A kind of seedling test-tube propagation method of broad-leaved orchid
A technology of test tube propagation and orchid orchid, applied in the field of plant tissue culture, can solve the problems of difficult germination, long cycle, high cost, etc., and achieve the effects of high seed germination rate, good seedling quality, and fast seedling growth
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Embodiment 1
[0040] (1) Collection of explants: the mature and uncracked fruits of Orchid latifolia were collected from the field as explants.
[0041] (2) Aseptic germination: the fruit described in step (1) was soaked in 75% ethanol for 2 minutes, rinsed with sterile water for 3 times, and then sterilized in 0.1% mercuric solution for 10 minutes, rinsed with sterile water for 5 times Afterwards, the fruit is cut open, and the powdered embryo is inoculated on a sterile germination medium and cultured for 93 days to obtain a protocorm, the contamination rate is 5.9%, and the seed germination rate is 100%. The sterile germination medium is: 1.0g / L Huabao No. 1 (HYPONeX 1)+0.5mg / L NAA+18g / L sucrose+4.0g / L agar+55ml / L coconut juice, pH is 5.6.
[0042](3) Protocorm proliferation: the protocorm obtained in step (2) was inoculated into the protocorm proliferation medium and cultivated for 30 days to proliferate to obtain a large number of protocorm-like protocorms, and the multiplication factor...
Embodiment 2
[0048] (1) Collection of explants: the mature and uncracked fruits of Orchid latifolia were collected from the field as explants.
[0049] (2) Aseptic germination: the fruit described in step (1) is soaked in 75% ethanol for 1 minute, rinsed with sterile water for 3 times, then disinfected in 0.1% mercuric solution for 10 minutes, rinsed with sterile water for 5 times The fruit is cut open, the powdered embryo is inoculated on the aseptic germination medium and cultivated for 100 days to obtain the protocorm, the pollution rate is 10%, and the germination rate of the seed reaches more than 95%. The sterile germination medium is: 1.5g / L Huabao No. 1 (HYPONeX 1)+0.4mg / L NAA+30g / L sucrose+5.0g / L agar+75ml / L coconut juice, pH is 5.4.
[0050] (3) Protocorm proliferation: the protocorm obtained in step (2) was inoculated into the protocorm proliferation medium and cultivated for 38 days to proliferate to obtain a large number of protocorm-like protocorms, and the multiplication coe...
Embodiment 3
[0056] (1) Collection of explants: the mature and uncracked fruits of Orchid latifolia were collected from the field as explants.
[0057] (2) Aseptic germination: the fruit described in step (1) was soaked in 75% ethanol for 5 minutes, rinsed with sterile water for 5 times, and disinfected in 0.1% mercuric solution for 30 minutes, rinsed with sterile water for 7 times The fruit is cut open, the powdered embryo is inoculated on the aseptic germination medium and cultivated for 110 days to obtain the protocorm, the pollution rate is lower than 3%, and the seed germination rate reaches more than 90%. The sterile germination medium is: 2.0g / L Huabao No. 1 (HYPONeX 1)+1.0mg / L NAA+30g / L sucrose+6.0g / L agar+100ml / L coconut juice, pH is 5.8.
[0058] (3) Protocorm proliferation: inoculate the protocorm obtained in step (2) into the protocorm proliferation medium and cultivate it for 42 days to proliferate and obtain a large amount of protocorm-like protocorms with a multiplication fa...
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