Agrobacterium-mediated dendrocalamus latiflorus transformation method

An Agrobacterium-mediated, Agrobacterium-based technology, applied in the field of forest tree genetics and breeding, can solve problems such as difficulties in transgenic hemp bamboo, and achieve the effect of increasing transformation efficiency

Active Publication Date: 2017-09-22
FUJIAN AGRI & FORESTRY UNIV
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Problems solved by technology

The present invention utilizes this method to solve the long-standing problem of difficulty in gene transfer o

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  • Agrobacterium-mediated dendrocalamus latiflorus transformation method

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Embodiment

[0028] A kind of Agrobacterium-mediated Ma bamboo transformation method specifically comprises the following steps:

[0029] (1) Preparation of explants: Cut off the young stems of Bamboo japonica growing in the greenhouse, soak them in detergent for 60 minutes, and rinse them under tap water for 2 hours. Soak in 75% ethanol for 1 minute under sterile conditions, rinse with sterile water for 5 times, then sterilize with 0.1% raw mercury for 8 minutes, and rinse with sterile water for 6 times. The sterilized stem ends were cut into 1cm segments, and the nodules were located at the end wounds; the cut stem ends were placed on callus induction medium for dark culture at 26°C; 1.5 months Afterwards, after the callus was induced, it was transferred to the callus proliferation medium and cultured in the medium for 7 months to produce light yellow embryogenic callus ( figure 1 (b)-i) in, and take it as the target of Agrobacterium infection;

[0030] (2) Preparation of Agrobacteriu...

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Abstract

The invention discloses an agrobacterium-mediated dendrocalamus latiflorus transformation method which comprises the steps of preparing explants, preparing an agrobacterium infected bacteria solution, infecting, screening, inducing of adventive buds, and inducting and rooting. agrobacterium is utilized to infect explant at stem apex of dendrocalamus latiflorus, co-culture is performed in a NB culture medium containing 2 mg/L, 2,4-D and 200 mu M acetosyringone, and steps such as screening are performed to finally obtain transgenic dendrocalamus latiflorus. The agrobacterium-mediated dendrocalamus latiflorus transformation method utilizes an agrobacterium-mediated efficient transformation system to firstly establish a transgenic system, which takes dendrocalamus latiflorus nutritive organs as start, of dendrocalamus latiflorus, and the system can overcome the obstacle that the conventional breeding method cannot be applied to bamboo, and provides premise for molecular breeding of dendrocalamus latiflorus through the transgenic technology. Through the method, technical support is provided for researching genetic functions of bamboos in the basic research field.

Description

technical field [0001] The invention belongs to the field of forest tree genetics and breeding, and in particular relates to an Agrobacterium-mediated transformation method of bamboo. Background technique [0002] Bamboo is a subgenus of Bamboo, Bamboo subfamily of Poaceae. It is naturally distributed in Fujian, Taiwan, Guangdong, Guangxi, Guizhou, Yunnan and other parts. High economic value and social value. [0003] With the development of social economy, people have higher and higher requirements on the output and quality of bamboo, but correspondingly, the bamboo breeding lags far behind. Traditional genetic breeding methods cannot be carried out in bamboo. The main reason is that the flowering of bamboo is long-term and uncertain, and hybridization between bamboo species cannot be carried out. Transgenic breeding is an important means in crop breeding. Precise gene regulation through transgenic can greatly improve the yield and quality of bamboo. [0004] At present,...

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Application Information

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IPC IPC(8): C12N15/82A01H4/00A01H5/00
Inventor 朱强叶善汶蔡昌杨唐晓珊朱彩萍尹腾飞张丽
Owner FUJIAN AGRI & FORESTRY UNIV
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