Triticum aestivum PITP TaSec14 gene and application thereof

A technology of phosphatidylinositol and transporter, which is applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve undiscovered problems and achieve the effect of improving resistance and improving resistance breeding

Inactive Publication Date: 2020-05-12
TIANJIN NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, no information has been found about Sec14 A report on the ...

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  • Triticum aestivum PITP TaSec14 gene and application thereof
  • Triticum aestivum PITP TaSec14 gene and application thereof
  • Triticum aestivum PITP TaSec14 gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] TaSec14 Gene sequence acquisition

[0039] 1 Experimental materials and methods

[0040] 1.1 Plant material and strains

[0041] Cultivated wheat Jing 411 is a semi-dwarf wheat variety with strong cold resistance, strong tillering ability, high spike rate, etc. The phenotype is hypersensitive. Brock is a wheat variety with strong resistance to powdery mildew, which was introduced from the UK as a disease-resistant genetic resource. Jing 411×Brock powdery mildew resistant near-isogenic line BJ-1, the material is the recurrent parent of the susceptible wheat Jing 411 and the resistant wheat Brock as the disease-resistant gene donor, and obtained F 1 After that, continuous backcrossing with the susceptible variety Maijing 411 was carried out for 6 generations. In each generation, disease-resistant plants were selected under the stress of powdery mildew, and finally obtained after one generation of self-crossing (for the preparation method, please refer to Zhang Rong et...

Embodiment 2

[0065] TaSec14 Analysis of Gene Expression Patterns in Different Resistance Wheat

[0066] 1 Experimental materials and methods

[0067] 1.1 Plant material and strain (same as Example 1)

[0068] 1.2 TaSec14 Gene Expression in Different Resistance Wheat

[0069] When the seedlings of Jing 411, BJ-1 and Brock reached the stage of one leaf and one heart, fresh powdery mildew spores were evenly inoculated on the surface of the first wheat leaf by shaking Buddha method. The first leaves of different varieties of wheat were collected at different time points (0, 2, 4, 8, 12, 24 and 48 h) respectively, and 3 cm of the leaf tip was cut off, and placed in a sterilized and Store in a 2 mL EP tube treated with liquid nitrogen and store in a -80 °C ultra-low temperature freezer. Obtain wheat cDNA according to the method in embodiment 1, and design primer SecQ-F / R according to cDNA non-conserved region sequence, take wheat Actin The gene was used as an internal reference, and the r...

Embodiment 3

[0073] Evaluation using VIGS technology TaSec14 Gene Responses to Wheat and Powdery Mildew Stress

[0074] 1 Materials and methods

[0075] 1.1 Experimental materials

[0076] Plant material: Wheat cultivar Jing 411

[0077] Virus: Barley stripe mosaic virus (BMV) used in the construction of VIGS vector is a positive-strand RNA virus with α, β and γ genomes.

[0078] 1.2 TaSec14 VIGS analysis of genes

[0079] according to TaSec14 The specific silencing primer SecV-F / R was designed for the sequence of the non-conserved region of the gene, and the silencing fragment was amplified using the cDNA of the susceptible wheat Jing 411 as a template. Use 1% agarose gel electrophoresis to analyze the PCR product, recover the silent fragment and combine it with BSMV γ : Vector backbone construction of recombinant vector BSMV γ : TaSec14 .

[0080] Extract the plasmid BSMV containing the viral vector α 、BSMV β 、BSMV γ:GFP 、BSMV γ:PDS and BSMV γ:TaSec14 , after linearization...

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Abstract

The invention discloses a cDNA sequence and an amino acid sequence of a PITP (Phosphatidylinositol Transfer Protein) gene TaSec14 cloned in powdery mildew infected triticum aestivum variety of Jing 411. The cDNA full length of the gene is 1546 bp; the DNA full length is 2186 bp; an open reading frame is 1212 bp; 403 amino acids are coded; 80.45-percent homology is realized with PITP3(XP_010236107.1) in Bd (Brachypodium Distachyon); SEC14 conserved domains are contained; and the gene is named as TaSec14. After inoculation with the powdery mildew, the TaSec14 has the consistent expression trendin Jing 411 and a near-isogenic line BJ-1 thereof, and the expression amount is respectively higher than that of a disease-resistant triticum aestivum variety of Brock. A virus induced gene silencingtechnology is used for knocking down the Tasec14 gene in Jing 411; through the expression reduction of the TaSec14 gene, the resistance of the diseased triticum aestivum Jing 411 on the powdery mildewcan be improved to a certain degree; and the TaSec14 gene has the regulation and control effects in the interaction process of the triticum aestivum with the powdery mildew. The invention provides anovel genetic resource for the triticum aestivum powdery mildew resistance mechanisms and disease resistance breeding.

Description

[0001] National Natural Science Foundation of China (No. 31071671), Tianjin Municipal Science and Technology Commission Key Project (No. 17JCZDJC34100) Science and Technology Support Project (No. 18YFZCNC01100). technical field [0002] The invention belongs to the technical field of agricultural biological genetic engineering, in particular to the phosphatidylinositol transporter gene in cultivated wheat Jing 411 TaSec14 and its application in wheat disease resistance breeding. Background technique [0003] Wheat (Triticum aestivum L.) is one of the most widely planted gramineous grain crops in the world, but the occurrence of various diseases and insect pests has seriously affected the yield of wheat, among which the wheat caused by Blumeria graminis f. sp.tritici Powdery mildew is one of the most serious diseases (Gao Hongyun et al., 2015; Morgounov et al., 2012). [0004] Sec14p is a phosphatidylinositol transfer protein (PITPs) widely present in eukaryotes (Mao Huaying...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46
CPCC07K14/415C12N15/8218C12N15/8282
Inventor 王振英刘晓颖范宝莉尚云涛张双力
Owner TIANJIN NORMAL UNIVERSITY
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