Method for promoting bud differentiation in Qijian white cymbidium sinense tissue culture process
A technology for tissue culture and bud differentiation, applied in the field of plant tissue culture, can solve the problems of low efficiency of factory propagation, difficult rhizome differentiation of intermediate propagules, and inability to meet commercial production, and achieves the technology of promoting bud differentiation and promoting expression. Effect
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Embodiment 1
[0042] Example 1 Yucasin combined with 6-BA promotes the differentiation of rhizomes and shoots of Qijian Baimolan
[0043] (1) Take the rhizome of Qijian Baimolan with a length of 1.0~1.5cm, and inoculate it in 6 treatments of CK, IAA, 6-BA, Yucasin, (Yucasin+IAA) and (Yucasin+6-BA) , each bottle is connected with 10 rhizomes, and each treatment is connected with 21 bottles. After inoculation, place them in an incubator for cultivation, temperature: 26±1°C, light: 2000lux, 12h / d. After photographing at 6 sampling points (5 days, 10 days, 15 days, 20 days, 25 days and 30 days), the differentiation rate of rhizome shoots was counted. The formula for calculating the bud differentiation rate is:
[0044] Bud differentiation rate=(number of rhizomes differentiated from buds / number of connected rhizomes)×100%.
[0045] (2) Culture medium and treatment conditions:
[0046] CK: MS medium + 25.0g / L sucrose + 7.0g / L carrageenan, pH=5.8;
[0047] IAA: MS medium + 25.0g / L sucrose + 7....
Embodiment 2
[0053] Example 2 Yucasin combined with 6-BA promotes the expression of hormone-related genes in the rhizome of Qijian Baimolan
[0054] (1) Take the rhizomes of Qijian Baimolan with a length of 1.0-1.5cm, and inoculate them in the four treatments of CK, 6-BA, Yucasin and (Yucasin+6-BA), and insert 10 roots into each bottle. Each treatment received 7 bottles; after inoculation, they were placed in an incubator (temperature: 26±1°C, light: 2000lux, 12h / d) and cultured at 7 sampling points (0 days, 5 days, 10 days, 15 days, 20 days, 25 days and 30 days) each process randomly gets 1 bottle, totally 10 rhizomes, and is divided into 3 groups at random, every group 3 rhizomes (i.e. a repeat sample) about 0.3 grams, Immediately after sampling, the samples were quickly frozen in liquid nitrogen, and stored in a -80°C refrigerator if no subsequent operations were performed.
[0055] (2) Culture medium and treatment conditions:
[0056] CK: MS medium + 25.0g / L sucrose + 7.0g / L carragee...
Embodiment 3Y
[0070] Example 3 Effects of Yucasin combined with 6-BA on the content of endogenous hormones in the rhizome of Qijian Baimolan
[0071] (1) Take the rhizomes of Qijian Baimolan with a length of 1.0-1.5cm, and inoculate them in the four treatments of CK, 6-BA, Yucasin and (Yucasin+6-BA), and insert 10 roots into each bottle. Shaped stems, each treatment received 3 bottles. After inoculation, place them in an incubator (temperature: 26±1°C, light: 2000lux, 12h / d) for cultivation.
[0072] (2) Media and treatment conditions
[0073] CK: MS medium + 25.0g / L sucrose + 7.0g / L carrageenan, pH=5.8;
[0074] 6-BA: MS medium + 7.0g / L carrageenan + 1.0mg / L 6-BA, pH=5.8;
[0075] Yucasin: MS medium+25.0g / L sucrose+7.0g / L carrageenan+50μM Yucasin, pH=5.8;
[0076] Yucasin+6-BA: MS medium+25.0g / L sucrose+7.0g / L carrageenan+200μM Yucasin+1.0mg / L6-BA, pH=5.8.
[0077] (3) After culturing for 30 days, 3 bottles of materials were taken for each treatment at the sampling point for the deter...
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