Aquilaria sinensis hairy root induction and genetic transformation method
A method of genetic transformation and a technology of genetic transformation, applied in the field of induction of hairy roots of A. chinensis and its genetic transformation, can solve the problems of no induction of hairy roots of A. argentina and no genetic transformation of A. argentina
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Embodiment 1
[0090] The embodiment of the present invention provides a method for inducing the efficiency of the hairy root of A. sinensis, comprising the following steps:
[0091] 1. Acquisition of exogenous
[0092] 1. Rinse the seeds with distilled water and peel off the seed coat;
[0093] 2. Soak the seeds in 75% ethanol for 30-50 sec, 20% sodium hypochlorite (NaClO) solution for 15 min, rinse with sterile water for 3-5 times, and dry the water with sterile filter paper on a clean bench;
[0094] 3. The treated seeds were placed on solid MS medium and cultured at 25°C under 8h light / 16h dark conditions;
[0095] 4. After being cultivated for 3 days, the seeds will start to germinate and be used as explant materials for subsequent infection.
[0096] 2. Agrobacterium rhizogenes infection (hairy root induction)
[0097] 1. Agrobacterium rhizogenes was cultured at 28°C in LB liquid medium containing 50mg / L Rif.
[0098] 2. Collect the bacteria by centrifugation, and resuspend the pel...
Embodiment 2
[0125] The embodiment of the present invention provides a method for inducing the efficiency of the hairy root of A. sinensis, comprising the following steps:
[0126] 1. Acquisition of exogenous
[0127] 1. Rinse the seeds with distilled water and peel off the seed coat;
[0128] 2. Soak the seeds in 75% ethanol for 30-50 sec, 20% sodium hypochlorite (NaClO) solution for 15 min, rinse with sterile water for 3-5 times, and dry the water with sterile filter paper on a clean bench;
[0129] 3. The treated seeds were placed on solid MS medium and cultured at 25°C under 8h light / 16h dark conditions;
[0130] 4. After being cultivated for 3 days, the seeds will start to germinate and be used as explant materials for subsequent infection.
[0131] 2. Agrobacterium rhizogenes infection (hairy root induction)
[0132] 1. Agrobacterium rhizogenes was cultured at 28°C in LB liquid medium containing 50mg / L Rif.
[0133] 2. Collect the bacteria by centrifugation, and resuspend the pel...
Embodiment 3
[0143] The embodiment of the present invention provides a method for inducing the efficiency of the hairy root of A. sinensis, comprising the following steps:
[0144] 1. Acquisition of exogenous
[0145] 1. Rinse the seeds with distilled water and peel off the seed coat;
[0146] 2. Soak the seeds in 75% ethanol for 30-50 sec, 20% sodium hypochlorite (NaClO) solution for 15 min, rinse with sterile water for 3-5 times, and dry the water with sterile filter paper on a clean bench;
[0147] 3. The treated seeds were placed on solid MS medium and cultured at 25°C under 8h light / 16h dark conditions;
[0148] 4. After being cultivated for 3 days, the seeds will start to germinate and be used as explant materials for subsequent infection.
[0149] 2. Agrobacterium rhizogenes infection (hairy root induction)
[0150] 1. Agrobacterium rhizogenes was cultured at 28°C in LB liquid medium containing 50mg / L Rif.
[0151] 2. Collect the bacteria by centrifugation, and resuspend the pel...
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