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Method for establishing perennial rye-grass high efficiency gene gun conversion system and its use

A gene gun transformation, ryegrass technology, applied in the fields of plant biology and crop breeding, can solve the problems of low transformation frequency and no transgenic plants

Inactive Publication Date: 2006-01-04
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In terms of biolistic transformation of perennial ryegrass, there is no successful report on using embryogenic callus as the recipient in China, and only a few laboratories abroad have obtained transgenic plants, but the transformation frequency is low, and many laboratories have obtained transgenic plants after transformation. Transgenic plants not obtained

Method used

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  • Method for establishing perennial rye-grass high efficiency gene gun conversion system and its use
  • Method for establishing perennial rye-grass high efficiency gene gun conversion system and its use
  • Method for establishing perennial rye-grass high efficiency gene gun conversion system and its use

Examples

Experimental program
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Effect test

Embodiment 1

[0018] Transforming DREB1A gene into perennial ryegrass by particle gun method to obtain drought-resistant, salt-tolerant and low-temperature-tolerant transgenic plants

[0019] The expression of plant genes is induced by drought, high salt and low temperature stress. According to the function of gene products, there are two main categories. The first category includes functional proteins that directly protect cells from water stress, synthetases of osmotic regulators, and Toxic-degrading enzymes; the second class includes transcription factors that transmit signals and regulate gene expression, protein kinases that sense and transduce stress signals, and proteases that play an important role in signal transduction. Both choline monooxidase (CMO) and betaine aldehyde dehydrogenase (BADH), which control the synthesis of betaine, belong to the synthase of osmotic regulators, and CMO-BADH is a double gene constructed by integrating the two into the same expression vector Expressi...

Embodiment 2

[0036] The gene gun method transfers the CMO-BADH double gene into perennial ryegrass to obtain drought-resistant and salt-tolerant transgenic plants. The acquisition of acceptor materials, the determination of the concentration and time of the screening agent, the coating of DNA, and the bombardment of the gene gun are the same as in Example 1. Transformation The receptor is the callus of perennial ryegrass Aishon. During PCR amplification, the CMO primer sequence is:

[0037] Primer F: ATG ATG GCA GCA AGC GCA AGC GCA AC

[0038] Primer R: TTA CTT CAA AGT TTG TTG CAA CCA GCA GTG G

[0039] The sequence of the BADH primer is:

[0040] Primer F: aac GGA TCC ATG GCG TTC CCA ATT CCT GC

[0041] Primer R: acc GAG CTC TCA AGG AGA CTT GTA CCA TCC CC Take the DNA of PCR-positive plants for Southern blot hybridization identification, and use the PCR product of the target gene as a probe. A total of 50 resistant plants were detected, including 7 positive plants, and the transformat...

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Abstract

The present invention provides a method of establishing efficient perennial ryegrass gene gun conversion system and its application. By means of regulating the effect factors of gene gun bombardment process, including the size of the bombarded callus block, gold powder coated matter, gold powder diameter, bombardment height, bombardment times, etc. as well as screening pressure and time of the screening agent, the present invention establishes the perennial ryegrass genetic transformation system successively. The genetic transformation system is applied in establishing transgenic perennial ryegrass and obtaining drought resistant, salt resistant and low temperature resistant transgenic perennial ryegrass plant.

Description

Technical field: [0001] The invention belongs to the field of plant biotechnology or the field of crop breeding. Specifically, it involves perennial ryegrass tissue culture technology, gene gun transformation technology and molecular detection technology of transformed plants, so as to efficiently obtain perennial ryegrass transgenic plants. Background technique: [0002] Turfgrass industry is a rapidly developing new industry in our country and has a very broad market prospect; however, there is a certain gap in our country's turfgrass scientific research compared with developed countries. Perennial ryegrass has many tillers, high yield, soft texture, long green period, well-developed root system, strong suitability for growth, and resistance to trampling; it has a certain potential for salt-alkali tolerance, can increase soil organic matter, improve soil structure, and prevent soil erosion. green fodder, and is a good lawn grass. Due to the high degree of self-sterility ...

Claims

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Application Information

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IPC IPC(8): C12N15/87C12N15/89C12N15/82A01H4/00
Inventor 韩烈保李雪刘君曾会明信金娜
Owner BEIJING FORESTRY UNIVERSITY
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