Inducing method for improving cell synchronization of tendril-leaved fritillary bulbs

Abstract

The invention discloses an inducing method for improving the cell synchronization of tendril-leaved fritillary bulbs, which belongs to the field of biotechnologies. The method disclosed by the invention is mainly implemented by carrying out synchronized inducing on the mitosis of tendril-leaved fritillary bulbs by using a helium gas blocking method and low-temperature induction. The inducing method can effectively improve the follow-up polyploidy inductivity thereof, solve the problem of variety degeneration occurring in the process of planting tendril-leaved fritillary bulbs, can obviously shorten the planting cycle, and then improve the yield and medicinal component contents of tendril-leaved fritillary bulbs. The method is simple in steps, stable in process, and suitable for industrialized large-scale production.

Description

technical field

[0001] The invention relates to a biotechnology method of Fritillaria sichuanensis, in particular to an induction method for effectively improving the synchronization of Fritillaria sichuanensis cells. Background technique

[0002] Fritillaria cirrhosa D.Don is a perennial herb of Liliaceae, whose bulbs are used as medicine. Fritillaria fritillata plants have strict requirements on the growth environment. It likes cold and cloudy climate conditions, and has the characteristics of cold resistance, humidity, shade, and high temperature. When the temperature reaches 30°C or the ground temperature exceeds 25°C, the plants will wither and cannot survive in areas with low altitude and high temperature. Therefore, Fritillaria can grow normally at an altitude of 3500m to 4500m, which makes the wild resources of Fritillaria very difficult. Scarce, can not meet the needs of the market.

[0003] Polyploid medicinal plants generally have the characteristics of giant ro...

Examples

Embodiment 1

[0019] Embodiment 1: a kind of induction method that improves the synchronization of fritillary mother cells, it comprises the following steps:

[0020] S1: Cut the callus induced by the scale leaves of Fritillaria sichuanensis into 0.5-2cm 2 The size of the callus is transferred to the culture medium that produces loose callus, the pH value is 5.6-6.5, and the culture conditions are: temperature 20-25°C, light for 8-12 hours per day, light intensity 1000-1800lx;

[0021] S2: The loose callus tissue obtained in step S1 was pressed by 5-20g·L -1 The amount of inoculation into MS+KT 0.5 ~ 1.0mg L -1 +2,4-D 0.5~2.0mg·L -1 + sucrose 10~25g·L -1 Suspension culture is carried out in liquid culture medium, pH value is 5.5-6.0, culture conditions: temperature 18-22 ℃, light 6-10h per day, light intensity 600-1000lx, shaker speed 80-100r / min , subculture once every 20-30 days;

[0022] S3: During the suspension culture process of step S2, choose to add 1-3mmol / L adenosine to the t...

Embodiment 2

[0024] Embodiment 2: a kind of induction method that improves the synchronization of fritillary mother cells, it comprises the following steps:

[0025] S1: Cut the callus induced by the scale leaves of Fritillaria sichuanensis into 0.5-2cm 2 Transfer to medium MS+6-BA 0.1~0.5mg·L for loose callus -1 +NAA 0.1~1.0mg·L -1 +2,4-D 1.0~4.0mg·L -1 + 30~50g·L sucrose -1 + Agar 4.5~6.0g·L -1 Cultivate in a medium with a pH value of 5.6-6.5, culture conditions: temperature 20-25°C, light for 8-12 hours per day, and light intensity of 1000-1800lx;

[0026] S2: The loose callus tissue obtained in step S1 was pressed by 5-20g·L -1 The amount of inoculation into MS+KT 0.5 ~ 1.0mg L -1 +2,4-D 0.5~2.0mg·L -1 + sucrose 10~25g·L -1 Suspension culture is carried out in liquid culture medium, pH value is 5.5-6.0, culture conditions: temperature 18-22 ℃, light 6-10h per day, light intensity 600-1000lx, shaker speed 80-100r / min , subculture once every 20-30 days;

[0027] S3: During the ...

Embodiment 3

[0029] Embodiment 3: a kind of induction method that improves the synchronization of fritillary mother cells, it comprises the following steps:

[0030] S1: Cut the callus induced by the scale leaves of Fritillaria sichuanensis into 0.5-2cm 2 Transfer to medium MS+6-BA 0.1mg·L for loose callus -1 +NAA 0.1mg·L -1 +2,4-D 1.0mg·L -1+ sucrose 30g·L -1 + Agar 4.5g·L -1 Cultivate in a medium with a pH value of 5.6-6.5, culture conditions: temperature 20°C, light for 8 hours a day, light intensity 1000lx;

[0031] S2: The loose callus obtained in step S1 was pressed by 5g·L -1 The amount of inoculation into MS+KT 0.5mg L -1 +2,4-D 0.5mg·L -1 + sucrose 10g·L -1 Suspension culture was carried out in liquid medium with a pH value of 5.5-6.0. Culture conditions: temperature at 18°C, light for 6 hours per day, light intensity of 600lx, and shaker speed of 80r / min, subculture once every 20 days ;

[0032] S3: During the suspension culture process of step S2, choose to add 1mmol / L...

technical field

[0001] The invention relates to a biotechnology method of Fritillaria sichuanensis, in particular to an induction method for effectively improving the synchronization of Fritillaria sichuanensis cells. Background technique

[0002] Fritillaria cirrhosa D.Don is a perennial herb of Liliaceae, whose bulbs are used as medicine. Fritillaria fritillata plants have strict requirements on the growth environment. It likes cold and cloudy climate conditions, and has the characteristics of cold resistance, humidity, shade, and high temperature. When the temperature reaches 30°C or the ground temperature exceeds 25°C, the plants will wither and cannot survive in areas with low altitude and high temperature. Therefore, Fritillaria can grow normally at an altitude of 3500m to 4500m, which makes the wild resources of Fritillaria very difficult. Scarce, can not meet the needs of the market.

[0003] Polyploid medicinal plants generally have the characteristics of giant ro...

Examples

Embodiment 1

[0019] Embodiment 1: a kind of induction method that improves the synchronization of fritillary mother cells, it comprises the following steps:

[0020] S1: Cut the callus induced by the scale leaves of Fritillaria sichuanensis into 0.5-2cm 2 The size of the callus is transferred to the culture medium that produces loose callus, the pH value is 5.6-6.5, and the culture conditions are: temperature 20-25°C, light for 8-12 hours per day, light intensity 1000-1800lx;

[0021] S2: The loose callus tissue obtained in step S1 was pressed by 5-20g·L -1 The amount of inoculation into MS+KT 0.5 ~ 1.0mg L -1 +2,4-D 0.5~2.0mg·L -1 + sucrose 10~25g·L -1 Suspension culture is carried out in liquid culture medium, pH value is 5.5-6.0, culture conditions: temperature 18-22 ℃, light 6-10h per day, light intensity 600-1000lx, shaker speed 80-100r / min , subculture once every 20-30 days;

[0022] S3: During the suspension culture process of step S2, choose to add 1-3mmol / L adenosine to the t...

Embodiment 2

[0024] Embodiment 2: a kind of induction method that improves the synchronization of fritillary mother cells, it comprises the following steps:

[0025] S1: Cut the callus induced by the scale leaves of Fritillaria sichuanensis into 0.5-2cm 2 Transfer to medium MS+6-BA 0.1~0.5mg·L for loose callus -1 +NAA 0.1~1.0mg·L -1 +2,4-D 1.0~4.0mg·L -1 + 30~50g·L sucrose -1 + Agar 4.5~6.0g·L -1 Cultivate in a medium with a pH value of 5.6-6.5, culture conditions: temperature 20-25°C, light for 8-12 hours per day, and light intensity of 1000-1800lx;

[0026] S2: The loose callus tissue obtained in step S1 was pressed by 5-20g·L -1 The amount of inoculation into MS+KT 0.5 ~ 1.0mg L -1 +2,4-D 0.5~2.0mg·L -1 + sucrose 10~25g·L -1 Suspension culture is carried out in liquid culture medium, pH value is 5.5-6.0, culture conditions: temperature 18-22 ℃, light 6-10h per day, light intensity 600-1000lx, shaker speed 80-100r / min , subculture once every 20-30 days;

[0027] S3: During the ...

Embodiment 3

[0029] Embodiment 3: a kind of induction method that improves the synchronization of fritillary mother cells, it comprises the following steps:

[0030] S1: Cut the callus induced by the scale leaves of Fritillaria sichuanensis into 0.5-2cm 2 Transfer to medium MS+6-BA 0.1mg·L for loose callus -1 +NAA 0.1mg·L -1 +2,4-D 1.0mg·L -1+ sucrose 30g·L -1 + Agar 4.5g·L -1 Cultivate in a medium with a pH value of 5.6-6.5, culture conditions: temperature 20°C, light for 8 hours a day, light intensity 1000lx;

[0031] S2: The loose callus obtained in step S1 was pressed by 5g·L -1 The amount of inoculation into MS+KT 0.5mg L -1 +2,4-D 0.5mg·L -1 + sucrose 10g·L -1 Suspension culture was carried out in liquid medium with a pH value of 5.5-6.0. Culture conditions: temperature at 18°C, light for 6 hours per day, light intensity of 600lx, and shaker speed of 80r / min, subculture once every 20 days ;

[0032] S3: During the suspension culture process of step S2, choose to add 1mmol / L...