Method for inducing generation of dactylis glomerata L.hairy roots by agrobacterium rhizogenes

A technology of Agrobacterium rhizogenes and hairy roots, applied in the biological field, can solve the problems of high cost and achieve the effects of short production cycle, strong physiological metabolic vitality, and strong proliferation

Active Publication Date: 2020-03-24
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The traditional tissue culture method mostly uses the young flower spikes, stem nodes or seedlings of Dactylis chinensis as explants to induce callus, which is time-consuming and costly.

Method used

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  • Method for inducing generation of dactylis glomerata L.hairy roots by agrobacterium rhizogenes
  • Method for inducing generation of dactylis glomerata L.hairy roots by agrobacterium rhizogenes
  • Method for inducing generation of dactylis glomerata L.hairy roots by agrobacterium rhizogenes

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Embodiment 1

[0058] A method utilizing Agrobacterium rhizogenes to induce the production of duckweed hairy roots, comprising the following steps:

[0059] (1) Activation of Agrobacterium rhizogenes:

[0060] I. Agrobacterium rhizogenes was streak inoculated to a concentration of 50 mg·L －1 Rif YEB solid medium, cultured at 28°C for 2-3 days;

[0061] II. Pick a single colony of Agrobacterium rhizogenes and inoculate it in YEB liquid medium, 28°C, 220r min －1 Shake culture for 12-16h;

[0062] III. Take 1ml of the bacterial solution obtained in step II and inoculate it with 50mg·L －1 Rif 50mLYEB liquid medium, 28°C, 220r min －1 Oscillate to OD 600 =0.5, packed in sterile EP tubes, 5000r·min －1 Centrifuge for 5min, remove the supernatant;

[0063] IV, with 200umol L －1 Resuspended in MS liquid medium of AS, 28°C and dark conditions, 220r·min －1 Shaking and culturing for 30 min, the activated Agrobacterium rhizogenes bacterial liquid was obtained.

[0064] (2) Infection: use sterile...

Embodiment 2

[0080] The difference between this embodiment and embodiment 1 is only OD 600 = 0.6.

Embodiment 3

[0082] The difference between this embodiment and embodiment 1 is only OD 600 =0.7.

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Abstract

The invention discloses a method for inducing generation of dactylis glomerata L.hairy roots by agrobacterium rhizogenes. The method comprises the following steps of (1) activating the agrobacterium rhizogenes; (2) performing infection; (3) inducing the dactylis glomerata L.hairy roots; (4) performing degerming and culturing; and (5) performing subculturing, so as to realize high efficiency induction and scaleable culture of the dactylis glomerata L.hairy roots. The production cycle is short, and the production cost is low.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for inducing the hairy roots of Dactylis chinensis by using Agrobacterium rhizogenes. Background technique [0002] Dactylis glomerata L. is a perennial herbaceous plant belonging to the Gramineae family. Duckgrass is soft, and cattle, horses, sheep, rabbits, etc. like to eat it. Young and fashionable can be fed to pigs. The amount of leaves is abundant, leaves account for 60%, and stems account for about 40%. Dactylis can be used for grazing or making hay, and it can also be harvested for green feed or making silage. Dactylis chinensis has excellent characteristics such as rapid growth, strong tillering ability, high yield, and tolerance to barrenness. It is a grass species widely used in grassland agricultural production in southern China. [0003] With the development of molecular biology, many experiments involving gene function verification require genetic transform...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12Q1/6895A01H5/06A01H6/46
CPCC12N15/8205C12Q1/6895C12N15/8241C12Q2600/13
Inventor 张新全韩佳婷许蕾冯光燕陈诚张欢黄琳凯周美亮
Owner SICHUAN AGRI UNIV
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