Method for obtaining transgenic Malus hupehensis rehd plant without selectable marker genes
A selection marker-free, Pingyi sweet tea technology, applied in the fields of botanical equipment and methods, horticultural methods, genetic engineering, etc.
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[0042] 1. Acquisition of transgenic Pingyi sweet tea plants without selection marker gene
[0043]Obtain the tissue culture seedlings (tissue culture seedlings) of Pingyi sweet tea according to the following steps: pick the annual branches of Pingyi sweet tea in early spring, cultivate them in water for about 10 days, take the full buds on the branches, wash them with running water for 3 hours, and use 0.1% mercury chloride (HgCl 2 ) for 6 minutes, rinsed with sterile water for 6 times, blotted the residual water with sterile absorbent paper, and inoculated the buds on the primary culture medium. The primary culture medium is the addition of 30 mg·L MS basic medium -1 Sucrose, 6.5mg·L -1 Agar powder, 1.0mg·L -1 6-benzyl adenine and 0.2mg·L -1 Indolebutyric acid, pH 5.6 medium. On the second day after inoculation, some materials began to brown. Once browned materials were found, they were immediately replaced with new primary medium until the browning was eliminated. 10 d...
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