Method for Tissue Culture Regeneration of Mature Bromegrass Seeds
A technique for tissue culture of brome, applied in the field of agricultural biology, can solve problems such as blanks, and achieve the effects of low cost, simple culture process and large number of plants
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Embodiment 1
[0016] Example 1: figure 1 As shown, the method for tissue culture regeneration of mature brome awnless seeds adopts the following specific process steps.
[0017] (1) Sterilization treatment of explants: select full-bodied brome seeds, shell them and place them in a 50ml triangular flask. Disinfect the surface with 75% ethanol by volume on the ultra-clean workbench for 1 min, rinse with sterile water once; then further disinfect with 0.1% mercuric chloride for 15 min, shaking continuously during this period, and finally rinse with sterile water Rinse 5 times, soak in sterile water at room temperature for 12 hours, let the seeds fully imbibition.
[0018] (2) Induction of callus: Inoculate sterilized brome awn seeds on callus induction medium for culture. The culture conditions are: temperature 26°C, light time 12h / d, light intensity 1500Lux; induction medium: add 1mg / L 2,4-D, 0.5mg / L 6-BA, 25g / L to MS medium L of sucrose and 7g / L of agar, pH 5.8, induction time 30d.
[00...
Embodiment 2
[0025] Embodiment 2: The method for tissue culture regeneration of mature brome awnless seeds adopts the following specific process steps.
[0026] (1) Sterilization treatment of explants: same as in Example 1.
[0027] (2) Callus induction: culture conditions: temperature 25°C, light time 13h / d, light intensity 2000Lux; induction medium: MS+0.8mg / L2,4-D+1mg / L6-BA+25g / L sucrose+7g / L agar, pH value 6.0, induction time 35d;
[0028] (3) Transfer the callus to subculture medium 1 for culture: culture conditions: temperature 27°C, light time 13h / d, light intensity 2000Lux; subculture medium 1: MS+1.5mg / L2,4 -D+0.5mg / L6-BA+25g / L sucrose+7g / L agar, pH value 5.9, culture time 35d.
[0029] (4) Transfer the callus to the differentiation medium for differentiation culture: the culture conditions are: temperature 25°C, light time 15h / d, light intensity 2000Lux; differentiation medium: MS+1.5mg / LNAA+0.8mg / L6- BA+30g / L sucrose+7g / L agar, pH value 6.0, culture time 32d.
[0030] (5) I...
Embodiment 3
[0033] Embodiment 3: The method for tissue culture regeneration of mature brome awnless seeds adopts the following specific process steps.
[0034] (1) Sterilization of explants: same as in Example 1.
[0035] (2) Callus induction: culture conditions: temperature 27°C, light time 14h / d, light intensity 1000Lux; induction medium: MS+0.5mg / L2,4-D+0.7mg / L6-BA+25g / L sucrose +7g / L agar, pH value 5.9, induction time 33d;
[0036] (3) Transfer the callus to subculture medium 1 for culture: culture conditions: temperature 25°C, light time 14h / d, light intensity 1000Lux; subculture medium 1: MS+2mg / L2, 4-D+0 .8mg / L6-BA+25g / L sucrose+7g / L agar, pH value 5.9, culture time 32d.
[0037] (4) Transfer the callus to the differentiation medium for differentiation culture: the culture conditions are: temperature 27°C, light time 16h / d, light intensity 1000Lux; differentiation medium: MS+1.8mg / LNAA+0.5mg / L6- BA+30g / L sucrose+7g / L agar, pH value 5.9, culture time 35d.
[0038] (5) Insert the...
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