Method for genetic transformation of zoysia matrella mediated by agrobacterium tumefaciens

A method of genetic transformation, the technology of Zoysia ditchleaf, applied in the fields of biochemical equipment and methods, horticultural methods, botanical equipment and methods, etc., can solve the problems of incomplete genetic transformation system and limit the development of molecular breeding, etc., and achieve simplification Experimental process, effect of reducing workload

Inactive Publication Date: 2019-12-17
INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
View PDF0 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, at present, there are very few reports on the genetic transformation of Zoysia rugosa, only in the preliminary research on the genetic transformation of Zoysia rugosa for resistance to glyphosate, using the 7-year-old callus of Zoysia rugosa as material, After being infected by Agrobacterium, they were regene...

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 genetic transformation of zoysia matrella mediated by agrobacterium tumefaciens
  • Method for genetic transformation of zoysia matrella mediated by agrobacterium tumefaciens
  • Method for genetic transformation of zoysia matrella mediated by agrobacterium tumefaciens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0022] The present invention will be further described below in conjunction with the examples, but not as a limitation of the present invention.

[0023] For the test methods that do not indicate the specific experimental conditions in the following examples, usually follow the routine experimental conditions or the experimental conditions suggested by the manufacturer.

[0024] 1.1 Method

[0025] 1.1.1 Induction and subculture of callus from Zoysia spp.

[0026] Embryogenic callus was induced from the stolons of Zoysia rugosa, and the callus induction and subculture were carried out according to the methods of Chai Mingliang et al.

[0027] Table 1 Basic medium for plant regeneration and Agrobacterium culture

[0028]

[0029] 1.1.2 Antibiotic susceptibility test

[0030] Determination of the inhibitory concentration of Timentin: the calli soaked in the Agrobacterium solution for 30 min were inoculated in the callus containing 0, 100, 150, 200, 250 and 300 mg / L Timenti...

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 genetic transformation of zoysia matrella mediated by agrobacterium tumefaciens. The method includes the steps of the concentration of a bacterial solution, the infection time, the co-culture time, the antibiotic concentration, acclimatization and transplantation, PCR detection of transgenic plants and the like. The method includes the following steps that an optimal genetic transformation system is that an OD600 value of the bacterial solution is 0.4, infection is conducted for 30 min, and selective culture is conducted after co-culture is conducted for 3d; the optimal bacteriostatic concentration of Timentin in a selective culture phase is 250 mg/L, the optimum selection pressure for hygromycin callus screening is 40 mg/L, and the optimum concentration of root taking screening is 15 mg/L; and histochemical analysis and PCR identification of GUS activity show that a target gene is successfully transferred into a genome of the zoysia matrella. Theabove research provides important technical support for gene function research and molecular breeding of the zoysia matrella. The method has the advantages that a stable and high-efficiency zoysia matrella genetic transformation system is obtained, the convenience is provided for genetic improvement of the zoysia matrella, and meanwhile, a key preliminary foundation is laid for verification of functional genes of the zoysia matrella.

Description

technical field [0001] The invention relates to a genetic transformation method in the biological field, in particular to a method for the genetic transformation of Zoysia sativa mediated by Agrobacterium. Background technique [0002] Zoysia ditchleaf ( Zoysia matrela ) is Poaceae ( Gramineae ) Perennial warm-season turfgrass of the genus Zoysia in the subfamily Choridoideae. It has the characteristics of slender leaves, dense grass layer, well-developed root system and long green period. It is also one of the most salt-tolerant halophytes. It is mostly used in urban greening, sports field lawns and water and soil conservation, and is widely used in tropical and subtropical regions of the world. [0003] With the rapid development of biotechnology, transgenic technology has gradually matured, and specific target genes can be genetically transformed to shorten the breeding cycle. Among many plant transgenic methods, the Agrobacterium-mediated method has become the pref...

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): C12N15/84A01H4/00A01H5/00A01H6/46
CPCA01H4/00C12N15/8205
Inventor 王凯曲爱爱刘建秀李建建郭海林李晓慧宗俊勤
Owner INST OF BOTANY JIANGSU PROVINCE & CHINESE ACADEMY OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap