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Efficient Induction and Cultivation Method of Uncaria Hairy Roots

A cultivation method and high-efficiency technology, applied in botany equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems such as the decrease of the positive rate of transgenic hairy roots, and achieve the effect of improving the induction efficiency

Active Publication Date: 2021-11-19
GUIZHOU UNIV
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  • Abstract
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Problems solved by technology

[0005] Wu Shun et al. reported the induction of Uncaria hairy roots, but the induction efficiency was only 54% after optimization (Wu Shun et al., Induction of Uncaria Hairy Roots and Determination of Rhynchophylline Content, Northern Horticulture, 2019)
This method can be used to obtain hairy root culture, but if it is used as a genetic transformation system of exogenous genes, the positive rate of transgenic hairy roots will drop sharply due to the addition of screening pressure

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  • Efficient Induction and Cultivation Method of Uncaria Hairy Roots

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Embodiment 1

[0025] Embodiment 1, efficient induction and culture method of Uncaria hairy root

[0026] The efficient induction and cultivation method of Uncaria hairy root comprises the following steps:

[0027] (1) Activation of Agrobacterium rhizogenes C58C1

[0028] Get the Agrobacterium rhizogenes C58C1 preserved in the ultra-low temperature refrigerator and streak it on the YEP solid medium (YEP+40mg L -1 Rifampicin + 7.2‰ agar), 28°C, cultivated for 24-48h, picked a single colony and inoculated it in 50ml selection medium (YEP+40mg·L -1 rifampicin), 28°C, 200r / min shaker culture for 24h for the first activation, then inoculate 1ml of a viable bacterial solution into 50ml of the above-mentioned selection medium at a ratio of 1:50, 28°C, 200r min -1 The second activation culture was carried out on the shaking table, and the OD of the bacterial solution was measured 600 =0.4-0.6, collect the bacterial liquid.

[0029] (2) Determination of bacterial concentration and dipping time

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Abstract

The invention discloses a method for high-efficiency induction and cultivation of Uncaria hairy roots. The specific method is to use activated Agrobacterium rhizogenes C58C1 to impregnate pre-cultivated Uncaria explants, and then inoculate them on a co-cultivation medium for dark culture 3d, and then transferred to the hairy root induction medium for induction culture; this method has a high induction efficiency of Uncaria hairy roots, which lays the foundation for the genetic transformation of exogenous genes.

Description

technical field [0001] The invention relates to the field of tissue culture, in particular to a high-efficiency induction and culture method for uncaria hairy roots. Background technique [0002] Uncaria rhynchophylla is an authentic medicinal material in Guizhou Province. It has a wide planting area and a high demand. It has the functions of clearing blood and calming the liver, calming wind and convulsions, and is used for wind-heat headaches, colds, convulsions and other symptoms. Its medicinal active ingredients are rhynchophylline and isomorphine, which have the effect of lowering blood pressure and can effectively control a variety of cardiovascular diseases. They are widely used in clinical medicine, and their content has been used as an indicator to evaluate the quality of Uncaria medicinal materials. standard. In recent years, due to the high frequency of cardiovascular diseases, the application and demand of Uncaria in the market have increased significantly, and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 强玮张明生谭艾娟路星星付维
Owner GUIZHOU UNIV