Method for effectively inducing cotton somatic embryo and regenerating plants and culture medium thereof
A technology of liquid culture medium and culture medium, applied in plant regeneration, botany equipment and methods, gardening methods, etc., can solve the problems of long cycle, poor repeatability, high frequency of deformed embryos, etc.
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Embodiment 1
[0075] Example 1. Efficiently Obtaining Somatic Embryos and Regenerated Plants
[0076] (1) Acquisition of sterile cotyledon seedlings
[0077] 1. Delinting of cotton seeds
[0078] Take 50g of cotton variety Kezimian 201 seeds, add about 15ml of concentrated sulfuric acid, stir quickly for 2-3 minutes, then neutralize with lime water, scrub continuously, and rinse with running water for 10-12 minutes to obtain clean delint seeds.
[0079] 2. Germination of cotton seeds
[0080] The depilled seeds were subjected to 0.1% (w / v) HgCl 2 The solution was sterilized for 10 minutes, then the seeds were washed 5-6 times with sterilized water, and finally 150ml of sterilized water was added, and germinated in the dark at 28°C for 24 hours to obtain germinated seeds.
[0081] 3. Germination of cotton seeds
[0082] Peel off the husks of the germinated seeds in an aseptic environment, plant the husk-removed cotton seed embryos on a germination medium, and culture them in the dark at ...
Embodiment 2
[0107] Example 2. Efficiently Obtaining Somatic Embryos and Regenerated Plants
[0108] (1) Acquisition of sterile cotyledon seedlings
[0109] 1. Delinting of cotton seeds
[0110] Take 50g of cotton variety Jiwu 2031 (purchased from Hebei Jifeng Seed Industry Co., Ltd., a variety approved by Hebei Provincial Crop Variety Approval Committee) seeds, add about 15ml of concentrated sulfuric acid, stir quickly for 2-3min, then neutralize in lime water, scrub continuously, and run water Rinse for 10-12 minutes to obtain clean depilled seeds.
[0111] 2. Germination of cotton seeds
[0112] The depilled seeds were subjected to 0.1% (w / v) HgCl 2 The solution was sterilized for 10 minutes, then the seeds were washed 5-6 times with sterilized water, and finally 150ml of sterilized water was added, and germinated in the dark at 28°C for 24 hours to obtain germinated seeds.
[0113] 3. Germination of cotton seeds
[0114] Peel off the husks of the germinated seeds in a sterile envi...
Embodiment 3
[0135] Example 3. Efficiently Obtaining Somatic Embryos and Regenerated Plants
[0136] (1) Acquisition of sterile cotyledon seedlings
[0137] 1. Delinting of cotton seeds
[0138] Take 50g of cotton variety Nongda 58 (purchased from Hebei Jifeng Seed Industry Co., Ltd., a variety approved by the Hebei Provincial Crop Variety Approval Committee) seeds, add about 15ml of concentrated sulfuric acid, stir quickly for 2-3min, then neutralize with lime water, scrub continuously, and rinse with running water 10-12min to obtain clean fluffed seeds.
[0139] 2. Germination of cotton seeds
[0140] The depilled seeds were subjected to 0.1% (w / v) HgCl 2 The solution was sterilized for 10 minutes, then the seeds were washed 5-6 times with sterilized water, and finally 150ml of sterilized water was added, and germinated in the dark at 28°C for 24 hours to obtain germinated seeds.
[0141] 3. Germination of cotton seeds
[0142] Peel off the husks from the germinated seeds in a steri...
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