A method for constructing a rapid propagation system of Phnom Penh Boxwood

Abstract

The invention belongs to the technical field of plant cell engineering and discloses a method for performing tissue culture and rapid propagation by using Euonymus japonicus L.f. aureo-marginatus Rehd stem tips and segments as explants. The method comprises the steps of conducting cluster bud induction and rooting culture on the stem tips and the stem segments with lateral buds to obtain complete regenerated plants, and sequentially conducting callus induction, callus proliferation and differentiation and rooting culture on the stem segments without lateral buds so as to obtain regenerated plants. The regenerated plants can be obtained in the two ways, operation is simple, the pollution rate and browning rate are low in the culturing process, a large amount of seedlings can be obtained within a short time without seasonal and environmental limits, and the shortcoming of cutting propagation is overcome, meanwhile conditions are provided for genetic transformation and somaclone variation breeding of the seedlings, and the method has significant practical value on theoretical research and landscape application.

Description

technical field

[0001] The invention belongs to the technical field of plant cell engineering, and in particular relates to a method for tissue culture and rapid propagation by utilizing the stem tips and stem segments of Boxwood Phnom Penh. Background technique

[0002] Euonymus Japonicus cv. Aureo-ma is a plant of the genus Euonymus Japonicus, also known as Euonymus Japonicus euonymus, one of the varieties of Euonymus Japonicus cv. Aureo-ma. , the surface is dark green, the leaf edge is golden yellow, shiny, very ornamental value. Phnom Penh boxwood is light-loving, slightly shade-tolerant, strong adaptability, strong germination and branching ability, and resistant to pruning. It is an excellent landscaping tree species. It is widely planted in major cities, parks, universities and residential areas, and the demand for seedlings is large. . At present, Buxus Phnom Penh is often propagated by cuttings, but the propagation of cuttings is limited by seasons and materials, ...

Examples

Embodiment 1

[0031] 1. Explant pretreatment and disinfection:

[0032] Select healthy, disease-free branches from the nursery, and select young shoot tips and stem segments with axillary buds as explants. Remove the mature leaves of the explants, soak them in 4°C washing powder water for 40 minutes, then rinse them with running water at 4°C for 1 hour, soak the explants in 75% alcohol for 30 seconds on an ultra-clean table, and rinse them with sterile water for 1 hour. Twice, and then soaked in 0.1% mercuric chloride solution at 4°C. The soaking time was set in five gradients of 4min, 6min, 8min, 15min and 25min. 1mL Tween-80 and sterile water were added to the mercuricl solution. Rinse 4 to 5 times, blot excess water with sterile filter paper; transfer to primary culture medium for culture.

[0033] In order to select the most suitable pretreatment conditions, the explants treated with different disinfection time were all inoculated on MS+6-BA2.5mg / L+NAA0.5mg / L+30g / L sucrose+8g / L agar+0....

Embodiment 2

[0047] 1. Explant pretreatment and disinfection:

[0048] Select the young stems of Buxus Phnom Penh without axillary buds as explants, soak them in 4°C washing powder water for 30 minutes, then wash them with 4°C running water for 1 hour, and place the explants on a super-clean bench with a mass fraction of 75% Soak in alcohol for 30s, rinse with sterile water once or twice, and then soak in 0.1% mercuric acid solution at 4°C. The soaking time is set to five gradients of 5min, 8min, 12min, 15min, and 25min, and 1mL of mercuricl solution needs to be added Tween-80, rinse with sterile water 4 to 5 times, and blot the excess water with sterile filter paper. The explants were all inoculated on the medium with the composition of MS+6-BA2.0+NAA0.5mg / L+30g / L sucrose+6g / L agar+0.2g / L activated carbon. The culture conditions were the same as in Example 1, and the pollution rate, browning rate and healing rate were counted in two weeks.

[0049] Table 3 Effects of different times of ...

technical field

[0001] The invention belongs to the technical field of plant cell engineering, and in particular relates to a method for tissue culture and rapid propagation by utilizing the stem tips and stem segments of Boxwood Phnom Penh. Background technique

[0002] Euonymus Japonicus cv. Aureo-ma is a plant of the genus Euonymus Japonicus, also known as Euonymus Japonicus euonymus, one of the varieties of Euonymus Japonicus cv. Aureo-ma. , the surface is dark green, the leaf edge is golden yellow, shiny, very ornamental value. Phnom Penh boxwood is light-loving, slightly shade-tolerant, strong adaptability, strong germination and branching ability, and resistant to pruning. It is an excellent landscaping tree species. It is widely planted in major cities, parks, universities and residential areas, and the demand for seedlings is large. . At present, Buxus Phnom Penh is often propagated by cuttings, but the propagation of cuttings is limited by seasons and materials, ...

Examples

Embodiment 1

[0031] 1. Explant pretreatment and disinfection:

[0032] Select healthy, disease-free branches from the nursery, and select young shoot tips and stem segments with axillary buds as explants. Remove the mature leaves of the explants, soak them in 4°C washing powder water for 40 minutes, then rinse them with running water at 4°C for 1 hour, soak the explants in 75% alcohol for 30 seconds on an ultra-clean table, and rinse them with sterile water for 1 hour. Twice, and then soaked in 0.1% mercuric chloride solution at 4°C. The soaking time was set in five gradients of 4min, 6min, 8min, 15min and 25min. 1mL Tween-80 and sterile water were added to the mercuricl solution. Rinse 4 to 5 times, blot excess water with sterile filter paper; transfer to primary culture medium for culture.

[0033] In order to select the most suitable pretreatment conditions, the explants treated with different disinfection time were all inoculated on MS+6-BA2.5mg / L+NAA0.5mg / L+30g / L sucrose+8g / L agar+0....

Embodiment 2

[0047] 1. Explant pretreatment and disinfection:

[0048] Select the young stems of Buxus Phnom Penh without axillary buds as explants, soak them in 4°C washing powder water for 30 minutes, then wash them with 4°C running water for 1 hour, and place the explants on a super-clean bench with a mass fraction of 75% Soak in alcohol for 30s, rinse with sterile water once or twice, and then soak in 0.1% mercuric acid solution at 4°C. The soaking time is set to five gradients of 5min, 8min, 12min, 15min, and 25min, and 1mL of mercuricl solution needs to be added Tween-80, rinse with sterile water 4 to 5 times, and blot the excess water with sterile filter paper. The explants were all inoculated on the medium with the composition of MS+6-BA2.0+NAA0.5mg / L+30g / L sucrose+6g / L agar+0.2g / L activated carbon. The culture conditions were the same as in Example 1, and the pollution rate, browning rate and healing rate were counted in two weeks.

[0049] Table 3 Effects of different times of ...

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