Begonia masoniana leaf in-vitro regeneration system establishment method
A technology for in vitro regeneration and begonias, applied in plant regeneration, horticultural methods, botanical equipment and methods, etc., can solve the problems of low overall proliferation rate, large variety morphological differences, long cycle, etc.
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Embodiment 1
[0025] Explant sampling time screening
[0026] 1. Comparison of sampling in different seasons
[0027] At the five time points of December 1, February 1, March 1, April 1 and May 1, the expanded leaves were selected for disinfection treatment and primary induction. Disinfection methods are "first shake and wash with an appropriate amount of detergent solution for 15 minutes, then rinse with tap water for 40 minutes, take it out, put it on the aseptic operating table, disinfect it with 75% alcohol for 10 seconds, and immediately transfer it to a solution containing 0.1% mercuric chloride for disinfection. 25min, then rinse with sterile water 5-6 times, 1min each time, sterilize the filter paper to dry the surface moisture, cut off the edge of the leaf and cut into 3-4cm 2 square pieces. "Pollution control is shown in Table 1. It can be seen that the best sampling time is from March to April, when the leaves of that year have just unfolded.
[0028] Table 1 Effect of seasona...
Embodiment 2
[0031] Explant Screening:
[0032] 2. Explant cutting size screening
[0033] The difference in cutting size of the explant leaves after disinfection treatment also leads to different culture effects. Cut each piece into 3~4cm 2 The explants germinate early, the number is large, the growth is extremely fast, and the effect of strong seedlings is good in the later stage; and each piece is cut to 0.5~1cm 2 The leaves of the leaves germinated late, and the effect of strong seedlings in the later stage was poor, and gradually tended to brown or wilt.
[0034] Table 2 Effect of Explant Cut Size
[0035] Explant cut size Number of explants The number of buds produced in 20 days Bud development 0.5~1cm 2
Embodiment 3
[0037] Lighting condition filter:
[0038] 3. Comparison of light conditions in the first induction
[0039]During the induction process of the first generation leaves of Begonia japonica, the effects of different light levels on the induction of adventitious buds from leaf explants were compared. The light conditions were set to four gradients of low light 1000lx, strong light 1500lx, extremely strong light 2000lx, and normal light after 1 day of dark cultivation. After 20 days of inoculation into the culture medium, observe and count the situation of leaf induction of adventitious buds.
[0040] 1000lx has a lot of buds, and each leaf can induce 200 times the number of adventitious buds. 1500lx buds are less, only about 10 times, and very strong light 2000lx basically no adventitious buds appear, after 1 day of dark culture, normal light culture buds are about 70 times, it can be seen that the light conditions have a very significant impact on the induction of adventitious...
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