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

Agrobacterium tumefaciens mediated transformation system for efficiently obtaining transgenic plants of paspalum vaginatum and application thereof

An Agrobacterium-mediated, seaside paspalum technology, applied in application, genetic engineering, plant peptides, etc., can solve problems such as difficulties in genetic transformation of warm-season turfgrass

Inactive Publication Date: 2016-08-24
NANJING AGRICULTURAL UNIVERSITY
View PDF1 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention is an Agrobacterium-mediated transformation system for efficiently obtaining coastal paspalum transgenic plants and its application. Infection time, infection conditions, co-cultivation conditions and other factors, to provide a genetic transformation system suitable for Agrobacterium-mediated seashore paspalum

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
  • Agrobacterium tumefaciens mediated transformation system for efficiently obtaining transgenic plants of paspalum vaginatum and application thereof
  • Agrobacterium tumefaciens mediated transformation system for efficiently obtaining transgenic plants of paspalum vaginatum and application thereof
  • Agrobacterium tumefaciens mediated transformation system for efficiently obtaining transgenic plants of paspalum vaginatum and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Agrobacterium-mediated transfer of the PpCBF gene into seashore paspalum to obtain cold-resistant transgenic plants

[0027] 1) Acquisition of recipient material: callus was induced with mature embryos of Paspalum ‘Seaspray’ as explants, and the germination medium was MS+2, 4-D 2.0 mg L -1 +AHC 0.5g L -1 ; Callus induction medium is MS+2,4-D 3.0mg L -1 +CuSO 4 10.0 mg L -1 ; Embryogenic callus induction medium is MS+2,4-D 3.0mg L -1 +CuSO 4 15.0mgL -1 +AHC 1.0g L -1 ; Subculture medium is MS+2,4-D 3.0mg L -1 +KT 0.1mg L -1 ; Culture in dark at 30°C, subculture once every 30 days, and select 2-3 months old embryogenic callus to be infected.

[0028] 2) Agrobacterium engineering strain cultivation: the Agrobacterium EHA105 strain containing the PpCBF gene expression vector was plated (containing 100 μg L -1 Kanamycin + 50 μg L -1rifampicin), cultured in the dark at 28°C for 3 days, picked a single colony from the plate, and inoculated it into LB liqu...

example 2

[0036] Example 2: Agrobacterium-mediated method to transfer PvGF14 gene overexpression vector and RNAi expression vector into seashore paspalum to obtain transgenic plants

[0037] In the early stage, the team obtained a 14-3-3 family protein member PvGF14 with salt tolerance function from the seaside paspalum expression library through the yeast screening system. Using this technical system, the seaside finches with overexpression of PvGF14 gene and RNAi inhibition expression vector were completed. The genetic transformation of barnyardgrass and the acquisition of transgenic lines have laid a solid foundation for further analysis of the downstream regulation mechanism of seashore paspalum PvGF14 salt tolerance and the analysis of salt tolerance functions. The specific implementation process is as follows:

[0038] 1) Receptor material preparation: use the mature seeds of seashore paspalum ‘Seaspray’ as explants to induce callus, and the germination medium is MS+2, 4-D 2.0mg L...

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 relates to an agrobacterium tumefaciens mediated transformation system for efficiently obtaining transgenic plants of paspalum vaginatum and application thereof. The agrobacterium tumefaciens mediated transformation system adopts the following steps of: by an efficient regeneration system, obtaining an embryonic callus receptor material; culturing donor agrobacterium tumefaciens strains; determining ablastin cephalosporin concentration; optimizing agrobacterium tumefaciens transformation conditions; carrying out screening culture and plant generation of resistant calli and carrying out molecular detection of the transgenic plants. By optimizing influence factors of bacteria concentration, infection time, infection conditions, co-culture conditions and the like in the agrobacterium tumefaciens transformation process, the paspalum vaginatum agrobacterium tumefaciens mediated transformation system is established. The invention overcomes the defect that an existing agrobacterium tumefaciens mediated technology is difficult to apply to genetic transformation of warm-season turfgrass, and establishes the efficient paspalum vaginatum agrobacterium tumefaciens transformation system which uses embryonic calluses as the receptor material.

Description

Technical field: [0001] The invention belongs to the field of plant genetic engineering or crop bioengineering breeding. It involves the tissue culture technology of seashore paspalum, the technology of Agrobacterium-mediated transformation and the molecular detection technology of transformed plants, and establishes an efficient method of obtaining seashore paspalum transgenic plants. Background technique: [0002] With the development of the economy, lawns, as an important part of urban landscaping, have become one of the important symbols to measure the environmental quality and civilization of modern cities. Traditional breeding methods such as hybrid breeding, mutation breeding and directional selection have made important contributions to turfgrass breeding, but the required period is long and the available genetic materials are limited, which is difficult to meet the requirements of modern turfgrass breeding. Require. With the development of modern biotechnology, tr...

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): C12N15/82A01H5/00A01H4/00
CPCC12N15/8205A01H4/008C07K14/415C12N15/8273
Inventor 刘君杨志民陈煜庄黎丽于景金
Owner NANJING 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