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Genetic transformation method for populus

A genetic transformation method and technology of poplar, applied in the field of genetic transformation of poplar, can solve the problems of cumbersome process, low transformation efficiency of poplar, etc.

Active Publication Date: 2018-07-31
BEIJING FORESTRY UNIVERSITY
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  • Claims
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Problems solved by technology

However, in the past, the conversion efficiency of poplar was relatively low, and the process was cumbersome

Method used

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  • Genetic transformation method for populus
  • Genetic transformation method for populus
  • Genetic transformation method for populus

Examples

Experimental program
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Effect test

Embodiment 1、84

[0058] Example 1, the rapid and efficient genetic transformation process of overexpressing miRNA156 in 84K poplar

[0059] The flow chart of the rapid and efficient genetic transformation process of overexpressing miRNA156 in 84K poplar is shown in figure 1 .

[0060] 1. First, construct the vector of the target gene to be transformed, transform it into Agrobacterium after successful construction, and shake the bacteria with liquid LB. Wherein the final concentration of kanamycin is 50 mg / L, and the final concentration of rifampicin is 25 mg / L, so that the OD600 value is about 0.6. Centrifuge the bacterial solution in a 50mL centrifuge tube at 18°C ​​and 3500rpm for 15min, add the resuspension liquid (liquid WPM medium) (that is, add sucrose to the WPM medium shown in Table 1 to a final concentration of 20g / L) and resuspend , the resuspension was finally added to a volume of about 100 mL (OD600 about 0.3), and then 100 μL of 100 mM AS was added to infect.

[0061] Among the...

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Abstract

The invention discloses a genetic transformation method for populus. The genetic transformation method for populus comprises the steps of 1, adopting an agrobacterium infection solution containing a target gene expression vector for infecting populus sterile leaves, the infected populus leaves are placed on a co-culture medium (containing 100 microM AS) for co-culture; 2, after the leaves on the co-culture medium are torn open, the leaves are inoculated on a screening culture medium for culture, and callus is obtained; 3, the callus is inoculated to a differential medium for culture, and adventive buds are obtained; 4, the adventive buds are inoculated to a rooting medium for culture, and rooting seedlings are obtained; 5, the rooting seedlings are identified. According to the method, thepopulus transformation period is shortened, and the populus transgenosis efficiency is improved, the method has the important technology supporting effect on populus transgenosis breeding and gene function identification, and the certain theoretical and practical significance in forest resistant variety improvement and new variety breeding is achieved.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering and relates to a method for genetic transformation of poplar. Background technique [0002] Poplar is the general designation of Populus tree species, which is widely distributed. At present, there are more than 100 species of poplar in the world. Because poplar has biological characteristics such as fast growth, fast seed maturation, and small genome, it is widely used in basic biological research. And because poplar is the natural host of Agrobacterium tumefaciens, it is convenient for Agrobacterium-mediated genetic transformation, so poplar is considered as a model plant for forest genetic engineering research. [0003] In recent years, the research on gene function of poplar has attracted more and more people's attention, mainly focusing on trait improvement, tree species transformation and breeding of new resistant varieties. However, in the past, the conversion efficiency of poplar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H4/00A01H5/00A01H6/00
CPCA01H4/001A01H4/008C12N15/8205
Inventor 林金星王鑫伟崔亚宁付春祥曹英萍吴振映
Owner BEIJING FORESTRY UNIVERSITY
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