Tissue culture and rapid propagation method for firmiana major

[0051] Induction and Differentiation Media Screening Methods

[0052] Take the young terminal buds and side buds of sycamore yunnanensis as explants, soak them in 1% soapy water for 10 minutes, rinse them with running water, put them on the ultra-clean table, cut them into stems with single nodes, and use 75% Disinfect with alcohol for 10 seconds, rinse with sterile water for 3 times, disinfect the surface with 0.1% mercuric chloride solution for 8 minutes, wash with sterile water for 3 to 5 times, and inoculate in the prepared sycamore yunnanensis induction medium, 1 node per bottle.

[0053] Inoculate the sterilized stem segments in MS+0.1mg / L 6-BA+0.5mg / L IAA, MS+0.5mg / L 6-BA+0.5mg / L IAA, MS+1mg / L 6-BA +0.5mg / L IAA, MS+1.5mg / L 6-BA+0.5mg / L IAA, MS+2mg / L6-BA+0.5mg / L IAA medium, and the medium also includes 30g / L sucrose, Agar 5g / L, pH 5.8. The light time is 10 hours a day, the light intensity is 1500 Lux, the temperature is 26±3°C, and the culture period is 60 days to obta...

[0058] The nodal buds obtained in Example 1 were inoculated onto the proliferation and subculture medium, and the MS medium containing sucrose 30g / L+agar 5g / L was used as the basic medium for proliferation and subculture hormone screening, and the cytokinins were 6-BA, the concentration range is 0.1mg / L, 0.2mg / L, 0.5mg / L, 0.8mg / L, 1.0mg / L, 1.2mg / L (6 concentration gradients), auxin IAA, the concentration is 0.5mg / L, using the uniform design method. The light time is 10 hours per day, the light intensity is 1500 Lux, the temperature is 26±3°C, and the culture period is 60 days to obtain the breeding seedlings.

[0059] The results are shown in Table 2.

[0060] Table 2 Proliferation and Subculture Medium Hormone Screening

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[0062] As can be seen from Table 2, MS medium+0.8mg / L 6-BA (6-benzylpurine)+0.5mg / L IAA (indole acetic acid)+sucrose 30g / L+agar 5g / L, pH value 5.8, culture period For 60 days, the light intensity is 1500Lux, and the temperature is 23-30°C. Bu...

[0064] The proliferating seedlings prepared in Example 2 are inoculated onto strong seedlings and rooting medium. Strong seedlings and rooting medium are based on the MS medium containing sucrose 30g / L+agar 5g / L, and the concentration range of IAA is 0.1mg / L. L, 0.2mg / L, 0.3mg / L, 0.5mg / L, 0.8mg / L, 1.0mg / L (6 concentration gradients), the concentration range of IBA is 0.1mg / L, 0.2mg / L, 0.3mg / L, 0.5mg / L, 0.8mg / L, 1.0mg / L. The light time is 10 hours per day, the light intensity is 1500 Lux, the temperature is 26° C., and the culture period is 60 days to obtain rooted seedlings. Statistical rooting rate.

[0065] The results are shown in Table 3.

[0066] Table 3 Hormone Screening Results of Strong Seedling and Rooting Medium

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[0068]

[0069] As can be seen from Table 3, MS medium+0.3mg / L IBA (indole butyric acid)+0.3mg / L IAA (indole acetic acid)+30g / L sucrose+5g / L agar, pH value 5.8, culture cycle 30 days , The rooting rate reaches 91%.