Method for cultivating bletilla striata seedlings by using intermittent immersion bioreactor
A technology of bioreactor and bletilla striata, which is applied in the field of cultivating bletilla striata seedlings using intermittent immersion bioreactors, can solve the problems of unreported and public application of bletilla striata seedlings, shorten the cultivation period, reduce the pollution rate, and improve the automation level Effect
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Embodiment 1
[0049] Embodiment 1: as figure 1 as shown,
[0050] (1) Pre-preparation steps: Sterilization of reactors and transparent containers: bandage the reactors and transparent containers, sterilize them with high temperature and high pressure and moist heat for later use; the reactor is a gap immersion bioreactor, and the transparent container is an Erlenmeyer flask;
[0051] (2) Seed sterilization step: perform aseptic treatment on fine seed pods of Bletilla striata;
[0052] Surface decontamination: rinse with tap water for 30 minutes; surface disinfection: then soak the cleaned Bletilla striata seed pods in 70~75% (v / v) alcohol for 30 (s) in an ultra-clean workbench, and then soak them in sterile water Rinse 2-3 times; surface sterilization: soak in mercuric chloride with a concentration of 0.1% (g / v) for 8 minutes, rinse 5-7 times with sterile water; Operation Cut a small mouth from one end of the bletilla striata seed pod;
[0053] (3) Seed germination steps: According to th...
Embodiment 2
[0061] Utilize intermittent immersion bioreactor to cultivate bletilla striata seedlings, concrete steps are as shown in embodiment 1, what differ with it is that semi-solid culture is adopted during transition multiplication culture, and the selected medium formula is 1 / 2MS+NAA. .5mg / L+50g / L potato liquid+30g / L sucrose+agar 3g / L, adjust the pH to 5.9, the formula of proliferation and rooting culture medium is 1 / 2MS+NAA1.0mg / L+50g / L potato liquid+30g / L sucrose, adjust the pH to 5.9, the culture conditions are light intensity 1500~2000lx, light time 12h / d, temperature 24±1℃. In one culture cycle, the adaptation period of Bletilla striata seedlings in the intermittent immersion bioreactor was shortened to 0 days, so that the proliferation efficiency of the reactor reached more than 1:30.
Embodiment 3
[0063] Utilize the bioreactor to cultivate Bletilla striata seedlings, the specific steps are as shown in Example 1, the difference is that it is a semi-solid culture that is adopted during the transitional multiplication culture, and the selected medium formula is 1 / 2MS+NAA0.5mg / L+50g / L potato liquid+30g / L sucrose+agar 2g / L, adjust the pH to 6.0, the culture medium for pseudobulb expansion is MS+NAA1.0mg / L+50g / L potato liquid+banana juice 40g / L+straw Juice 60g / L, adjust pH6.0, the adaptation period of Bletilla striata seedlings in the intermittent immersion bioreactor is shortened to 0.3 days, in a culture period, the pseudobulbs of Bletilla striata seedlings expand about 2 ~ 3 times, and the seedling survival rate obtained Above 90%.
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