Bletilla striata germplasm resource in-vitro preservation method
A technology for in vitro preservation and resources, which is applied in the field of in vitro preservation of bletilla germplasm resources, can solve the problems of not being able to meet the needs of in vitro preservation of bletilla and germplasm resources, and shorten the storage time of bletilla and germplasm resources in vitro, so as to prolong the in vitro preservation time , Solve the problem of germplasm resource preservation and large-scale seedling breeding, and the effect of high survival rate
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Embodiment 1
[0023] A method for in vitro preservation of P. chinensis germplasm resources, comprising the following steps:
[0024] Step 1, place Pseudobulb in the first culture medium as explant, obtain clump bud, before placing Pseudomonas Pseudobulb in the first medium, first place Pseudobulb at 75 ℃. Soak in ethanol for 45s, then rinse 3 times with sterile water, and put into 0.1% Hgcl with 2% Tween-20 2 The solution was soaked for 4 minutes, then rinsed with sterile water for 3 times, and then put into 0.1% Hgcl with 2% Tween-20 again. 2 The solution was soaked for 4min, finally used sterile water for 3 times, then peeled off the 3 layers of outer skin of Pseudobulb, and inoculated in the first medium for cultivation;
[0025] Step 2, treat the clump buds to grow to 2~3cm, place the clump buds in the second medium and cultivate to have 4~6 leaves (the test tube seedlings have taken root at this moment), obtain the test tube seedlings;
[0026] Step 3: Inoculate the test tube seedli...
Embodiment 2
[0034] A method for in vitro preservation of P. chinensis germplasm resources, comprising the following steps:
[0035] Step 1, place Pseudobulb in the first culture medium as explant, obtain clump bud, before placing Pseudomonas Pseudobulb in the first medium, first place Pseudobulb at 75 ℃. Soak in ethanol for 45s, then rinse 3 times with sterile water, and put into 0.1% Hgcl with 2% Tween-20 2 The solution was soaked for 4 minutes, then rinsed with sterile water for 3 times, and then put into 0.1% Hgcl with 2% Tween-20 again. 2 The solution was soaked for 4min, finally used sterile water for 3 times, then peeled off the 3 layers of outer skin of Pseudobulb, and inoculated in the first medium for cultivation;
[0036] Step 2, treat the clump buds to grow to 2~3cm, place the clump buds in the second medium and cultivate to have 4~6 leaves (the test tube seedlings have taken root at this moment), obtain the test tube seedlings;
[0037] Step 3: Inoculate the test tube seedli...
Embodiment 3
[0045] A method for in vitro preservation of P. chinensis germplasm resources, comprising the following steps:
[0046] Step 1, place Pseudobulb in the first culture medium as explant, obtain clump bud, before placing Pseudomonas Pseudobulb in the first medium, first place Pseudobulb at 75 ℃. Soak in ethanol for 45s, then rinse 3 times with sterile water, and put into 0.1% Hgcl with 2% Tween-20 2The solution was soaked for 4 minutes, then rinsed with sterile water for 3 times, and then added 0.1% Hgcl with 2% Tween-20 2 Soak in the solution for 4 minutes, and finally use sterile water for 3 times, then peel off the three layers of skin of the white and pseudobulbs, and inoculate them in the first medium for cultivation;
[0047] Step 2, when the clustered buds grow to 2-3 cm, the clustered buds are placed in the second culture medium and cultivated until there are 4-6 leaves (the test-tube plantlets have taken root at this time), to obtain the test-tube plantlets;
[0048] S...
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