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Genetic transformation method for insect-resistant gene transgenic Narrow-crown Populus Nigra No.11

A method of genetic transformation and insect-resistant gene technology, which is applied in the field of genetic transformation of Narrow Crown Heiyang No. 11 with insect-resistant gene, can solve the problems that long-beetle control does not achieve the expected effect, shorten the transformation time, and improve the genetic transformation. Efficiency, conversion system stability effect

Active Publication Date: 2020-11-17
SHANDONG AGRICULTURAL UNIVERSITY
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Problems solved by technology

However, through experiments, it was found that after the Bt886Cry3A gene was transformed into poplar, the control of long beetles did not achieve the expected effect
So far, there has been no report on how to improve its ability to resist borer pests by transferring the insect resistance gene into Narrow Crown Black Populus 11

Method used

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  • Genetic transformation method for insect-resistant gene transgenic Narrow-crown Populus Nigra No.11
  • Genetic transformation method for insect-resistant gene transgenic Narrow-crown Populus Nigra No.11
  • Genetic transformation method for insect-resistant gene transgenic Narrow-crown Populus Nigra No.11

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Experimental program
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Effect test

Embodiment 1

[0047] Example 1: Genetic Transformation Method of Narrow Crown Black Populus No. 11 Transplanted with Insect-resistant Gene

[0048] (1) Select the leaves of the aseptic seedling No. 11 Narrow Crown Black Populus, and the selected aseptic seedling leaves should be the 3rd to 6th leaves from the terminal bud down to the number of leaves that grow robustly and have a flat leaf shape. Remove the petiole and one-third of the front end, and cut 3-4 cuts perpendicular to the main vein of the remaining part, put it on the pre-cultivation medium, and cultivate it in the dark at 24°C for 24 hours.

[0049] (2) Freezing and thawing the recombinant plasmid ( figure 1 ) into the Agrobacterium to obtain the Agrobacterium infection solution;

[0050] Poplar stem borer resistance gene P2-Bt886Cry3Aa-P1 is a fusion gene. Among them, Bt886Cry3Aa is derived from Bacillus thuringiensis Bt strain, P 1 (LPPNPTK) and P 2 (TPHRSPL) is a cellulase-specific affinity short peptide coding sequence ...

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Abstract

The invention discloses a genetic transformation method for insect-resistant gene transgenic Narrow-crown Populus Nigra No.11, and belongs to the technical field of plant genetic engineering. The genetic transformation method comprises the processes of explant selection and pre-culture, agrobacterium infection, tissue culture seedling formation through induction culture and the like. A large number of high-quality trunk borer-resistant Narrow-crown Populus Nigra No.11 new strains can be cultivated in a short time by utilizing the genetic transformation method, the transformation time can be shortened, and the genetic transformation efficiency can be improved; and moreover, the transformation system is stable, is not influenced by time and seasons, and can be used for cultivating and producing excellent populus nigra strains with trunk borer resistance for northern China on a large scale.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to a method for genetic transformation of Narrow Crown Black Populus No. 11 with an insect-resistant gene. Background technique [0002] Poplar is one of the fast-growing timber tree species with the largest cultivation area in the mid-latitude plains in the world. It has the characteristics of fast growth, high yield and easy regeneration, and it is also the forest tree species that contributes the most to carbon sequestration. In terms of cultivation area, the area of ​​poplar plantations in my country has reached 7 million hectares, ranking first in the world. [0003] With the continuous expansion of poplar afforestation area, the overuse of a single strain has led to the intensification of poplar plant diseases and insect pests. According to statistics, the annual loss of my country's forests due to pests and diseases is about 140 million mu, and the direct e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/32A01H4/00A01H5/00A01H6/00
CPCC12N15/8205C12N15/8286C07K14/325A01H4/001A01H4/008Y02P60/40Y02A40/22
Inventor 李际红邢世岩桑亚林王锦楠王如月国浩平侯丽丽王宝锐
Owner SHANDONG AGRICULTURAL UNIVERSITY
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