A method for rapid propagation and detoxification of grapefruit stem tips

Abstract

The present invention relates to the technical field of detoxification and propagation of fruit trees, in particular to a method for rapid propagation and detoxification of pomelo stem tips, comprising the following steps: (1) selection and disinfection of explants, taking pomelo tubers indoors in spring or autumn After germination, spray fungicides regularly after the buds grow up. After two weeks, take the buds again. After the buds are heat-treated at 35-40 ° C, they are placed in 5% sodium hypochlorite solution for 8-10 minutes, and then placed in the dissecting mirror on the ultra-clean table for 8-10 minutes. Carry out shoot apex stripping under 40 times, select the shoot apex or nucellus tissue of terminal bud as standby; (2) primary culture, put the selected shoot apex or nucellus tissue into 5% sodium hypochlorite solution again after 3-5min, Put it into the first-generation culture medium and grow up quickly; the present invention establishes a method for rapid propagation and detoxification of grapefruit stem tips through the sterilization of explants, the selection of medium and hormones in each stage of tissue culture, and is a method for grapefruit stem tip rapid propagation and detoxification. The industrial production of tissue culture seedlings provides theoretical basis and technical support, and provides a new way for new breed selection.

Description

technical field

[0001] The invention relates to the technical field of detoxification and propagation of fruit trees, in particular to a method for rapid propagation and detoxification of pomelo stem tips. Background technique

[0002] Pomelo fruit has high nutritional value and sweet taste. It is a very distinctive local characteristic agricultural product. Planting pomelo is of great significance to increase farmers' income and improve people's living standards.

[0003] Huanglongbing is a representative disease of pomelo, which has caused the destruction of millions of pomelo orchards in my country. Virus-free treatment of fine varieties is currently the main measure to control these diseases. The stem tip microbud grafting detoxification technology is mainly a rapid propagation and detoxification technology developed by utilizing the principle that the plant shoot apex meristem has no virus and the cell division speed is fast.

[0004] Since Murashige et al. proposed m...

Examples

Embodiment 1

[0025] 1) Primary medium: MS medium + 0.1 mg / L NAA and / or 0.1 mg / L BA, pH 5.6.

[0026] 2) Secondary medium: MS medium + 0.1 mg / L NAA and / or 0.1 mg / L BA + 0.6 mg / L GA, pH 5.6.

[0027] 3) Basic medium: MS medium + 6-BA 0.2mg / L + NAA 0.2mg / L + sucrose 28g / L + agar 6g / L, pH 5.6.

[0028] Taking the configuration of 1L primary medium as an example, the specific preparation process is as follows: weigh the parts of MS medium required for 1L medium to dissolve and mix in sequence; then weigh NAA0.1mg and / or BA0.1mg, respectively added to MS solution to dissolve in turn, constant volume, adjust the pH value to 5.6; finally subpackage, sterilize at high temperature 121℃, high pressure 0.1MPa for 15-20 minutes, cool the culture medium before use.

Embodiment 2

[0030] 1) Primary medium: MS medium + 0.3 mg / L NAA and / or 0.3 mg / L BA, pH 5.8.

[0031] 2) Secondary medium: MS medium + 0.3 mg / L NAA and / or 0.3 mg / L BA + 0.8 mg / L GA, pH 5.8.

[0032] 3) Basic medium: MS medium + 6-BA 0.3mg / L + NAA 0.3mg / L + sucrose 30g / L + agar 7g / L, pH 5.8.

[0033] Taking the configuration of 1L primary medium as an example, the specific preparation process is as follows: Weigh the parts of MS medium required for 1L medium to dissolve and mix in turn; then weigh NAA0.3mg and / or BA0.3mg, respectively added to MS solution to dissolve in turn, constant volume, adjust the pH value to 5.8; finally subpackage, sterilize at high temperature 121°C, high pressure 0.1MPa for 15-20 minutes, cool the culture medium before use.

Embodiment 3

[0035] 1) Primary medium: MS medium + 0.5 mg / L NAA and / or 0.5 mg / L BA, pH 6.0.

[0036] 2) Secondary medium: MS medium + 0.5 mg / L NAA and / or 0.5 mg / L BA + 1.0 mg / L GA, pH 6.0.

[0037] 3) Basic medium: MS medium + 6-BA 0.5 mg / L + NAA 0.5 mg / L + sucrose 32 g / L + agar 8 g / L, pH 6.0.

[0038] Taking the configuration of 1L primary medium as an example, the specific preparation process is: weigh the parts of MS medium required for 1L medium to dissolve and mix in sequence; then weigh NAA0.5 mg in the primary medium formula in sequence And / or BA0.5mg, respectively added to MS solution to dissolve in turn, constant volume, adjust the pH value to 6.0; finally subpackage, sterilize at high temperature 121℃, high pressure 0.1MPa for 15-20 minutes, and use the medium after cooling .

[0039] Table 1

[0040]

[0041] It can be seen from Table 1 that the choice of medium components has little effect on tissue culture.

[0042] (2) Rapid propagation and detoxification of grapefruit...

technical field

[0001] The invention relates to the technical field of detoxification and propagation of fruit trees, in particular to a method for rapid propagation and detoxification of pomelo stem tips. Background technique

[0002] Pomelo fruit has high nutritional value and sweet taste. It is a very distinctive local characteristic agricultural product. Planting pomelo is of great significance to increase farmers' income and improve people's living standards.

[0003] Huanglongbing is a representative disease of pomelo, which has caused the destruction of millions of pomelo orchards in my country. Virus-free treatment of fine varieties is currently the main measure to control these diseases. The stem tip microbud grafting detoxification technology is mainly a rapid propagation and detoxification technology developed by utilizing the principle that the plant shoot apex meristem has no virus and the cell division speed is fast.

[0004] Since Murashige et al. proposed m...

Examples

Embodiment 1

[0025] 1) Primary medium: MS medium + 0.1 mg / L NAA and / or 0.1 mg / L BA, pH 5.6.

[0026] 2) Secondary medium: MS medium + 0.1 mg / L NAA and / or 0.1 mg / L BA + 0.6 mg / L GA, pH 5.6.

[0027] 3) Basic medium: MS medium + 6-BA 0.2mg / L + NAA 0.2mg / L + sucrose 28g / L + agar 6g / L, pH 5.6.

[0028] Taking the configuration of 1L primary medium as an example, the specific preparation process is as follows: weigh the parts of MS medium required for 1L medium to dissolve and mix in sequence; then weigh NAA0.1mg and / or BA0.1mg, respectively added to MS solution to dissolve in turn, constant volume, adjust the pH value to 5.6; finally subpackage, sterilize at high temperature 121℃, high pressure 0.1MPa for 15-20 minutes, cool the culture medium before use.

Embodiment 2

[0030] 1) Primary medium: MS medium + 0.3 mg / L NAA and / or 0.3 mg / L BA, pH 5.8.

[0031] 2) Secondary medium: MS medium + 0.3 mg / L NAA and / or 0.3 mg / L BA + 0.8 mg / L GA, pH 5.8.

[0032] 3) Basic medium: MS medium + 6-BA 0.3mg / L + NAA 0.3mg / L + sucrose 30g / L + agar 7g / L, pH 5.8.

[0033] Taking the configuration of 1L primary medium as an example, the specific preparation process is as follows: Weigh the parts of MS medium required for 1L medium to dissolve and mix in turn; then weigh NAA0.3mg and / or BA0.3mg, respectively added to MS solution to dissolve in turn, constant volume, adjust the pH value to 5.8; finally subpackage, sterilize at high temperature 121°C, high pressure 0.1MPa for 15-20 minutes, cool the culture medium before use.

Embodiment 3

[0035] 1) Primary medium: MS medium + 0.5 mg / L NAA and / or 0.5 mg / L BA, pH 6.0.

[0036] 2) Secondary medium: MS medium + 0.5 mg / L NAA and / or 0.5 mg / L BA + 1.0 mg / L GA, pH 6.0.

[0037] 3) Basic medium: MS medium + 6-BA 0.5 mg / L + NAA 0.5 mg / L + sucrose 32 g / L + agar 8 g / L, pH 6.0.

[0038] Taking the configuration of 1L primary medium as an example, the specific preparation process is: weigh the parts of MS medium required for 1L medium to dissolve and mix in sequence; then weigh NAA0.5 mg in the primary medium formula in sequence And / or BA0.5mg, respectively added to MS solution to dissolve in turn, constant volume, adjust the pH value to 6.0; finally subpackage, sterilize at high temperature 121℃, high pressure 0.1MPa for 15-20 minutes, and use the medium after cooling .

[0039] Table 1

[0040]

[0041] It can be seen from Table 1 that the choice of medium components has little effect on tissue culture.

[0042] (2) Rapid propagation and detoxification of grapefruit...

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