A method for rapidly obtaining tissue-cultured seedlings of Zoysia japonica
A technology of zoysia and tissue culture seedlings, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems that endophytes are difficult to eliminate, affect gene transformation efficiency, etc., and achieve the effect of prolonging the sterilization time
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Embodiment 1Z
[0042] Embodiment 1 ZJ-SM medium formulation concentration screening
[0043] In the ZJ-SM medium, the function of kinetin is to promote tissue differentiation and sprouting, and 2,4-D plays the role of auxin, which promotes the formation of adventitious roots and promotes growth. The screening experiment of kinetin concentration and 2,4-D concentration in ZJ-SM medium is as follows:
[0044] Screen for kinetin concentration:
[0045] The concentration of kinetin is 1.0 μmol / L, 2.0 μmol / L, and 3.0 μmol on a solid MS medium containing 2% sucrose mass percentage and 0.3% (w / v) plant gel content. / L, 4.0 μmol / L, 5.0 μmol / L, 6.0 μmol / L, 7.0 μmol / L, 8.0 μmol / L and 0.0 μmol / L (control). Zoysia japonica explants were inoculated on the above medium, and the germination rate was counted.
[0046] The method for obtaining the explants is as follows: Germinate the sterilized Zoysia japonica seeds and cultivate them in a solid MS medium until the height of the shoot is 3 cm. At this ti...
Embodiment 2
[0056] Example 2 Seed Culture Mode Screening
[0057] Soak the sterilized Zoysia japonica seeds in water at 4°C for 3 days (72 hours), dry the seeds until the surface turns white, and then inoculate them directly on MS medium and culture them in water just submerged in the seeds In both ways, the culture temperature is room temperature culture.
[0058] It has been observed that a small amount of seeds will germinate within 2 days when the seeds are soaked in water, and most of the seeds will germinate on the 3rd day. For seeds sown directly on MS medium, the fastest time to germinate is the 3rd day, and a large number of seeds will germinate in 4-5 days.
Embodiment 3
[0059] Embodiment 3 seeding density screening
[0060] Prepare solid MS medium and distribute it in tissue culture bottles with a diameter of 8cm, about 30ml per bottle.
[0061] Dry the seeds after soaking at 4°C for 72 hours until the surface turns white, and culture them with water that has just submerged the seeds until 1-2mm buds germinate. 10 grains, 20 grains, 30 grains, 40 grains, 50 grains per bottle The sowing density was inoculated into the above-mentioned tissue culture bottle equipped with solid MS medium, and cultivated for 15 days. After observation, it was not found that the sowing density had a significant impact on the growth of Zoysia japonica.
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