Method for obtaining conifer embryoid through liquid culture and its culture medium
A technology of liquid medium and embryoid body is applied in the field of cultivation of plant explants, and can solve the problems of vitrification, hypocotyl failure, abnormal cotyledon development, etc.
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Embodiment 1
[0178] 1. Experimental materials and methods
[0179] 1. Experimental materials
[0180] Immature zygotic embryos of Larix principis, Japanese larch, Larix olgensis, Japan × North China hybrid larch, and Japan × Landrace hybrid larch.
[0181] Larix kaempferi were selected from the 30-year-old seed garden of the Dagujia Forest Farm in Qingyuan County, Liaoning Province, including Riyong 8, Riyong 85, Rifu 27, and Ricao 103 clones; Larix olgensis was collected from the Hada Forest Farm in Fushun City, Liaoning Province The 30-year-old white larch seed orchard includes the Chang 3, Chang 14, Chang 11, Chang 4C, and Chang 33 clones; Larix principis is collected from the Qiuqiangou Forest Farm of Guancheng Forestry Bureau in Shanxi Province. 6. Hua 7 and other clones. The hybridization techniques of the Japanese, Landrace, and Larix principis-rupprechtii combinations are carried out using open controlled pollination.
[0182] 2. Experimental method
[0183] 1) Inoculation, induction c...
Embodiment 2
[0199] The culture was carried out in the same manner as in Example 1, and the medium used was as shown in Table 3. The somatic embryo culture of coniferous species was carried out, and the same results as in Example 1 were obtained.
[0200] The difference is that the proliferation culture is carried out in a fermenter, and the inoculation amount is 0.5-1% by weight to volume, that is, 0.5-1 g of fresh, free embryonic cell lines are inoculated in 100 ml of liquid medium. Cultivation is carried out at 23±5°C and dark culture conditions.
[0201] Table 3 Medium formula (mg / L)
[0202] Compound
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