In-vitro preservation method of radix tetrastigme test-tube plantlets
A technology of in vitro preservation and cloverleaf, applied in the field of plant cultivation, can solve the problem that the genetic evaluation of the in vitro preservation technology has not been reported, the normal subculture preservation of tissue culture is frequently transferred, and the planting of cloverleaf germplasm resources is difficult. and other problems, to achieve the effect of preventing species degradation and virus infection, prolonging subculture time, and ensuring purity
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Embodiment 1
[0030] A method for in vitro preservation of green clover test-tube plantlets, comprising the following steps:
[0031] (1) Select the shoot tip of Clover as an explant, soak it in 75% alcohol in mass concentration, wash it twice with sterile water, add dropwise 0.1% mercury liter in mass concentration, rinse it with sterile water for 8 times, and then add it dropwise The mass concentration is 0.1% mercuric chloride, washed 8 times with sterile water, cut into 0.5 cm long and inoculated in the first culture medium; the first culture medium includes 500mg / L NH 4 NO 3 、700mg / LKNO 3 , 150mg / L MgSO 4 ·7H 2 O, 70mg / L KH 2 PO 4 , 0.1mg / L BA, 22g / L sucrose, 8g / L agar; 20±2℃, light intensity 1500lx, light time 16h / d, subculture once every 280d;
[0032] (2) Under aseptic conditions, the isolated stem tip with a diameter of 1mm is cut from the vigorously grown aseptic seedling as material, and the isolated shoot tip is placed in the second medium for cultivation, and the second m...
Embodiment 2
[0039] A method for in vitro preservation of green clover test-tube plantlets, comprising the following steps:
[0040] (1) Select the stem tips of Clover as explants, soak them in 75% alcohol in mass concentration, rinse them twice with sterile water, add dropwise 0.1% mercury liter in mass concentration, rinse them with sterile water for 7 times, and then add them dropwise The mass concentration is 0.1% mercuric chloride, washed 8 times with sterile water, cut into 0.8 cm long and inoculated in the first culture medium; the first culture medium includes 650mg / L NH 4 NO 3 、600mg / LKNO 3 , 120mg / L MgSO 4 ·7H 2 O, 100mg / L KH 2 PO 4 , 0.1mg / L BA, 20g / L sucrose, 10g / L agar; 20±2℃, light intensity 1800lx, light time 16h / d, subculture once every 280d;
[0041] (2) Under aseptic conditions, the isolated stem tip with a diameter of 1mm is cut from the vigorously grown aseptic seedling as material, and the isolated shoot tip is placed in a second medium for cultivation, and the s...
Embodiment 3
[0048] A method for in vitro preservation of green clover test-tube plantlets, comprising the following steps:
[0049] (1) Select the stem tips of A. trifoliate as explants, soak them in 75% alcohol by mass concentration, rinse them twice with sterile water, add 0.1% mercury liter dropwise, rinse them with sterile water for 6 times, and then add them dropwise The mass concentration is 0.1% mercuric chloride, rinsed with sterile water for 6 times, cut into 1.0 cm lengths and inoculated in the first culture medium; the first culture medium includes 600mg / L NH 4 NO 3 、700mg / LKNO 3 , 150mg / L MgSO 4 ·7H 2 O, 60mg / L KH 2 PO 4 , 0.1mg / L BA, 20g / L sucrose, 5g / L agar; 20±2℃, light intensity 1800lx, light time 16h / d, subculture once every 300d;
[0050] (2) Under aseptic conditions, the isolated stem tip with a diameter of 1mm is cut from the vigorously grown aseptic seedling as material, and the isolated shoot tip is placed in a second medium for cultivation, and the second medi...
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