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Method for cultivation of transgenic wheat resisting take-all and sharp eyespot and related biological materials thereof

A technology for transgenic wheat and take-all disease, which is applied in botany equipment and methods, biochemical equipment and methods, genetic engineering, etc., and can solve problems such as poor resistance and slow breeding progress

Inactive Publication Date: 2013-11-20
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, at present, the large-scale popularized varieties and high-generation lines in production are generally poor in resistance to take-all and sheath blight; the conventional disease-resistant breeding of these two soil-borne diseases is progressing slowly

Method used

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  • Method for cultivation of transgenic wheat resisting take-all and sharp eyespot and related biological materials thereof
  • Method for cultivation of transgenic wheat resisting take-all and sharp eyespot and related biological materials thereof
  • Method for cultivation of transgenic wheat resisting take-all and sharp eyespot and related biological materials thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1, the acquisition of AcAMP-sn gene and the construction of recombinant expression vector

[0046] The coding sequence of AcAMP-sn gene was artificially synthesized, and the recognition sites of Sma I and Sac I restriction endonucleases were added at its 5' end and 3' end, respectively. The nucleotide sequence of the AcAMP-sn gene is shown in sequence 1 in the sequence listing, consisting of 411 nucleotides, and its coding sequence is shown in the 7th to 405th nucleotides of sequence 1 in the sequence listing, its encoding The protein sequence of the protein is shown as sequence 2 in the sequence listing, and consists of 132 amino acid residues.

[0047] The AcAMP-sn gene was connected to pMD18-T vector, the intermediate vector pMD18-AcAMP-sn was constructed, and cloned into Escherichia coli cells.

[0048] The intermediate vector plasmid pMD18-AcAMP-sn was digested with restriction endonucleases Sma I and Sac I, and the AcAMP-sn gene fragment was recovered....

Embodiment 2

[0049] Embodiment 2, the acquisition of transgenic AcAMP-sn gene wheat plant and its disease resistance analysis

[0050] 1. Acquisition of transgenic AcAMP-sn gene wheat plants

[0051]1. The young embryo callus of 1600 pieces of Yangmai 18 was used as the acceptor bombarded by the gene gun, and the recombinant plasmid pAHC25::AcAMP-sn of Example 1 was bombarded into the callus with the gene gun, and the specific method was as follows:

[0052] 1) The immature embryo callus of Yangmai 18 was osmotically treated on a high-osmotic medium (SD2 medium supplemented with 0.2mol / L mannitol and 0.2mol / L sorbitol) for 4-6h (25°C, dark culture) : the calli are placed within the diameter of about 2.5cm in the center of the petri dish, and 50 calli are placed in each petri dish. Among them, the formula of SD2 medium is MS medium + 1mg / L VB 1 +150mg / L asparagine +2mg / L2,4-D.

[0053] 2) The pAHC25::AcAMP-sn plasmid was used to wrap gold powder with a diameter of 1 μm, and the PDS-1000 / ...

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Abstract

The invention discloses a method for cultivation of transgenic wheat resisting take-all and sharp eyespot and related biological materials thereof. The method for cultivation of transgenic wheat of disease resistance includes: importing an related gene which has a code sequence composed of 7th-405th nucleotides in the sequence 1 of a sequence table into recipient wheat so as to obtain the transgenic wheat of disease resistance with disease resistance higher than that of the recipient wheat. The transgenic wheat of disease resistance is the wheat of a1 or a2: a1. transgenic wheat resisting take-all and sharp eyespot, with the disease resistance being resistance to take-all and sharp eyespot; and a2. anti-take-all transgenic wheat, with the disease resistance being resistance to take-all. Experiments prove that, compared with Yangmai 18, the anti-take-all effect of the transgenic wheat with the imported anti-take-all related gene is improved by 60%-80%, and the anti-sharp eyespot effect is improved by 20%-33%.

Description

technical field [0001] The invention relates to a method for cultivating transgenic wheat resistant to take-all and sheath blight and related biological materials. Background technique [0002] In recent years, the occurrence of soil-borne fungal diseases in my country's wheat production areas has been increasing. Wheat take-all (take-all), also known as "blackfoot" and "blight", is a worldwide soil-borne wheat disease caused by the fungus Gaeumannomyces graminis var. tritici (Ggt). Diseases (Gutteridge R J, Bateman G L, Todd A D. Variation in the effects of take‐all disease on grain yield and quality of winter cereals in field experiments. Pest management science, 2003, 59(2): 215-224). Ggt infects the root and stem base of wheat in the form of mycelium, causing most of the root and stem base of the diseased plant to turn black, the conduction tissue is blocked, the number of tillers is reduced, the earing rate is low, the thousand-grain weight is significantly reduced, th...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00C12N15/29C12N15/63C12N1/21C12N1/19C12N1/15C12N1/13C12N5/10
Inventor 张增艳杨坤刘欣杜丽璞徐惠君
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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