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

Method for increasing amount of plant root nodules on the basis of gene negative regulation and control

A plant root system, plant technology, applied in the field of genetic engineering, can solve unseen problems

Active Publication Date: 2019-12-31
HUAZHONG AGRI UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the role of RR in regulating soybean nodulation has not yet been seen.
Whether the type B response factor RR11 plays a regulatory role in soybean root nodulation and symbiotic nitrogen fixation has not been reported

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
  • Method for increasing amount of plant root nodules on the basis of gene negative regulation and control
  • Method for increasing amount of plant root nodules on the basis of gene negative regulation and control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1. Analysis of the tissue expression pattern of the target gene and its expression pattern in response to rhizobia treatment.

[0023] 1) Material acquisition: The material used in the experiment was Williams 82 (W82); the material was carried out according to the following process: soybean seeds were sterilized with 70% alcohol for 30 sec, sown in vermiculite matrix soaked in low-nitrogen nutrient solution, They were cultivated in a culture room with 16 h light / 8 h dark, light intensity of 7000 LUX, temperature of 26 °C, and relative humidity of 70%. Ten days after sowing, USDA110 with OD600=0.08 was used for inoculation treatment, and samples were taken at corresponding time points to detect gene expression. For the expression patterns in roots, leaves and tumors, samples were taken at inoculation 28DAI.

[0024] 2) Isolation of mRNA: Trizol method was used to extract soybean total RNA. ①Put the tissue into a grinder and grind it with liquid nitrogen for 3 tim...

Embodiment 2

[0030] Example 2. Using an overexpression vector to up-regulate the expression of a target gene to change the number of nodules.

[0031] 1) Extraction of total RNA from root tissue of Wilimas82 soybean

[0032] The mortar has been treated at 180 ℃ for 8 hours or burned to eliminate RNase contamination; reagents such as chloroform, isopropanol, and ethanol should be freshly opened and uncontaminated; other equipment such as pipette tips, centrifuge tubes and reagents such as ultrapure water , NaAc, were treated with 1 ‰ DEPC water overnight, and then sterilized at 121 ℃ for 30 minutes, and the pipette tip and centrifuge tube were dried at 65 ℃ for later use; Trizol method was used to extract soybean total RNA:

[0033] (1) Take 100 mg of soybean root material and grind it with liquid nitrogen, add 1 mL of TRI pure reagent, after fully homogenizing, inhale the homogenate into a 1.5 mL centrifuge tube, and place it at room temperature for 5 min;

[0034] (2) Add 200 μl chlorofo...

Embodiment 3

[0064] Example 3. Using a knockout vector to down-regulate the expression of a target gene to change the number of nodules.

[0065] 1) Extraction of total RNA from root tissue of Wilimas82 soybean

[0066] The mortar has been treated at 180 ℃ for 8 hours or burned to eliminate RNase contamination; reagents such as chloroform, isopropanol, and ethanol should be freshly opened and uncontaminated; other equipment such as pipette tips, centrifuge tubes and reagents such as ultrapure water , NaAc, were treated with 1 ‰ DEPC water overnight, and then sterilized at 121 ℃ for 30 minutes, and the pipette tip and centrifuge tube were dried at 65 ℃ for later use; Trizol method was used to extract soybean total RNA:

[0067] (1) Take 100 mg of soybean root material and grind it with liquid nitrogen, add 1 mL of TRI pure reagent, after fully homogenizing, inhale the homogenate into a 1.5 mL centrifuge tube, and place it at room temperature for 5 min;

[0068] (2) Add 200 μl chloroform, v...

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 increasing an amount of plant root nodules on the basis of gene negative regulation and control. The method comprises the following steps of: firstly, extracting atotal RNA (Ribonucleic Acid) in a plant tissue; through reverse transcription PCR (Polymerase Chain Reaction), synthesizing a PCR template; constructing a recombinant expression vector used for knocking out a corresponding gene as a knockout carrier; and through an agrobacterium tumefaciens-mediated hairy root transformation method, transforming a receptor plant so as to down-regulate the expression of the corresponding gene in the receptor plant and increase the nodulation number of the root system of the receptor plant. The recombinant expression vector constructed by the method disclosed by the invention is used for transforming the receptor soybean plant to obtain a transgenosis plant, and the number of root nodules of a soybean root system can be obviously improved.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for cultivating plants with high nodulation ability. Background technique [0002] Soybean is an important grain and oil economic crop. Through root nodules, nitrogen in the air can be converted into ammonia nitrogen that can be absorbed and utilized by plants, thus providing the necessary nitrogen nutrition for soybean growth and development. The symbiotic nitrogen fixation process mediated by root nodules not only affects the normal growth and yield of soybean, but also helps to save energy and reduce environmental chemical pollution. At present, improving the efficiency of symbiotic nitrogen fixation has become one of the important ways to increase soybean yield and ensure the sustainable development of agriculture. Understanding the molecular mechanism of soybean nodulation and root nodule development at the molecular level is a more reasonable way to use...

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 Applications(China)
IPC IPC(8): C12N15/82C12N15/29A01H5/06A01H6/54
CPCC07K14/415C12N15/8218C12N15/8261
Inventor 李霞王利祥胡阳阳陈嘉欢
Owner HUAZHONG AGRI UNIV
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