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Embryoid induction method taking polyphylla bud axis as explant

A technology of embryoid body and explant, which is applied in the field of tissue culture of Prunus polyphylla, can solve the problems of low germination rate of Pricklywood seeds and the inability to meet the needs of large-scale planting of Pricklywood plants, and achieve the effect of low cost

Inactive Publication Date: 2014-02-05
CHENGDU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the obvious "double dormancy" characteristics of incomplete embryo development, hard endosperm, post-ripening hypocotyl, etc., the seeds of Chonglou usually need two winters and one summer to germinate into seedlings after sowing, which is a typical "biennial seed". In the natural state, even after a long period of dormancy, the germination rate of the seeds of the plant is not high, so the current sexual reproduction of the seeds of the plant can not meet the needs of large-scale planting of the plant

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] (1) Pre-cultivation: Take the buds that grow vigorously on the rhizomes of P. chinensis, the diameter of the buds is more than 0.5cm, and the buds that have not germinated are washed with tap water, dried in the shade, and then disinfected with 70% alcohol on the ultra-clean bench. 10s, then sterilized with 0.1% mercuric chloride for 4 minutes, then washed with sterile water for 3 times, and repeatedly blotted the water on the surface of the sprouts with sterilized filter paper, and then inoculated in the pre-medium MS+6- BA0.5mg·L -1 + Salicylic acid 20mg·L -1 In this method, the pre-cultivation was carried out under the conditions of a culture temperature of 16°C, 6 hours of light per day, and a light intensity of 1000 lx. After 20 days of culture, the bud expansion rate was 78.93%, and the bacterial infection rate was 20.23%;

[0018] (2) Callus induction of bud axis cells: select buds that have been pre-cultured and obviously expanded, first cut off the bud sheath ...

Embodiment 2

[0023] (1) Pre-cultivation: Take the buds that grow vigorously on the rhizomes of P. chinensis, with a diameter of more than 0.5cm, and have not germinated. First, rinse them with tap water and dry them in the shade. Then use 75% alcohol on the ultra-clean bench Sterilize for 40 seconds, then sterilize with 0.1% mercury chloride for 8 minutes, then shake and wash with sterile water for 5 times, use sterilized filter paper to dry the water on the surface of the sprouts repeatedly, and then inoculate in the pre-treated medium with a pH value of 5.8. Medium MS+6-BA2.0mg·L -1 + Salicylic acid 60mg·L -1 + sucrose 40g·L -1 +Agar 6.0g·L -1 Among them, pre-cultivation was carried out under the condition that the culture temperature was 20°C, 8 hours of light per day, and the light intensity was 1500 lx. After 20 days of culture, the bud expansion rate was 90.3%, and the bacterial contamination rate was 0%;

[0024] (2) Callus induction of bud axis cells: select buds that have been ...

Embodiment 3

[0028] (1) Pre-cultivation: Take the buds that grow vigorously on the rhizomes of P. chinensis and have a diameter of more than 0.5cm and have not germinated. Rinse them with tap water first, dry them in the shade, and then disinfect them with 75% alcohol on an ultra-clean bench 60s, then sterilized with 0.15% mercuric chloride for 8 minutes, then washed with sterile water for 6 times, and repeatedly blotted the water on the surface of the sprouts with sterilized filter paper, and then inoculated in the pre-medium MS+6- BA3.0mg·L -1 + Salicylic acid 80mg·L -1 Among them, pre-cultivation was carried out under the condition that the culture temperature was 22°C, the light was 14 hours a day, and the light intensity was 2000 lx. After 20 days of culture, the bud expansion rate was 82.53%, and the bacterial contamination rate was 0%;

[0029] (2) Callus induction of bud axis cells: select buds that have been pre-cultured and obviously expanded, first cut off the bud sheath on the...

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Abstract

The invention discloses an embryoid induction method taking a polyphylla bud axis as an explant. The method comprises the following steps: firstly, carrying out disinfection and pre-cultivation on an un-germinated bud on polyphylla rhizome, and then cutting off coleoptile of the bud of which the volume obviously expands after pre-cultivation; inserting the bud axis into a callus-inducing medium to cultivate from the center after cutting in length, and then cultivating the callus tissue generated by the bud axis to obtain polyphylla embryonic tissues; finally carrying out induction cultivation on the embryonic tissues, so as to generate white spherical embryoids. By adopting the embryoid induction method, a new path is supplied for cultivation of excellent variety of the polyphylla and massive and rapid propagation of tissue culture seedlings; mass production of polyphylla plants at low cost within a short period of time can be achieved.

Description

technical field [0001] The invention relates to a method for tissue culture of a polyphylla plant, in particular to a method for inducing an embryoid body using the bud axis of a polyphylla plant as an explant. Background technique [0002] Chinese Pharmacopoeia stipulates that Paris polyphylla var. yunnanensis is the rhizome of Paris polyphylla var. yunnanensis and Paris polyphylla var. chinensis. The former is mainly produced in Yunnan, and the latter is mainly produced in Sichuan. The earliest record in "Shen Nong's Materia Medica" is Huachonglou. Zhonglou has the effects of clearing heat and detoxifying, reducing inflammation and relieving pain, promoting blood circulation and removing stasis, cooling liver and relieving convulsions. [0003] In recent years, due to the discovery that Chonglou has anti-virus and anti-tumor effects, the demand has increased greatly. Traditional Chinese medicinal materials of Chonglou have been provided with wild resources for medicinal ...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 王跃华张珏王金鹏陈燕刘银花宋超段茂华任鹏飞杨霞
Owner CHENGDU UNIV
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