Method for promoting jatropha curcas explant to regenerate adventitious buds
A technology of explants and Jatropha curcas, applied in the field of plant biology, can solve the problems of long regeneration cycle, low regeneration rate, poor quality of regeneration buds, etc., and achieves the effects of simple method, reduced negative impact, and shortened culture cycle.
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Embodiment 1
[0039] S1. Preparation of Vessels for Short-Term Cytokinin Solution Treatment of Explants
[0040] Select a petri dish with a diameter of 9 cm, place it in a high-pressure steam sterilizer, and sterilize it at 121 °C and 0.1 MPa for 20 min.
[0041] S2. Preparation of phenylthiadiazolyl urea (TDZ) treatment solution.
[0042] Accurately weigh a certain amount of TDZ powder, fully dissolve it with 1 mol / l NaOH solution, and then dilute it with deionized water to prepare 0, 10, 20, 30, 40, 50, 60 mg / l TDZ treatment solutions ( Control group 0 mg / l TDZ solution is sterile deionized water). Use 1 mol / l HCl solution to adjust the pH of the TDZ treatment solution to 5.8~6.0; filter and sterilize the prepared TDZ treatment solution with a 0.22 micron water filter before use.
[0043] S3. Acquisition of Jatropha curcas sterile hypocotyl explants
[0044] Jatropha curcas seeds soaked for 2 days were dehulled, sterilized by 0.1% mercuric chloride solution, and the embryos wer...
Embodiment 2
[0059] Example 2 The influence of the time of soaking in TDZ solution to treat Jatropha curcas sterile hypocotyl explants on the effect of direct regeneration of adventitious buds
[0060] S1. Preparation of the container for short-term cytokinin solution treatment of explants: same as in Example 1.
[0061] S2. Prepare a 20 mg / l phenylthiadiazolyl urea (TDZ) solution: the same as in Example 1.
[0062] S3. Acquisition of Jatropha curcas sterile hypocotyl explants: same as in Example 1
[0063] S4. Jatropha curcas sterile hypocotyl explants treated with cytokinin (TDZ) solution: the cut sterile hypocotyl explants were placed in a container treated with cytokinin (TDZ) solution, and the Pour the TDZ solution with a concentration of 20 mg / l into the wide-mouth glass bottle of the container until all the hypocotyl explants are submerged (0 mg / l TDZ solution of the control group is sterile deionized water), and cover the bottle cap , after soaking the hypocotyl explants ...
Embodiment 3
[0069] S1. Preparation of the container for short-term cytokinin solution treatment of explants: same as in Example 1.
[0070] S2. Prepare a 20 mg / l phenylthiadiazolyl urea (TDZ) solution, and the specific preparation method is the same as in Example 1.
[0071] S3. Acquisition of sterile hypocotyl explants of Jatropha curcas and hypocotyl explants of seedlings: the specific method is the same as that in Example 1.
[0072] S4. Cytokinin (TDZ) solution treated Jatropha curcas sterile hypocotyl explants and seedling hypocotyl explants respectively: the cut 2 types of hypocotyl explants were placed in cell division In the container treated with TDZ solution, pour TDZ solution with a concentration of 20 mg / l into the wide-mouth glass bottle of the container until all hypocotyl explants are submerged, cover the bottle cap, and test the hypocotyl explants. After soaking the axis explants for 20 min, pour off the TDZ solution, keep the hypocotyl explants, take out ...
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