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

A kind of Agrobacterium-mediated botrytis gene transformation method

A technology mediated by Botrytis and Agrobacterium, applied in the field of microorganisms, can solve the problem of no gene transformation and achieve the effect of ensuring the regeneration rate

Inactive Publication Date: 2017-05-10
ZHEJIANG UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For Botrytis, a fungus with important economic significance, so far, we have not seen any reports on how to carry out gene transformation for this fungus

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
  • A kind of Agrobacterium-mediated botrytis gene transformation method
  • A kind of Agrobacterium-mediated botrytis gene transformation method
  • A kind of Agrobacterium-mediated botrytis gene transformation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0040] The method of the present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments.

[0041] 1. Preparation of Botrytis protoplasts

[0042] (1) Preparation of reagents and medium

[0043] ① 0.7M sodium chloride solution: Dissolve 40.95g sodium chloride (NaCl) in 1000mL water, and perform conventional high-temperature sterilization (1.1 atmospheres, 121°C for 20 minutes) after constant volume.

[0044] ②1M Tris-Cl, pH=7.5: Tris(hydroxymethyl)aminomethane (Tris(hydroxymethyl)aminomethane, generally abbreviated as Tris) 121.14 was dissolved in 600mL double-distilled water, adjusted to pH=7.5 with concentrated hydrochloric acid, and then adjusted to volume with double-distilled water to 1 L, perform conventional high-temperature sterilization (1.1 atmospheres, sterilize at 121°C for 20 minutes), and set aside.

[0045] ③Cell wall degrading enzyme solution: Weigh 100mg of Driselase and 100mg of Lysing Enzyme on a ba...

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
diameteraaaaaaaaaa
Login to View More

Abstract

The invention discloses an agrobacterium tumefaciens mediated gene transformation method of botryosphaeria dothidea. The method comprises the steps of preparing the protoplast of botryosphaeria dothidea, and carrying out renewal culture on the protoplast; transforming a plasmid vector containing a target gene into agrobacterium tumefaciens; mixing agrobacterium tumefaciens with the protoplast, co-culturing agrobacterium tumefaciens and the protoplast on an induction plate, and then transferring agrobacterium tumefaciens and the protoplast to a screening plate to be cultured, thus obtaining a potential transformant; confirming whether a potential transformant genome contains the target gene, thus obtaining a genetically stable transformant. The method has the beneficial effects that the protoplast is used as a transformation material and the target gene is transferred into botryosphaeria dothidea by utilizing agrobacterium tumefaciens mediation and is integrated into the genome; before co-culture, the protoplast needs renewal culture; when the renewal culture time is 2-6 hours, transformation materials in each tube can obtain 20-30 potential transformants; when the renewal culture time is more than 6 hours or less than 2 hours, the transformation efficiency is quickly reduced, and only few potential transformants can be obtained and even the potential transformants are difficult to obtain.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to an Agrobacterium-mediated gene transformation method for Botrytis spp. Background technique [0002] Botryosphaeria dothidea is a common and important fungus with global distribution and has important development and utilization value. Some of these fungi can saprophyte on a variety of decaying wood, some parasitize on a variety of woody plants, causing diseases of woody plants, and some form symbionts with plants, which are endophytic fungi of many plants. Botrytis, which lives saprophytically on rotting wood, can decompose woody plant residues and plays an important role in the material cycle and energy cycle of the ecosystem; at the same time, this type of fungus can also be developed to produce lignin enzymes and Cellulase has good prospects for economic development. The strains belonging to endophytic fungi can be used as bioreactors. For example, a fungus iso...

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): C12N15/80C12N1/21C12R1/645
Inventor 王洪凯林福呈
Owner ZHEJIANG 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