A kind of rapid propagation method of Xueluo cherry tissue culture
A tissue culture and rapid technology, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve problems such as reproductive barriers and no commercial seedlings, and achieve the effects of large proliferation, long growth cycle, and cost reduction.
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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 1.5 cm stems with 1-2 buds, and inoculate them in vertical and horizontal ways. MS, 1 / 2MS, 1 / 4MS, and improved MS (ammonium nitrate halved) were used as the basic medium, and 6-BA 1.0 mg / L+IBA 0.01 mg / L was used as the starting medium of the hormone combination; 6.4 g / L agar and 30 g / 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 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, and use 6-BA (0.5, 1.0,...
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 h. Wash with 75% alcohol for 30 s, wash with sterile water for 3-5 times, and then wash with 0.1% HgCl 2 7 min, washed 5-8 times with sterile water; cut into 1 cm 2 Leaflets of the same size, with cuts on the edges, main veins and petioles, were inoculated on MS, B5, N6, WPM as the basic medium, and the hormone ratio was 6-BA 1.0 mg / L+NAA 1.0 mg / L +2,4-D 2.0mg / L medium, add 6.8 g / L agar and 30g / L sucrose to the medium, pH5.8, 121℃, 20 kg / cm 2High temperature and high pressure sterilization for 20 minutes, culture temperature (25±2) ℃, culture in the dark and light, subculture once every 25 days. The results showed that after 10 days in MS medium, callus appeared in the wound of explants, which grew healthy and fast, and was light milky white; the callus rate of B5 medium ...
Embodiment 3
[0045] The healthy pale milky white callus induced by Example 2 was inoculated into MS+6-BA 1.0mg / L+NAA 0.6mg / 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, 20 mg / L), CA (10, 50, 100 mg / L), AC (100, 500, 800 mg / L), PVP (50, 100, 200 mg / L) L).
[0047] The results showed that the anti-browning effect of Vc 20 mg / 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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