Chorispora bungeana suspended embryogenic callus transformation method

A technology of alpine mustard and embryogenic callus, applied to cells modified by introducing foreign genetic material, using vectors to introduce foreign genetic material, recombinant DNA technology, etc., can solve problems such as unreported alpine mustard and achieve reproduction Fast, easy to obtain, high conversion efficiency effect

Inactive Publication Date: 2013-03-13
LANZHOU UNIVERSITY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] In recent years, embryogenic callus has attracted much attention as a receptor system for transformation, and it has been successful in crops such as citrus, tea tree, cotton, cocoa, and alfalfa, but it has not been reported in alpine mustard.

Method used

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  • Chorispora bungeana suspended embryogenic callus transformation method
  • Chorispora bungeana suspended embryogenic callus transformation method
  • Chorispora bungeana suspended embryogenic callus transformation method

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Embodiment Construction

[0032] Below, in conjunction with accompanying drawing and embodiment, the present invention is described in further detail again:

[0033] A method for transformation of alpine ion mustard suspension cells, the steps of which are carried out in five parts:

[0034] (1) Culture of suspension embryogenic callus

[0035] Sampling of alpine ion mustard. The proposed candidate criteria are no wax-secreting glands, no epidermis villi, and no thickening of the cuticle; excavate alpine mustard plants near the Tianshan glacier with a collection pole, collect plants with soil in polyethylene bags, and remove the polyethylene bags during transportation Transplant into plastic foam transport boxes.

[0036]Select robust leaves of Alpine mustard, cut off the middle part of the leaves, about 2 / 3 of the leaves, and inoculate them on the callus induction medium with sterilized forceps to induce the generation of callus. The solid basic medium for callus induction is MS, and the exogenous ...

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Abstract

The invention relates to a chorispora bungeana transformation method adopting a suspended embryogenic callus. The chorispora bungeana transformation method adopting a suspended embryogenic callus comprises the following steps of 1, suspended embryogenic callus culture, 2, bacterial strain activation and strain preparation, 3, agrobacterium tumefaciem invasion, and 4, suspended embryogenic callus and agrobacterium tumefaciem co-culture. The expression of an exogenous gene by a transgenic callus culture is identified through a GUS dye technology. After a suspended embryogenic callus obtained by co-culture in a medium containing 50 micromoles per liter of acetosyringone is dyed and then is decolored, the suspended embryogenic callus shows a large area of blue cells under an anatomical lens, and transformation efficiency is about 80%. The chorispora bungeana transformation method adopting a suspended embryogenic callus has the advantages of simple and easy operation, relaxed implementation conditions and good repeatability.

Description

technical field [0001] The invention relates to a method for transforming periglacial plants, in particular to a method for transforming a suspension embryogenic callus mediated by Agrobacterium tumefaciens. Background technique [0002] Alpine Ion mustard is a perennial plant of the genus Ion mustard in the family Cruciferae, distributed in high-altitude subalpine meadows and gravel hillsides. Its environmental conditions are characterized by cold climate, thin air, strong radiation and strong wind. The sampling site of this experiment is at 43.05N, 86.49E at Liushidu, the source area of ​​Urumqi River, at an altitude of 3600-3900m. The annual average temperature in this area is between -5 and -7°C, and the average temperature from June to August is 3.6°C, with a large daily temperature difference. It is an alpine plant accompanied by repeated freezing and thawing environments. By means of biotechnology, introducing exogenous genes into the suspended embryogenic callus of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12N5/10
Inventor 安黎哲尚琛晶张华樊小川
Owner LANZHOU UNIVERSITY
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