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Sterilization method for medium-large size plant explant

A sterilization method and explant technology, which is applied in the field of sterilization of medium and large plant explants, can solve the problems of incomplete surface disinfection, low sterilization efficiency, and difficulty in thorough disinfection, and achieve a wide range of applications and save reagents , the effect of easy operation

Active Publication Date: 2015-12-16
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Some medium and large explants, such as lily fruit, young solanaceous fruit, young fruit of Cucurbitaceae, tubers of Dioscoreaceae, etc., are relatively large in size, and generally have a length or a long diameter of more than 4cm. Using the current general disinfection technology in the industry, there are the following: Disadvantages: 1. Due to the limitation of sterilization equipment, only a few explants can be sterilized at a time, and the sterilization efficiency is low; 2. For explants with large internal cavities, such as lily fruit, the volume of the internal cavity Large, light specific gravity, low density, floating on the surface of the liquid during the whole disinfection process, which may easily lead to incomplete disinfection of the surface, resulting in pollution; 3. Dioscoreaceae tubers grow in the soil and carry a variety of fungi, so it is not easy to thoroughly Disinfection, the sterilization efficiency of traditional sterilization methods is low

Method used

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  • Sterilization method for medium-large size plant explant
  • Sterilization method for medium-large size plant explant
  • Sterilization method for medium-large size plant explant

Examples

Experimental program
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Embodiment 1

[0033] The explant of the present embodiment is the lily fruit of 45-60 days after pollination, and long diameter is 6-9 centimetre; It has larger cavity inside, specific gravity is light, density is little, as adopting traditional sterilization method to sterilize, Then it floats on the surface of the liquid during the whole disinfection process, which may easily lead to incomplete disinfection of the surface, thereby causing pollution.

[0034] The sterilization method of the present embodiment comprises the following steps:

[0035] (1) Cleaning: Pick the lily fruit 45-60 days after pollination, wash off the floating dust on the surface with water, and absorb the water with absorbent paper.

[0036] (2) Soaking: In the aseptic operating table, clamp the fruit with tweezers and rinse it back and forth three or five times in an ethanol solution with a volume percentage concentration of 95% (reusable for multiple times) for about 1-3 seconds to obtain Soaked fruit.

[0037] ...

Embodiment 2

[0049] The explants of this embodiment are tomato young fruits 15-25 days after pollination. According to the size of the fruit, tomatoes are usually divided into three types: small fruit, medium fruit and large fruit. The method of this embodiment has outstanding advantages when dealing with medium fruit, especially large fruit type. Generally, the long diameter of the medium fruit is 4-6 centimeters, and the long diameter of the large fruit is 5-7 centimeters.

[0050] The sterilization method of the present embodiment comprises the following steps:

[0051] (1) Cleaning: Pick young tomato fruits 15-25 days after pollination, wash off the floating dust on the surface with water, and absorb the water with absorbent paper.

[0052] (2) Soaking: In the aseptic operating table, the young fruit is immersed in a 500ml beaker of 95% ethanol solution (reusable for many times) with 200ml volume percentage concentration, and rolls for 1-3 seconds to ensure The surface of the young ...

Embodiment 3

[0066] The explant of the present embodiment is the eggplant young fruit 20-35 days after pollination, and the long diameter is 8-10cm.

[0067] The sterilization method of the present embodiment comprises the following steps:

[0068] (1) Cleaning: pick the young eggplant fruits 20-35 days after pollination, wash off the floating dust on the surface with water, and absorb the water with absorbent paper.

[0069] (2) Soaking: In the aseptic operating table, put the young fruit into a large beaker or an iron lunch box that is equipped with a volume percentage concentration of 95-99% ethanol solution (reusable) for 1-3 minutes. second, ensure that the surface of the young fruit is covered with ethanol solution, and obtain the young fruit after soaking.

[0070] (3) Burning and cooling: Clamp the soaked young fruit with tweezers or clips, and burn until the flame disappears completely. Burn twice in total, with a total time of 20-30 seconds, to obtain the sterilized young fruit....

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Abstract

The invention belongs to the field of plant tissue culturing, and particularly relates to a sterilization method for medium-large size plant explant. The sterilization method comprises a soaking step: the explant is soaked in an ethanol solution, and the soaked explant is obtained; a burning step: the soaked explant is burnt, and the sterilized explant is obtained. According to the sterilization method, a sodium hypochlorite solution, a mercuric chloride solution and the like are not required for sterilization of the explant, large amounts of distilled water for high-pressure sterilization is not required, accordingly, the sterilization method is simple in operation, low in cost and environment-friendly, saves reagents and has a wide use range and high application value.

Description

technical field [0001] The invention belongs to the field of plant tissue culture, and in particular relates to a method for sterilizing medium and large plant explants. Background technique [0002] Effective sterilization of explants is an important step in all plant tissue cultures. Plant tissue culture refers to intercepting the required cells, tissues or organs from the plant, and cultivating them under artificial control through aseptic operation. It is widely used in the long-term preservation of endangered and rare plants, the rapid propagation of economic plants, and distant hybridization. The rescue and cultivation of deformed embryos or immature embryos, the creation of new germplasm resources (DH, genetic engineering, cell fusion), detoxification and rejuvenation, etc. [0003] Different from disinfection in medical equipment, food processing and other industries, keeping explants still have high vitality after disinfection is the principle that must be adhered ...

Claims

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Application Information

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IPC IPC(8): A61L2/18A61L2/04A61L101/34
Inventor 赵泓王贤王永勤
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES