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

Wheat disease resistance proteins and coding genes and their application in regulating plant disease resistance

A protein and amino acid technology, applied in applications, plant peptides, genetic engineering, etc., can solve the problems of lack of easy utilization and slow progress of wheat resistant to sheath blight

Active Publication Date: 2019-03-26
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, conventional breeding methods have been slow in breeding sheath blight-resistant wheat varieties due to the lack of readily available germplasm resources for wheat 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
  • Wheat disease resistance proteins and coding genes and their application in regulating plant disease resistance
  • Wheat disease resistance proteins and coding genes and their application in regulating plant disease resistance
  • Wheat disease resistance proteins and coding genes and their application in regulating plant disease resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Embodiment 1, the cloning of wheat disease resistance protein TaZnF2 and its coding gene

[0094] The inventors of the present application isolated and cloned a wheat disease resistance protein from sheath blight resistant wheat CI12633, and named it TaZnF2. The specific cloning method of TaZnF2 gene is as follows:

[0095]The total RNA of wheat CI12633 leaf sheath was extracted, and the extracted RNA sample was reverse-transcribed to synthesize the first-strand cDNA according to the procedure of Invitrogen’s first-strand cDNA synthesis kit, which was used as a template for gene cloning, using TaZnF2-3R1:5′-ATTCGGACGAGGAACCCC -3′, 3RACE-Outer Primer: 5′-TACCGTCGTTCCACTAGTGATTT-3′ (Takara), for the first round of PCR amplification. The amplification program was as follows: pre-denaturation at 94°C for 3 minutes; then 35 cycles at 94°C for 45 seconds, 52°C for 45 seconds, and 72°C for 1 minute; extension at 72°C for 10 minutes. The first round of PCR amplification produ...

Embodiment 2

[0105] Example 2, the acquisition of sheath blight resistant transgenic wheat and the identification of disease resistance

[0106] 1. Construction of overexpressed transgene vector

[0107] The complete ORF sequence of the TaZnF2 gene was constructed on the monocotyledonous plant expression vector pAHC25-cMYC, and the construction process was as follows: figure 2 As shown, the specific steps are as follows:

[0108] 1. Preparation of linearized plasmid: Digest the plant expression vector pAHC25-cMYC with Spe I and EcoICR I (is isolytic enzyme with SacI, but produce blunt ends after digestion with EcoICR I), perform 1% agarose gel electrophoresis, agar Glycogel DNA Purification and Recovery Kit to recover the linearized pAHC25-cMYC vector backbone.

[0109] 2. Acquisition of the target gene TaZnF2 containing restriction sites: Design a pair of primers TaZnF2-OT-F: 5′-TAA according to the ORF sequence of the TaZnF2 gene ACTAGT ATGTCGCCGGCGGAGATGGA-3' (the underlined sequen...

Embodiment 3

[0157] Example 3, Breeding Gene-Silenced Wheat with Reduced Sheath Blight Resistance

[0158] 1. The two ends of the DNA fragment shown in the 756th-955th nucleotides of sequence 1 in the sequence listing are respectively provided with the restriction endonuclease NheI recognition sequence. After NheI digestion, the DNA fragment (200bp) shown in the 756-955th nucleotide of sequence 1 in the sequence listing is inserted into the BSMV-γ (the γ vector of BSMV virus) after the NheI enzyme linearization by the reverse insertion method ), the DNA molecule (named antiTaZnF2) that is reverse complementary to the DNA fragment shown in the 756-955 nucleotides of sequence 1 in the sequence listing is driven by the T7 promoter of the gamma vector to obtain the recombinant vector BSMV-γ : anti TaZnF2.

[0159] 2. At the two-leaf one-heart stage, use the recombinant vector BSMV-γ: anti TaZnF2 to transfect the second leaf of the wheat sheath blight-resistant material——wheat CI12633. The spe...

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 disease resistance protein, an encoding gene and an application of the wheat disease resistance protein and the encoding gene in regulation of plant disease resistance. The wheat disease resistance protein TaZnF2 is a protein in A1), A2) or A3) as follows: A1) a protein with the amino acid sequence shown as sequence 2, A2) a protein related with the plant disease resistance, wherein the amino acid sequence shown in the sequence 2 in a sequence table is subjected to substitution and / or deletion and / or addition of one or more amino acid residues for acquisition of the protein, and A3) a fusion protein obtained by connecting labels to an N end or / and a C end of A1) or A2). Experiments prove that the resistance, to wheat sharp eyespot, of wheat genetically modified with TaZnF2 and having TaZnF2 gene overexpression is improved remarkably; while TaZnF2 gene expression in disease resistance wheat CI12633 is inhibited, resistance of plants to the wheat sharp eyespot is reduced, and the result that the TaZnF2 gene is a necessary disease resistance gene for the wheat sharp eyespot resistant reaction is proved.

Description

technical field [0001] The invention relates to wheat disease resistance protein and coding gene in the field of biotechnology and their application in regulating plant disease resistance. Background technique [0002] Wheat is one of the most important food crops in the world and plays a pivotal role in ensuring food security. With the improvement of fertilizer and water conditions, the increase of planting density and the change of farming system, wheat sheath blight has developed into a major disease of wheat production in my country, and has become an important factor limiting the high and stable yield of wheat in my country. Wheat sheath blight, also known as wheat sharp eyespot, is caused by the saprophytic trophic fungi Rhizoctonia cerealis CAG-1 and Rhizoctonia solani AG4, A worldwide soil-borne fungal disease of wheat caused by the AG5 fusion group. The main pathogen of wheat sheath blight in my country is Rhizoctonia cerealis. Sheath blight can generally reduce ...

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): C07K14/415C07K19/00C12N15/29C12N15/62C12N5/10C12N15/82A01H6/46A01N65/44A01P3/00
CPCA01N65/44C07K14/415C07K2319/20C07K2319/21C07K2319/22C07K2319/41C07K2319/43C12N15/8216C12N15/8282C12N2800/60
Inventor 张增艳刘鑫祝秀亮魏学宁杜丽璞徐慧君
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