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Bacteriostatic agent for plant tissue culture and application of bacteriostatic agent in anoectochilus formosanus tissue culture

A plant tissue culture and bacteriostatic agent technology, applied in the direction of plant growth regulators, applications, fungicides, etc., can solve the problems of high cost of sterile seedlings, pollution, and high cost of tissue culture, etc., to improve the seedling rate , reduce pollution, and reduce production costs

Active Publication Date: 2019-03-12
ZHEJIANG JIAXING AGRI SCI ACADEMY INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The large-scale production of clematis tissue culture (hereinafter referred to as "tissue culture") is developing rapidly in my country, and the tissue culture technology is becoming more and more perfect, but it often presents different degrees of pollution during tissue culture.
At the stage of primary culture, due to pollution problems, the cost of obtaining sterile vaccines is high, or although sterile vaccines are obtained in the primary culture, a large amount of pollution occurs during subculture, which makes the cost of tissue culture remain high

Method used

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  • Bacteriostatic agent for plant tissue culture and application of bacteriostatic agent in anoectochilus formosanus tissue culture
  • Bacteriostatic agent for plant tissue culture and application of bacteriostatic agent in anoectochilus formosanus tissue culture
  • Bacteriostatic agent for plant tissue culture and application of bacteriostatic agent in anoectochilus formosanus tissue culture

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Experimental program
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Effect test

Embodiment 1

[0045] The preparation of embodiment 1 plant tissue culture antibacterial agent

[0046] 1. Preparation of plant extract:

[0047] (1) Get 40g garlic bulbs, 20g ginger rhizomes, 20g balsam pear peel and 20g pomegranate peel, after cleaning, add in 100ml of 95% ethanol, stir and pulverize to obtain a mixed solution;

[0048] (2) Ultrasonic treatment is performed on the above mixed solution. The temperature of ultrasonic treatment is 60°C, the frequency is 40KHz, and the time is 30min. After filtering to remove solid impurities, use a rotary evaporator to remove ethanol to obtain a concentrated solution, and use pure water to dilute the concentrated solution. Dilute to 100ml to obtain plant extract.

[0049] 2. Preparation of bacteriostatic agent for plant tissue culture:

[0050] Take 20ml of plant extract, 2g of chitosan oligosaccharide, 1ml of co-solvent tween20 and 60ml of pure water, put them into a 100ml volumetric flask, mix evenly, set the volume to 100ml with deionize...

Embodiment 2

[0075] The bacteriostatic property of embodiment 2 plant tissue culture bacteriostatic agent to bacterium and fungus

[0076] The plant tissue culture bacteriostatic agent that adopts embodiment 1 and comparative example 1~4 to prepare is to 3 strains of bacteria (pseudomonas, bacillus and water-dwelling Enterobacter) and 3 strains of fungi (alternaria fungus) in tissue culture pollution , Rhodotorula colloides and Penicillium) to study the antibacterial effect.

[0077] The specific content is as follows:

[0078] Bacterial inhibition test: Take an Oxford cup with a specification of Φ7.8mm×6mm×10mm (outer diameter×inner diameter×height), put it in 121°C for autoclaving, then dry and cool it for later use. Spread Bacillus, Pseudomonas and Enterobacter aquaticus on the surface of LB plate (diameter 90mm) in ultra-clean workbench, each plate contains 200μl concentration 10 6 CFU / ml of test bacteria. Put the Oxford cups on the surface of each plate, absorb 200 μl of the bacter...

Embodiment 3

[0087] Embodiment 3 Plant tissue culture uses bacteriostatic agent to the inhibitory effect of penicillium

[0088] Penicillium is smeared on the surface of clematis seedlings (tissue cultured seedlings obtained by tissue culture) in the ultra-clean workbench, and inoculated respectively in the culture medium containing or not containing the bacteriostatic agent for plant tissue culture prepared in Example 1 , placed at 25°C, lighted 12h / d, light intensity 2000lx and cultured for 3-5d.

[0089] Test results: if image 3 As shown, the contamination degree of tissue culture seedlings containing antibacterial agents was significantly lower.

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Abstract

The invention discloses a bacteriostatic agent for plant tissue culture and application of bacteriostatic agent in anoectochilus formosanus tissue culture. The bacteriostatic agent for the plant tissue culture comprises the following components in parts by calculating at 1L: 100 to 500 ml of plant extraction liquid, 5 to 20 g of chitosan oligosaccharide, 5 to 20 ml of surfactant and the balance ofwater; and the plant extraction liquid is an ethanol extract solution of a mixture of garlic bulbs, ginger roots, bitter gourd peels and pomegranate peels, and the plant extract liquid and the chitosan oligosaccharide are mixed to obtain the bacteriostatic agent. The bacteriostatic agent can effectively reduce the pollution rate of bacteria and fungus in the tissue culture, can significantly increase the seedling forming rate of the anoectochilus formosanus tissue culture, can reduce the production cost, can be widely applied to the technical field of plant biology such as tissue culture rapid propagation, seedling detoxification, regenerated plant system establishment and the like.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a bacteriostatic agent for plant tissue culture and its application in tissue culture of Aurantia chinensis. Background technique [0002] Clematis is a traditional precious medicinal material in my country, also known as golden clematis, also known as golden silkworm, golden stone pine, tree grass lotus, bird ginseng, golden thread tiger head banana, golden thread into the bone, and belongs to the genus Orchidaceae (Orchidaceae) (Anoectochilus) perennial herb. "Fujian Drugs" records that its nature and taste are sweet and flat, clearing heat and cooling blood, dispelling wind and dampness, and mainly treating hemoptysis, bronchitis, tuberculous meningitis, nephritis, cystitis, urinary tract stones, rheumatoid arthritis, and minor emergency Convulsions, Tetanus in children. "Zhonghua Yaohai" records that its nature and taste are sweet and flat, and it enters the fou...

Claims

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Application Information

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IPC IPC(8): A01N65/48A01P1/00A01P3/00A01H4/00
CPCA01H4/001A01H4/008A01N65/00A01N65/08A01N65/42A01N65/48
Inventor 李白李军王蕾
Owner ZHEJIANG JIAXING AGRI SCI ACADEMY INST
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