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A method of genetic transformation of lily scales based on spray plant reactor

A genetic transformation method and reactor technology, applied in the field of genetic transformation of lily scales based on spray-type plant reactors, can solve the problems of not being able to meet the large-scale requirements of genetic transformation, the small number of roots of small bulbs, and being easily restricted by natural conditions. Achieve the effects of promoting the adsorption of receptor cells, preventing excessive sugar content, and promoting plant elongation and sprout formation

Inactive Publication Date: 2018-03-27
BEIJING AGRO BIOTECH RES CENT
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Problems solved by technology

[0004] The existing propagation methods of lily mainly include scale propagation, seed ball propagation and bulbil propagation in production, which have the advantages of slow propagation efficiency, low ball formation rate or small number of small bulbs rooting, low propagation coefficient, and are susceptible to natural conditions. The disadvantage of limitation, with the development of biotechnology, tissue culture technology has improved the reproduction efficiency to a certain extent, but it still has the disadvantages of complex operation, high cost input, pollution rate and so on
Cannot meet the large-scale demand in the process of genetic transformation

Method used

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  • A method of genetic transformation of lily scales based on spray plant reactor
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  • A method of genetic transformation of lily scales based on spray plant reactor

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Embodiment 1

[0047] Such as Figure 1 ~ Figure 3 Shown, a kind of lily scale genetic transformation method based on spray type plant reactor comprises the steps:

[0048] (A) Carry out Agrobacterium culture in the first liquid culture medium; The first liquid culture medium is LB+Rifamycin50mg / L+Km 50mg / L, volume is 250mL, culture condition is: 28 ℃, 200rpm, OD600=0.6-1.0 between.

[0049] (B) Centrifuge the bacterial liquid, collect the bacterial cells, and resuspend the bacterial cells with the second liquid medium; centrifuge the bacterial liquid at 6000rpm, collect the bacterial cells, and use 1 / 2MS+6-BA1.0mg / L+ NAA0.2mg / L+As0.2mg / L re-suspend the bacteria until OD600=0.6-1.0.

[0050] (C) Take the washed lily scales and immerse them in the resuspended thalline, take out the lily scales, immerse the base in the second liquid medium, and carry out dark culture, i.e. co-cultivation;

[0051] Take the washed lily scales and immerse them in the resuspended bacteria for 15 minutes, take ...

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Abstract

The invention discloses a method for genetic transformation of lily scales based on a spray-type plant reactor, comprising the following steps: (A) cultivating Agrobacterium in a first liquid culture medium; (B) centrifuging the bacterial liquid to collect the bacterial cells , using the second liquid medium to resuspend the thalline; (C) take the washed lily scales and immerse them in the resuspended thallus, take out the lily scales, immerse their base in the second liquid medium, and carry out dark Cultivation, that is, co-cultivation; (D) taking out the co-cultured lily scales, placing them in a spray plant reactor, inducing lily bulb buds, spraying the lily bulbs with a spray solution, and obtaining induced small bulbs; (E) Transplanting the induced bulblets for screening of transgenic plants. The method of the invention has short conversion period, low pollution rate, low cost and simple and convenient operation.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for genetic transformation of lily scales based on a spray-type plant reactor. Background technique [0002] Lily is a perennial herbaceous bulbous plant of the genus Lilium in the family Liliaceae. It is native to China and is mainly distributed in temperate regions of the northern hemisphere such as eastern Asia, Europe, and North America. At least 120 species have been found in the world, 55 of which are produced in China. In recent years, many new species have been produced through artificial hybridization, such as Asian lily, perfume lily, and fire lily. The bulbs are rich in starch, edible and also used as medicine. Lily can be eaten fresh or dried. Lily is a traditional Chinese export specialty. [0003] Lily has been a world-famous flower loved by people since ancient times. Lily has elegant posture, green leaves and graceful stems. It is a rare and precious cut f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H5/00A01H4/00C12N5/04C12N13/00A01H6/56
Inventor 张秀海杜运鹏陈绪清王璐邢礼军张倩毕彧
Owner BEIJING AGRO BIOTECH RES CENT
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