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

Gene 4-CL for Chinese white poplar lignose monomer synthesis and application thereof

A technology for synthesizing genes and poplar tomentosa, applied in the field of molecular biology

Inactive Publication Date: 2008-10-08
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
View PDF0 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although there are many genes or cDNAs encoding 4-CL proteins isolated from various plants, before the application of the present invention, no one in China has disclosed or published the 4-CL gene sequence of Populus tomentosa lignin monomer synthesis gene and its Uses in Brassica napus

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
  • Gene 4-CL for Chinese white poplar lignose monomer synthesis and application thereof
  • Gene 4-CL for Chinese white poplar lignose monomer synthesis and application thereof
  • Gene 4-CL for Chinese white poplar lignose monomer synthesis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 (cloning and sequencing of genes)

[0036] Retrieve the published poplar 4-CL gene sequence in the NCBI database, and use the Oligo6.0 software (purchased through commercial channels, the same below) to design primers on both sides of the gene coding sequence (forward primer: [5'-ATGAATCCACAAGAAGAATTCATC -3']; reverse primer: [5'-TTATATGCCTGCCAACTTTTCTTTCAG-3']), used to amplify the corresponding sequence of 4-CL from Populus tomentosa.

[0037] 1. Extract Populus tomentosa mRNA.

[0038] Extraction of RNA (TRIZOL TM Kit for RNA extraction)

[0039] Liquid nitrogen was used to triturate 100 mg of material.

[0040] A. Add 1ml TRIZOL and place at room temperature (20-25°C, the same below) for 5min.

[0041] B. Add 200ul chloroform, shake vigorously for 30s, and place at room temperature for 2min.

[0042] C.12000g, 15min, 4 degrees Celsius. Take the supernatant to a new tube, add 500ul of isopropanol, mix well and place at room temperature for 15min.

...

Embodiment 2

[0048] Embodiment 2 (vector construction)

[0049] Use PCR reaction to introduce HindIII and BamHI restriction site sequences at the 5' end and 3' end of the C4H promoter, respectively, and then use HindIII / BamHI to double-enzyme digest the C4H promoter PCR product and PBI121 plasmid respectively, and digest the PCR fragment and plasmid After enzyme digestion and ligation of large fragments, transform into competent cells DH5α (obtained from commercial sources) for amplification, and then introduce the 4-CL fragment into the vector (PBI121) and replace the original GUS sequence in the vector (PBI121), The 35S promoter sequence in the vector is replaced by the C4H promoter sequence (such as figure 2 shown), and its insert was identified by DNA sequencing.

Embodiment 3

[0050] Embodiment 3 (transformation and screening of plant)

[0051] Transformation of Rapeseed with Agrobacterium LBA4404

[0052] A. Preparation of sterile seedlings: Seeds are treated with 70% ethanol for 1min, mercuric chloride (HgCl 2 ) Soak for 13-15min, ddH 2 After O washing 5 times, spread in MS medium (PH 5.8), agar concentration 0.8%. Rapeseed aseptic seedlings for use.

[0053] B. Rapeseed petiole transformation: Inoculate Agrobacterium tumefaciens LBA4404 (containing 4-CL gene) on solid medium LB, pick a single colony in 50ml YEP liquid medium two days later (tryptone 10g+yeast extract 10g+NaCl5g+ MgSO 4 0.5g). Take 4-5 day old sterile seedling petioles, soak in the bacterial solution for 5-8 minutes, shake gently during this period, then pour out the bacterial solution, and absorb the residual bacterial solution on the explants with sterile filter paper. Placed in co-culture medium (MS+0.2mg / L 6-benzyl adenine (6-BA)+1mg / L 2,4-dichlorophenoxyacetic acid (2,...

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 Chinese white poplar lignin monomer synthesis gene 4-CL and an application thereof. The gene is responsible to catalyzing each cinnamic acid and each corresponding coenzyme A ester generated by derivative thereof, the synthesis of each downstream secondary product is participated, and the gene is the essential gene of controlling the lignin synthesis. The findings indicates that the infection of the plant resistance pathogen is reinforced through the increase of the lignin content, the rape stem strength is enhanced, and thus the lodging resistance is obviously enhanced. The concrete result shows that the lignin content of the transgenic rape increases over 10 percents than the acceptor compared with the lignin content; the transgenic plant leaf reduces about 30 percents than the acceptor compared with the disease spots expanding area; and the cane strength of the transgenic plant comparably increases about 20 percents.

Description

technical field [0001] The invention belongs to the field of molecular biology. Specifically, the invention relates to a 4-CL sequence of the monomeric lignin synthesis gene of Populus tomentosa, and at the same time, the invention also relates to the 4-CL synthesis gene of the monomeric lignin in Brassica napus use in . Background technique [0002] As a complex phenylpropanoid polymer in plants, lignin is one of the main components of plant mechanical tissues and cell walls, and plays an important role in plant growth and stress resistance. During the lignification process of the cell wall, lignin penetrates into the cell wall and fills the cell wall structure, which enhances the hardness of the cell wall and strengthens the mechanical support and compressive strength of the cell. In the interaction with pathogenic bacteria, the lignification of the host cell wall is one of the characteristics of the disease resistance response. Lignin, as an important physical antibacter...

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
IPC IPC(8): C12N15/52C12N15/82A01H1/00A01H5/00
CPCC12N15/8255C12N15/8282
Inventor 王汉中杨向东刘贵华王新发华玮刘静杨庆郑元本
Owner INST OF OIL CROPS RES 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