Induction medium for green spherical bodies of hemionitis arifolia
A technology of green spheroids and induction medium, applied in the field of plant tissue culture, to achieve the effects of short induction time, high induction rate and improved reproduction efficiency
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Embodiment 1
[0020] (1) Acquisition of aseptic explants of Alismatis fern
[0021] Inoculate the aseptic spores of Alismatis fern on 1 / 2MS + 30.0g / L sucrose + 7.0g / L agar, pH 5.80 culture medium, the culture conditions are: 20~25℃, the daily light time is 12 hours, the light intensity It is 2000lx, and after cultivating for 6-7 months, young and tender aseptic seedlings can be obtained, and the young and tender aseptic seedlings are used as Alisma fern aseptic explants.
[0022] The method for obtaining sterile spores of Erythronia fern is to place 10-15 mg of spores in a 1.5 ml centrifuge tube and pre-wash them three times with sterile water, centrifuge at 6000r / min for 2min to obtain the spore precipitate, and add a solution with a volume fraction of 5% NaClO Sterilize for 4 minutes, and wash with sterile water for 3-5 times to obtain the aseptic spores of Alismatis fern.
[0023] (2) Induction of green spheroids of Alisma fern
[0024] Inoculate the sterile explants of Alisma fern obt...
Embodiment 2
[0044] Embodiment 2 is the same as Embodiment 1 except that the following steps are different, and will not be repeated here.
[0045] (2) Induction of green spheroids of Alisma fern
[0046] Green spheroid induction medium is: 1 / 6MS+TDZ0.6mg / L+NAA0.2mg / L+hydrolyzed casein 600mg / L+sucrose 30.0g / L+agar 7.0g / L, pH 5.70.
[0047] After 2 weeks of cultivation, the shoot tip and petiole swollen base of the aseptic seedlings will grow into green spheroids, and the average induction rate can reach 95%.
[0048] The average survival rate of the plantlets of Alisma alisma in step (6) is 96.67%.
Embodiment 3
[0050] Embodiment 3 is the same as Embodiment 1 except that the following steps are different, and will not be repeated here.
[0051] (2) Induction of green spheroids of Alisma fern
[0052] Green spheroid induction medium is 1 / 5MS+TDZ 1.0mg / L+NAA0.3mg / L+hydrolyzed casein 800mg / L+sucrose 30.0g / L+agar concentration 7.0g / L, pH is 5.90.
[0053] After 2 weeks of cultivation, the shoot tips and petiole bases of sterile seedlings swelled to form green spheroids, and the average induction rate could reach 98.33%.
[0054] In step (6), the average survival rate of the tissue-cultured seedlings of Alisma alizima is 100%.
[0055] One Alisma fern green spheroid induced by the above embodiments can reproduce 1.5 to 2 million high-quality Alisma fern seedlings within one year. Preservation of green spheroids of Alisma fern: use the proliferation medium of green spheroids for preservation, and replace the medium every 3 to 4 weeks. The growth medium of green spheroids is: 1 / 6MS+TDZ0.2mg / ...
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