Induction culture method of radix astragali callus tissue and application of radix astragali callus tissue prepared by induction culture method

A technology of callus induction and callus, applied in the field of plant tissue culture, to achieve the effect of preventing browning, simple operation and fast proliferation

Active Publication Date: 2020-06-19
内蒙古自治区生物技术研究院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, there are many research reports on the tissue culture of Astragalus membranaceus, but about the rapid propagation of Astragalus membranaceus in vitro with sterile seedlings, and the problem of inhibiting browning in the process of tissue culture , and how to increase the content of astragaloside IV in tissue-cultured Astragalus membranaceus

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  • Induction culture method of radix astragali callus tissue and application of radix astragali callus tissue prepared by induction culture method
  • Induction culture method of radix astragali callus tissue and application of radix astragali callus tissue prepared by induction culture method
  • Induction culture method of radix astragali callus tissue and application of radix astragali callus tissue prepared by induction culture method

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preparation example Construction

[0040] (1) Preparation of aseptic seedlings of Astragalus membranaceus: soak Astragalus seeds in tap water for 24 hours, soak them in 75% ethanol for 3 minutes, and rinse them with sterile water for 5 times; Rinse 5 times; inoculate on pre-sterilized MS medium, cultivate at 25°C and 60% humidity to obtain Astragalus aseptic vaccine;

[0041] (2) Under sterile conditions, cut the stems, leaves, cotyledons and hypocotyls of aseptic seedlings of Astragalus membranaceus into small pieces of 0.6 cm, and inoculate them on medium supplemented with different growth hormones for culture, 10 pieces per bottle, each group 3 bottles, calculate the callus induction rate after 20 days, induction rate (%)=number of pieces of callus / total number of pieces×100%.

[0042] Table 1

[0043] serial number 6-BA(mg / L) NAA(mg / L) Chinese medicine extract (g / L) induction rate 1 0.5 0.2 10 96.7 2 0.7 0.2 10 100 3 1.0 0.2 10 93.3 4 0.8 0.2 20 93.3 5 ...

Embodiment 1

[0046] A method for cultivating Astragalus callus, comprising the following steps:

[0047] Step 1. Preparation of Astragalus aseptic vaccine

[0048] Soak Astragalus mongolica seeds in tap water for 24 hours, soak in 75% ethanol for 3 minutes, rinse with sterile water for 5 times; pour 1% mercuric chloride solution for 10 minutes; rinse with sterile water for 5 times; inoculate until pre-sterilized cultured on MS medium at a temperature of 25°C and a humidity of 60%, to obtain aseptic Astragalus seedlings;

[0049] Step 2. Induction culture of callus

[0050] The stem of the Astragalus aseptic seedling of step 1 is cut into 1.0cm long segment and inoculated on the callus induction medium for dark cultivation for 20d,

[0051] The callus induction medium MS medium, 6-benzylaminopurine 6-BA mass concentration 0.7mg / L, naphthaleneacetic acid NAA mass concentration 0.2mg / L, sucrose mass concentration 45g / L, agar mass concentration 10g / L L, the mass concentration of activated car...

Embodiment 2

[0056] A method for cultivating Astragalus callus, comprising the following steps:

[0057] Step 1. Preparation of Astragalus aseptic vaccine

[0058] Soak the Astragalus membranaceus seeds in tap water for 24 hours, soak them in 75% ethanol for 3 minutes, rinse them with sterile water for 5 times, pour them into 1% mercuric chloride solution and soak them for 10 minutes, rinse them with sterile water for 5 times, and inoculate them until they are extinguished in advance. Bacteria on the MS culture medium, temperature 25 ℃, humidity 60% culture, obtain Astragalus membranaceus sterile vaccine;

[0059] Step 2. Induction culture of callus

[0060] Cut the blade of the Astragalus aseptic seedling of step 1 into 0.6 cm long segments and inoculate them on the callus induction medium for dark cultivation for 20 days,

[0061] The callus induction medium MS medium, 6-benzylaminopurine 6-BA mass concentration 0.8mg / L, naphthalene acetic acid NAA mass concentration 0.4mg / L, sucrose m...

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Abstract

The invention relates to an induction culture method of radix astragali callus tissue and an application of radix astragali callus tissue prepared by the induction culture method. Radix astragali seeds are selected for germination for preparing bacteria-free radix astragali seedlings, then the bacteria-free radix astragali seedlings are subjected to subsequent callus tissue culture, additional disinfection is not needed, and being free from influence of disinfecting agents, the generated callus tissue is high in vitality and can be subcultured for a long term; besides, stems, leaves, cotyledons and hypocotyls of the radix astragali bacteria-free seedlings are subjected to induction culture, 90% or above of inductivity can be obtained; and when activated carbon and traditional Chinese medicine extracting solutions are applied to radix astragali callus tissue induction culture mediums, the inductivity can be obviously increased, and the content of astragaloside IV in the callus tissue can be increased. The method has the advantages that the inductivity is high, the growth of the callus tissue is vigorous, the proliferation speed is high, the culture cycle is short, the content of theastragaloside IV is high, the operation is simple, and browning can be effectively prevented.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a method for inducing culture of Astragalus membranaceus callus and its application. Background technique [0002] Astragalus is a perennial herb belonging to the leguminous family, and its roots have been used as medicine with a history of more than 2,000 years. The main pharmacological effects of Astragalus include invigorating qi and strengthening the surface, diuresis, detoxification and expulsion of pus, and promoting muscle growth. It is mainly used to treat shortness of breath, collapse, palpitations, spontaneous sweating, physical weakness and edema, chronic nephritis, dehydration, chronic diarrhea, qi deficiency, and insufficient qi and blood. , Sinking of Qi, has antibacterial and anti-inflammatory, antihypertensive and diuretic, detoxifying and choleretic effects, and is a commonly used Chinese herbal medicine. [0003] With the strengthening of people's ...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/005A01H4/001A01H4/008Y02A50/30
Inventor 白朕卿张秀娟随洋吴佳文田雨高新新焦巍陈启渊
Owner 内蒙古自治区生物技术研究院
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