Method for inducing ardisia crenata to generate hairy roots by efficiently utilizing agrobacterium rhizogenes

A technology of Agrobacterium rhizogenes and hairy roots, which is applied in the field of genetic transformation and plant tissue culture, can solve the problems of shortening the culture time and achieve the effect of high induction rate and simple operation

Pending Publication Date: 2019-11-29
SICHUAN AAS HORTICULTURE RES INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, no new method has been proposed to obtain cinnabar root hairy roots with high content of cinnabar root saponin, and can significantly shorten its culture time

Method used

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  • Method for inducing ardisia crenata to generate hairy roots by efficiently utilizing agrobacterium rhizogenes
  • Method for inducing ardisia crenata to generate hairy roots by efficiently utilizing agrobacterium rhizogenes
  • Method for inducing ardisia crenata to generate hairy roots by efficiently utilizing agrobacterium rhizogenes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A method for efficiently utilizing Agrobacterium rhizogenes to induce cinnabar roots to produce hairy roots, comprising the steps of:

[0034] 1) Activation of Agrobacterium rhizogenes ( figure 1 ):

[0035] Agrobacterium rhizogenes strain ATCC 15834 stored at -80°C in YMB+Amp 50mg·L -1 Stretch on the solid medium, place in an electric thermostat incubator at 28°C and culture in the dark until a single plaque grows, pick a single plaque in YMB+Amp 50mg L -1 In the liquid culture medium, place it in an air bath constant temperature shaker at 180rpm, 28°C and cultivate in the dark until the liquid is turbid, then streak again, pick a single plaque, and culture in liquid. This step is repeated twice. After liquid culture, suck 1mL of bacteria solution in a 100mL conical flask (containing 50mLYMB+Amp 50mg·L -1 +AS 100μmol·L -1 liquid medium) to OD 600 When the value is 0.5-0.8 (spectrophotometer), the Agrobacterium rhizogenes competent cells are prepared for dip-mediat...

experiment example 1

[0043] The induction dynamic parameter and the morphological feature statistics of embodiment 1 induction gained cinnabar root hairy root and tissue culture seedling root (control group), statistical method and result are as follows:

[0044] Table 1

[0045]

[0046] Table 2

[0047]

[0048]

[0049] Note: ACRC is the abbreviation of cinnabar root tissue culture seedling root, and ACHR is the abbreviation of cinnabar root hairy root induced by Agrobacterium rhizogenes ATCC 15834;

[0050] The data statistics method is as follows:

[0051] Induction rate (%)=number of rooted explants / number of inoculated explants×100%;

[0052] Browning rate (%) = number of browned explants / number of inoculated explants × 100%;

[0053] Callus rate (%) = number of callus explants / number of inoculated explants × 100%;

[0054] Pollution rate (%) = number of polluted explants / number of inoculated explants × 100%;

[0055] Mortality rate (%)=number of dead explants / number of inocul...

experiment example 2

[0064] The content of cinnabar root saponins in different tissues of cinnabar root was detected, and the results are shown in Table 3.

[0065] Table 3 Cinnabar root saponin content in different tissues of cinnabar root

[0066]

[0067] Therefore it can be seen that the removal of Amp 50mg L in Comparative Example 1 -1 The induction rate of cinnabar root hairy roots will be significantly reduced. In comparative example 2, no addition of cefotaxime sodium Cef can not achieve a good sterilization effect, and the explants will die soon due to contamination, and hairy roots cannot be induced. Do not get any cinnabar root saponins.

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Abstract

The invention provides a method for inducing ardisia crenata to generate hairy roots by efficiently utilizing agrobacterium rhizogenes and belongs to the technical field of plant tissue culture. The method comprises the following steps: (1) activation of the agrobacterium rhizogenes: (2) induction of the hairy roots of sterile leaves in tissue culture of ardisia crenata; (3) hairy root subculture.The method has the advantages that the method is simple in operation, capable of saving time and high in efficiency, the induction rate of the hairy roots of the ardisia crenata is high, the hairy roots of the ardisia crenata with high ardisia crenata saponin content can be obtained, the culture time of the ardisia crenata can be obviously shortened, technical support and a scientific basis are provided for industrial production, and ecological environment protection is facilitated.

Description

technical field [0001] The invention relates to the technical field of genetic transformation and plant tissue culture, and provides a method for inducing hairy roots of cinnabar roots, in particular, a method for efficiently using Agrobacterium rhizogenes to induce hairy roots from cinnabar roots. Background technique [0002] Cinnabar root (Ardisia crenata Sims) is a small evergreen shrub belonging to the genus Ardisia of the family Myrsinaceae. It is a traditional Chinese ornamental medicinal plant. In "Chinese Pharmacopoeia" (1997 edition), Cinnabar root is one of the two recorded medicinal plants of the genus Aurora. Its germplasm resources are abundant and widely distributed. In my country, it is mainly distributed in the provinces and regions south of the Yangtze River Basin where it is humid, warm and shaded. Cinnabar root is evergreen in all seasons and has a long period of fruit viewing. It can be used as indoor fruit viewing and foliage potted plants. Because of it...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00C12N15/82
CPCA01H4/005C12N15/8205
Inventor 胡菊陈睿鲜小林马明东田效琴
Owner SICHUAN AAS HORTICULTURE RES INST
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