Tissue culture rapid propagation method for Cerasus xueluoensis C.H.Nan & X.R.Wang
A tissue culture and rapid technology, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of non-commercial seedlings, reproductive obstacles, etc., and achieve the effects of long growth cycle, large proliferation, and optimal cultivation system.
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Embodiment 1
[0028] A method for rapid propagation of Xueluo cherry tissue culture, comprising the following steps:
[0029] 1) Induction culture of stem buds: 1) Cut the young stems of Xueluo cherry blossoms in spring, disinfect and wash them, cut them into 1.5cm stems with 1-2 buds, and inoculate them in the way of vertical insertion and horizontal placement MS, 1 / 2MS, 1 / 4MS, and improved MS (ammonium nitrate halved) were used as the basic medium, and 6-BA1.0mg / L+IBA0.01mg / L was used as the starting medium for the hormone combination; 6.4 g / L agar and 30g / L sucrose. The results showed that the germination rates of MS, 1 / 2MS, 1 / 4MS, and improved MS medium were 75.35%, 88.46%, 97.75%, and 84.92%, respectively, and the germination time of 1 / 4MS medium was the shortest, about one week, and the other 3 The germination time of the seed medium is about 13 days, such as figure 1 shown.
[0030] 2) Proliferate bud clusters (3-5 buds) that grow robustly as explants, use MS as the basic medium,...
Embodiment 2
[0034] A method for rapid propagation of Xueluo cherry tissue culture, comprising the following steps:
[0035] 1) Callus induction culture: Take the young leaves and wash them under running water for 1-3 hours. Wash with 75% alcohol for 30s, wash with sterile water for 3~5 times, then 0.1%HgCl 2 7min, washed 5-8 times with sterile water; cut into 1cm 2 Leaflets of the same size, cut on the edge, cut on the main vein and petiole, inoculated on MS, B5, N6, WPM as the basic medium, and the hormone ratio is 6-BA1.0mg / L+NAA1.0mg / L In the medium of +2,4-D2.0mg / L, 6.8g / L agar and 30g / L sucrose were added to the medium, pH5.8, 121℃, 20kg / cm 2 Sterilize under high temperature and high pressure for 20 minutes, culture at (25±2)°C, in the dark and light, and subculture once every 25 days. The results showed that after 10 days in MS medium, callus tissue appeared in the wound of explants, which grew healthy and fast, and was light milky white; the callus rate of B5 medium was second o...
Embodiment 3
[0045] The healthy pale milky white callus induced by Example 2 was inoculated into MS+6-BA1.0mg / L+NAA0.6mg / L+sucrose 30g / L+agar 6.4g / L callus containing different anti-browning agents. A total of 12 combinations of wound proliferation medium were cultured in dark conditions and subcultured every 25 days. Each treatment was repeated 5 times, counted once every 7 days, and counted 3 times in total.
[0046] The types of anti-browning agents are as follows: VC (5, 10, 20mg / L), CA (10, 50, 100mg / L), AC (100, 500, 800mg / L), PVP (50, 100, 200mg / L).
[0047] The results showed that the anti-browning effect of Vc20mg / L was better than that of sodium citrate (CA); comparing the two adsorbents, the anti-browning effect of PVP was better than that of activated carbon (AC).
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