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Ammopiptanthus mongolicus (Maxim. ex kon) Chengf. hairy root system induction and culture method for improving yield of formononetin

A technology of formononetin and a culture method, applied in the field of genetic engineering, can solve the problem of no hairy root strain activation method, report, etc., and achieve the effects of being conducive to protection and sustainable utilization, rapid growth, and not limited by seasons

Active Publication Date: 2022-01-07
GANSU ACAD OF SCI INST OF BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no relevant research on the induction and cultivation methods of the hairy roots of Ilex chinensis, and there is no relevant report on the strain activation method during the hairy root cultivation process.

Method used

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  • Ammopiptanthus mongolicus (Maxim. ex kon) Chengf. hairy root system induction and culture method for improving yield of formononetin
  • Ammopiptanthus mongolicus (Maxim. ex kon) Chengf. hairy root system induction and culture method for improving yield of formononetin
  • Ammopiptanthus mongolicus (Maxim. ex kon) Chengf. hairy root system induction and culture method for improving yield of formononetin

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Embodiment 1

[0055] Embodiment 1, a kind of induction and culture method of the hairy root system of Ilex mongolica that improves formononetin output

[0056] 1. Experimental method

[0057] (1) Obtaining Ilex mongolica explants: select plump, uniform-sized Ilex mongolica seeds, and disinfect the surface. Put the sterilized seeds on the germination medium to obtain sterile seedlings, such as figure 1 As shown, six different explant parts of epicotyl, hypocotyl, cotyledon, true leaf, stem and root were selected according to the experiment for subsequent hairy root induction experiments;

[0058] (2) Selection and activation of Agrobacterium: Select the strain of Agrobacterium, pick a single clone from the culture plate, place it in 5 mL of LB liquid medium containing the corresponding antibiotic, culture it with shaking at 28°C and 200 r / min for 20 hours; absorb 5 mL of the above bacterial liquid Inoculate in 50mL LB liquid medium for expansion culture, shake culture at 28°C and 240r / min ...

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Abstract

The invention relates to the technical field of gene engineering, in particular to an Ammopiptanthus mongolicus (Maxim. ex kon) Chengf. hairy root system induction and culture method for improving the yield of formononetin. The Ammopiptanthus mongolicus (Maxim. ex kon) Chengf. hairy root system induction and culture method comprises the following steps of (1) obtaining an Ammopiptanthus mongolicus (Maxim. ex kon) Chengf. explant; (2) carrying out Agrobacterium rhizogenes selection and activation; (3) carrying out secondary activation of Agrobacterium rhizogenes; (4) infecting the explant with the Agrobacterium rhizogenes; (5) carrying out hairy root induction and propagation; and (6) carrying out amplification culture. By means of the method, hairy roots capable of producing the formononetin can be obtained. The content of the formononetin produced by the hairy roots is remarkably increased and is increased by 14 times if being compared with that of wild plants. The Ammopiptanthus mongolicus (Maxim. ex kon) Chengf. hairy root has the advantages of high growth speed, short periods, high relative yield of secondary metabolites and the like, is not limited by seasons, and can be used for industrial production of the Ammopiptanthus mongolicus (Maxim. ex kon) Chengf. formononetin.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for inducing and cultivating the hairy root system of Ilex mongolica for increasing the yield of formononetin. Background technique [0002] Sand holly (Ammopiptantus mongolicus (Maxim.ex kon) Chengf.), also known as Mongolian sand holly and Dasha holly, belongs to the genus Ammopiptantus mongolicus of the Fabaceae Papilionaceae, and is the only super-xeric evergreen broad-leaved shrub in the desert area of ​​my country. The tree species was included in the first batch of rare and endangered plant list by the state. According to records in "Chinese Medicine Cihai", "National Chinese Herbal Medicine Compilation", and "Inner Mongolia Plant Medicine Chronicle", holly holly is a traditional national medicinal material (Mongolian medicine), named "Menghe Harigana", its branches and leaves can be used as medicine, and has the functions of dispelling rheumatism, It ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H4/00A01H5/06A01H6/54
CPCC12N15/8205C12N15/8243A01H4/005
Inventor 王沛雅方彦昊郭琪张军李鑫赵疆杨晖
Owner GANSU ACAD OF SCI INST OF BIOLOGY
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