Cryopreservation method for in vitro shoot tips of petunia hybrida
A technology for cryopreservation and petunia, which is applied in the field of cryopreservation of plant germplasm resources, can solve the problems of different procedures and technologies, and the cryopreservation method has not yet been established, and achieves the effect of restoring good growth.
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Embodiment 1
[0025] Example 1 Petunia test-tube plantlet isolated stem tip droplet vitrification cryopreservation technology and plant regeneration culture method
[0026] Experimental materials: Petunia (Petunia×hybrida Vilm.) "Panmei Dream" series of 4 varieties of test tube seedlings: cow 1 (white), cow 2 (blue), cow 3 (red), cow 4 (rose red) .
[0027] The specific steps of the experimental method are as follows:
[0028] 1. Material proliferation
[0029] 1) Inoculate petunia test-tube plantlets on MS solid medium (containing 9.0g / L agar) containing 0.5mg / L BA and 30g / L sucrose for propagation;
[0030] 2) Transfer the multiplied test-tube plantlets to the hormone-free MS medium for rooting and rejuvenation. After 20 days, transfer the rooted plantlets to MS medium containing 30g / L sucrose for cultivation; the culture temperature is 25±2°C, and the light 14h / d, the light intensity is 700lux.
[0031] 2. Stem tip stripping
[0032] Under aseptic conditions, the isolated shoot tip ...
Embodiment 2
[0046] Example 2 Effect of sucrose concentration in preculture solution on cryopreservation of petunia shoot tips
[0047] Only the sucrose concentration in the pre-culture solution was changed, and the rest of the steps were the same as in Example 1 to study the effect of the sucrose concentration in the pre-culture solution on petunia shoot tip cryopreservation. The result is as image 3 shown.
[0048] From image 3 It can be seen that the survival rate and regeneration rate of petunia shoot tips are more sensitive to the change of sucrose concentration in the preculture solution after cryopreservation by droplet vitrification method. With the increase of the sucrose concentration in the pre-culture solution, the shoot tip survival rate and regeneration rate showed a trend of rising first and then decreasing. Too high or too low concentration of sucrose may lead to a decline in the survival rate and regeneration rate, which contains 0.2~0.3mol / L sucrose pre-culture solu...
Embodiment 3
[0049] The influence of embodiment 3 precultivation time on petunia shoot tip cryopreservation
[0050] Only the pre-cultivation time was changed, and the rest of the steps were the same as in Example 1 to study the effect of the pre-culture time on petunia shoot tip cryopreservation. The result is as Figure 4 shown.
[0051] From Figure 4 It can be seen that the survival rate and regeneration rate of petunia shoot tips are more sensitive to the change of preculture time after cryopreservation by droplet vitrification method. The shoot tip survival rate and regeneration rate increased slowly when the pre-culture time was extended from 0d to 2 days, and the shoot tip survival rate and regeneration rate were the highest when the pre-culture time was 2-3 days, reaching more than 80% and 70%. The shoot tip survival rate and regeneration rate decreased after 3~5 days of pre-culture.
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