Method for in-vitro induction of lobed kudzuvine root microtuber
A test-tube potato and in vitro technology is applied in the field of pueraria test-tube potato in vitro induction, and can solve the problems of no report on pueraria test-tube potato, long research and observation time, and inconvenience to obtain materials.
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[0023] In the in vitro induction method of kudzu root tuber tuber of the present invention, kudzu root tissue culture seedlings are inoculated in the rooting medium for rooting culture to obtain rooting shoots; the rooting medium uses 1 / 2MS medium as the basic medium, and also Including: 0.02-0.05mg / L NAA, 25-36g / L sucrose and 4-8g / L agar. The preparation method of kudzu root tissue culture seedling of the present invention preferably comprises:
[0024] a) Take the tender tissue of kudzu root as the explant, inoculate it in the primary culture medium after disinfection, and carry out primary culture to obtain sterile seedlings; the primary culture medium uses MS medium as the basic medium, and also includes: 0.1-0.3mg 6-BA / L, 0.02-0.1mg / L NAA, 25-38g / L sucrose and 4-8g / L agar;
[0025] b) Inoculate the sterile seedlings obtained in step a) into a proliferation medium for proliferation culture to obtain clustered seedlings; the proliferation medium uses MS medium as the basic...
Embodiment 1
[0035] Select young stems and vines that are growing robustly and without pests and diseases, cut off their leaves, soak in water with detergent for 15 minutes, clean the dirt on the surface with a soft brush, and rinse in running water for 20 minutes.
[0036] On the ultra-clean bench, use 0.1% HgCl 2 Soak for 8 minutes, then rinse with sterile water for 5 times, and dry the surface moisture of the explants with sterile filter paper.
[0037] Under sterile conditions, cut the stem section into a 1.2cm-long stem section with an axillary bud, and insert the biological lower end vertically into the primary medium MS+0.2mg / L 6-BA+0.05mg / L NAA+30.0g / L sucrose+6.0g / L agar (pH, 5.8) for cultivation. Culture conditions: culture temperature (25±1)°C, light intensity 1800lx, light time 16h / d.
[0038] After 4 weeks of culture, transfer the sterile single-bud stem section of the primary culture to the subculture medium MS+0.03mg / L 6-BA+0.05mg / L NAA+30.0g / L sucrose+6.0g / L agar (pH, 5...
Embodiment 2
[0043] Select young stems and vines that are growing robustly and without pests and diseases, cut off their leaves, soak in water with detergent for 20 minutes, clean the dirt on the surface with a soft brush, and rinse in running water for 20 minutes.
[0044] On the ultra-clean bench, use 0.1% HgCl 2 Soak for 6 minutes, then rinse with sterile water for 4 times, and dry the surface moisture of the explants with sterile filter paper.
[0045] Under sterile conditions, cut the stem section into a 1cm-long stem section with an axillary bud, and insert the biological lower end vertically into the primary medium MS+0.1mg / L 6-BA+0.05mg / L NAA+30.0g / Cultured in L sucrose+6.0g / L agar (pH, 5.8). Culture conditions: culture temperature (25±1)°C, light intensity 1500lx, light time 16h / d.
[0046] After 5 weeks of culture, transfer the single-bud stem section of the sterile seedlings cultured in the first generation to the subculture medium MS+0.01mg / L 6-BA+0.05mg / L NAA+30.0g / L sucros...
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