Sterilization and inoculation method of pteridophyte spores

A technology for fern spores and spores, which is applied in the field of fern spore sterilization and inoculation, can solve the problems of insufficient sterilization, prolonged sterilization time, inactivation of spores, etc. Inoculation, the effect of improving inoculation efficiency

Inactive Publication Date: 2017-05-10
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the fern spores are too small, when multiple species are inoculated at the same time, the encapsulation method is easy to leak the spores and cause mixed species, and the multi-layer filter paper encapsulation may lead to insufficient sterilization; Difficult to remove waste liquid, easy to suck away spores, or potentially prolong the sterilization time due to residual disinfectant, resulting in inactivation of spores

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Material: Golden Retriever Cibotiumbarometz (Linnaeus) J.Smith.

[0026] 2. Experimental equipment: ultra-clean table, centrifuge, centrifuge tube rack, DNA purification column, pipette gun, 1ml tip, sterilizer

[0027] 3. Operation steps:

[0028] Step 1: Collect the spores, put the spore-bearing leaves of the Golden Retriever into a sulfuric acid paper bag, mark the code and place it in a ventilated and dry place, and wait for the spores to fall off freely.

[0029] Step 2: Sterilize the DNA purification column (specification: adsorption column 1ml, collection tube 2ml), 1ml pipette tip, and purified water at 121°C for 20-30min for later use.

[0030] Step 3: Spread a layer of lens-cleaning paper on the folded sulfuric acid paper, pour the spores that fall freely in the sulfuric acid paper bag on the lens-cleaning paper, shake the lens-cleaning paper slightly for simple filtration, the spores can pass through the lens-cleaning paper Pores, while impurities such a...

Embodiment 2

[0039] 1. Material: Pterisaspericaulis var.tricolor (Linden) T.Moore exE.J.Lowe.

[0040] 2. Experimental equipment: ultra-clean table, centrifuge, centrifuge tube rack, DNA purification column, pipette gun, 1ml tip, sterilizer

[0041] 3. Operation steps:

[0042] Step 1: Collect the spores, put the spore leaves of Pteris tricolor into a sulfuric acid paper bag, mark the code and place it in a ventilated and dry place, and wait for the spores to fall off freely.

[0043] Step 2: Sterilize the DNA purification column (specification: adsorption column 1ml, collection tube 2ml), 1ml pipette tip, and purified water at 121°C for 20-30min for later use.

[0044] Step 3: Spread a layer of lens-cleaning paper on the folded sulfuric acid paper, pour the spores that fall freely in the sulfuric acid paper bag on the lens-cleaning paper, shake the lens-cleaning paper slightly for simple filtration, the spores can pass through the lens-cleaning paper Pores, while impurities such as brok...

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PUM

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Abstract

The invention discloses a sterilization and inoculation method of pteridophyte spores. One set of sterilization and inoculation method applicable to trace pteridophyte spores is designed by combining a sterilization method of a tissue culture technology with tools including a DNA (Deoxyribonucleic Acid) purification column and the like, and is applicable to an in-vitro preservation process. According to the technology disclosed by the invention, the problems of insufficient sterilization and interbreeding and disinfection liquid residues, which easily occur in a disinfection process, are solved; the pollution rate can be effectively reduced and the inoculation efficiency is improved. The sterilization and inoculation method disclosed by the invention has the advantages that the trace pteridophyte spores can be efficiently disinfected and inoculated; experimental apparatuses are easy to obtain; the cost is low, the operation is convenient, the inoculation efficiency is high and the like.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a fern spore sterilization method and inoculation suitable for in vitro preservation. Background technique [0002] my country is very rich in fern resources, about 2,600 species, accounting for 1 / 5 of the world's total, many of which are endemic to China. Ferns not only play an important role in the study of biological evolution and plant phylogeny, but are also closely related to human life. Many ferns are of great value in many aspects such as medicine and ornamental. It can be seen that the development of commercial production and cultivation of ferns has broad prospects. In the process of fern tissue culture, tissue culture with spores as explants has the advantages of not damaging the mother plant and having a large reproduction coefficient, and is the most widely used propagation method for fern production. Fern spores are very small and dust-lik...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/00
Inventor 何俊李村富孟静杨俊波李德铢
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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