Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Breeding method of disease-resistant transgenic tam2k wheat and related biological materials

A disease resistance, tam2k technology, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve problems such as slow progress and lack of

Active Publication Date: 2021-06-29
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, progress in conventional breeding methods for wheat breeding for sheath blight resistance has been slow due to the lack of readily available wheat germplasm resources for sheath blight resistance

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
  • Breeding method of disease-resistant transgenic tam2k wheat and related biological materials
  • Breeding method of disease-resistant transgenic tam2k wheat and related biological materials
  • Breeding method of disease-resistant transgenic tam2k wheat and related biological materials

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0073] The preparation method of bacterial wheat kernels: soak the wheat kernels for 5-6 hours, boil them for 20 minutes, fill up the triangular flask, inoculate the mycelia block of the pathogenic bacteria of wheat sheath blight into the triangular flask after sterilization, and cultivate at a constant temperature of 25°C until The mycelia are densely covered with grains.

[0074] The preparation method of bacterial toothpick: put the toothpick segment upright and fill it with a small beaker, pour liquid MS medium into it, inoculate the mycelia block of wheat sheath blight pathogenic bacteria into the small beaker after sterilization, and cultivate it at a constant temperature of 25°C until the mycelia Densely strewn with toothpicks.

Embodiment 1

[0075] Embodiment 1, sheath blight resistant wheat kinase TaM2K and cloning of its coding gene

[0076] 1. Cloning of TaM2K gene

[0077] The inventors of the present invention isolated and cloned a wheat disease resistance-related protein from sheath blight resistant wheat CI12633, as shown in sequence 1 of the sequence listing, and named it TaM2K protein, which belongs to wheat kinase. The gene encoding TaMK protein is named TaM2K gene, and its cDNA is shown in sequence 2 of the sequence listing. In sequence 2 of the sequence listing, nucleotides 39-1145 are open reading frames.

[0078] Specific cloning steps: extract the total RNA from the stem of wheat CI12633 inoculated with the pathogenic bacteria of wheat sheath blight, and reverse-transcribe the RNA to synthesize the first-strand cDNA according to the procedure of the first-strand cDNA synthesis kit of Tiangen Biochemical Company, as The template for gene cloning was amplified with primers TaM2K-OF: 5'-GGAGCCAGCCTCG...

Embodiment 2

[0093] Embodiment 2, acquisition and disease resistance identification of transgenic TaM2K wheat resistant to sheath blight

[0094] 1. Construction of recombinant expression vector

[0095] 1. Extract the total RNA from the stem of wheat CI 12633 and reverse transcribe to obtain cDNA.

[0096] 2. Using the cDNA obtained in step 1 as a template, using a primer pair composed of TaM2K-O-F1 and TaM2K-O-R1, carry out PCR amplification under the action of the high-fidelity amplification enzyme PrimeSTAR, and recover the PCR amplification product.

[0097] TaM2K-O-F1: 5'-cG GGATCC ATGCGTCCGGGCGGGCCG-3';

[0098] TaM2K-O-R1: 5'-c GAGCTC gtggtggtggtggtggtgGCACGGCGGGGCGGCGAGGG-3'.

[0099] PCR reaction program: 94°C pre-denaturation for 5 minutes; 35 cycles of 98°C for 10s, 60.8°C for 30s, 68°C for 1.5min; 68°C for 10min.

[0100] 3. Take the PCR amplification product obtained in step 2, perform double digestion with restriction endonucleases BamHI and SacI, and recover the dige...

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 method for cultivating disease-resistant transgenic TaM2K gene wheat and related biological materials. The present invention first protects an important kinase derived from wheat sheath blight resistance, named TaM2K protein, which is a protein composed of the amino acid sequence shown in sequence 1 or sequence 3 in the sequence listing. Nucleic acid molecules encoding TaM2K protein also belong to the protection scope of the present invention. The invention also protects a method for cultivating transgenic plants, which includes the following steps: introducing TaM2K gene into target plants to obtain transgenic plants with higher disease resistance than the target plants. The method for cultivating transgenic plants with improved disease resistance of the present invention has important theoretical and practical significance, and will play an important role in the genetic improvement of plants.

Description

technical field [0001] The invention relates to a method for cultivating disease-resistant transgenic TaM2K gene wheat and related biological materials. Background technique [0002] More than 40% of the world's population uses wheat as a staple food crop, so wheat plays a pivotal role in ensuring food security. With the increase of planting density, changes in fertilizer and water conditions and farming systems, wheat sheath blight has developed into the largest soil-borne disease in my country's wheat production, and has become an important limiting factor for high and stable wheat yields in my country. [0003] Wheat sheath blight, also known as wheat sharp eyespot. Wheat sheath blight in my country is mainly caused by the saprotrophic fungus Rhizoctonia cerealis. Sheath blight can generally reduce wheat production by 10% to 30%, and severe plots can reduce wheat production by more than 50%. According to the report of the National Agricultural Technology Promotion Stat...

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): C12N9/12C12N15/54C12N15/82A01H5/00A01H6/46
CPCC12N9/12C12N15/8282
Inventor 张增艳王开
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com