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Induction method of Rabdosia rubescens Hemsl. Hara hairy roots, and induction expression method of oridonin

A technology of dongbeongrass, hairy roots, applied in the field of biology

Inactive Publication Date: 2016-11-30
GUANGDONG PHARMA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As for Rubescensus, there is no report on the synthesis of Rubescensin A using the hairy root induction culture system.

Method used

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  • Induction method of Rabdosia rubescens Hemsl. Hara hairy roots, and induction expression method of oridonin
  • Induction method of Rabdosia rubescens Hemsl. Hara hairy roots, and induction expression method of oridonin
  • Induction method of Rabdosia rubescens Hemsl. Hara hairy roots, and induction expression method of oridonin

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Experimental program
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Effect test

Embodiment 1

[0028] Cultivation and activation of Agrobacterium rhizogenes: spread Agrobacterium rhizogenes liquid on YNB plate, culture at 28°C for 2 days, pick a single colony, inoculate into YNB medium, 28°C, 160 r min -1 After culturing for 1 day, the activated Agrobacterium rhizogenes was inoculated into conical flasks with an inoculum volume of 1%, at 28°C, 160 r min -1 After 30 h of incubation, it can be used for infection experiments.

[0029] Induction of hairy roots: Cut the young leaves, petioles and stems from the plants of Rubescens chinensis, and wash them with 75% alcohol for 5 seconds; 0.1% mercury liter for 5 minutes; rinse with sterile single distilled water for 7 to 8 times After treatment, they were inoculated in pre-culture solid medium, and pre-cultured at 25°C for 1 day in the dark. Afterwards, soak the 1-day-pre-cultured Rubescens explants in the Agrobacterium rhizogenes solution diluted 5 times with MS liquid medium, take them out for 10, 20, and 30 min, and wipe ...

Embodiment 2

[0034] The influence of different media on the growth of the hairy root of Rubescens chinensis: four basic media were prepared respectively: MS, 1 / 2 MS, N6, B5, and the formulations of MS, N6, and B5 were referred to from "Plant Cell Engineering", 1 / 2 MS means that a large number of elements in MS are halved, and the rest remain unchanged. Sterilize at 121°C for 25 minutes, and cool to room temperature for later use.

[0035] Suspension culture of hairy roots of Rubex japonicus: Take vigorously growing and white hairy roots, inoculate them at a ratio of 0.2 g (fresh weight) hairy roots per 100 mL medium, 25°C, 75 r min -1, and the roots were taken out after 45 days of dark cultivation and weighed. For the condition of hairy roots cultured for 20d and 45d, see image 3 , 4 .

[0036] The multiplication multiples of the hairy roots of Rubescens japonicus in different basic media were B5>1 / 2 MS>MS>N6, and the fresh weight of the hairy roots in B5 reached the maximum value (se...

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Abstract

The invention belongs to the biological technology field, and more specifically discloses an induction method of Rabdosia rubescens Hemsl. Hara hairy roots, and an induction expression method of oridonin. According to the induction expression method, Rabdosia rubescens Hemsl. Hara petiole explants are infected with agrobacterium ATCC15834 for 20min, so that induction of generation of hairy roots is realized; the Rabdosia rubescens Hemsl. Hara hairy roots are cultured in B5 culture medium for 45d, and then oridonin is obtained via extraction. The induction method of Rabdosia rubescens Hemsl. Hara hairy roots is disclosed for the first time, high efficiency oridonin production method is established based on method improvement, and foundation of application of hairy root culturing technology in oridonin batch production is provided.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for inducing the hairy root of Rubescens oridinus and a method for inducing the expression of Rubescensine A. Background technique [0002] Rabdosia rubescens (Rabdosia rubescens Hemsl. Hara), also known as broken rice stalks, is a plant of the Lamiaceae genus Fragrantia, and was included in the "Pharmacopoeia of the People's Republic of China" in 1977. The dried part above the stem is used as medicine. It has the effects of clearing away heat and detoxification, promoting blood circulation and relieving pain. It is used by folks to treat hiccups (ie, esophageal cancer or cardia cancer). Studies have shown that Rubescensin A, the main active component of Rubescens oridoides, has a broad-spectrum anti-tumor effect, can inhibit the proliferation of various tumor cells and induce tumor cell apoptosis, and is effective against leukemia, esophageal cancer, gastric cancer, and col...

Claims

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Application Information

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IPC IPC(8): C12N15/82C07D493/10A01H5/00
CPCC12N15/8205C07D493/10C12N15/8243
Inventor 张鸿陆幸妍黄泽波林晓文陈俊杰陈婉琪
Owner GUANGDONG PHARMA UNIV
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