Method for quick breeding Tianshan Saussurea involucrata by application of phenyl-N'-1,2,3-thiadiazol-5-urea

Abstract

An application of the plant growth regulator TDZ (N-phenyl-N'-1,2,3-thiadiazine- 5-urea) in fast reproduction of Tianshan mountain snow lotus and control its morphogenesis is disclosed.

Description

technical field

[0001] The invention belongs to the field of biotechnology, in particular to the use of novel plant growth regulator TDZ (N-phenyl-N'-1,2,3-thiadiazole-5-urea) in Tianshan Snow Lotus (Saussurea involucrata Kar.et Kir.exMaxim ) application method in rapid propagation. Background technique

[0002] Tianshan Snow Lotus belongs to Compositae (Comppositae) Trib.Cynareae Less. (Sausgurea sp.) plant, is a kind of precious Chinese herbal medicine commonly used by folks in high mountain areas of our country. Snow lotus is warm in nature and slightly bitter in taste. It has the functions of dispelling cold and dehumidification, strengthening tendons and yang, anti-inflammatory and analgesic, and shrinking the uterus. . Its main active ingredients are flavonoids, alkaloids, and polysaccharides.

[0003] Tianshan Saussurea is a perennial herb that grows slowly in an environment above the snow line at an altitude of 2500-3000 meters; Tianshan Saussurea is a one-off flo...

Examples

Embodiment 1

[0018] 1. Take the seeds of Snow Lotus from Tianshan, sterilize the surface with 10% sodium hypochlorite solution for 20 minutes, rinse with sterile water for 3 to 5 times, and then connect them to hormone-free MS solid medium. After the seedlings grow for 30 days, take the leaves and cut them into 0.5cm 2 The small pieces were placed on MS medium containing 0.5 μM TDZ to induce dedifferentiation, the culture temperature was 25° C., the light intensity was 4000 Lux, and the photoperiod was 16 hours.

[0019] 2. After culturing for 35 days, transfer the generated callus to a medium containing 0.1 μM TDZ for subculture, and treat at 4° C. for 2 weeks every 3 generations. Then transfer to MS medium containing 0.5 μM TDZ to continue culturing, culture temperature is 25±2°C, light intensity is 4000 Lux, and photoperiod is 16 hours.

[0020] 3. The callus cultured for 25 days was transferred to MS hormone-free medium to continue culturing, the culture temperature was 25±2°C, the li...

Embodiment 2

[0024] 1. Take the seeds of Snow Lotus from Tianshan, sterilize the surface with 10% sodium hypochlorite solution for 20 minutes, rinse the excess sterilizing solution with sterile water, and then connect it to the hormone-free MS solid medium. After the seedlings grow for 30 days, take the leaves and cut them into 0.5cm 2 The small pieces were cultured on the culture medium containing 1 μM TDZ, the culture temperature was 25° C., the light intensity was 3500 Lux, and the photoperiod was 16 hours.

[0025] 2. After 35 days of culture, the callus produced was transferred to MS medium containing 0.25 μM TDZ for subculture, and treated at 4° C. for 1 week every 3 generations. Then transfer to culture medium containing 1 μM TDZ, culture temperature is 25±2°C, light intensity is 4000 Lux, and photoperiod is 16 hours.

[0026] 3. The callus cultured for 25 days was transferred to MS hormone-free medium to continue culturing, the culture temperature was 25±2°C, the light intensity w...

Embodiment 3

[0030] 1. Take the seeds of Snow Lotus from Tianshan, sterilize the surface with 0.1% mercuric chloride solution for 12 minutes, rinse with sterile water for 3 to 5 times, and then connect to hormone-free MS solid medium. After the seedlings grow for 30 days, take the leaves and cut them into 0.5cm 2 The small pieces were placed on MS medium containing 0.5 μM TDZ to induce dedifferentiation, the culture temperature was 25° C., the light intensity was 4000 Lux, and the photoperiod was 16 hours.

[0031] 2. After culturing for 35 days, transfer the generated callus to a medium containing 1 μM TDZ for subculture, and treat it at 4° C. for 5 weeks every 3 generations. Then transfer to MS medium containing 0.5 μM TDZ to continue culturing, culture temperature is 25±2°C, light intensity is 4000 Lux, and photoperiod is 16 hours.

[0032] 3. The callus cultured for 25 days was transferred to MS hormone-free medium to continue culturing, the culture temperature was 25±2°C, the light i...

technical field

[0001] The invention belongs to the field of biotechnology, in particular to the use of novel plant growth regulator TDZ (N-phenyl-N'-1,2,3-thiadiazole-5-urea) in Tianshan Snow Lotus (Saussurea involucrata Kar.et Kir.exMaxim ) application method in rapid propagation. Background technique

[0002] Tianshan Snow Lotus belongs to Compositae (Comppositae) Trib.Cynareae Less. (Sausgurea sp.) plant, is a kind of precious Chinese herbal medicine commonly used by folks in high mountain areas of our country. Snow lotus is warm in nature and slightly bitter in taste. It has the functions of dispelling cold and dehumidification, strengthening tendons and yang, anti-inflammatory and analgesic, and shrinking the uterus. . Its main active ingredients are flavonoids, alkaloids, and polysaccharides.

[0003] Tianshan Saussurea is a perennial herb that grows slowly in an environment above the snow line at an altitude of 2500-3000 meters; Tianshan Saussurea is a one-off flo...

Examples

Embodiment 1

[0018] 1. Take the seeds of Snow Lotus from Tianshan, sterilize the surface with 10% sodium hypochlorite solution for 20 minutes, rinse with sterile water for 3 to 5 times, and then connect them to hormone-free MS solid medium. After the seedlings grow for 30 days, take the leaves and cut them into 0.5cm 2 The small pieces were placed on MS medium containing 0.5 μM TDZ to induce dedifferentiation, the culture temperature was 25° C., the light intensity was 4000 Lux, and the photoperiod was 16 hours.

[0019] 2. After culturing for 35 days, transfer the generated callus to a medium containing 0.1 μM TDZ for subculture, and treat at 4° C. for 2 weeks every 3 generations. Then transfer to MS medium containing 0.5 μM TDZ to continue culturing, culture temperature is 25±2°C, light intensity is 4000 Lux, and photoperiod is 16 hours.

[0020] 3. The callus cultured for 25 days was transferred to MS hormone-free medium to continue culturing, the culture temperature was 25±2°C, the li...

Embodiment 2

[0024] 1. Take the seeds of Snow Lotus from Tianshan, sterilize the surface with 10% sodium hypochlorite solution for 20 minutes, rinse the excess sterilizing solution with sterile water, and then connect it to the hormone-free MS solid medium. After the seedlings grow for 30 days, take the leaves and cut them into 0.5cm 2 The small pieces were cultured on the culture medium containing 1 μM TDZ, the culture temperature was 25° C., the light intensity was 3500 Lux, and the photoperiod was 16 hours.

[0025] 2. After 35 days of culture, the callus produced was transferred to MS medium containing 0.25 μM TDZ for subculture, and treated at 4° C. for 1 week every 3 generations. Then transfer to culture medium containing 1 μM TDZ, culture temperature is 25±2°C, light intensity is 4000 Lux, and photoperiod is 16 hours.

[0026] 3. The callus cultured for 25 days was transferred to MS hormone-free medium to continue culturing, the culture temperature was 25±2°C, the light intensity w...

Embodiment 3

[0030] 1. Take the seeds of Snow Lotus from Tianshan, sterilize the surface with 0.1% mercuric chloride solution for 12 minutes, rinse with sterile water for 3 to 5 times, and then connect to hormone-free MS solid medium. After the seedlings grow for 30 days, take the leaves and cut them into 0.5cm 2 The small pieces were placed on MS medium containing 0.5 μM TDZ to induce dedifferentiation, the culture temperature was 25° C., the light intensity was 4000 Lux, and the photoperiod was 16 hours.

[0031] 2. After culturing for 35 days, transfer the generated callus to a medium containing 1 μM TDZ for subculture, and treat it at 4° C. for 5 weeks every 3 generations. Then transfer to MS medium containing 0.5 μM TDZ to continue culturing, culture temperature is 25±2°C, light intensity is 4000 Lux, and photoperiod is 16 hours.

[0032] 3. The callus cultured for 25 days was transferred to MS hormone-free medium to continue culturing, the culture temperature was 25±2°C, the light i...

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