In-vitro sticktight regeneration system establishing method

An in vitro regeneration and ghost needle grass technology, applied in the field of plant tissue culture, can solve the problems of low content of medicinal ingredients, affecting processing and utilization, and easy degradation of quality, so as to overcome large quality differences, ensure seedlings, and improve stress resistance effect of ability

Active Publication Date: 2014-04-30
ZHENGZHOU NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Ghost needle grass is a good traditional Chinese medicinal material. However, because ghost needle grass is usually propagated by seeds, there are problems such as low reproduction rate, easy degradation of quality, low and unstable medicinal ingredients, etc., which affect the Its industrialized and standardized processing and utilization establishes a fast and efficient method for in vitro regeneration of P. punctatus, which can obtain whole plants with consistent properties, solve the above problems, ensure the uniformity and effective application of P. sp. It can lay the foundation for further genetic transformation research to improve its traits, but so far, there have been no reports on the establishment of tissue culture and in vitro regeneration systems of P.

Method used

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  • In-vitro sticktight regeneration system establishing method

Examples

Experimental program
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Effect test

Embodiment 1

[0017] 1. Acquisition of sterile materials: Soak the fruit of P. japonica in 75% ethanol for 15 seconds to sterilize, take it out, rinse it with sterile water for 3 times, add 0.1% high mercury chloride to sterilize, and control the time to 8 minutes. Rinse with sterile water for 4 times, dry the water with sterile filter paper, plan the fruit, take out the seeds and inoculate on MS solid medium, put them into the light incubator after inoculation, the whole operation is carried out on the ultra-clean workbench, and the cultivation conditions are as follows: The temperature is 23±2°C, the light time is 12h / d, the light intensity is 2000Lux, and it is kept dark at night. After 15 days, a sterile material with true leaves—sterile seedlings are obtained.

[0018] 2. Adventitious bud induction culture: take the explants of sterile seedlings—cotyledons and hypocotyls, see figure 1 , cotyledons cut into 0.5cm 2 Size, the leaf back is placed on the induction medium, the hypocotyl i...

Embodiment 2

[0023] 1. Acquisition of sterile materials: Soak the fruit of P. japonica in 75% ethanol for 15 seconds to sterilize, take it out, rinse it with sterile water 5 times, add 0.1% high mercury chloride to sterilize, and control the time to 8 minutes. Rinse with sterile water for 5 times, dry the water with sterile filter paper, plan the fruit, take out the seeds and inoculate on MS solid medium, put them into the light incubator after inoculation, the whole operation is carried out on the ultra-clean workbench, and the cultivation conditions are as follows: The temperature is 23±2°C, the light time is 12h / d, the light intensity is 3000Lux, and it is kept dark at night. On the 13th day, a sterile material with true leaves - sterile seedlings are obtained.

[0024] 2. Adventitious bud induction culture: take the explants of sterile seedlings—cotyledons and hypocotyls, and cut the cotyledons into 0.5cm 2Size, the leaf back is placed on the induction medium, the hypocotyls are cut in...

Embodiment 3

[0029] 1. Acquisition of sterile materials: Soak the fruit of Elephant needles in 75% ethanol for 15 seconds to sterilize, take it out, wash it with sterile water 4 times, add 0.1% high mercury chloride to sterilize, the time is controlled at 8 minutes, Rinse with sterile water for 4 times, dry the water with sterile filter paper, plan the fruit, take out the seeds and inoculate on MS solid medium, put them into the light incubator after inoculation, the whole operation is carried out on the ultra-clean workbench, and the cultivation conditions are as follows: The temperature is 23±2°C, the light time is 12h / d, the light intensity is 2500Lux, and it is kept dark at night, and a sterile material with true leaves—sterile seedlings are obtained on 14 days.

[0030] 2. Adventitious bud induction culture: take the explants of sterile seedlings—cotyledons and hypocotyls, and cut the cotyledons into 0.5cm 2 Size, the leaf back is placed on the induction medium, the hypocotyls are cut...

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Abstract

The invention relates to an in-vitro sticktight regeneration system establishing method. The In-vitro sticktight regeneration system establishing method comprises the following five stages of: cultivating aseptic seedlings, inducing and cultivating to generate adventives buds, carrying out multiplication subculture and differentiation to obtain multiple shoots, carrying out the rooting culture on the multiple shoots to obtain rooted seedlings, and transplanting the seedlings to form regeneration plants. The entire period is carried out in the controllable conditions, and is not limited by the climatic conditions and the natural sticktight growth period, the cotyledon induction ratio is 90%-95%, and the hypocotyls induction ratio is 100%. According to the in-vitro sticktight regeneration system establishing method, the reproductive cycle is greatly shortened, a large amount of seedlings can be rapidly reproduced, the harm of viruses, fungi and bacteria can be eliminated, the robust growth of the seedlings can be ensured, and a large amount of sticktights, which are consistent in shape and high in genetic stability, can be obtained.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for establishing an in vitro regeneration system of ghost needle grass. Background technique [0002] ghost needle grass ( Bidens bipinnata L.) is an annual herbaceous plant belonging to the family Asteraceae, which is distributed in most parts of the country. The stems are slightly square or cylindrical, and the young stems are sparsely pubescent, especially at the nodes. Leaves are papery, yellow-green, brittle, often shrunken or broken, and often fall off. After flattening, the intact leaves are 2-pinnately parted, lobes lanceolate, glabrous above, and sparsely hairy on the main veins below. There is often a flat disc-shaped receptacle on the top of the stem, bearing more than 10 needle-shaped, four-sided fruits, and occasionally yellow flower heads. The whole herb of ghost needle grass is used as medicine, which has the effect of clearing...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 崔波李长看蒋素华田云芳王默霏马杰
Owner ZHENGZHOU NORMAL UNIV
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