1. Mediated Transformation Agrobacterium is a gram-negative bacterium commonly found in soil that can chemotactically infect most dicotyledonous wounds under natural conditions and induce crown gall tumors or Hairy roots. The Agrobacterium tumefaciens and A. rhizogenes cells contain Ti and Ri plasmids, respectively, which contain a piece of T-DNA. After Agrobacterium infects the plant wounds and enters cells, the T-DNA can be inserted into the plant genome. Therefore, Agrobacterium is a natural plant genetic transformation system. People insert the target gene into the modified T-DNA region and use Agrobacterium infection to transfer and integrate foreign genes into plant cells. Then, the transgenic plants are regenerated through cell and tissue culture techniques.
The Agrobacterium-mediated method was initially used only in dicotyledons. In recent years, Agrobacterium-mediated transformation has also been widely used in some monocotyledons (especially rice).
2. The gene gun-mediated transformation method utilizes a gunpowder explosion or high-pressure gas acceleration (this acceleration device is called a gene gun) to directly send a high-speed microprojectile containing a DNA solution with a target gene into intact plant tissues and cells, and then Through cell and tissue culture techniques, plants are regenerated and the transgene-positive plants are selected as transgenic plants. One of the major advantages of the gene gun method transformation compared to Agrobacterium transformation is that it is not limited by the range of the recipient plants. Moreover, the construction of its vector plasmid is also relatively simple, so it is also a widely used method in the current transgenic research.
3. The pollen tube channel method injects a DNA solution of the desired gene into the ovary after pollination, and uses the pollen tube channel formed during flowering and fertilization of the plant to introduce the exogenous DNA into the fertilized egg cell and is further integrated into the recipient cell. In the genome, it becomes a new individual with a transgene as the fertilized egg develops. This method was proposed by Zhou Guangyu, a scholar of China in the early 1980s. The most widely used transgenic insect-resistant cotton in China is cultivated using the pollen tube channel method. The greatest advantage of this method is that it does not rely on tissue culture to artificially regenerate plants. The technique is simple, and well-equipped laboratories are not required. Conventional breeding workers are easy to master.
The Agrobacterium-mediated method was initially used only in dicotyledons. In recent years, Agrobacterium-mediated transformation has also been widely used in some monocotyledons (especially rice).
2. The gene gun-mediated transformation method utilizes a gunpowder explosion or high-pressure gas acceleration (this acceleration device is called a gene gun) to directly send a high-speed microprojectile containing a DNA solution with a target gene into intact plant tissues and cells, and then Through cell and tissue culture techniques, plants are regenerated and the transgene-positive plants are selected as transgenic plants. One of the major advantages of the gene gun method transformation compared to Agrobacterium transformation is that it is not limited by the range of the recipient plants. Moreover, the construction of its vector plasmid is also relatively simple, so it is also a widely used method in the current transgenic research.
3. The pollen tube channel method injects a DNA solution of the desired gene into the ovary after pollination, and uses the pollen tube channel formed during flowering and fertilization of the plant to introduce the exogenous DNA into the fertilized egg cell and is further integrated into the recipient cell. In the genome, it becomes a new individual with a transgene as the fertilized egg develops. This method was proposed by Zhou Guangyu, a scholar of China in the early 1980s. The most widely used transgenic insect-resistant cotton in China is cultivated using the pollen tube channel method. The greatest advantage of this method is that it does not rely on tissue culture to artificially regenerate plants. The technique is simple, and well-equipped laboratories are not required. Conventional breeding workers are easy to master.
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