Method for realizing gene transient transfection in capitulum
A flower head and transient transfection technology, applied in the field of genetic engineering, can solve the problems of chimera, low transformation efficiency, and long transformation cycle, and achieve the effect of short time, high transformation efficiency, and short transformation cycle
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[0055] 1) Preparation of plant material:
[0056] On the adult plant of Chrysanthemum chrysanthemum, the cuttings with young bud diameters of 2.5-4mm are selected, and the length of the cuttings is 6-8cm. In plug trays, cultivate them for 20-30 days at 20° C. under humid, short-day (light / dark=8h / 16h) conditions, keep the soil moist, and wait for them to take root and bloom.
[0057] 2) Preparation of infection solution:
[0058] On the first day, pick a single colony of Agrobacterium pCAMBIA1305.2 carrier to 50mL YEB liquid medium + 50μL 50mg / mL-1 Kan + 50μL 50mg / mL-1 Rif, shake the bacteria overnight at 28°C and 200rpm;
[0059] On the second day, draw 10mL of the first-day bacterial solution to 500ml YEB liquid medium + 5ml EMS (1M) + 200μl 1% AS + 500μl 50μg / mL Kan + 500μl 50mg / ml Rif, shake the bacteria overnight at 28°C and 200rpm;
[0060] On the third day, prepare the infection solution 500mL ddH 2 O+5ml EMS(1M)+2mL 1%AS+5ml MgCl 2 . Centrifuge the overnight shake...
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