Method for quickly inducing carthami flos hairy roots

A hairy root and safflower technology, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of low transformation efficiency, no reported successful induction of safflower, long growth cycle, etc., and achieve fast analysis speed, Inexpensive, fast induction effect

Inactive Publication Date: 2020-05-22
GUIZHOU UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Studies have shown that Arabidopsis thaliana or induced callus regeneration are generally used in the method of gene function verification of safflower, the growth cycle is relatively long and the transformation efficiency is relatively low
The hairy root has the advantages of short growth cycle, easy cultivation, easy transformation, and high conversion rate. Especially, there is no report on the successful induction and related application of safflower in hairy root at home and abroad.

Method used

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  • Method for quickly inducing carthami flos hairy roots
  • Method for quickly inducing carthami flos hairy roots

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A method for rapidly inducing safflower hairy roots, including the steps:

[0023] (1) Infection solution A: Centrifuge with 50ml of OD value (600) 0.4-0.6 Agrobacterium K599, resuspend with 2.0 times volume of MS liquid to obtain mixing for use.

[0024] (2) Infection solution B: 50ml OD value (600) 0.4-0.6 Agrobacterium K599 with GUS reporter gene was centrifuged, resuspended with 2.0 times volume of MS liquid to obtain mixing for later use.

[0025] (3) Take two bottles of sterile safflower seedlings that are 15-20 days old.

[0026] (4) One group was added to 50 ml of infection solution A, so that the hypocotyls and cotyledons were completely immersed in the solution and infected for 10 minutes. The other group was added to 50 ml of infection solution B.

[0027] (5) Add to MS medium at 25°C and cultivate in the dark for about 24h.

[0028] (6) After dark culture is over, transfer the MS solid medium containing 200mg / ml cephalosporin for 10-25 days and then the hypocotyls and...

Embodiment 2

[0033] A method for rapidly inducing safflower hairy roots, including the steps:

[0034] (1) Infection solution A: Centrifuge with 50 ml of OD value (600) 0.8 Agrobacterium K599 and resuspend with an equal volume of MS liquid to obtain a mixture for use.

[0035] (2) Infection solution B: 50ml OD value (600)0.8 Agrobacterium K599 with GUS reporter gene, centrifuged and resuspended with an equal volume of MS liquid to get mixed for use.

[0036] (3) Take two bottles of sterile safflower seedlings about 25-30 days old.

[0037] (4) One group was added to 50 ml of infection solution A, so that the hypocotyls and cotyledons were completely immersed in the solution, and the infection was performed for 15 minutes. The other group was added to 50 ml of infection solution B.

[0038] (5) Add to MS medium at 25°C and cultivate in the dark for about 48h.

[0039] (6) After dark culture is over, transfer the MS solid medium containing 200mg / ml cephalosporin for 10-25 days and then the hypocotyls ...

Embodiment 3

[0044] A method for rapidly inducing safflower hairy roots, including the steps:

[0045] (1) Infection solution A: Centrifuge with 50ml of OD value (600) 1.0 Agrobacterium K599 and resuspend it with 0.5 times MS liquid to obtain mixing for use.

[0046] (2) Infection solution B: 50ml OD value (600) about 1.0. Agrobacterium K599 with GUS reporter gene was centrifuged and resuspended with 0.5 times volume of MS liquid to obtain mixing for use.

[0047] (3) Take two bottles of 35-42 days old sterile safflower seedlings.

[0048] (4) One group was added to 50 ml of infection solution A, so that the hypocotyls and cotyledons were completely immersed in the solution and infected for 15 minutes. The other group was added to 50 ml of infection solution B.

[0049] (5) Add it to MS medium at 25°C and incubate in the dark for about 72 hours.

[0050] (6) After dark culture is over, transfer the MS solid medium containing 200mg / ml cephalosporin for 10-25 days and then the hypocotyls and cotyledon...

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Abstract

The invention discloses a method for quickly inducing carthami flos hairy roots. The method comprises the following steps of: independently infecting an aseptic seedling of 3-4 weeks in an infection liquid A and an infection liquid B for 8-15 minutes; then, after co-culture is carried out for 24-72 hours, transferring, screening and culturing; after 10-25 days, cutting off a hairy root which quickly grows from an explant, and transferring to a subculture medium; after 15 days, transferring to a liquid culture medium for expanded shaking culture; and after 30-45 days, obtaining a great quantityof hairy roots, wherein the infection liquid A is wild agrobacterium rhizogenes K599, and the infection liquid B is K599 agrobacterium rhizogenes of a GUS ([beta]-glucuronidase) reporter gene. The method quickly induces the carthami flos hairy roots, is suitable for inducing the hairy roots of various carthami flos varieties, and has the beneficial effects of convenience in drawing materials, high applicability and a high induction speed.

Description

Technical field [0001] The invention relates to a method for inducing safflower hairy roots, in particular to a method for rapidly inducing safflower hairy roots. Background technique [0002] Safflower is a dry flower of safflower of the Compositae family, also known as grass safflower, safflower, and safflower. Its pungent and warm nature, Guixin and Liver Channel, is a good medicine for promoting blood circulation, clearing blood stasis and relieving pain. The chemical components of safflower mainly include flavonoids, alkaloids, polyacetylenes, spermidines, lignans, sesquiterpenes, organic acids, sterols, alkyl glycols, polysaccharides and Other ingredients, etc. The compounds in safflower have a wide range of pharmacological activities. They not only have certain effects on human cardiovascular, cerebrovascular, nervous system, immune system, etc., but also have various physiological activities such as anti-inflammatory and analgesic, anti-tumor, and antioxidant. Safflowe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/005
Inventor 程剑平范昱赖弟利陈星宇欧阳子泽何艾玲薛国兴周月霞
Owner GUIZHOU UNIV
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