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

Construction and application of an anti-sclerotinia gene gmgst1 and gmgst1 transgenic plant

A technology for sclerotinia and plants, applied in the field of plant genetic engineering, can solve the problems of scarcity of genetic resources, restriction of efficiency and accuracy of new variety breeding, complicated mechanism of soybean anti-sclerotinia, and achieve the effect of improving resistance

Active Publication Date: 2022-06-28
NORTHEAST AGRICULTURAL UNIVERSITY
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the limitations of traditional breeding methods, the use of molecular methods to screen and identify effective disease-resistant genes is particularly important for the breeding of disease-resistant varieties. efficiency and accuracy
[0003] In the study of soybean sclerotinia resistance genes, it is limited to the application of the oxalate oxidase gene Germin, oxalate decarboxylase and respiratory burst oxidase genes. There are no related reports on other types of genes. However, the mechanism of soybean anti-sclerotinia resistance is complex and there are a large number of loci and key genes to be discovered and utilized

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
  • Construction and application of an anti-sclerotinia gene gmgst1 and gmgst1 transgenic plant
  • Construction and application of an anti-sclerotinia gene gmgst1 and gmgst1 transgenic plant
  • Construction and application of an anti-sclerotinia gene gmgst1 and gmgst1 transgenic plant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1. Obtaining the antibacterial sclerotinia gene GmGST1.

[0031] 1) Using the soybean antibacterial sclerotinia cultivar Maple Arrow as the material, when the first group of three-leaf compound leaves grows, the material is taken and the total RNA is extracted, and the results are as follows figure 1 shown, and reverse transcribed to synthesize the first strand of cDNA.

[0032] 2) According to the GmGST1 gene sequence on Phytozome, utilize Primer 5 software to design gene cloning primers,

[0033] Primer 1-S: 5'-AGATCTATGCCGTTACCTTCTCAAAT-3', the sequence is as shown in SEQ ID NO.2;

[0034] Primer 1-a: 5'-GGTTACCTTAGATTTTGTTGAATGCAAC-3', the sequence is shown in SEQ ID NO.3.

[0035] Using cDNA as template PCR reaction, the reaction system is as follows:

[0036]

[0037] Reaction program: 94 °C for 5 min; 37 cycles: 94 °C for 30 s, 54 °C for 30 s, 72 °C for 40 s; 72 °C for 10 min, and 4 °C storage. After the reaction, the PCR products were taken for 1%...

Embodiment 2

[0039] Example 2. Construction of antibacterial sclerotinia transgenic soybean plants.

[0040] (1) construct overexpressing GmGST1 gene recombinant vector, the concrete steps are as follows:

[0041] The plasmid pCambia3301 and the pGM-GmGST1 prepared in Plasmid Example 1 were digested with BstEII and BglII, respectively. After detection by agarose gel electrophoresis, the pCambia3301 vector backbone and the GmGST1 gene digested fragment were recovered and purified, and the two fragments were connected and transformed. The picked single spot was identified by BstEII and BglII double enzyme digestion, and the recombinant vector pCambia3301-GmGST1 carrying the GmGST1 gene was obtained. The enzyme digestion identification results are as follows image 3 shown.

[0042] (2) The recombinant vector constructed in step (1) is introduced into Agrobacterium to obtain the recombinant bacteria with GmGST1 gene, and the specific steps are as follows:

[0043] The pCambia3301-GmGST1 exp...

Embodiment 3

[0052] Example 3. Construction of antibacterial sclerotinia transgenic soybean plants.

[0053] Example 2 was repeated, and the difference from Example 2 was that the seeds used in 2) in step (3) of this example were soybean variety Hefeng 25. In this example, a total of 16 T0 generation positive plants were obtained.

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

PropertyMeasurementUnit
concentrationaaaaaaaaaa
Login to View More

Abstract

The invention relates to the construction and application of an anti-sclerotinia gene GmGST1 and a GmGST1 gene-transferred plant, belonging to the technical field of plant genetic engineering. In order to solve the problem that currently available gene resources are scarce in the aspect of soybean sclerotinia resistance, the present invention provides an antibacterial sclerotinia gene GmGST1, the sequence of which is shown in SEQ ID No.1. On the basis of the obtained transgenic soybean plants, through subsequent phenotypic identification, it was determined that GmGST1 can participate in the response to soybean sclerotinia resistance, and can significantly improve the level of soluble pigment in the stem of the recipient soybean germplasm and its resistance to Sclerotinia sclerotiorum , to provide effective molecular markers and genetic resources for soybean anti-sclerotinia molecular design breeding.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, in particular to the construction and application of an antibacterial sclerotinia gene GmGST1 and a GmGST1 gene-transformed plant. Background technique [0002] Soybean sclerotinia is a fungal disease caused by Sclerotinia sclerotiorum (Lib.) de Bary. It is a serious worldwide disease that directly affects the quality and yield of soybeans. % to 90%, or even aborted. Due to the limitations of traditional breeding methods, it is particularly important to screen and identify effective disease-resistant genes by molecular means, and the selection of disease-resistant varieties is particularly important. Efficiency and accuracy. [0003] The research on soybean sclerotinia resistance genes is limited to the application of oxalate oxidase gene Germin, oxalate decarboxylase and respiratory burst oxidase genes. There are no related reports on other types of genes, and the mechanism o...

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): C12N15/54C12N15/11C12N15/84A01H5/00A01H6/54
CPCC12N9/1088C12N15/8282C12Y205/01018
Inventor 赵雪张倩宋伟姜海鹏韩英鹏李文滨
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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