Method for establishing genetic transformation system of dendrobium nobile
Dendrobium nobile, transgenic technology, applied in the field of genetic transformation
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[0078] Induction of Dendrobium nobile callus: Transfer the aseptic seedlings of Dendrobium nobile to sucrose 30g / L+agar 7.5g / L+1 / 2MS+1.4mg / L 6-BA+0.2mg / L NAA medium . After 3 weeks, green protrusions appeared around the rhizoids, which was the germination of callus tissue. After 4 weeks, green protrusions were seen to expand to form green callus tissue. At this time, subculture was carried out again to induce proliferation of callus. The culture temperature is 25±1°C, and the culture is dark.
[0079] Screening of Spectinomycin resistance of Dendrobium nobile callus: inoculate the callus of Dendrobium nobile into medium supplemented with different concentrations of spectinomycin, the inoculation amount of each concentration is not less than 10, every 5 days The survival rate of the callus was counted, and the counting was stopped after 30 days. Medium formula: sucrose 30g / L+agar 7.5g / L+1 / 2MS+1.0mg / L 6-BA+0.1mg / L NAA. Spectinomycin concentrations were 50, 100, 150, 200, 250...
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