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Method for generating large-scale sympodial bamboos through Agrobacterium mediation

An Agrobacterium-mediated method and Agrobacterium-mediated technology are applied in the field of producing large-scale clustered bamboos by using the Agrobacterium-mediated method, which can solve the problems of restricting gene expression and regulating application, and achieve the advantages of increasing probability, broad application prospects, and inhibiting pollution. Effect

Inactive Publication Date: 2012-06-13
胡尚连
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Problems solved by technology

The reason is that a large-scale industrial pulp bamboo genetic transformation system has not been established, which restricts the research on related gene expression and regulation and its application in bamboo genetic improvement.

Method used

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Embodiment Construction

[0030] Below in conjunction with embodiment the present invention is further described.

[0031] Agrobacterium-mediated method to produce the method for large cluster bamboo, comprising the steps:

[0032] a. callus induction: take mature embryos of bamboo seeds and place them on the callus induction medium MS2 to culture them for 2-3 weeks to obtain callus tissue, and after 3-4 weeks, obtain bamboo embryogenic callus tissue;

[0033] b. Pre-cultivation: the bamboo embryogenic callus in the late stage of induction is transferred to fresh callus induction medium MS2 for pre-cultivation for one week;

[0034] c. Activation and resuspension of Agrobacterium containing the target gene: Pick the Agrobacterium containing the target plasmid, streak it on the Agrobacterium medium YEB containing 50mg / L rifampicin and 100mg / L kanamycin, and store it at 28°C Cultivate in the incubator for two days; pick a single colony and continue to culture it on another fresh Agrobacterium medium YEB...

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Abstract

The invention discloses a method for generating large-scale sympodial bamboos through Agrobacterium mediation and belongs to the field of plant transgenic technology. The method comprises the following steps of: callus induction, pre-cultivation, activation and re-suspension for Agrobacterium containing target genes, infection and co-cultivation, choosing for resistant callus, differentiation andgrowing of the cluster buds, identification of transformed plant, cultivation for transformed plant, transplanting for seedlings, and the like. The invention also discloses the specific culture medium for transforming processes and the prescription of induction solution. The invention solves the problem that genetic transformation is difficult to perform on the large-scale sympodial bamboos, efficiently restrains the pollution on explant of the plant caused by Agrobacterium, shortens the period of obtaining the transgenic plant and promotes the probability of obtaining the transgenic plant. After choosing for one month, the resistant callus can be obtained and two to four months later, the regenerated transgenic plant can be obtained. The invention has the advantages of high transforming efficiency, better repeatability, significant function on bamboo hereditary improvement for large-scale sympodial bamboos pulp and bamboo functional gene research, wide prospect, and application on generating large-scale sympodial bamboos such as sinocalamus affinis, D.farinosus, and the like.

Description

Technical field [0001] The invention relates to a method for producing clustered bamboos by using a transgenic method, in particular to a method for producing large clustered bamboos by using an agrobacterium-mediated method, and belongs to the technical field of plant transgenics. Background technique [0002] At present, research on bamboo gene cloning and its genetic improvement is still in its infancy and exploration stage. In terms of gene cloning, although the genes regulating bamboo lignin-related synthetases have been cloned in Cizhu, Lvzhu and Phyllostachys edulis successively, there is no research report on the successful genetic transformation in industrial pulp bamboo. The reason is that a large-scale genetic transformation system of bamboo for industrial pulp has not been established, which restricts the research on related gene expression and regulation and its application in bamboo genetic improvement. Contents of the invention [0003] Aiming at the above ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82C12Q1/68A01H4/00A01H5/00
Inventor 胡尚连曹颖李晓瑞段宁卢学琴任鹏
Owner 胡尚连
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