Agrobacterium mediated transformation system for obtaining perennial rye-grass transgenic plant in high efficiently and its use

An Agrobacterium-mediated, ryegrass technology, which is applied in crop bioengineering breeding and plant genetic engineering, and can solve problems such as the difficult application of genetic transformation of gramineous plants

Inactive Publication Date: 2006-01-04
BEIJING FORESTRY UNIVERSITY
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Problems solved by technology

[0010] The present invention is an Agrobacterium-mediated transformation system for efficiently obtaining transgenic plants of perennial ryegrass and its application. Infection time, infection conditions, co-cultivation time, and the added concentration of bacteriostatin provide an Agrobacterium-mediated transformation system suitable for perennial ryegrass

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  • Agrobacterium mediated transformation system for obtaining perennial rye-grass transgenic plant in high efficiently and its use
  • Agrobacterium mediated transformation system for obtaining perennial rye-grass transgenic plant in high efficiently and its use
  • Agrobacterium mediated transformation system for obtaining perennial rye-grass transgenic plant in high efficiently and its use

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Embodiment 1

[0020] Agrobacterium-mediated transfer of DREB1A gene into perennial ryegrass to obtain drought-resistant and salt-alkali tolerant transgenic plants

[0021] Acquisition of recipient material: Callus was induced with mature embryos of perennial ryegrass Aishente as explants, and both callus induction and subculture medium were MS+6mg / L 2, 4-D+0.2mg / L 6 -BA+0.1g / L hydrolyzed casein. Incubate in the dark at 25°C. The embryogenic calli cultured for 2-3 months were picked and pre-cultured for 3 days to be infected.

[0022] Determination of the concentration of bacteriocin: Agrobacterium strain LBA4404 was cultured in YEP liquid medium containing different concentrations of cephalosporin in the dark at 28°C and 220rmp, and the OD of the bacterial solution was measured. 600 value (Table 1). Cephalosporin above 250mg / L can basically inhibit the growth of Agrobacterium LBA4404. When Agrobacterium grows to the logarithmic growth phase, dilute the bacterial solution with 1 / 2MS cult...

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Abstract

The present invention includes the following steps: obtaining embryonic callus acceptor material via high frequency regeneration system, culturing donor strain, determining concentration of ablastin, optimizing agrobacterium transforming condition, screening and culturing transformed callus and molecular detection of plant regeneration and resisting plant. By means of optimizing the conditions for transforming agrobacterium, including bacterium liquid concentration, infection time, infection condition, co-culture time, ablastin concentration, etc., the present invention provides agrobacterium mediated transformation system suitable for perennial ryegrass. The present invention solves the technological problem of applying agrobacterium mediating technology in genetic transformation of gramineous plant and establishes efficient agrobacterium mediated transformation system with embryonic callus as acceptor material for perennial ryegrass.

Description

Technical field: [0001] The invention belongs to the field of plant genetic engineering or crop bioengineering breeding. Specifically, it involves tissue culture technology of perennial ryegrass, Agrobacterium-mediated transformation technology and molecular detection technology of transformed plants, so as to efficiently obtain transgenic plants of perennial ryegrass. Background technique: [0002] Turfgrass seeds are the second most traded seeds in the world, but the grass seeds currently used in my country mainly rely on imports, and the breeding work is particularly weak. Perennial ryegrass is a turfgrass species that is widely used in the north and south of my country and has strong adaptability. It has fast lawn formation, strong resistance to diseases and insect pests and tillering ability. However, it still has the disadvantages of high water consumption, inability to tolerate the extreme values ​​of cold, heat, and drought, and the need for water in the germination...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N5/04A01H4/00
Inventor 韩烈保李雪刘君曾会明信金娜
Owner BEIJING FORESTRY UNIVERSITY
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