Method for carrying out gene transformation on manioca by using Agrobacterium tumefaciens mediated transformation
A technique of Agrobacterium-mediated and gene transformation, applied in the tissue culture of Jatropha curcas and in the field of genetic engineering, can solve the problems that breeding techniques cannot be genetically improved, and achieve the effects of avoiding molecular detection work, high rooting rate, and avoiding a large number of escapes
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[0082] This example uses Agrobacterium to transfer the GUS gene into wild Jatropha curcas in Yuanmou, Yunnan. The method created by the present invention is described in detail. Please refer to Table 1 for the total number of samples and the number of survivors after each processing step. Each process step is as follows:
[0083] Step 1: Collect wild Jatropha seeds from Yuanmou, Yunnan. Pick full seeds, peel off the shell, soak in 75% ethanol for 30 seconds, rinse with sterile water for 2-3 times, then disinfect in 0.1% mercury chloride for 3-5 minutes, rinse with sterile water for 5-7 times, Then soak in sterile water for 2-4 hours and peel off the endosperm. Take complete embryos and cotyledons, inoculate them in MS medium, pick 2-3 seeds per bottle, culture in dark at 23-26°C for 2-4 days, then transfer to light culture for 10-12 days to obtain sterile seedlings for experiments.
[0084] Step 2: pick the single clone of Agrobacterium EHA105 (containing the expression vect...
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