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Method for establishing perennial ryegrass inheritance and transformation receptor system through embryoid way

A technology of genetic transformation and embryoid bodies, applied in the fields of biotechnology and modern agricultural biology, can solve problems such as unsatisfactory regeneration systems

Inactive Publication Date: 2006-02-22
INST OF FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Unfortunately, so far, the establishment of a high-frequency stable regeneration system using mature embryos as explants is still not ideal

Method used

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  • Method for establishing perennial ryegrass inheritance and transformation receptor system through embryoid way
  • Method for establishing perennial ryegrass inheritance and transformation receptor system through embryoid way
  • Method for establishing perennial ryegrass inheritance and transformation receptor system through embryoid way

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The mature seeds of perennial ryegrass (Lolium perenne L.) were soaked in tap water for 0.5 hours; mixed with detergent and water at a ratio of 1:50, soaked for 1 hour, rinsed with running tap water for 12 hours, soaked in 70% alcohol for 30s, sterile water Rinse 5 times; then soak in 0.1% mercuric chloride for 15 minutes, and rinse with sterile water for 5 to 10 times. Under sterile conditions, the seeds were cross-sectioned with a scalpel and then inoculated into the callus-induced medium, and 5.0 mg / L of 2,4-dichlorophenoxyacetic acid 2,4-D,6-benzylaminoadenine 6 was added. - BA 0.5mg / L, naphthalene acetic acid NAA 0.5mg / L and hydrolyzed casein CH 500mg / L medium, cultured in the dark for 30 days. Among them, the preparation of the medium is to directly add hormones to N 6 other components of the medium and then autoclaved.

[0024] The instruments and equipment used are the well-known ultra-clean working machines and automatic autoclave sterilizers for plant tissue...

Embodiment 2

[0030] The culture procedures were all carried out as in Example 1, except that 2,4-dichlorophenoxyacetic acid 2,4-D was added at 8.0 mg / L, and 6-benzylaminoadenine 6 was added to the callus induction medium. -BA is 0.6mg / L, naphthaleneacetic acid NAA is 0.6mg / L, 2,4-dichlorophenoxyacetic acid 2,4-D is added to the callus subculture medium is 8.0mg / L, kinetin KT is 0.6mg / L, add 2,4-dichlorophenoxyacetic acid 2,4-D to the embryoid body induction medium to 0.7mg / L, kinetin KT to 1.0mg / L, in the embryoid body regeneration plant medium Add 6-benzylaminoadenine 6-BA to 0.4mg / L and kinetin KT0.4mg / L.

[0031] In the regeneration system established by the above method, the callus induction rate reaches 85.25%, the embryoid body induction rate reaches 75.04%, the regeneration rate reaches 85.72%, and the transplant survival rate reaches 97.5%.

Embodiment 3

[0033] The culture procedures were all carried out according to Example 1, except that 2,4-dichlorophenoxyacetic acid 2,4-D was added to the callus induction medium at 6.0 mg / L, 6-benzylaminoadenine 6 -BA is 0.8mg / L, naphthaleneacetic acid NAA is 0.3mg / L, 2,4-dichlorophenoxyacetic acid 2,4-D is added to the callus subculture medium is 6.0mg / L, kinetin KT is 0.8mg / L, add 2,4-dichlorophenoxyacetic acid 2,4-D to the embryoid body induction medium to 0.5mg / L, kinetin KT to 1.5mg / L, in the embryoid body regeneration plant medium Add 6-benzylaminoadenine 6-BA to 0.8mg / L and kinetin KT0.4mg / L.

[0034] In the regeneration system established by the above method, the callus induction rate reaches 86.06%, the embryoid body induction rate reaches 75.33%, the regeneration rate reaches 85.2%, and the transplant survival rate reaches 98%.

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Abstract

The invention discloses a method for establishing perennial ryegrass genetic conversion receptor system through embryoid generation approach, which mainly consists of using perennial ryegrass' mature seeds as material, obtaining callus through medium induced culture, carrying out subculture, inducing embryoid and regenerated strains in callus induced medium and regenerated medium, charging 2,4-D,6-BA,NAA and hydrolyse casein CH of a predetermined proportion into the induced callus medium, so as to effectively increase the inductivity of the callus.

Description

Technical field: [0001] The invention relates to a method for establishing a genetic transformation receptor system of perennial ryegrass through embryoid body generation, and belongs to the fields of biotechnology and modern agricultural biotechnology. Background technique: [0002] The perennial ryegrass is a perennial herb of the Poaceae family. Native to southern Europe, northern Europe and southwestern Asia. It is an important gramineous forage widely cultivated in areas with high rainfall in the temperate zone of the world. Because of its well-developed fibrous root system, fine and soft leaves, green leaves, fast planting speed, strong tillering ability and strong wear resistance, it is often used as a pioneer plant in soil and water conservation mixed sowing, and is an important and excellent turfgrass. . [0003] However, the root system of perennial ryegrass is not deep into the soil, and its cold resistance is poor. The growth in semi-arid regions is also not ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/04A01H4/00
Inventor 孙振元冯霞彭镇华韩蕾钱永强
Owner INST OF FORESTRY CHINESE ACAD OF FORESTRY
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