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Transgenic method of pollen plant of populus simonii x p.nigra

A transgenic and plant technology, applied in the fields of plant genetic improvement, botanical equipment and methods, genetic engineering, etc., can solve the problems of low transgenic efficiency of poplar pollen plants, and achieve easy-to-master, simple operation, and improved genetic transformation efficiency. Effect

Inactive Publication Date: 2013-01-09
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to solve the problem of low transgenic efficiency of the existing poplar pollen plants, and provides a transgenic method for poplar pollen plants

Method used

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  • Transgenic method of pollen plant of populus simonii x p.nigra
  • Transgenic method of pollen plant of populus simonii x p.nigra
  • Transgenic method of pollen plant of populus simonii x p.nigra

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specific Embodiment approach 1

[0009] Specific embodiment one: the transgenic method of poplar small black poplar pollen plant of this embodiment, carry out according to the following steps: one, get the rootless seedling that is 1~2cm that the height that cultivates in the stem-taking medium is moved into the rooting medium to induce rooting, wait for the seedling When growing to 5~8cm, get the top of 2~3cm on the seedling and move it into the rooting medium to carry out secondary rooting, choose the plant after the secondary rooting of seedling height 4~5cm as explant; For the newly born leaves with a length of 1-3cm on the implant, cut a knife at the base of the leaf and the middle of the leaf, discard the tip of the leaf, and inoculate it on the differentiation medium for pre-culture for 2-3 days; 3. Pick and carry exogenous Genetic Agrobacterium EHA105 was inoculated into LB medium containing 50mg / L rifampicin and 50mg / L kanamycin and cultured for 16-24h, the culture temperature was 28°C, the shaker spe...

specific Embodiment approach 2

[0013] Specific embodiment 2: the difference between this embodiment and specific embodiment 1 is that in step 2, the newborn leaves with a length of 2 cm on the explant of step 1 are taken. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0014] Embodiment 3: This embodiment differs from Embodiment 1 or Embodiment 2 in that: the exogenous gene in the Agrobacterium EHA105 carrying the exogenous gene described in Step 3 is Populus trichocarpa AP2 gene or Tamarix bZIP gene. Others are the same as in the first or second embodiment.

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Abstract

The invention discloses a transgenic method of a populus simonii * p.nigra pollen plant and relates to the transgenic method of the populus simonii * p.nigra pollen plant. The transgenic method is used for solving the problem of low transgenic efficiency of the existing populus simonii * p.nigra pollen plant. The method comprises the following steps: taking a rootless seedling in a stem culture medium, transferring into a rooting culture medium and taking the plant as an explant; taking blades on the explant, respectively cutting the base part and the middle part once, abandoning blade tips and inoculating into a differentiation culture medium; picking agrobacterium tumefaciens EHA 105 carrying exogenous genes, inoculating into an LB (Luria-Bertani) culture medium for culture so as to getbacterial liquid and diluting the bacterial liquid with sterile water; placing the blades after pre-culture into the bacterial liquid for soaking, taking out the blades and sucking away the surplus bacterial liquid; inoculating the blades into the differentiation culture medium for culture; and taking out the blades, sucking dry the water, transferring the blades to a selection culture medium forculture, producing calli, placing the calli into the selection culture medium for culture and producing resistant buds for completion. By adopting the transgenic method, the more resistant buds can be obtained and the transgenic efficiency is high.

Description

technical field [0001] The invention relates to a transgenic method for poplar small black poplar pollen plants. Background technique [0002] Poplar is an important tree species for afforestation and industrial use in my country. Especially in the ecological construction and commercial forest construction in northern my country, it occupies an irreplaceable position. Conventional poplar breeding has a long cycle, slow results, and poor operability. The application of genetic engineering technology in poplar breeding improves the purpose and operability of breeding, and opens up an attractive new way for poplar breeding. However, there are significant differences in the regeneration efficiency and transformation efficiency of different genotypes of poplar, so it is necessary to explore the best transgenic system for different genotypes of poplar. [0003] Small black poplar is the main tree species for greening and afforestation in northern my country. At present, transge...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84
Inventor 李慧玉刘玲李园园姜静刘桂丰
Owner NORTHEAST FORESTRY UNIVERSITY