Unlock instant, AI-driven research and patent intelligence for your innovation.

Wheat powdery mildew resistance related protein taeds1-d1 and its coding gene and application

A technology for powdery mildew and wheat, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as loss of resistance of production varieties

Inactive Publication Date: 2019-07-26
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Emergence of new races often leads to loss of resistance in mass-produced varieties

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Wheat powdery mildew resistance related protein taeds1-d1 and its coding gene and application
  • Wheat powdery mildew resistance related protein taeds1-d1 and its coding gene and application
  • Wheat powdery mildew resistance related protein taeds1-d1 and its coding gene and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Example 1. Cloning and sequence structure analysis of wheat TaEDS1-D1 gene

[0090] 1. Acquisition of the cDNA and genomic DNA sequences of the coding region of the wheat TaEDS1-D1 gene

[0091] 1. Extraction of wheat genomic DNA and total RNA

[0092] (1) Extraction of wheat genomic DNA

[0093] Wheat genomic DNA was extracted according to the conventional CTAB method. The specific steps are as follows: take about 0.2 g of fresh leaves of 7-day-old Piedea chinensis seedlings, grind them into fine powder in liquid nitrogen, and transfer them to a sterilized 2 ml centrifuge tube. Add 1.0ml CTAB extraction buffer (1.3%CTAB, 133mM Tris / HCl pH8.0, 13mM EDTA, 0.93M NaCl, 0.66%PVP3600, 0.18Mβ-mercaptoethanol), and mix well. Incubate at 65°C for 1.0 h, during which the samples are mixed from time to time to fully extract. After the sample was cooled to room temperature, 1ml of chloroform / isoamyl alcohol (v / v=24:1) was added, and mixed by inversion for 1h. Centrifuge at 12...

Embodiment 2

[0111] Example 2. Using VIGS technology to verify the powdery mildew resistance function of TaEDS1-D1 gene

[0112]1. Obtaining of TaEDS1-D1 Gene Silenced Plants

[0113] 1. Construction of BSMV-VIGS vector system for induction of TaEDS1-D1 gene

[0114] (1) Acquisition of Silent Sequence

[0115] 1) Obtaining the silence sequence TaEDS1-S1

[0116] Using the recombinant plasmid pGEM-T-TaEDS1-D1 prepared in Example 1 as a template, PCR amplification was performed on TaEDS1-S1F and TaEDS1-S1R using primers to obtain a PCR amplification product of 292bp (corresponding to sequences 1 to 5 'Terminal 1142-1433 nucleotide sequence), it is named as the silent sequence TaEDS1-S1.

[0117] The nucleotide sequences of the above-mentioned primers to TaEDS1-S1-F and TaEDS1-S1-R are as follows (the sequence shown in the underline is the restriction endonuclease NheI enzyme cutting recognition site):

[0118] TaEDS1-S1-F:5'-TAC GCTAGC GTGGCCTTGAACTGAG-3';

[0119] TaEDS1-S1-R:5'-TAC ...

Embodiment 3

[0160] Example 3. Verification of the function of TaEDS1-D1 using the transient expression technology of wheat leaf epidermal cells

[0161] The function of wheat powdery mildew resistance is an autonomous behavior of single cells, and powdery mildew only infects the epidermal cells of the host leaves. Therefore, transient expression in epidermal cells is an effective technique to verify the function of wheat powdery mildew resistance genes. The target gene was randomly introduced into the epidermal cells of isolated wheat leaves through a gene gun, and a reporter gene was co-transformed at the same time to mark the positively transformed cells. Effect of disease-associated gene expression on powdery mildew infection. The specific implementation steps are as follows:

[0162] 1. Construction of transient expression vectors in epidermal cells of wheat leaves

[0163] 1) Using the pGEM-T-TaEDS1-D1 plasmid prepared in Example 1 as a template, using TaEDS1-D166-NcoI-f3 and TaED...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a wheat powdery-mildew-resistant related protein TaEDS1-D1 and an encoding gene and application thereof. An EDS1 gene is cloned form a wheat variety Shangeda and is named as TaEDS1-D1. Experiments prove that 1, after the expression quantity of the TaEDS1-D1 gene in the powdery-mildew-resistant wheat Shangeda is reduced by utilizing a BSMV-VIGS method, then sphaerotheca fuliginea is inoculated, a disease-resistant plant is changed into a diseased plant, and it is indicated that the resistance of the plant is lost; 2, after the TaEDS1-D1 is instantaneously expressed in leaves of the wheat variety Chancellor with powdery mildew, then sphaerotheca fuliginea is inoculated, the haustorium index of the plant is reduced, and it is indicated that the resistance of the plant is improved; 3, the TaEDS1-D1 gene overexpressed in arabidopsis thaliana eds1 can be partially and mutually complemented with the function of an arabidopsis thaliana EDS1 gene to change disease infection into disease resistance, and it is indicated that the TaEDS1-D1 gene and encoded protein TaEDS1-D1 have a powdery mildew resistant function.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a wheat powdery mildew resistance-related protein TaEDS1-D1 and its coding gene and application. Background technique [0002] Wheat powdery mildew is an important fungal disease caused by Blumeria graminis f.sp. Tritici (Bgt). Wheat powdery mildew can infect all above-ground organs of wheat plants, but it mainly damages leaves, and in severe cases, it can infect leaf sheaths, stems and ears. Powdery mildew mainly affects leaf photosynthesis and plant metabolism. When the onset is earlier and heavier, the plants will not head or the ears taken out will be short, reducing the number of ears per mu, the number of grains per ear and the thousand-grain weight. Generally, the yield can be reduced by 5%-10%. In severe epidemic years, the yield reduction can reach more than 30% (Johnson, J.W., Baenziger, P.S., Yamazaki, W.T., Smith, R.T. Effects of powdery mildew on yield and ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C07K19/00C12N15/29A01H5/00A01H6/46
CPCC07K14/415C07K2319/20C07K2319/21C07K2319/41C07K2319/43C12N15/8203C12N15/8282
Inventor 张相岐陈桂平范仁春卫波
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI