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Plant disease resistance-related protein and coding gene and application thereof

A gene and plant technology, applied in the field of plant disease resistance-related proteins and their coding genes and applications, can solve the problems of slow breeding progress and lack of resistance sources

Inactive Publication Date: 2010-12-15
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

However, scab and sheath blight resistance are quantitative traits controlled by multiple genes, and there is a lack of sources of resistance within crop species, and the progress of conventional breeding is slow

Method used

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  • Plant disease resistance-related protein and coding gene and application thereof
  • Plant disease resistance-related protein and coding gene and application thereof
  • Plant disease resistance-related protein and coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, discovery of TaBS108 protein and its full-length cDNA sequence

[0033] 1. Discovery of TaBS108 protein and its full-length cDNA sequence

[0034] 1. The leaves of the wheat variety Sumai No. 3 seedlings grown for four weeks were inoculated with scab;

[0035] 2. After 48 hours, use the TRIZOL method to extract the total RNA of the leaves, synthesize the first strand of cDNA according to the method provided by Dalian Bao Biological Company, and use it as a template for PCR amplification and cloning of TaBS108;

[0036] 3. A pair of PCR amplification primers (WBS108-U and WBS108-L) designed.

[0037] WBS108-U: 5'-CCCATCTCACCATCTCCA-3';

[0038] WBS108-L: 5'-TCCTCTCGCTGTCACGCA-3'.

[0039] 4. Using the cDNA in step 2 as a template, perform PCR amplification with the primer pair in step 3 to obtain an amplified product.

[0040] The total volume of the PCR reaction was 25 μL, including 2 μL cDNA (50 ng), 100 μmol / L dNTP, 0.4 μmol / L each primer, 1U LA-Taq ...

Embodiment 2

[0047] Embodiment 2, the acquisition of transgenic TaBS108 wheat plant and its disease resistance analysis

[0048] 1. Construction of recombinant expression vector

[0049] 1. Extract the RNA from the leaves of Sumai No. 3 seedlings and reverse transcribe it into cDNA;

[0050] 2. Using the cDNA in step 1 as a template, perform PCR amplification with a primer pair composed of WBSO-SMF and WBSO-SAR to obtain a PCR amplification product (full-length ORF of TaBS108 gene carrying SmaI and SacI sites).

[0051] WBSO-SMF: 5'-AT CCCGGG ATGGCTCGCGGTGCAGCTA-3' (the SmaI enzyme recognition site is underlined);

[0052] WBSO-SAR: 5'-TT GAGC TC AACGAATCTTAGAACAGTCCA-3' (SacI enzyme recognition site is underlined).

[0053] PCR reaction program: first 95°C pre-denaturation for 3min; then 95°C for 30s, 60°C for 1min, 72°C for 1min, 35 cycles; finally 72°C for 10min to blunt the end.

[0054] 3. The PCR amplified product was recovered and connected with the pMD18-T vector (Dalian Ba...

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Abstract

The invention discloses a plant disease resistance-related protein and a coding gene and application thereof. The protein is (a) or (b): (a) a protein consisting of an amino acid sequence shown as the sequence 1 in a sequence table, and (b) a protein which is derived from the sequence 1, related to the plant disease resistance and obtained by substituting and / or losing and / or adding one or more amino acid residues in the amino acid sequence of the sequence 1. When the coding gene of the protein is transferred into a plant, a transgenic plant with obviously improved disease resistance can be obtained. The plant disease resistance-related protein and the coding gene and application thereof have great significance for the cultivation of disease-resistant plants, particularly sheath blight-resistant and gibberella-resistant plants.

Description

technical field [0001] The invention relates to a plant disease resistance-related protein, its coding gene and application. Background technique [0002] Lipid transfer proteins (lipid transfer proteins, LTPs) are an important class of active proteins in plant life activities. They are basic, small molecular proteins rich in cysteine ​​amino acids that can bind lipids. LTPs are secreted proteins. located in the cell wall. Lipid transfer protein is involved in membrane biosynthesis, regulation of intracellular fatty acid pool and plant defense response, and is a member of PR14 family of pathogenesis-related proteins (PR) (Kirubakaran S.Isaac, Begum S.Mubarak, Ulaganathan K ., Sakthivel N.2008, Characterization of a new antifungal lipid transfer protein from wheat, Plant Physiology and Biochemistry 46: 918-927). Studies have shown that some LTPs can inhibit the growth of some fungal pathogens in vitro, and have the ability to synergistically enhance the resistance of defens...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
Inventor 张增艳李钊辛志勇徐惠君杜丽璞周淼平
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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