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

Efficient genetic transformation method of agrobacterium-mediated dendrobium candidum

A genetic transformation method and technology mediated by Agrobacterium, applied in the field of genetic transformation of Dendrobium officinale mediated by Agrobacterium, can solve the problems of low transformation efficiency, imperfect genetic transformation system of Dendrobium officinale, etc., and achieve the improvement and enhancement of genetic transformation efficiency. effect of sensitivity

Active Publication Date: 2017-10-03
SICHUAN AGRI UNIV
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the above-mentioned problems existing in the prior art, the present invention provides an efficient Agrobacterium-mediated genetic transformation method of Dendrobium officinale, which can effectively solve the problem of the incomplete genetic transformation system of Dendrobium officinale mediated by Agrobacterium in the prior art and the low transformation efficiency. high problem

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
  • Efficient genetic transformation method of agrobacterium-mediated dendrobium candidum
  • Efficient genetic transformation method of agrobacterium-mediated dendrobium candidum
  • Efficient genetic transformation method of agrobacterium-mediated dendrobium candidum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A highly efficient Agrobacterium-mediated genetic transformation method for Dendrobium officinale comprises the following steps:

[0036] (1) Activation culture of Agrobacterium

[0037] Pick -80°C Agrobacterium transformants in 5ml liquid medium, place them at 28°C, 200r / min, and activate them. bottle), placed in 28°C and 200r / min to expand the culture until OD600=0.8, pour it into a 50ml centrifuge tube, centrifuge at 4000r / min for 10min, discard the supernatant, and collect the bacteria; among them, the liquid Medium includes: 1% yeast extract, 1% tryptone, 0.5% sodium chloride, 20mg / L rifampicin, 50mg / L kanamycin and 50mg / L gentamicin;

[0038] (2) Preparation of liquid dye

[0039] Add the bacterial cells obtained in step (1) into MS liquid medium, place them on ice to completely dissolve the bacterial cells in the medium, then adjust the bacterial liquid to OD600=0.4, 0.6, 0.8 and 1.0 with MS liquid medium, and then Add 100 μmol / L acetosyringone (AS) and surfac...

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 provides an efficient genetic transformation method of agrobacterium-mediated dendrobium candidum. The efficient genetic transformation method comprises the following steps: (1) performing activating culture on agrobacteria; (2) performing a dip dyeing liquid: adding the agrobacteria obtained in the step (1) into an MS liquid culture medium, putting on ice so as to enable the agrobacteria to be completely dissolved in the culture medium, adjusting bacterial liquid by using the MS liquid culture medium till OD600 is 0.4-0.8, then adding 100[mu]mol / L of acetosyringone(AS) and 0.001-0.01% of Triton-100, and uniformly shaking; (3) performing ultrasonic treatment and dip dyeing: under the conditions of 280-320W and 36-45KHz, performing ultrasonic treatment for 1-3min; (4) performing co-culture; (5) screening; (6) performing plant regeneration. A transformation system is complete; the survival rate of protocorms of dendrobium candidum is high; through optimization of the steps and parameters, plants with excellent resistance are obtained through transformation; the transformation rate is high, and the positive transformation rate is as high as 69.7%.

Description

technical field [0001] The invention belongs to the technical field of plant transgenesis, and in particular relates to an efficient Agrobacterium-mediated genetic transformation method of dendrobium officinale. Background technique [0002] Dendrobium candidum, as an important traditional Chinese medicinal material with both ornamental and medicinal value, is mainly distributed in Yunnan, Guizhou, Zhejiang, Sichuan and other places in my country. It contains multiple biologically active components such as dendrobium polysaccharides, dendrobine, amino acids, etc. It is sweet in taste and slightly cold in nature. However, due to its special requirements for the growth environment, it mostly grows in high-temperature and humid tropical and subtropical high mountains and cliffs, with slow growth and extremely low self-reproductive ability. Under the destruction of people's long-term predatory mining and living conditions, wild resources are on the verge of extinction, and the ...

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): C12N15/82A01H4/00A01H5/00
CPCA01H4/00C12N15/8205
Inventor 田孟良陈稷刘晨王丽黄进刘帆梁欢
Owner SICHUAN 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