Method for establishing tissue culture regeneration system of tea tree

A technology of tea tree and system, which is applied in the field of tea tree tissue culture regeneration system, can solve the problems of somatic embryo induction and multiplication difficulty, and achieve the effect of high callus induction rate, simple multiplication method and stable genetics

Inactive Publication Date: 2012-10-17
ANHUI AGRICULTURAL UNIVERSITY
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Problems solved by technology

Experiments have proved that somatic embryos are ideal objects for genetic transformation of tea trees, Mondal [11] Even using this method to successfully obtain the first transgenic plant of tea tree, but in this method, the induction and proliferation of somatic embryos are technical difficulties
Although induction of somatic embryos is more difficult, induction of embryogenic callus is simpler

Method used

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  • Method for establishing tissue culture regeneration system of tea tree
  • Method for establishing tissue culture regeneration system of tea tree
  • Method for establishing tissue culture regeneration system of tea tree

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Experimental program
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Embodiment 1

[0029] Main equipment: ultra-clean workbench; constant temperature light incubator; high temperature and high pressure sterilization pot; pH meter

[0030] Reagents and materials:

[0031] 1. Explant material: seeds of common tea tree

[0032] 2. Main reagents:

[0033] (1) 75% ethanol, the medical 75% alcohol purchased on the market, mainly used for the disinfection of explants;

[0034] (2) 1% mercuric chloride (HgCl 2 ), weigh 1 gram of mercuric chloride and dissolve it in 1 liter of water for explant disinfection;

[0035] (3) Phytohormones: dichlorophenoxyacetic acid (2,4-D), indole butyric acid (IBA), 6-benzyladenine (6-BA), 6-furfurylaminopurine purchased on the market (KT), phenylthiadiazolyl urea (TDZ);

[0036] (4) Vermiculite and Danish Pinstop peat soil were purchased from the market.

[0037] A kind of concrete operation step that tea tree tissue culture regeneration system establishes is as follows:

[0038] 1.1 Induction of callus

[0039] Pick tea seeds...

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Abstract

The invention relates to a method for establishing tissue culture regeneration system of tea tree. The method comprises the steps of inducing callus; performing callus subculture and proliferation; inducing adventitious bud; proliferating adventitious bud; hardening seedlings; rooting and transplantation. The method includes two proliferation stages, i.e., callus proliferation and adventitious bud proliferation. The callus proliferation manner has the characteristics of simple proliferation manner and stable inheritance. After subculture for 4 years, cell activity and differentiation ability have no obvious change. The callus with low phenols content can be taken as genetic transformation object. The technology establishes solid experimental foundation for establishment of genetic transformation system of tea tree.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for establishing a tea tree tissue culture regeneration system. Background technique [0002] Plant tissue culture is an in vitro culture technology based on the theory of cell totipotency. Since its establishment in the early 20th century, it has been widely used in the rapid propagation of plants, variety improvement, genetic engineering breeding, etc. [1] . The tea tree regeneration system or tissue culture technology is the basis for establishing the tea tree genetic transformation system, which is an important technical platform for gene function verification, tea tree engineering gene breeding, and tea tree secondary metabolism research. Unfortunately, due to the lack of a suitable regeneration system, the tea tree genetic transformation system has not been established so far, which limits the development of tea tree genetic engineering a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00A01G31/00
Inventor 高丽萍王婕刘亚军贡年娣
Owner ANHUI AGRICULTURAL UNIVERSITY
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