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Tomato fruit gene transformation method based on agrobacterium injection

A technology of Agrobacterium injection and gene transformation, applied in botany equipment and methods, using vectors to introduce foreign genetic material, angiosperms/flowering plants, etc., can solve the problems that cannot fully reflect the changes in the fruit growth and development process, and achieve The effect of simplifying the operation process

Active Publication Date: 2015-11-25
JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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  • Description
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Problems solved by technology

The initial fruit injection method is to inject some drugs to observe the growth and development of the fruit, but these injection methods mostly use isolated fruit injection, which cannot fully reflect the growth of the fruit and the changes in the development process.

Method used

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  • Tomato fruit gene transformation method based on agrobacterium injection
  • Tomato fruit gene transformation method based on agrobacterium injection
  • Tomato fruit gene transformation method based on agrobacterium injection

Examples

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Embodiment

[0025] The test object of this embodiment selects the fruit of the tomato variety 'Micro-Tom' as the plant receptor for Agrobacterium injection, and plants it in a solar greenhouse. Micro-Tom is a tomato mutant, which retains the basic characteristics of ordinary tomato as a model plant, but the plant is short and has a shorter life cycle, which has great advantages of saving research space and shortening research time, and can make the target The gene was expressed in tomato fruit and the related gene function was studied and analyzed. During the test, the same test was repeated at least 5 times to ensure the stability and repeatability of the test.

[0026] The three Agrobacterium tumefaciens selected in this example are C58, AGLO and EHA105. These three strains are Agrobacterium strains commonly used in tomato transgenics. The expression levels of related genes after injection can determine which strain is the best for Agrobacterium injection in tomato fruits. All three st...

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Abstract

The invention discloses a tomato fruit gene transformation method based on agrobacterium injection. The method comprises the steps of conducting agrobacterium liquid cultivation, conducting agrobacterium injection on fruits in different mature stages, picking off the injected fruits at the third day, the fifth day and the seventh day after injection respectively for gus histological stain examination, and finally examining gus gene expression level. By the adoption of the method based on agrobacterium injection, gene expression can be achieved within a short period of time in the tomato fruits, and gene expression can be achieved more rapidly and conveniently compared with traditional gene transformation methods. Vital injection is adopted in the method. By the adoption of conventional methods, gene expression level can not be changed fundamentally, and the fruit development condition can not be observed through interference with certain genes or by increasing the number of certain genes beyond the standard in the growth and development process of the fruits. By the adoption of the method based on agrobacterium injection, operation processes are simplified, the change of the tomato fruits in the maturation and development process can be observed by changing the expression levels of certain genes during fruit development, and people can better learn what genes control the maturation and development of the tomato fruits.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a tomato fruit gene transformation method. Background technique [0002] Tomato (Fragaria×ananassa) is a crop with high economic value and is widely planted all over the world. It is also a common vegetable in our country and is indispensable to people's life. In recent years, with the development of tomato genetic engineering, the research on some gene functions of tomato has gradually become a hot spot, and an important way to verify the function of genes is to carry out transgenic operations. Tomato transgenic research was carried out earlier in horticultural crops, and tomato transgenic plants were successfully obtained as early as the 1960s. However, using the conventional leaf disc transformation method to carry out transgenes to study the function of genes requires obtaining transgenic plants, which can be observed and analyzed after flowering and fruiting. Ho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/08
Inventor 王媛花张立伟史红林王全智
Owner JIANGSU POLYTECHNIC COLLEGE OF AGRI & FORESTRY
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