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Tomato transgenic method

A transgenic, tomato technology, applied in the field of agricultural biology, can solve the problems of high cost, easy pollution, and long time.

Inactive Publication Date: 2015-04-01
FRUIT TREE INST OF CHINESE ACAD OF AGRI SCI
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  • Abstract
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  • Application Information

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Problems solved by technology

[0004] To sum up, conventional transgenic methods require specialized equipment (autoclave, ultra-clean workbench, light incubator / culture room, etc.), and are time-consuming, cumbersome, easily polluted, and costly

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Embodiment Construction

[0018] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0019] The embodiment of the present invention provides a tomato transgenic method, such as figure 1 As shown, the method includes the following steps:

[0020] 101. Activate the Agrobacterium to OD600=0.6.

[0021] First pick Agrobacterium in a 50mL Erlenmeyer flask, and shake overnight in the liquid medium. In the next morning, transfer 0.5mL of the bacteria solution to a 150mL Erlenmeyer flask, add 25mL of liquid medium and continue shaking culture until O...

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Abstract

The embodiment of the invention provides a tomato transgenic method and relates to the technical field of agricultural biotechnologies. The method can be carried out not in a sterile environment, is simple, is easy to operate and is low-cost. The method comprises the following steps: agrobacterium is activated to OD600=0.6; four lateral branches are cut off from a tomato plant which has grown for 2.5-3.5 months so as to obtain four shear mouths; the shear mouths undergo impregnation by the use of the OD600=0.6 agrobacterium at the day of shear mouth cutting; a vector for transgenosis is Pcambia3301-n, and the vector contains beta-galactosidase (GUS) reporter gene; when adventitious buds which germinate at the shear mouths grow to 6-10cm, petiolule of the adventitious buds is cut for PCR / GUS dyeing; and the petiolule is dyed to blue adventitious buds which are identified as transgenic adventitious buds, and the transgenic adventitious buds are removed so as to guarantee nutrition supply for the transgenic adventitious buds.

Description

technical field [0001] The invention relates to the field of agricultural biotechnology, in particular to a tomato transgenic method. Background technique [0002] As a model plant for genetic engineering research, tomato plays an important role in the functional identification of genes related to fruit development, quality and other traits. [0003] The conventional tomato transgenic method is as follows: Step 1: Disinfect tomato seeds with alcohol and sodium hypochlorite under aseptic conditions, and then wash them with sterile water; Step 2: Sow the treated tomato seeds in pre-prepared MS medium Cultivate for 3 to 4 days at about 23°C in the dark; step 3, after the seeds germinate, transfer them to light and continue to cultivate until the cotyledons are fully expanded; step 4, cut the cotyledons under sterile conditions, Remove the two ends, keep about 0.5cm long part in the middle (the leaf back is down) and inoculate it in the pre-made differentiation medium, and pre-...

Claims

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Application Information

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IPC IPC(8): C12N15/84A01H5/00
Inventor 田义丛佩华安秀红张彩霞李武兴张利义王强杨玲
Owner FRUIT TREE INST OF CHINESE ACAD OF AGRI SCI
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