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Tomato genetic transformation method

A technology of genetic transformation and tomato, applied in the field of genetic engineering, can solve the problems of long transformation period, cumbersome operation process, and low efficiency of tomato genetic transformation, and achieve the effects of improving transformation efficiency, simplifying the experimental process, and reducing the workload of identification

Inactive Publication Date: 2013-05-01
JILIN NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In view of the above-mentioned deficiencies, the object of the present invention is to overcome the difficulties of low tomato genetic transformation efficiency, long transformation period and cumbersome operation process, and provide a method for overall infection of tomato seeds, thereby improving the transformation efficiency of tomato

Method used

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Embodiment Construction

[0035] Below in conjunction with implementation case the present invention will be further described:

[0036] 1. Preparation and sterilization of culture medium

[0037] 1. Preparation of LB liquid medium: Weigh 10 g NaCl, 10 g peptone and 5 g yeast extract respectively into a 1 L Erlenmeyer flask, add distilled water to make the volume to 1 L, dissolve, adjust the pH value to 7.0, Seal with parafilm, then use 121 o C autoclave for 20 minutes, cool and place at 4 o C in the refrigerator, spare;

[0038] 2. Preparation of LB solid medium: Weigh 10 g NaCl, 10 g peptone, 5 g yeast extract and 15 g agar into a 1 L Erlenmeyer flask, add distilled water to 1 L, dissolve, and adjust the pH Value to 7.0, seal with parafilm, then use 121 o C autoclave with moist heat for 20 minutes, pour into a sterile petri dish, cool and place at 4 o C in the refrigerator, spare;

[0039] 3. Preparation of MS medium:

[0040] (1) Preparation of MS medium mother solution

[0041] 1) Prepara...

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Abstract

The present invention relates to a tomato genetic transformation method, belongs to the technical field of genetic engineering, and specifically discloses a whole tomato seed infection method. The method comprises the following steps: measuring a physiological imbibition curve of mature tomato seeds; culturing target gene-containing agrobacterium rhizogenes; transforming disinfected and sterilized tomato seeds into the agrobacterium rhizogenes liquid with an OD600 value of 0.8-1.3 through combining the physiological imbibition curve of the tomato seeds to carry out oscillation infection for 7-9 h; carrying out co-culture for 3 days under a dark condition and at a temperature of 25 DEG C; screening a resistant plant; carrying out rooting and cultivation on the resistant plant; and carrying out plant transforming, seedling exercising, transplantation and the like. According to the present invention, agrobacterium rhizogenes liquid infection is adopted at the tomato seed imbibition stage so as to substantially improve transformation efficiency; pressure culture medium screening is adopted to screen, such that an experimental period is shortened by 2-3 months, an experimental process is simplified, and identification workload after transformation is reduced; and important significances are provided for theoretical researches and genetic breeding practices of tomato genetic engineering.

Description

technical field [0001] The invention relates to a method for transforming tomato mediated by Agrobacterium, belonging to the technical field of genetic engineering, in particular to a method for whole tomato seed infection and transformation. Background technique [0002] Tomato is one of the most important fresh and processed raw materials grown worldwide. Its small genome makes it an ideal model plant. In addition, tomato also has the advantages of easy homozygosity for self-pollination, short growth cycle, and annual growth in the greenhouse, so its genetic transformation research has attracted much attention. Since McCormick first obtained transgenic tomato plants by Agrobacterium-mediated method in 1986, the research on genetic transformation of tomato has been continuously developed, but the transformation efficiency varies greatly. [0003] There are many factors that affect Agrobacterium-mediated genetic transformation of tomato, such as tomato genotype, infection ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00
Inventor 徐洪伟周晓馥吕杰
Owner JILIN NORMAL UNIV
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