Method for generating grape somatic embryo
A technology for somatic embryos and grapes, applied in horticultural methods, botanical equipment and methods, plant regeneration, etc., can solve the problems of complex operation procedures, high technical difficulties, only suitable for individual varieties, etc., and achieve uniform traits and easy genetic engineering. The effect of simple operation and operation steps
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Embodiment 1
[0019] (1) During the seedless flowering period of the grape variety Moritani, collect flower buds from healthy plants 5 days before flowering, store the collected flower buds at 4°C for 2 days, and use detergent (such as washing powder, detergent, soapy water, laundry detergent, etc.) for 5 minutes, rinsed with tap water for 20 minutes, sterilized with 75% volume percent ethanol for 30 seconds, 2% volume percent sodium hypochlorite solution for 6 minutes, washed 4 times with sterile water, and then blotted dry with sterile filter paper Reserve the excess water on the buds;
[0020] (2) Place the flower buds obtained in step (1) under a dissecting microscope, puncture the petals with a sterile dissecting needle, take out a single stamen and inoculate it on the medium for cultivation, the cultivation condition is dark cultivation, the temperature is 28±1°C ;
[0021] The medium formula is NN basic medium and add 1.0mg·L -1 2,4-Dichlorophenoxyacetic acid, 2.0mg·L -1 6-Benzyla...
Embodiment 2
[0027] (1) During the full flowering period of the grape variety Cabernet Sauvignon, collect flower buds from healthy plants 7 days before flowering, store the collected flower buds at 4°C for 3 days, and wash them with detergent (such as washing powder, detergent, soap, etc.) water, laundry detergent, etc.) for 10 minutes, rinsed with tap water for 30 minutes, sterilized with 75% volume percent ethanol for 30 seconds, 2% volume percent sodium hypochlorite solution for 6 minutes, washed 4 times with sterile water, and then dried the buds with sterile filter paper Add excess water for later use;
[0028] (2) Place the flower buds obtained in step (1) under a dissecting microscope, puncture the petals with a sterile dissecting needle, take out a single stamen and inoculate it on the medium for cultivation, the cultivation condition is dark cultivation, the temperature is 28±1°C ;
[0029] The medium formula is NN basic medium and add 1.2mg·L -1 2,4-Dichlorophenoxyacetic acid, ...
Embodiment 3
[0035] (1) is consistent with the step (1) in embodiment 2, and different varieties are replaced by snake dragon ball;
[0036] (2) Place the flower buds obtained in step (1) under a dissecting microscope, puncture the petals with a sterile dissecting needle, take out a single stamen and inoculate it on the medium for cultivation, the cultivation condition is dark cultivation, the temperature is 28±1°C ;
[0037] The medium formula is NN basic medium and add 1.0mg·L -1 2,4-Dichlorophenoxyacetic acid, 1.5mg·L -1 6-Benzylaminoadenine, 5.0mg·L -1 AgNO 3 , 60g·L -1 Sucrose and 6g·L -1 Agar; if the pH value of the prepared medium is not 5.8-6.0, it is also necessary to adjust the pH value of the medium to 5.8-6.0 with 1N hydrochloric acid or sodium hydroxide;
[0038] (3) consistent with step (3) in embodiment 2;
[0039] (4) consistent with step (4) in embodiment 2;
[0040] (5) is consistent with step (4) in embodiment 2.
[0041] Schedule 1:
[0042] NN Basic Medium ...
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