Chestnut somatic embryo development synchronization method and tissue culture seedling rooting method
A somatic embryo, synchronization technology, applied in the field of plant tissue culture, can solve the problems of difficult rooting, low synchronization rate, and inability to achieve large-scale asexual reproduction of fine varieties, and achieve the effect of promoting rooting and improving rooting rate.
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Embodiment 1
[0112] (1) Plant material
[0113] Yanshan red chestnut (C. mollissima cv. Yanshanhongli) was collected from Chestnut Experimental Station, Huairou District, Beijing. The sampling period of chestnut is 45-54 days after flowering, and immature young embryos are selected.
[0114] (2) Induction of chestnut embryogenic callus
[0115] Take the immature embryo tips of chestnut, sterilize them with 3% sodium hypochlorite and 75% alcohol, and inoculate them on the IMM solid medium. When the callus grows to 5 mm, put the callus in the liquid IMM medium for 7 days. weeks, and finally transferred to E2 solid medium for culture until embryogenic callus was formed.
[0116] (3) Culture of Chestnut Somatic Embryo Synchronization
[0117] a. Inoculate 0.1 g of embryogenic callus tissue into a 250 mL sterile Erlenmeyer flask, and add 100 mL of E1 liquid medium. After cultivating on a shaker for 7 days, filter the suspension culture with 20-mesh and 40-mesh sieves successively, Retain th...
Embodiment 2
[0137] The cotyledon-shaped embryos in Example 1 are induced to obtain tissue cultured plantlets, and then the Chinese chestnut tissue cultured plantlets with good growth status are inoculated into the rooting medium for cultivation, wherein the components of the rooting medium include: 2.3g / L WPM , 30g / L sucrose, 0.1mg / L IBA and 6.5g / L agar (pH 5.5). Chestnut tissue culture seedlings were cultured at 23-25°C with a photoperiod of 16h / 8h. Repeat three times, count the rooting situation and analyze the data after 25 days of cultivation.
Embodiment 3
[0139] The difference with Example 2 is that the concentration of IBA in the rooting medium is 0.2mg / L.
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