Tissue culture and rapid propagation method for pinellia tuber plant
A technology for tissue culture and rapid propagation of Pinellia, which is applied in the field of plant cell engineering, can solve problems such as increasing labor input, and achieve the effects of high automation level, large biological output, and reduced mutation frequency
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Embodiment 1
[0044] The specific steps for multiplying the petiole explants of the aseptic seedlings of Pinellia trifoliata by using a batch immersion submerged culture reactor are as follows:
[0045] (1) Selection of intermittent immersion culture reactors: use the patent No. ZL 200920116327.X of "Cultivation reactors for intermittent immersion of plant tissues and organs". The volume size is 3L.
[0046] (2) Material selection and treatment: The petiole of the sterile Pinellia trifoliate seedlings obtained by conventional tissue culture methods was used as explants, and the petiole that grew vigorously was taken, and the middle part of the petiole was cut into lengths of 0.5, 1, 1.5, and 2 cm, respectively. Several kinds, later used as explant material for inoculation in the next step.
[0047] (3) Proliferation culture: the specific method is to inoculate the treated explants into the pre-sterilized batch immersion culture reactor under aseptic condition. The parameters of the batch ...
Embodiment 2
[0052] Concrete steps for multiplying the clustered bud explants of Pinellia aseptic seedlings by using a batch submerged culture reactor:
[0053] (1) Selection of intermittent immersion culture reactors: use the patent No. ZL 200920116327.X of "Cultivation reactors for intermittent immersion of plant tissues and organs". The volume size is 5L.
[0054] (2) Material selection and processing: 2) Material selection and processing: The differentiated cluster buds of sterile Pinellia trifoliate tissue cultured seedlings obtained by conventional tissue culture methods are used as explants, and the cluster shoots with unexpanded leaves are taken in a good state , cut into a single state and used as explant material for inoculation.
[0055] (3) Proliferation culture: the specific method is to inoculate the treated explants into the pre-sterilized batch immersion culture reactor under aseptic condition. The parameters of the batch immersion culture reactor were set as follows: imm...
Embodiment 3
[0059] The specific steps for multiplying the leaf explants of the aseptic seedlings of Pinellia trifoliata by using a batch submerged culture reactor are as follows:
[0060] (1) Selection of intermittent immersion culture reactors: use the patent No. ZL 200920116327.X of "Cultivation reactors for intermittent immersion of plant tissues and organs". The volume size is 1L.
[0061] (2) Material selection and treatment: The leaves of sterile Pinellia tissue-cultured seedlings obtained by conventional tissue culture methods were used as explants, and the fully expanded leaves were taken, and three wounds were cut across them for the next inoculation explant material.
[0062](3) Proliferation culture: the specific method is to inoculate the treated explants into the pre-sterilized batch immersion culture reactor under aseptic condition. The parameters of the batch immersion culture reactor were set as follows: immersion frequency: 8-20 min / 2h; culture time: 15-30 d. The cultu...
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