A method for overcoming the vitrification phenomenon of sisal tissue culture seedlings
A technique of tissue culture and vitrification, which is applied to overcome the vitrification phenomenon of sisal tissue culture seedlings. In the field of sisal tissue culture, it can solve the problems of different, not universally applicable, abnormal physiological functions of vitrified seedlings, etc., and achieve loose implementation conditions , The effect of simple implementation steps and easy implementation steps
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Embodiment 1
[0028]Take 15 bulbil seedlings with a height of 8-10cm stored in the sisal germplasm nursery of the South Subtropical Crops Research Institute, remove the roots and old leaves, wash them with running water, treat them with potassium permanganate solution 1000 times for 30 minutes, and then soak them in 75% alcohol for 1 minute , then soak in 0.1% mercuric chloride solution for 15-30 min, and finally rinse with sterile water 3-5 times. After drying, excise part of the explants, inoculate them on the improved SH+6-BA3.0-5.0mg / L medium after longitudinal cutting, and the culture conditions are 12-14 hours of light per day, 10-12 hours of dark culture, light intensity 2000lux, culture temperature is 28±2℃. After 45 days of culture, it was transferred to the bud induction and proliferation medium. The basic medium for bud induction and proliferation was improved SH medium, and the exogenous hormones were 6-BA, NAA and IBA; the concentration of 6-BA was 1.0-5.0 mg / L; the concentrat...
Embodiment 2
[0030] Take 85 clusters (5-8 plants per cluster) of partially or fully vitrified regenerated plants and inoculate them on the modified SH medium, add exogenous hormones as 6-BA, NAA and IBA; the concentration of 6-BA is 1.0-5.0mg / L; the concentration of NAA is 0-1.5mg / L, the concentration of IBA is 0-1.0mg / L, the pH is 5.8-6.0; the light is 12-14h, the dark is 10-12h, the light intensity is 2000Lx, and the temperature is 28±2 ℃. After 1 month of cultivation, 23 clusters had fully recovered to normal growth, 34 clusters had partially recovered, and the recovery rate of vitrified seedlings was 67.06%.
Embodiment 3
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