Eucalyptus urophylla*grandis embryoid induction seedling raising method
A technology of embryoid body and Eucalyptus grandis, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problem of less chimera of transgenic plants, and achieve the effect of stable results and good repeatability
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Embodiment 1
[0021] On a sterile operating table, select robust seedlings of Eucalyptus grandis without seedlings, remove the terminal buds and axillary buds, cut the stem segments into 0.5 cm long segments, and inoculate the stem segments into embryogenic callus induction medium (MS Medium+ 30g·L -1 Sucrose + 7 g L -1 Agar + 100 mg·L -1 The volume ratio of Vc+ is 20% coconut water + 0.01 mg L -1 TDZ+0.1 mg·L -1 NAA, pH 5.8), cultured in the dark at 25°C, replaced with fresh medium every 2 weeks, and cultured in the dark for 4 weeks;
[0022] After induction of embryogenic callus, transfer the embryogenic callus to embryoid induction medium (MS medium + 30g·L -1 Sucrose + 7 g L -1 Agar + 100 mg·L -1 Vc+1 mg·L -1 Spermidine + volume ratio is 20% coconut water + 0.01 mg L -1 TDZ+0.05mg·L -1 NAA, pH 5.8), cultured in the dark at 25°C for 3 weeks to obtain embryoid bodies;
[0023] Transfer the induced embryoid bodies to embryoid body germination medium (MS medium + 30 g L -1 Sucr...
Embodiment 2
[0025] On a sterile operating table, select robust seedlings of Eucalyptus grandis without seedlings, remove the terminal buds and axillary buds, cut the stem segments into 0.6 cm long segments, and inoculate the stem segments in embryogenic callus induction medium (MS Medium+ 30g·L -1 Sucrose + 7 g L -1 Agar + 100 mg·L -1 The volume ratio of Vc+ is 20% coconut water + 0.01 mg L -1 TDZ+0.1 mg·L -1 NAA, pH 5.9), cultured in the dark at 25°C, replaced with fresh medium every 2 weeks, and cultured in the dark for 5 weeks;
[0026] After induction of embryogenic callus, transfer the embryogenic callus to embryoid induction medium (MS medium + 30g·L -1 Sucrose + 7 g L -1 Agar + 100 mg·L -1 Vc+1 mg·L -1 Spermidine + volume ratio is 20% coconut water + 0.01 mg L -1 TDZ+0.05mg·L -1 NAA, pH 5.9), cultured in the dark at 25°C for 4 weeks to obtain embryoid bodies;
[0027] Transfer the induced embryoid bodies to embryoid body germination medium (MS medium + 30 g L -1 Sucros...
Embodiment 3
[0029] On a sterile operating table, select robust seedlings of Eucalyptus grandis without seedlings, remove the terminal buds and axillary buds, cut the stem segments into 0.8 cm long segments, and inoculate the stem segments into embryogenic callus induction medium (MS Medium+ 30g·L -1 Sucrose + 7 g L -1 Agar + 100 mg·L -1 The volume ratio of Vc+ is 20% coconut water + 0.01 mg L -1 TDZ+0.1 mg·L -1 NAA, pH 6.0), cultured in the dark at 25°C, replaced with fresh medium every 2 weeks, and cultured in the dark for 5 weeks;
[0030] After induction of embryogenic callus, transfer the embryogenic callus to embryoid induction medium (MS medium + 30g·L -1 Sucrose + 7 g L -1 Agar + 100 mg·L -1 Vc+1 mg·L -1 Spermidine + volume ratio is 20% coconut water + 0.01 mg L -1 TDZ+0.05mg·L -1 NAA, pH 6.0), cultured in the dark at 25°C for 3 weeks to obtain embryoid bodies;
[0031] Transfer the induced embryoid bodies to embryoid body germination medium (MS medium + 30 g L -1 Sucr...
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