Method for improving cotton transgenosis efficiency

A technology of transgenic cotton, applied in genetic engineering, plant genetic improvement, botanical equipment and methods, etc., can solve the problems of offspring not conforming to Mendelian inheritance rules, long transformation cycle, transient expression and chimera, etc., to achieve optimal stem Advanced conversion technology system, shortened conversion cycle, and reduced utilization value

Active Publication Date: 2014-08-27
HEBEI AGRICULTURAL UNIV.
View PDF3 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this technical solution is mainly an in vitro shoot tip transformation method, without the process of cotton cell and tissue culture, the probability of chimerism increases, and the inheritance cannot be stable, and the offspring do not conform to the Mendelian inheritance law
However, many cotton transgenic methods (such as by hypocotyl transformation) through cell and tissue culture, because the hypocotyl will be induced and differentiated through callus, the transformation cycle is longer, generally 6-9 months ( Liu Mingyue, Optimization of Agrobacteri

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 improving cotton transgenosis efficiency
  • Method for improving cotton transgenosis efficiency
  • Method for improving cotton transgenosis efficiency

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1 improves cotton transgenic efficiency

[0029] 1 Materials and methods

[0030] 1.1 Test material

[0031] The test material used in this experiment is the upland cotton (G.hirsutum L.) variety: Jimian 14.

[0032] Transformation strain: Agrobacterium tumefaciens LBA4404.

[0033] The transformed gene GbAPX was derived from the root tissue transcriptome library of sea-island cotton variety Pima90-53, which was stressed by Verticillium dahliae, constructed in our laboratory.

[0034] 1.2 Main Reagents and Drugs

[0035] Acetosyringone, phytagel, plant growth regulators (IAA and KT), kanamycin and cephalosporin.

[0036] PCR primers were mainly synthesized by Sangon Bioengineering Co., Ltd.

[0037] 1.3 Test method

[0038] 1.3.1 Culture medium preparation

[0039] Prepare the required media according to Table 1.

[0040] The medium used in the test in table 1

[0041]

[0042]

[0043] 1.3.2 Aseptic vaccine culture

[0044] Peel the cottonsee...

Embodiment 2

[0068] Example 2 Co-cultivation of transgenic cotton and optimization of screening medium

[0069] The operation method is the same as in Example 1, except that the difference from Example 1 is that in this example, the shoot tips of sterile cotton seedlings are infected with Agrobacterium and placed on co-cultivation medium and screening medium of different hormones (combinations) for growth. Specifically:

[0070] The shoot tips of sterile cotton seedlings were infected by Agrobacterium and placed on the co-cultivation medium and screening medium of different hormones (combinations), 2 bottles of each hormone (hormone combination), 46 bottles in total (including 2 bottles of control) , 5 stem tips per bottle.

[0071] The co-cultivation medium is: MSB 5 +hormone (combination)+200μmol AS+30g / L glucose+2.4g / L phytagel, pH5.4;

[0072] The screening medium is: MSB 5 +200mg / Lcef+80mg / L kan+glucose 15g / L+sucrose 15g / L+hormone (combination)+0.25% Phytagel, pH5.8;

[0073] The...

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

A provided method for improving cotton transgenosis efficiency comprises the following steps: a, employing an agrobacterium-mediated method to infect cotton aseptic-seedling shoot tips with a plant expression carrier containing a target gene, and placing the infected cotton shoot tips in a co-culture medium for culture; and b, after co-culture is performed, transferring the cotton shoot tips to a selective medium for culture. The co-culture medium and the selective medium both contain KT with a concentration of 0.3 mg/L and IAA with a concentration of 0.2 mg/L. By utilizing the method, the aseptic seedling culture time is 2-3 days, the co-culture time is 2 days, the selective culture time is 15-20 days, the rooting culture time is 20 days, the conversion period does not exceed 2 months, and the positive conversion rate reaches 8% after screening is performed.

Description

technical field [0001] The invention relates to the technical field of cotton transgenes, in particular to a method for improving the efficiency of cotton transgenes. Background technique [0002] Significant progress has been made in the genetic improvement of cotton using genetic engineering technology. In 1987, Umbeck et al first reported the transfer of kanamycin-resistant gene into cotton by Agrobacterium-mediated method (Umbeck P. Genetically transformed cotton (Gosspium hirsutum L.) Plants. Bio / Teehnology, 1987, 5: 263-266). The traditional genetic transformation method has a long transformation period, strong genotype dependence of plant regeneration, high deformity rate of regenerated seedlings, and low survival rate of transplanting, which has become a bottleneck restricting the research of cotton transgenic breeding. [0003] Cotton shoot tips have the characteristics of direct differentiation and regeneration, once transformed, the formation of leaves and roots ...

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): C12N15/84A01H5/00
Inventor 吴金华王省芬张桂寅吴立强李志坤张艳王国宁柯会锋马峙英
Owner HEBEI AGRICULTURAL UNIV.
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