CN116195513A - Disinfection method of orchid explants - Google Patents

Abstract

The invention discloses a method for disinfecting orchid explants. In a normal environment with bacteria, the freshly cut orchid explants are quickly and completely soaked in a transparent container of nano-silver sol, and then the transparent container is exposed to ultraviolet rays. Or after irradiating for 10-3000 seconds under visible light, the explants are taken out to complete the disinfection. The invention can carry out disinfection operation in a sterile environment, greatly improves the convenience and efficiency of operation, and overcomes the traditional cumbersome limitation of explant excision and disinfection in a sterile environment.

Description

Disinfection method of orchid explants

technical field

The invention belongs to the field of nanometer materials, and in particular relates to the application of nanometer silver sol in disinfection of orchid explants.

Background technique

The use of explants for orchid tissue culture propagation has become the most important propagation method in the orchid industry. However, in the above-mentioned propagation process, problems such as explant contamination caused by incomplete disinfection or damage to the explant caused by excessive use of disinfectants are prone to occur. Therefore, how to select a suitable disinfectant and its application method has become a key technical bottleneck restricting orchid tissue culture.

Correlative technology adopts disinfectants such as ethanol, sodium hypochlorite, hydrogen peroxide, and the antimicrobial spectrum of these disinfectants is relatively narrow, and the phenomenon of incomplete disinfection easily occurs; Simultaneously, the time required for the disinfection of these disinfectants is longer, and explants are easily caused damage, affect later reproduction. In order to overcome the above-mentioned defects of disinfectants, related technologies use mercuric chloride to disinfect explants. However, mercury chloride is a highly toxic and strong carcinogen, which is easy to pollute the environment and pose a serious threat to the health of operators. In addition, the above-mentioned related technologies all need excision, disinfection and subsequent propagation of explants in a sterile clean box, and the operation process is inconvenient and cumbersome.

To sum up, choosing an efficient, safe, and broad-spectrum disinfectant, and adopting a simple and easy disinfection method, has become a key technology that urgently needs to be solved in the field of orchid explant disinfection.

Contents of the invention

The main purpose of the present invention is to provide a flow regulator.

In order to solve the above problems, the invention provides a method for disinfecting orchid explants, which is specifically realized through the following technical solutions;

(1) Adjust the concentration of nano-silver sol to 1mg/L-1000mg/L (calculated in the same way as silver content) with sterile water or ultrapure water, and place it in a transparent vessel; the transparent Utensils can pass ultraviolet light;

(2) Under normal bacterium-containing environment, the orchid explant that is freshly cut is soaked rapidly and completely in the above-mentioned transparent container containing the nano-silver sol;

(3) After irradiating the above-mentioned quartz vessel under ultraviolet light or visible light for 10 seconds to 3000 seconds, the explants are taken out to complete the disinfection. For example, 20 seconds, 40 seconds, 60 seconds, 120 seconds, 180 seconds, 240 seconds, 300 seconds, 360 seconds, etc.

The normal environment with bacteria in the present invention refers to the indoor or outdoor environment where the air is normally circulated.

Preferably, the nano-silver sol refers to nano-silver particles and their composites with a particle size of 1nm-100nm;

Preferably, the ultraviolet radiation has a radiation wavelength of 10-400nm;

Preferably, the visible light has a radiation wavelength of 400-800nm;

Preferably, the transparent vessel comprises a quartz vessel.

Preferably, the light intensity of the ultraviolet light and visible light is 1.65×10 ¹³ -1.98×10 ¹⁸ photons/sec/cm ³ .

Preferably, it is irradiated under ultraviolet light or visible light for 60 seconds to 600 seconds.

The orchid explant disinfection method of the present invention can kill all endogenous or exogenous pathogenic microorganisms in the orchid explant. When the invention is used to sterilize the orchid explant, the bacteria-contaminated ratio of the explant in the later tissue culture is no more than 0.15%, which lays the foundation for large-scale orchid tissue culture.

Technical effect of the present invention and advantage are as follows:

(1) High-efficiency, short-term disinfection effect is achieved, while ensuring the disinfection effect, the damage to the explant is avoided;

(2) Overcome the environmental pollution and health threats of traditional mercuric chloride;

(3) The disinfection operation can be carried out in a sterile environment, which greatly improves the convenience and efficiency of the operation, and overcomes the traditional cumbersome limitation of explant removal and disinfection in a sterile environment.

Detailed ways

Hereinafter, the present invention will be described in detail in combination with examples. It should be noted that, in the case of no conflict, the embodiments in the present application and the features in the embodiments can be combined with each other.

Example 1

The present embodiment provides a method for disinfecting orchid explants. In the present embodiment, the nano-silver sol with a concentration of 50 mg/L is placed in a quartz vessel; Cut off, rinse with sterile water, place in the above-mentioned quartz vessel immediately, and irradiate the above-mentioned quartz vessel under ultraviolet light for 300 seconds (radiation light intensity is 1.65×10 ¹⁶ photons/sec/cm ³ ), take out the sprouts, The disinfection is now complete.

Example 2

This embodiment provides a method for disinfecting orchid explants. In this embodiment, the nano-silver sol with a concentration of 100mg/L is placed in a quartz vessel; Cut off, rinse with sterile water, place in the above-mentioned quartz vessel immediately, and irradiate the above-mentioned quartz vessel under ultraviolet light for 100 seconds (radiation light intensity is 3.3×10 ¹⁶ photons/sec/cm ³ ), take out the stem tip , which completes the disinfection.

Example 3

This embodiment provides a method for disinfecting orchid explants. In this embodiment, the nano-silver sol with a concentration of 200mg/L is placed in a quartz vessel; After rinsing with sterile water, place it in the above-mentioned quartz vessel immediately, and irradiate the above-mentioned quartz vessel with ultraviolet rays for 50 seconds (radiation light intensity is 1.65×10 ¹⁷ photons/sec/cm ³ ), take out the root, and complete disinfect.

Example 4

The present embodiment provides a method for disinfecting orchid explants. In the present embodiment, the nano-silver sol with a concentration of 400mg/L is placed in a quartz vessel; After being washed with sterile water, it was immediately placed in the above-mentioned quartz vessel, and the above-mentioned quartz vessel was irradiated with visible light for 50 seconds (the radiation intensity was 2.0×10 ¹⁸ photons/sec/cm ³ ), and the leaves were taken out, namely Complete disinfection.

Example 5

This embodiment provides a method for disinfecting orchid explants. In this embodiment, the nano-silver sol with a concentration of 500mg/L is placed in a quartz vessel; After being washed with sterile water, it was immediately placed in the above-mentioned quartz vessel, and the above-mentioned quartz vessel was irradiated with visible light for 200 seconds (the radiation intensity was 2.0×10 ¹⁷ photons/sec/cm ³ ), and the pedicels were taken out, namely Complete disinfection.

Example 6

This embodiment provides a method for disinfecting orchid explants. In this embodiment, the nano-silver sol with a concentration of 500mg/L is placed in a quartz vessel; After being washed with sterile water, it was immediately placed in the above-mentioned quartz vessel, and the above-mentioned quartz vessel was irradiated with ultraviolet light for 100 seconds (the radiation intensity was 2.0×10 ¹⁷ photons/sec/cm ³ ), and the pedicels were taken out. The disinfection is now complete.

Example 7

This embodiment provides a method for disinfecting orchid explants. In this embodiment, the nano-silver sol with a concentration of 600mg/L is placed in a quartz vessel; Cut off, rinse with sterile water, place in the above-mentioned quartz vessel immediately, and irradiate the above-mentioned quartz vessel under ultraviolet light for 150 seconds (radiation light intensity is 1.0×10 ¹⁸ photons/sec/cm ³ ), take out the pedicel , which completes the disinfection.

Example 8

This embodiment provides a method for disinfecting orchid explants. In this embodiment, the nano-silver sol with a concentration of 900mg/L is placed in a quartz vessel; Cut off, rinse with sterile water, immediately place in the above-mentioned quartz vessel, and irradiate the above-mentioned quartz vessel under ultraviolet light for 20 seconds (radiation light intensity is 3.0×10 ¹⁷ photons/sec/cm ³ ), take out the pedicel , which completes the disinfection.

Adopt above-mentioned embodiment, sterilized 1850 explants (embodiment 1-embodiment 7 each embodiment 250, embodiment 8 is 100), be inoculated in normal culture medium then, observe after 7 days, pollute There were only 2 cases of explants, and the contamination rate was 0.108%.

In the following comparative examples, 100 explants were taken for each comparative example.

Comparative example 1:

In a normal environment with bacteria, the young orchid embryos are cut off from the plant, washed with sterile water, soaked in 75% alcohol for 30 seconds, then disinfected with 10% sodium hypochlorite, and finally rinsed with sterile water for 3 to 5 times. Inoculated in the culture medium and observed after 7 days, the contamination rate was 57%.

Comparative example 2:

In a normal environment with bacteria, the orchid pedicel is cut off from the plant, rinsed with sterile water, soaked in 75% alcohol for 30 seconds, then disinfected with 0.5% mercury liter, and finally rinsed with sterile water for 3 to 5 times. Inoculated in the culture medium and observed after 7 days, the contamination rate was 43%.

Comparative example 2:

In a normal environment with bacteria, the orchid pedicel is cut off from the plant, rinsed with sterile water, soaked in 75% alcohol for 30 seconds, then disinfected with 10% hydrogen peroxide, and finally rinsed with sterile water for 3 to 5 times , inoculated in the culture medium, observed after 7 days, the contamination rate was 60%.

It should be noted that the parameters used in the preparations in the above examples are only for illustration and are not intended to limit the present application. For those skilled in the art, the preparation parameters can be flexibly selected according to actual needs. The nano-silver sol described in the present invention can adopt one of all nano-silver sols in the related art and its mixture. The core of the present invention is to fully integrate the photocatalytic properties and disinfection performance of the nano-silver sol.

The above is only a preferred embodiment of the present invention, so the scope of the present invention cannot be limited accordingly, that is, the equivalent changes and modifications made according to the patent scope of the present invention and the content of the specification should still be covered by the present invention within range.

Claims (8)

1. The orchid explant disinfection method is characterized in that under the normal bacteria environment, the freshly cut orchid explant is quickly and completely soaked in a transparent vessel containing nano silver sol, the transparent vessel can pass through ultraviolet rays, and then the transparent vessel is irradiated for 10 seconds to 3000 seconds under the ultraviolet rays or visible light, and the explant is taken out, so that disinfection is completed.

2. The method of sterilizing orchid explants according to claim 1, wherein the concentration of the nanosilver sol is 1mg/L to 1000mg/L calculated as silver content.

3. The method of sterilizing orchid explants according to claim 1, wherein the nano silver in the nano silver sol has a particle size of 1-100nm.

4. The method of claim 1, wherein the transparent vessel is a quartz vessel.

5. The method of sterilization of orchid explants according to claim 1, wherein the ultraviolet radiation has a wavelength of 10-400nm.

6. The method of sterilization of orchid explants according to claim 1, wherein the visible light radiation has a wavelength of 400-800nm.

7. The method of claim 1, wherein the ultraviolet or visible light has a light intensity of 1.65x10 ¹³ ～1.98×10 ¹⁸ photons/sec/cm ³ 。

8. The method of sterilization of orchid explants according to claim 1, wherein the irradiation is performed under ultraviolet or visible light for 60 seconds to 600 seconds.