Culture method for reducing pollution rate of transgenic tobacco seedlings and promoting rapid rooting of transgenic tobacco seedlings

A technology of transgenic seedlings and cultivation methods, which is applied in the field of tobacco transgenics, can solve the problems of long cycle and few transformants, and achieve the effects of shortening the transformation cycle, reducing the pollution rate, and improving the transformation efficiency

Inactive Publication Date: 2017-02-01
HENAN ACAD OF AGRI SCI
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Problems solved by technology

[0003] In order to solve the technical problems existing in the prior art, the object of the present invention is to provide a kind of cultivation method that reduces the contamination rate of tobacco transgenic seedlings and promotes its rapid rooting, overcomes the deficiencies in the prior art, and utilizes the method mediated by Agrobacterium tumefaciens , using to

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  • Culture method for reducing pollution rate of transgenic tobacco seedlings and promoting rapid rooting of transgenic tobacco seedlings

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[0021] Example

[0022] A culture method for reducing the pollution rate of tobacco transgenic seedlings and promoting its rapid rooting, including the following steps:

[0023] (1) Preparation and infestation of explants: Take a good-growing Nicotiana benthamiana leaf, avoid the main vein and leaf edge, cut into small pieces of 1cm*1cm, and place them on MS medium for pre-cultivation for 2-3 days . Infect the Agrobacterium suspension infection solution containing 200μmol / L acetosyringone for 20 minutes, shake the petri dish every 5 minutes during the process, then pour out the bacterial solution, and blot the explants with sterilized absorbent paper The surface bacteria liquid is transferred to the solid co-medium (the solid co-medium formula is MS macro elements, MS trace elements, MS iron salt, 0.5 mg / L thiamine hydrochloride, 0.2 mg / L pyridoxine hydrochloride, 0.2 mg / L niacin, 150 mg / L inositol, 200 μmol / L acetosyringone, 1 mg / L naphthoxyacetic acid, 4% sucrose and 0.25% Gel...

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Abstract

The invention relates to the technical field of tobacco transgene methods, and concretely discloses a culture method for reducing the pollution rate of transgenic tobacco seedlings and promoting rapid rooting of the transgenic tobacco seedlings. A resistant test material is obtained by using selective medium screening through an Agrobacterium tumefaciems mediation technology with tobacco leaves as a receptor in order to promote rapid rooting and seedling formation; the tobacco leaves and Agrobacterium tumefaciems are infected in an Agrobacterium tumefaciems infection solution containing 200 [mu]mol/L of acetosyringone for 20 min, so Agrobacterium tumefaciems adhered to calluses are smoothly removed without influencing the growth of the receptor; a liquid co-culture medium containing 1 g/L of cephalosporin and 200 mg/L of dithiothreitol is used after co-culture is carried out for 3 d in order to clean, so the pollution rate of the receptor is greatly reduced, and the conversion efficiency is improved; a selective differentiation medium containing 200 mg/L of cephalosporin and 50 mg/L of Kan can rapidly generate calluses and a large amount of bud points; and adventitious buds with the length of 1-3 cm are transferred to a rooting medium, so rapid formation of roots is facilitated.

Description

technical field [0001] The invention relates to the technical field of tobacco transgenic methods, and in particular discloses a cultivation method for reducing the pollution rate of tobacco transgenic seedlings and promoting their rapid rooting. Background technique [0002] Tobacco is an important model plant, which plays an important role in the study of gene function, pathogenic mechanism and disease resistance mechanism of transgenic plants. Tobacco genetic modification technology is widely used. The existing problems are severe pollution, slow rooting and low seedling rate. Reducing the pollution rate of tobacco transgenic seedlings and promoting their rapid rooting has very broad application prospects. Contents of the invention [0003] In order to solve the technical problems existing in the prior art, the object of the present invention is to provide a kind of cultivation method that reduces the contamination rate of tobacco transgenic seedlings and promotes its...

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 赵凤霞高相彬宋学立李海江王正平李海峰李小杰李彦平何嘉
Owner HENAN ACAD OF AGRI SCI
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