Method for fast obtaining Japanese lawngrass tissue culture seedlings
A technology for zoysia and tissue culture seedlings is applied in horticultural methods, botanical equipment and methods, horticulture and other directions, which can solve the problems of difficulty in eliminating endophytes, affecting gene transformation efficiency, etc., and achieve the effect of prolonging sterilization time.
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Embodiment 1Z
[0042] Example 1 ZJ-SM medium formula concentration screening
[0043] In the ZJ-SM medium, the role of kinetin is to promote tissue differentiation and budding, and 2,4-D plays the role of auxin to promote the formation of adventitious roots and promote growth. The screening experiment of kinetin concentration and 2,4-D concentration in ZJ-SM medium is as follows:
[0044] Screen kinetin concentration:
[0045] On a solid MS medium with a sucrose mass percentage of 2% and a plant gel content of 0.3% (w / v), the concentration of kinetin was set to 1.0 μmol / L, 2.0 μmol / L, and 3.0 μmol. / L, 4.0μmol / L, 5.0μmol / L, 6.0μmol / L, 7.0μmol / L, 8.0μmol / L and 0.0μmol / L (control). Zoysia japonica explants were inoculated on the above-mentioned medium, and the budding rate was counted.
[0046] The method for obtaining the explants is as follows: After germination of the sterilized Zoysia japonica seeds, they are cultivated on a solid MS medium to a height of 3 cm above the ground. At this time, rou...
Embodiment 2
[0056] Example 2 Screening of seed culture methods
[0057] Soak the sterilized Japanese Zoysia japonica seeds in water at 4°C for 3 days (72 hours), blow-dry the seeds until the surface becomes white, and then set them to inoculate directly on MS medium and cultivate them with water that has just submerged the seeds. In both methods, the culture temperature is room temperature culture.
[0058] According to observations, if the seeds are soaked in water, a few seeds will germinate within 2 days, and most of the seeds will germinate on the third day. For seeds directly sown in 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] Example 3 Seeding density screening
[0060] Configure the solid MS medium and distribute it into tissue culture bottles with a diameter of 8cm, each bottle is about 30ml.
[0061] Blow-dry the seeds after 72 hours of soaking at 4℃ until the surface becomes white, and cultivate with the water that has just submerged the seeds to germinate 1-2mm buds, according to 10, 20, 30, 40, 50 per bottle The seeding density was inoculated into the above-mentioned tissue culture bottle containing solid MS medium and cultured for 15 days. After observation, it was not found that the seeding density had a significant effect on the growth of Zoysia japonica.
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