The 326-FLAG expression vector was double digested with restriction enzymes XbaI and BamHI, and the vector backbone of about 3827 bp was r...

Embodiment 3

[0073] Embodiment 3, ATSAR25 gene is inducing PR1 gene promoter (Pro AtPR1 ) application in promoting gene expression

[0074] 1. Transient expression of recombinant plasmids in Arabidopsis leaf protoplasts

[0075] The recombinant plasmid 326-ATSAR25-FLAG and recombinant plasmid 326-Pro constructed in Example 2 AtPR1 ::GFP co-transformed Arabidopsis protoplasts (recombinant plasmid 326-T 7 -FLC is used as a negative control of the recombinant plasmid 326-ATSAR25-FLAG; the 326-FLAG expression vector is used as another negative control of the recombinant plasmid 326-ATSAR25-FLAG), the specific steps are as follows:

[0076] 1. Germinate Colombian ecotype Arabidopsis seeds on MS medium, transplant them into soil when the roots grow to 1-3 cm, and cultivate them in a greenhouse at 23° C. (12 hours of light per day, light intensity of 150 μE).

[0077] 2. Add 20ml of double distilled water to a 90mm petri dish, then add 1.82g of D-mannitol and dissolve it.

[0078] 3. Take the...

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Abstract

The invention discloses a plant resistance related protein ATSAR25, and encoding genes and application thereof. The protein provided in the invention is named ATSAR 25, originates from Arabidopsis thaliana, and is one selected from the following proteins: (a) a protein composed of an amino acid sequence represented by sequence 1; (b) a protein derived from sequence 1, wherein, the protein is obtained by substituting and / or deleting and / or adding one or more deoxynucleotide residue of the amino acid sequence of sequence 1 and has either the function of inducing promotion of ICS 1 gene promoters or the function of inducing promotion of PR 1 gene promoters; (c) a protein which is derived from sequence 1 and related to synthesis of salicylic acid in plants and to plant resistance. The ATSAR 25 protein provided in the invention can enhance disease resistance of plants and has a critical value to breeding of disease resistant plants.

Description

technical field [0001] The invention relates to a plant resistance-related protein ATSAR25, its coding gene and application. Background technique [0002] Plants often suffer from various diseases in the process of cultivation and production. Some diseases pose a great threat to plant growth and seriously damage the yield and quality of crops. For plant diseases, at present, the main measures are to control them with drugs. Recently, people have begun to use abiotic inducers to induce plant disease resistance, and have been widely used in many important crops such as tobacco, potato, tomato, cucumber, bean, and rice. Although the effect of this measure is ideal, it is costly and pollutes the environment. Therefore, there is an urgent need to develop new biological means to improve the plant's own disease resistance. [0003] Systemic acquired resistance is a plant defense response that is induced when plants are attacked by pathogens and pests and can quickly spread to ot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/21C12N15/82A01H5/00
Inventor 金京波蔡斌李媛
Owner FUJIAN SANAN SINO SCI PHOTOBIOTECH CO LTD
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