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.

Active Publication Date: 2012-10-17
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A variety of cryopreservation methods have been developed. However, for different species or even for different cultivars of the same species, the appropriate cryopreservation methods and procedures may be different. So far, no universal cryopreservation method has been established internationally. cryopreservation method, which is also a major bottleneck in the practical application of cryopreservation technology

Method used

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  • Cryopreservation method for in vitro shoot tips of petunia hybrida
  • Cryopreservation method for in vitro shoot tips of petunia hybrida
  • Cryopreservation method for in vitro shoot tips of petunia hybrida

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The present invention provides a cryopreservation method for in vitro shoot tips of petunia hybrida. The method, mainly based on droplet vitrification cryopreservation technology, includes the following steps of: expanding propagation of materials, stripping of shoot tips, performing pre-culture, loading, performing PVS2 processing and performing cryopreservation. Plant regeneration culture can be performed on the in vitro shoot tips of the petunia hybrida obtained by adopting the droplet vitrification cryopreservation technology in the invention, which indicates that the method in the invention is a method that can preserve resources of the species of the petunia hybrida with long-term security, stability, reliability, simplicity and effectiveness, regeneration seedlings being capable ofrecovering growth favorably, plant regeneration rate of the plant achieving more than 50%.

Description

technical field [0001] The invention relates to a method for cryopreservation of plant germplasm resources, in particular to a method for cryopreservation of isolated petunia shoot tips. Background technique [0002] Petunia (Petunia×hybrida Vilm.), aliased as Ganoderma lucidum peony, petunia, Zhuangyu Chaoyan, etc., is a flower of the genus Petunia in the family Solanaceae. Petunia is a perennial herbaceous dicotyledonous plant, often cultivated as one or two years, with a plant height ranging from 15 to 80 cm; the leaves are oval or oval; the vegetative growth period is short, and it can bloom in the same year, and the flowering period can last for several months The corolla is trumpet-shaped (similar to morning glory); petal edge types include flat petals, wavy petals, serrated petals, etc.; flower colors include purple, red, white, pink, inlaid, reticulated, striped, etc.; fruit is a pointed round capsule , the seeds are extremely small, and the thousand-grain weight is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00A01N3/00
Inventor 陈晓玲黄斌张金梅卢新雄辛霞
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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