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Efficient genetic transformation method of hybridized tulip tree

A genetic transformation method and technology of hybrid Liriodendron, applied in the directions of biochemical equipment and methods, botanical equipment and methods, angiosperms/flowering plants, etc., can solve the problems of long growth cycle of hybrid Liriodendron, difficulty in breeding, etc., To achieve the effect of reducing the generation of chimeras and good practicability

Active Publication Date: 2013-05-08
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the hybrid Liriodendron has a long growth cycle, and it is difficult to breed by conventional methods.

Method used

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  • Efficient genetic transformation method of hybridized tulip tree
  • Efficient genetic transformation method of hybridized tulip tree
  • Efficient genetic transformation method of hybridized tulip tree

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Establishment of the Transformation Receptor of Hybrid Liriodendron.

[0038] Successful gene transformation firstly depends on the establishment of a good plant receptor system. The so-called plant gene transformation receptor system means that the explants used for transformation can efficiently and stably regenerate clones through tissue culture or other non-tissue culture methods, and can accept foreign DNA integration, and are sensitive to transformation selection antibiotics regeneration system. Since the 1970s, a lot of work has been done on the plant gene transformation receptor system, and a variety of effective receptor systems have been established successively, which are suitable for the requirements of different transformation methods and different transformation purposes.

[0039] In this example, the hybrid Liriodendron suspension cell line was used as the recipient material. The establishment of suspension cell lines is related to some physic...

Embodiment 2

[0043] Example 2 Preparation of Agrobacterium.

[0044] The culture, growth state and purity of Agrobacterium play an important role in the transformation. If Agrobacterium itself grows poorly, its ability to infect is greatly reduced. If the gene of interest in Agrobacterium has been lost, it cannot be used for transformation. Therefore, the key to transformation is to prepare an Agrobacterium infection solution with high purity, vigorous growth and strong infectivity. This infecting bacteria solution is also called engineering bacteria solution.

[0045] The culture of Agrobacterium can be divided into solid plate culture and liquid shaking culture. Solid culture generally takes 2 to 3 days, and liquid culture grows faster and generally takes 1 to 2 days. Agrobacterium does not start to proliferate immediately after inoculation in liquid medium. It usually takes 1 to 2 hours to start splitting. When growth begins, the number of bacteria doubles at a constant exponential...

Embodiment 3

[0052] Example 3 Infestation.

[0053] The so-called infection is to inoculate the surface of the receptor material with engineering bacteria. The method is to add the prepared Agrobacterium bacteria solution into the culture medium of the receptor material, soak for a certain period of time, and carry out co-cultivation. During the infection process, mastering the infection time will help reduce the possible pollution during the later cultivation process and reduce the poisonous effect of bacteria on plants. The dipping time is too short to allow enough Agrobacterium to attach to the explant wound, thereby reducing the frequency of genetic transformation. If the dipping time is too long, it is easy to cause allergic reaction of the explants, and at the same time, it may cause Agrobacterium contamination in the subsequent culture process, which will eventually lead to browning and death of the explants. The specific operation is as follows:

[0054] Shake the bacterial solut...

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Abstract

The invention discloses an efficient genetic transformation method of a hybridized tulip tree. The efficient genetic transformation method comprises the following steps of: setting up a hybridized tulip tree transformation receptor, preparing agrobacterium, infecting, culturing without bacterium and culturing in a screening manner. According to the efficient genetic transformation method, unicell subjected to the suspension cultivation of the hybridized tulip tree serves as a genetic transformation acceptor system; the GUS gene is regarded as a report gene through a southern buddhism; and transient expression and long-term expression of the GUS gene are determined through a cytological method and efficient transformation of foreign gene is determined by detecting through a molecular biological method. As a somatic embryogenesis system generated by the unicell of the hybridized tulip tree is adopted, the efficient genetic transformation method of the hybridized tulip tree is good for leading in the foreign gene, and reduces generation of chimera.

Description

technical field [0001] The invention relates to a genetic transformation method of hybrid tulip tree, in particular to a highly efficient genetic transformation method of hybrid tulip tree. Background technique [0002] Hybrid Liriodendron, also known as Liriodendron chinensis (Mandarin Liriodendron), is the first artificial hybrid method by Professor Ye Peizhong from Nanjing Forestry University, which combines the Chinese hybrid Liriodendron distributed in Asia with the North American hybrid Liriodendron distributed in North America. Catalpa is a hybrid tree species bred by artificially controlled pollination. Hybrid Liriodendron has peculiar leaf shape and bright colors, and is an excellent garden cultivation and garden ornamental tree species. At the same time, the tree is tall, the trunk is round and straight, the wood structure is fine and uniform, the color is light, easy to dry, easy to process, and the fibers are long. It is an excellent wood species for pulp and pa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84A01H5/00
Inventor 陈金慧王丹施季森成铁龙王鹏凯
Owner NANJING FORESTRY UNIV
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