Rapid and efficient method for induction and propagation of bryophyllum hairy roots

A technology of rooting on the ground and hairy roots, applied in the field of agricultural biotechnology research, can solve the problems of long growth cycle and low transformation efficiency, and achieve the effects of fast analysis speed, simple operation, and simple and intuitive method

Inactive Publication Date: 2019-04-16
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that Arabidopsis thaliana or induced callus regeneration are generally used in the method of gene function verification in the ground root, which has a long growth cycle and low transformation efficiency.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: a kind of method for fast and efficiently inducing big leaves to fall to the ground and take root hairy roots, comprising steps:

[0023] (1) Infection solution A: obtained by centrifuging 50 ml of A4 Agrobacterium with an OD600 value of 0.8 and resuspending it with an equal volume of MS liquid, and mixing it for later use;

[0024] (2) Infection solution B: 50 ml of A4 Agrobacterium with a GUS reporter gene with an OD600 value of 0.8 was centrifuged and resuspended with an equal volume of MS liquid, and mixed for later use;

[0025] (3) Take two bottles of 2-week-old big leaves to take root, cut them into sections, add one group to 50 ml of infecting solution A, completely soak the explants in the liquid, and infect for 5-10 minutes. The other group was added to 50 ml of infection solution B, and infected for 5-10 minutes;

[0026] (4) Place on solid medium without anti-MS, and culture in dark at 25 °C for 48 hours;

[0027] (5) After co-cultivation, tr...

Embodiment 2

[0031] Embodiment 2: a kind of method for fast and efficiently inducing bar leaves to take root hairy roots, comprising steps:

[0032] (1) Infection solution A: obtained by centrifuging 50 ml of A4 Agrobacterium with an OD 600 value of 0.8 and resuspending it with an equal volume of MS liquid, and mixing it for later use.

[0033] (2) Infection solution B: 50 ml of A4 Agrobacterium with a GUS reporter gene with an OD 600 value of 0.8 was centrifuged and resuspended with an equal volume of MS liquid, and mixed well for later use.

[0034] (3) Take two bottles of 2-week-old stick leaves to take root, and cut them into sections. One group was added to 50 ml of infection solution A, so that the explants were completely immersed in the solution, and infected for 5-10 minutes. Another group was added to 50 ml of infection solution B, and infected for 5-10 minutes.

[0035] (4) Place on MS solid medium and culture in dark at 25 °C for 48 hours.

[0036] (5) After co-cultivation, ...

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Abstract

The invention provides a rapid and efficient method for induction and propagation of bryophyllum hairy roots. The method includes the following steps that aseptic explants of bryophyllum are infectedwith a wild type GUS-reporter-gene-containing A4 agrobacterium infection solution; 2, after infection, the explants are placed on a resistance-free MS solid culture medium for co-culture under dark conditions, and co-culture is performed in a resistance MS solid culture medium; 3, screening culture is performed on the resistance MS solid culture medium; 4, subculture is performed in the resistanceMS solid culture medium; 5, enlarge cultivation is performed in an MS liquid culture medium; 6, GUS dyeing is performed on the GUS-reporter-gene-containing hairy roots to explore the transformationefficiency. The method is suitable for inducing various bryophyllum hairy roots, and has the advantages that materials are convenient to obtain, the applicability is high, operation is simple and easy, the induction speed is high, the genetic stability is achieved, and the cost is saved.

Description

technical field [0001] The invention relates to the field of agricultural biotechnology research, and in particular provides a fast and accurate research method for rapidly inducing and multiplying hairy roots that take root, and for regulating the transformation of genes related to characteristics of the root. Background technique [0002] Bryohyllm pinnatum, also known as lantern flower, belongs to Crassulaceae (Crassulaceae) Bryohyllm. It is a perennial succulent herb. It is an important Chinese medicinal material. Pain relief, detoxification and muscle growth, which contains a large number of rich chemical components, in addition to having ornamental value, it also has a wide range of pharmacological effects, and has great development and application value. In the research on the breeding and development of the ground root, the cloning and gene function verification of the gene in the ground root are particularly important. Studies have shown that Arabidopsis thaliana o...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/06A01H6/32
CPCC12N15/8205C12N15/8261
Inventor 徐秉良李世娟李隆姚军胜
Owner GANSU AGRI UNIV
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