A method for detoxification of clover green vitrification with ultra-low temperature therapy
A technology of ultra-low temperature vitrification and clover, applied in horticultural methods, botany equipment and methods, plant regeneration, etc., can solve the problems of low plant tissue survival rate and low virus clearance rate, and improve the survival rate of recovery culture, damage reduction effect
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Embodiment 1
[0032] The steps for detoxification of clover green vitrification ultra-low temperature therapy are as follows:
[0033] (1) Sterile seedling culture: Take the tested stems with TMV clover with buds, rinse them with water, soak them in 0.1% mercuric chloride solution for 30 minutes under aseptic conditions, and then transfer them to sodium hypochlorite solution for disinfection. After washing with sterile water, use sterile paper to absorb the water on the surface of the stem section of cloverleaf with buds, and transfer it into the MS medium containing 6-BA 1.0mg / L and NAA 0.5mg / L. is 2000Lux, and the illumination time is 12h·d -1 , the temperature is 25 ± 1 ℃ under the conditions of 5 days of culture, to obtain three leaf green sterile seedlings.
[0034] (2) Pre-cultivation: the aseptic seedlings of Trifolium chinensis cultivated in step (1) were subjected to a light intensity of 2000Lux and a light time of 12h·d. -1 , and continued to culture for 7 days at a temperature ...
Embodiment 2
[0039] The steps for detoxification of clover green vitrification ultra-low temperature therapy are as follows:
[0040](1) Sterile seedling culture: Take the tested stems with TMV clover with buds, rinse them with water, soak them in 0.1% mercuric chloride solution for 30 minutes under aseptic conditions, and then transfer them to sodium hypochlorite solution for disinfection. After washing with sterile water, use sterile paper to absorb the water on the surface of the stem section of cloverleaf with buds, and transfer it into the MS medium containing 6-BA 1.0mg / L and NAA 0.5mg / L. is 2000Lux, and the illumination time is 12h·d -1 , the temperature is 25 ± 1 ℃ under the conditions of 5 days of culture, to obtain three leaf green sterile seedlings.
[0041] (2) Pre-cultivation: the aseptic seedlings of Trifolium chinensis cultivated in step (1) are 1800Lux in the light intensity and 13h·d for the light time. -1 , the temperature was 40 ± 2 ℃ for 8 days, and the medium was MS ...
Embodiment 3
[0046] The steps for detoxification of clover green vitrification ultra-low temperature therapy are as follows:
[0047] (1) Sterile seedling culture: Take the tested stems with TMV clover with buds, rinse them with water, soak them in 0.1% mercuric chloride solution for 30 minutes under aseptic conditions, and then transfer them to sodium hypochlorite solution for disinfection. After washing with sterile water, use sterile paper to absorb the water on the surface of the stem section of cloverleaf with buds, and transfer it into the MS medium containing 6-BA 1.0mg / L and NAA 0.5mg / L. is 2000Lux, and the illumination time is 12h·d -1 , the temperature is 25 ± 1 ℃ under the conditions of 5 days of culture, to obtain three leaf green sterile seedlings.
[0048] (2) Pre-cultivation: the aseptic seedlings of Trifolium chinensis cultivated in step (1) were subjected to a light intensity of 2000Lux and a light time of 12h·d. -1 , the temperature was 40 ± 2 ℃ and the culture was cont...
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