A kind of Melaleuca prothallus induction and propagation method
A technology for prothallus and Melaleuca tower, which is applied in the field of prothallus induction and proliferation of Melaleuca tower, can solve the problems of browning, explant contamination, inability to grow, differentiate and proliferate, etc., so as to avoid sterilization difficulties and overcome the effect of not growing
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Embodiment 1
[0025] (1) Selection of explants
[0026] Gather Melaleuca from the wild [ Huperzia serrata (Thunb.) Trev.] sporangia, rinsed with tap water;
[0027] (2) Surface sterilization of explants
[0028] Dip the cleaned sporangia in an ethanol solution with a mass fraction of 70% for 30-60s in an ultra-clean workbench; 2 ) solution for 5-8 minutes, and then rinsed with sterile water for 3-5 times, as explants induced by Melaleuca prothallus;
[0029] (3) Induction culture of prothallus
[0030] The explants are inoculated on the prothallus induction medium, the composition of the prothallus induction medium is: 0.2 mg / L-2 mg / L of 3-indolebutyric acid is added to the conventional MS medium. Cultivate for 10-12 weeks at a culture temperature of 22±2°C, a light intensity of 300 Lx, and a light time of 12 / d-16 / d, and grow green prothallus tissue from the explants. The induction frequency is 70%;
[0031] (4) Proliferation of the prothallus
[0032] Cut the prothallus tissue obta...
Embodiment 2
[0034] (1) Selection of explants
[0035] Gather Melaleuca from the wild [ Huperzia serrata (Thunb.) Trev.] sporangia, rinsed with tap water;
[0036] (2) Surface sterilization of explants
[0037] Dip the cleaned sporangia in an ethanol solution with a mass fraction of 70% for 30-60s in an ultra-clean workbench; 2 ) solution for 5-8 minutes, and then rinsed with sterile water for 3-5 times, as explants induced by Melaleuca prothallus;
[0038] (3) Induction culture of prothallus
[0039] The explants were inoculated on the prothallus induction medium (the composition of the prothallus induction medium was the same as in Example 1), the culture temperature was 22±2°C, the light intensity was 1500 Lx, and the light time was 12 / d~16 / Cultivate under the condition of d for 10-12 weeks, grow green prothallus tissue from the explant, and the induction frequency of prothallus is 15%;
[0040] (4) Proliferation of the prothallus
[0041] Cut the prothallus tissue obtained in s...
Embodiment 3
[0043] (1) Selection of explants
[0044] Gather Melaleuca from the wild [ Huperzia serrata (Thunb.) Trev.] sporangia, rinsed with tap water;
[0045] (2) Surface sterilization of explants
[0046] Dip the cleaned sporangia in an ethanol solution with a mass fraction of 70% for 30-60s in an ultra-clean workbench; 2 ) solution for 5-8 minutes, and then rinsed with sterile water for 3-5 times.
[0047] Squeeze the sterilized sporangia to release the spores from the sporangia, and then collect the spores by centrifugation as explants induced by the prothallus of Melaleuca;
[0048] (3) Induction culture of prothallus
[0049] Take the spores in step (2), first dilute the spores to 100 / ml suspension with conventional MS liquid medium, inoculate 1ml of the spore suspension evenly on the prothallus induction medium (prothallium induction medium The composition is the same as that in Example 1), cultured for 10-12 weeks under the condition that the culture temperature is 22±2°C...
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