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Disinfection method of bryophyte explants

The technology of a bryophyte and a disinfection method is applied in the field of disinfection of bryophyte explants, and can solve the problems of low bryophyte biomass, unsatisfactory disinfection effect, and great human injury.

Inactive Publication Date: 2019-04-09
SOUTHWEST UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Bryophytes generally grow in darker, moister soils, under forests, swamps, rocks and bark, etc., and absorb nutrients in the atmosphere through micropores such as gametophyte leaves. Therefore, a large number of microorganisms are attached to the explants, and it is not easy to remove
And because most of the moss explants are composed of single-layer cells, the cells are easy to die due to excessive disinfection, some scarce bryophytes are not easy to obtain sporangia, the biomass of moss is small, and it is not easy to collect, etc. These factors have greatly hindered the development of moss tissue. Development of cultivation, conservation and utilization of moss germplasm resources
Disinfection of explants is the first step in tissue culture. In the existing methods, mercuric chloride is mostly used for disinfection. If the concentration is too high, the explants will die easily, and it is easy to remain, difficult to clean and eliminate, and the disinfection effect is not ideal. Mercury chloride is a highly toxic pollutant, which is harmful to the human body, and it is easy to cause heavy metal residues in the soil when discharged into the environment

Method used

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  • Disinfection method of bryophyte explants
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  • Disinfection method of bryophyte explants

Examples

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example 1

[0039] A, select 120 robust, exuberant moss explants, after the rhizoid part of the explants is excised, put into running water and rinse for 5 minutes;

[0040] B. Then put it into the ultrasonic oscillator for cleaning for 1 hour, and thoroughly clean the dirt mixed in the junction of the stem and the leaves. The ultrasonic cleaning frequency of the ultrasonic oscillator is 40kHz;

[0041] C, then soak the cleaned explants in 500mg / L ampicillin solution, and soak for 1h under the condition of avoiding light and 4 degrees;

[0042] D. Then put the explants into the concentration of 75% alcohol and soak for 5s, use sterile water to wash for 5s, then put them into the sodium hypochlorite solution with a concentration of 1% and soak for 10s, use sterile water to wash for 5s and then put them into the concentration Soak in 75% alcohol for 5s, wash with sterile water for 5s, then soak in 1% sodium hypochlorite solution for 10s, and finally wash with sterile water for 3 times;

[...

example 2

[0047] A, select 120 robust, exuberant moss explants, after the rhizoid part of the explants is excised, put into running water and rinse for 5 minutes;

[0048] B. Then put it into the ultrasonic oscillator for cleaning for 2 hours, and thoroughly clean the dirt mixed in the junction of the stem and the leaves. The ultrasonic cleaning frequency of the ultrasonic oscillator is 40kHz;

[0049] C, then soak the cleaned explants in 500mg / L ampicillin solution, and soak for 2h under the condition of avoiding light and 4 degrees;

[0050] D. Then put the explants into the concentration of 75% alcohol and soak for 5s, use sterile water to wash for 5s, then put them into the sodium hypochlorite solution with a concentration of 1% and soak for 10s, use sterile water to wash for 5s and then put them into the concentration Soak in 75% alcohol for 5s, wash with sterile water for 5s, then soak in 1% sodium hypochlorite solution for 10s, and finally wash with sterile water for 3 times

[...

example 3

[0055] A, select 120 robust, exuberant moss explants, after the rhizoid part of the explants is excised, put into running water and rinse for 5 minutes;

[0056] B. Then put it into the ultrasonic oscillator for cleaning for 1 hour, and thoroughly clean the dirt mixed in the junction of the stem and the leaves. The ultrasonic cleaning frequency of the ultrasonic oscillator is 40kHz;

[0057] C, then soak the cleaned explants in 500mg / L ampicillin solution, and soak for 1h under the condition of avoiding light and 4 degrees;

[0058] D, then the explant is put into concentration and is soaked in 75% alcohol for 5s, utilizes sterile water to wash 5s and then puts into concentration and then soaks in 1% sodium hypochlorite solution for 10s, and finally utilizes sterile water to clean 3 times;

[0059] E, then put into the petri dish, put 4-5 explants in each petri dish, the bottom of each petri dish is equipped with sterile filter cotton, and add nutrient solution in the petri di...

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Abstract

The invention discloses a disinfection method of bryophyte explants. By use of the method, the sterilization effect is good, and disinfection waste liquor does not pollute environment, is harmless tothe human body, has low toxicity and cannot damage bryophytes easily. According to the disinfection method, the bryophyte explants are cleaned and then subjected to soaking disinfection treatment by alcohol, sodium hypochlorite, alcohol and sodium hypochlorite in sequence, next, the explants are cleaned up by sterile water and placed in a culture dish to be cultured for 14-21 days until protonemaand cormus are formed on the surface of a culture medium, the protonema and the cormus formed on the surface of the culture medium are sheared and subjected to soaking disinfection treatment by alcohol, sodium hypochlorite, alcohol and sodium hypochlorite in sequence, finally, the explants are cleaned up by sterile water, and the sterile bryophyte explants are obtained. The disinfection method issimple to operate, low in operation threshold, harmless to environment and good in test effect, requires a small quantity of explants, does not use high-pollution disinfectants and is suitable for promotion and application in the technical field of plant tissue culture.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a disinfection method for bryophyte explants. Background technique [0002] Bryophytes generally grow in darker, moister soils, under forests, swamps, rocks and bark, etc., and absorb nutrients in the atmosphere through micropores such as gametophyte leaves. Therefore, a large number of microorganisms are attached to the explants, and it is not easy to remove them. And because most of the moss explants are composed of single-layer cells, the cells are easy to die due to excessive disinfection, some scarce bryophytes are not easy to obtain sporangia, the biomass of moss is small, and it is not easy to collect, etc. These factors have greatly hindered the development of moss tissue. The development of cultivation, and the protection and utilization of moss germplasm resources. Disinfection of explants is the first step in tissue culture. In the existing methods, merc...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/00
Inventor 陈振华陈仕勇韦彦妃
Owner SOUTHWEST UNIVERSITY FOR NATIONALITIES
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