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Efficiency and rapid transgenic method for chrysanthemum

A transgenic, chrysanthemum technology, applied in the field of chrysanthemum transgenic system construction, can solve the problems of long time-consuming and low transformation efficiency

Inactive Publication Date: 2018-06-05
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0005] The purpose of the present invention is to provide a high-efficiency and fast transgenic method for chrysanthemums, solve the problems of low transformation efficiency and time-consuming in the traditional leaf disc transgenic process, and provide a new way for efficient transformation and gene function analysis of chrysanthemums

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  • Efficiency and rapid transgenic method for chrysanthemum
  • Efficiency and rapid transgenic method for chrysanthemum
  • Efficiency and rapid transgenic method for chrysanthemum

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Embodiment Construction

[0058] The present invention will be further described in detail in conjunction with specific implementation below.

[0059] Provided by the present invention utilizes the vacuum negative pressure method to carry out transgenic work to the chrysanthemum stem segment, and its specific implementation examples are as follows:

[0060] 1. Sampling and pre-cultivation

[0061] Taking the cultivated ground cover chrysanthemum variety 'Yuhua Luoying' as the experimental material, take tissue-cultured seedlings with a seedling age of about 30 days, remove the stem tip and leaves on the stem, and cut into about 0.5cm stem sections as explants for later use. It is cross-cut, and the lower part is obliquely cut, and the stem segment must contain undifferentiated axillary buds such as Figure 4 As shown in A. The lower mouth is obliquely cut to increase the wound area and ensure the efficiency of Agrobacterium infection. The selected explants were pre-cultured on the pre-medium for 3 da...

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Abstract

The invention discloses an efficiency and rapid transgenic method for chrysanthemum, belonging to the fields of genetic breeding of the chrysanthemum and molecular biology. The method comprises the following steps: (1) selecting a chrysanthemum stem as an explant, and carrying out pre-culturing on a pre-culturing medium; (2) activating and culturing recombinant expression vector transformed agrobacterium liquid, enriching thalli, and carrying out resuspension by virtue of an MS liquid culture medium, so as to obtain infection liquid; (3) soaking the explant pre-cultured in the step (1) into the infection liquid, and carrying out infection by virtue of a vacuum negative pressure method; and (4) putting the infected explant onto the pre-culturing medium, carrying out co-culturing in a dark condition, carrying out decarboxylation culture on a decarboxylation culture medium, finally, sequentially carrying out selective culture by virtue of a selective culture medium 1 and a selective culture medium 2, when determining that agrobacterium does not outbreak, carrying out subculture on a root culture medium, and carrying out further rooting screening, so as to obtain genetic chrysanthemumplants. Compared with a traditional leaf-disc-method transgenic method for the chrysanthemum, the method has the advantages that the transgenosis time is remarkably shortened, the conversion efficiency of transgenosis of the chrysanthemum is increaed, the workload of science researchers is reduced, a new way is provided for the efficient conversion and gene function analysis of the chrysanthemum,and a new thought is provided for the transgenosis of other plants.

Description

technical field [0001] The invention relates to the field of chrysanthemum transgenics. Different from the traditional leaf disc transgenic method, the present invention treats the chrysanthemum stems through vacuum negative pressure method, which solves the two problems of low transformation efficiency and time-consuming in the traditional leaf disc method, and realizes high efficiency. Construction of a chrysanthemum transgenic system that is simple and stable. Background technique [0002] Chrysanthemum is native to China. It is one of the top ten famous flowers in my country and one of the four major cut flowers in the world. The progress of chrysanthemum breeding and genetic research. [0003] Agrobacterium tumefaciens-mediated transgenic technology is one of the common methods to obtain genetically modified organisms, and it is also an important means to study gene functions. In the long-term practice of genetic engineering, people have noticed that due to factors suc...

Claims

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Application Information

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IPC IPC(8): C12N15/82
CPCC12N15/8205
Inventor 滕年军徐素娟赵静雅蒋甲福陈素梅陈发棣房伟民
Owner NANJING AGRICULTURAL UNIVERSITY
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