Agrobacterium-mediated method for culturing transgenic populus wutunensis plants

An Agrobacterium-mediated and transgenic technology, applied in the field of genetic engineering, can solve the problems such as the lack of Qingyang faction, and achieve the effect of convenient material acquisition.

Inactive Publication Date: 2012-01-25
DALIAN NATIONALITIES UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the establishment of the transgenic system of Populus wutunensis, especially Populus wutunensis

Method used

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  • Agrobacterium-mediated method for culturing transgenic populus wutunensis plants
  • Agrobacterium-mediated method for culturing transgenic populus wutunensis plants
  • Agrobacterium-mediated method for culturing transgenic populus wutunensis plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] a. will cut 3 incisions along the vertical main veins of the leaves of the aseptic seedlings of Wutun Yang as transformed explants, the incisions are deep to the main veins, and the front is upwards, and put into the pre-cultivation medium, totally 50 blades, each Inoculate 9 to 11 leaves in the petri dish, inoculate 5 petri dishes in total, and culture in light for 2 days, the culture temperature is 26°C, the light intensity is 3400 Lux, and the light time is 16 hours / day;

[0049] The above pre-culture medium is: MS+6-BA 1.0mg / L+NAA 0.2mg / L+sucrose 30g / L+agar powder 8g / L;

[0050] b. Pick the colony of Agrobacterium AGLO with the bar gene and inoculate it into 10ml YEP liquid medium containing 50mg / L of kanamycin, culture at 26°C with shaking at 200rpm for 24 hours, and transfer 100μl of the bacterial solution to 30ml of YEP liquid medium was cultured with shaking at 200rpm, when the cells were 0D 600 When it reaches 0.8, collect the bacteria by centrifugation at 120...

Embodiment 2

[0078] The determination of embodiment 2 herbicide critical concentration

[0079] Since the Agrobacterium AGLO used for gene transformation has the bar gene, the positive plants transformed by Agrobacterium AGLO have resistance to herbicides, and herbicides are used as screening reagents to select transformed cells. For herbicide resistance, the critical concentration test of leaf resistance to herbicide was carried out, and the uniform leaves were placed on the medium containing 0.5, 1.0, 1.5, 2.0, 2.5mg / L PPT: MS+ 6-BA 1.0mg / L+NAA 0.2mg / L+sucrose 30g / L+agar powder 8g / L, the inoculation density was 5 leaves per petri dish, the selection culture temperature was 26°C, the light intensity was 3400Lux, and the light time was 16 hours / days. Observe the response of the leaves to a series of concentration gradient herbicides, and make statistics after culturing for 30 days to determine the critical concentration of the herbicides. The control medium is: MS medium supplemented wit...

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Abstract

The invention relates to an agrobacterium-mediated method for culturing transgenic populus wutunensis plants, and relates to a method for culturing a new anti-reverse populus wutunensis variety by using a gene engineering technology. The method is convenient in material section; populus wutunensis leaves are directly used as explants; factors affecting gene transformation, namely antibiotics sensitivity, are optimized; and an efficient genetic transformation system for populus wutunensis is established. A method with higher transformation efficiency is provided for culturing the new anti-reverse populus wutunensis variety, the time for culturing the new anti-reverse populus wutunensis variety can be shortened, and a new path is provided for genetic improvement of forests.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for cultivating transgenic Populus wutun plants mediated by Agrobacterium. Background technique [0002] Poplar is an important fast-growing timber tree species. In previous studies, most of the research aimed at increasing the yield of poplar, but neglected the improvement of its stress resistance and other traits. At the same time, the environment continues to deteriorate, and poplar production is severely restricted by natural disasters such as drought and salinity. At present, poplar occupies a relatively large proportion in the world and my country's plantation forests, so it is of great practical significance to cultivate new varieties of anti-retroversion gene poplar. Since Fillatti et al first obtained herbicide-resistant poplar genetically engineered plants in 1987, research on poplar gene transformation has made great progress. Some hybrids of Pop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12Q1/68A01H4/00A01H5/00
Inventor 金华姜国斌郭鹏王颖刘磊
Owner DALIAN NATIONALITIES UNIVERSITY
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