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Establishment of transgenic system of Populus chinensis and propagation method of transgenic plants

A technology of transgenic Populus chinensis, applied in botany equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems of low survival rate of Populus chinensis afforestation, long tree growth cycle, and long time required for breeding, etc. Achieve the effect of short cycle, high reproduction rate and low cost

Inactive Publication Date: 2011-12-21
NORTHEAST FORESTRY UNIVERSITY
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0003] Most of the afforestation lands of Populus chinensis are lands returned to forests with dry and barren soil and serious soil decline, resulting in a low survival rate of Populus chinensis afforestation and a long growth cycle of trees. It takes a long time and slow results to select new varieties using conventional breeding techniques
In the past, the research on Populus chinensis mainly focused on conventional breeding such as provenance test, seed sowing, space breeding, and polyploidy breeding. With the rapid development of modern biotechnology, there have been no reports on the use of Agrobacterium-mediated transgenic Populus chinensis.

Method used

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  • Establishment of transgenic system of Populus chinensis and propagation method of transgenic plants
  • Establishment of transgenic system of Populus chinensis and propagation method of transgenic plants
  • Establishment of transgenic system of Populus chinensis and propagation method of transgenic plants

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specific Embodiment approach 1

[0016] Specific embodiment 1: The method for establishing the Populus chinensis transgenic system in this embodiment is realized according to the following steps: 1. The annual leaves and stems of Populus chinensis are soaked in alcohol with a volume concentration of 70% in an ultra-clean workbench for 30s to 1min , washed 2-3 times with sterilized water, then transferred to 1% NaClO solution for disinfection for 10-20 minutes, rinsed 3-5 times with sterilized water, and then inoculated with 0.5 mg / L 6-benzylaminopurine, 0.1mg / L naphthaleneacetic acid, 20g / L sucrose and 5g / L agar 1 / 2MS culture medium for one month to obtain tissue culture plantlets;

[0017] 2. Pick a single colony of engineering bacteria containing the LbDREB gene and inoculate it into 20ml of LB medium and cultivate it to the OD of the bacterial solution 600 If the value is 0.5-1.0, use a pipette to suck 3-5ml of bacterial solution into the centrifuge tube, centrifuge at 12000r / min for 1min, discard the supe...

specific Embodiment approach 2

[0027] Specific embodiment two: the difference between this embodiment and specific embodiment one is that in step one, the leaves and stems of Populus chinensis are soaked in alcohol with a volume concentration of 70% for 40s in the ultra-clean workbench, and rinsed with sterilized water for 2 Then transfer to a 1% NaClO solution for disinfection for 15 minutes, and rinse with sterilized water for 4 times. Other steps and parameters are the same as those in Embodiment 1.

specific Embodiment approach 3

[0028]Embodiment 3: The difference between this embodiment and Embodiment 1 is that the inoculation in step 1 is to place the stem segment directly on the surface of the culture medium; the back of the blade is placed on the culture medium. Other steps and parameters are the same as those in Embodiment 1.

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Abstract

The invention relates to the establishment of a transgenic system of Populus chinensis and the propagation method of the transgenic plant, which relates to the establishment of the transgenic system and the propagation method of the transgenic plant. System establishment: obtain tissue culture seedlings; activate engineering bacteria containing LbDREB gene and transform exogenous genes; obtain resistant buds after degerming; resistant buds take root and transplant; Positive plants are complete. Propagation: The transgenic regenerated plants of Populus chinensis obtained by establishing the transgenic system of Populus chinensis were differentiated and cultured to obtain tissue culture seedlings; PCR and RT-PCR were performed to detect positive roots, stems and leaves; 3. Repeat operation 2 times and you are done. The present invention can realize the rapid cultivation of transgenic Populus chinensis, and lays the foundation for the development of fine varieties of Populus chinensis; the produced offspring have good genetic stability, and are useful for realizing large-scale, short-period, high reproduction rate and low nutrient density of transgenic Populus chinensis seedlings. The cost of factory production is of great significance.

Description

technical field [0001] The invention relates to the establishment of a transgenic system and a method for multiplying transgenic plants. Background technique [0002] Daqing Yang (Populus ussuriensis kom.), also known as Han Dayang and Hada Yang, is mainly distributed in Heilongjiang, Jilin Province, Russia (Far East), and North Korea. It is a unique native tree species in the forest area of ​​Northeast China. The wood is light and soft, and the material is white, dense and durable. It is an excellent raw material for papermaking and plywood. [0003] Most of the afforestation lands of Populus chinensis are lands returned to forests with dry and barren soil and serious soil decline, resulting in a low survival rate of Populus chinensis afforestation and a long growth cycle of trees. It takes a long time and slow results to select new varieties using conventional breeding techniques . In the past, the research on Populus chinensis mainly focused on conventional breeding suc...

Claims

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Application Information

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IPC IPC(8): A01H4/00C12Q1/68
Inventor 李成浩靳春莲李开隆刘桂丰杨静莉周晨光刘立辉杜佳李春燕
Owner NORTHEAST FORESTRY UNIVERSITY
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