Simplified tilia cordata mill tissue culture method
A technology for tissue culture and induction medium, applied in the field of plant tissue culture, can solve the problems of affecting the growth of seedlings, virus accumulation, and high cost of grafting reproduction, and achieve the effects of easy promotion, low cost, high reproduction efficiency and versatility
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Embodiment 1
[0027] Embodiment 1, the selection of explant
[0028] On April 2nd, April 12th, April 22nd, May 2nd and May 12th, the healthy plants of Tilia microphylla 'Xinle' were collected as explants in the same year, and compared with different sampling times. Effect of implant contamination rate and induction rate.
[0029] After the above sprouts are collected, cut off the leaves and keep the petiole of about 1 cm, shake and soak the obtained stems in 2%-5% alkaline detergent solution for 20 minutes, rinse with running water for 2 hours, and use 75 % alcohol for 15 seconds, sterilized with 2% hypochlorous acid for 5 minutes, and washed with sterile water for 5 times, each time for 8 minutes.
[0030] The parietal leaves and axillary buds were respectively inoculated on the induction medium, the formula was: MS+6-BA 1.0mg / L+IBA 0.2mg / L+sucrose 30g / L+agar 5.8g / L, the pH value was 5.9; each treatment was inoculated 50 explants were used, and the contamination rate and induction rate w...
Embodiment 2
[0034] Embodiment 2, disinfection mode screening
[0035] Around April 22, collect the sprouting strips of the healthy plants of European small-leaved linden 'Xinle' in the same year, cut off the top 1-2 buds of the sprouting strips, and remove the leaves to keep about 1 cm of petiole, and the obtained stems are 2%-5% Shake and soak in alkaline detergent solution for 20 minutes, rinse with running water for 2 hours, and then use the following 4 disinfection methods to disinfect the surface of explants on the ultra-clean workbench, with 50 explants per treatment. The specific disinfection methods are as follows: (1) disinfect with 75% alcohol for 15 seconds, disinfect with 2% sodium hypochlorite for 5 minutes, wash with sterile water for 5 times, each time for 8 minutes; (2) disinfect with 75% alcohol for 15 seconds, Disinfect with 2% sodium hypochlorite for 10 minutes, wash 5 times with sterile water, 8 minutes each time; (3) Disinfect with 75% alcohol for 15 seconds, disinfec...
Embodiment 3
[0039] Embodiment 3, induction medium screening
[0040] Around April 22, the sprouting strips of the healthy plants of Tilia microphylla 'Xinle' were collected in the same year, 1-2 buds on the top of the sprouting strips were cut off, and the leaves were removed to keep about 1 cm of petiole. On the induction medium of ratio (sucrose 30g / L+agar 5.8g / L in each medium component, pH value is 5.9), every treatment explant is inoculated 100, the induction rate of adventitious bud and the multiplication coefficient of counting after 30d, The effects of different media on the induction of adventitious buds were compared, and the experimental results are shown in Table 3.
[0041] Table 3 Effects of different hormone components on the induction of adventitious buds of Tilia microphylla ‘Xinle’
[0042]
[0043]
[0044] It can be seen from Table 3 that when the hormone components are MS+6-BA 1.0mg / L+IBA 0.2mg / L, the effect of explant adventitious bud induction is the best, the ...
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