A kind of tissue culture method of Hedyotis diffusa
A technology of Hedyotis diffusa and tissue culture, applied in the field of plant tissue culture, can solve the problems of increasing the risk of seedling variation, reducing the quality and quality of seedlings, unable to guarantee the stability of traits and the like
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Embodiment 1
[0042] (1) Acquisition and induced culture of sterile materials: cut the axillary buds of Hedyotis diffusa that grow well and are free from diseases and insect pests, rinse them with tap water for 30 minutes, and then wash them in an aqueous solution of 1% detergent (volume ratio w / w) Shake in medium for 2 minutes, rinse with sterile water; soak in 70% volume concentration ethanol water solution for 30 seconds on the ultra-clean workbench, then soak in mixed disinfectant solution for 10 minutes, and then rinse with sterile water for 3 times; Blot the water on the top, then inoculate the axillary buds into a culture bottle filled with cluster bud induction medium, cover the bottle cap and seal the bottle mouth with a parafilm, place it in an incubator, and cultivate it for 15 days at 24°C and 1500 lx light The mixed disinfectant is a mixed solution of Tween 20 and 84 disinfectant with a volume ratio of 1:200; the cluster bud induction medium is added with 6-BA (ie 6-benzylaminoa...
Embodiment 2
[0046] The influence of different disinfectants of embodiment 2 on the growth of Hedyotis diffusa explants
[0047] 10% hydrogen peroxide (H 2 o 2 ) aqueous solution, mercuric chloride (HgCl) with a mass concentration of 0.1% 2 ) aqueous solution and the mixed disinfectant solution in Example 1 (i.e. Tween 20 and 84 disinfectant solution with a volume ratio of 1:200) to disinfect the explant material (i.e. axillary bud) of Hedyotis diffusa, and other operations are the same as in Example 1 . The axillary buds were inoculated into culture bottles filled with cluster bud induction medium, and the bottles were sealed with a parafilm and placed in an incubator. After culturing for 10 days at 24° C. under the condition of light of 1500 lx, the effects of different disinfectants on the induction and growth of Hedyotis diffusa buds were observed and counted. The results are shown in Table 1. The results showed that the mixed disinfectant and mass concentration of 0.1% HgCl 2 The...
Embodiment 3
[0050] Embodiment 3 different hormone combination is to the induction influence of Hedyotis diffusa bud
[0051] The axillary buds of Hedyotis diffusa were respectively inoculated in MS basic medium supplemented with different concentrations of 6-BA, 2,4-D and NAA. Other operations were the same as in Example 1. The growth results of Hedyotis diffusa adventitious buds are shown in Table 2. Show. The results showed that the medium with different combinations of hormones could induce the formation of shoots to varying degrees. Among them, the medium MS+6-BA1.0mg / L+2,4-D1.0mg / L+NAA 0.2mg / L+0.4g / L acid hydrolyzed casein is the best, the most regenerated shoots are induced, and the growth Fast, the average bud length is 2.55cm, the bud induction rate is 76%, the seedlings are large and strong, there is no vitrified seedling phenomenon, and there is little callus, and the incision parts of the medium without acid hydrolyzed casein show different degrees of browning.
[0052] Wher...
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