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Cryopreservation method for plant germplasm of whortleberry

A cryopreservation and plant technology, which is applied in the field of plant germplasm resource preservation, can solve the problems of different procedures and technologies, and achieve the effects of strong cell regeneration ability, convenient material extraction, and high genetic stability

Active Publication Date: 2017-06-13
JILIN AGRICULTURAL UNIV
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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 species of the same species, the appropriate cryopreservation methods and procedures may be different.
At present, there is no report on the research on the preservation of lingonberry germplasm resources by cryopreservation technology

Method used

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  • Cryopreservation method for plant germplasm of whortleberry
  • Cryopreservation method for plant germplasm of whortleberry
  • Cryopreservation method for plant germplasm of whortleberry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Cryopreservation technology of lingonberry stem tip droplet vitrification method and plant regeneration culture method

[0031] The specific operation of the experimental method is as follows:

[0032] 1. Seedling training: Select 4-week-old tissue-cultured seedlings, take 0.5 cm stem segments with terminal buds and inoculate them on WPM medium, and treat them in the dark at 4°C for 3 weeks;

[0033] 2. Bilberry stem tip stripping: Under sterile conditions, the terminal buds after hardening are stripped under a stereo microscope, and the stem tip length is 1.5-2.0mm;

[0034] 3. Pre-cultivation: Inoculate the stripped shoot tips on WPM solid culture with 0.3M sucrose added, and culture in the dark for 24 hours in the tissue culture room;

[0035] 4. Loading: Transfer the pre-cultured isolated shoot tips to the loading solution consisting of WPM+2M glycerol+1.0M sucrose, and load at room temperature for 30 minutes;

[0036] 5. Vitrification treatment: transfe...

Embodiment 2

[0041] Example 2 Effects of Seedling Hardening Time and Loading Time on Cryopreservation of Bilberry Stem Tips

[0042] The operation steps are the same as in Example 1, only the seedling hardening time is changed, the seedling hardening time is set to 0, 1, 2, 3, 4, 5 weeks, and the loading time is 0, 10, 20, 30, 40 min. The survival rate of the lingonberry shoot tip was counted after 7 days of recovery culture and the regeneration situation was counted after 30 days. The results showed that hardening time and loading time had important effects on the survival rate and regeneration rate of lingonberry stem tips after preservation. The survival rate and regeneration rate of the shoot tip showed a trend of rising first and then falling with the change of hardening time, as shown in figure 1 As shown, with the change of loading time, there is basically a trend of rising first and then falling, as shown in figure 2 shown. When the hardening time was 3 weeks, the survival and ...

Embodiment 3

[0043] Example 3 Effects of pre-cultivation time and vitrification treatment time on cryopreservation of lingonberry shoot tips

[0044] The operation steps are the same as in Example 1, only the pre-cultivation time is changed, inoculate the peeled lingonberry stem tip into the WPM solid medium containing 0.3 M sucrose, pre-cultivate in the dark in the tissue culture room for 0-48h, and set the vitrification time 0, 20, 40, 60, 80 minutes. The survival rate of the lingonberry shoot tip was counted after 7 days of recovery culture and the regeneration situation was counted after 30 days. The results showed that the pre-cultivation time and vitrification treatment time had important effects on the survival rate and regeneration rate of lingonberry stem tips after preservation. The survival rate and regeneration rate of the shoot tip basically showed a trend of increasing first and then decreasing with the change of pre-culture time, as shown in image 3 As shown, with the cha...

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Abstract

The invention discloses a droplet vitrification cryopreservation method for stem tip of whortleberry, which is a simple and effective method for saving high-quality germplasm resources of whortleberry with long-term security, stability, and reliability. The method in the invention takes terminal bud stem tip as the cryopreservation material, which, on one hand, is convenient in obtaining materials, and has high hereditary stability on the other hand because the stem tip contains stem tip meristem with strong cell regeneration capacity; after cryopreservation, the survival rate of the stem tip of the whortleberry reaches 100%, and the rate of regeneration of the stem tip is 91.7%.

Description

technical field [0001] The invention relates to the field of preservation of plant germplasm resources, in particular to a method for cryopreservation of lingonberry plant germplasm and a method for restoring and cultivating the cryopreservation bilberry plant germplasm. Background technique [0002] Lingonberry belongs to the Vaccinium spp. plant of the Rhododendron family (Ericaceae), and is a perennial deciduous or evergreen shrub or small shrub species. There are about 400 species of Vaccinium plants in the world, and there are about 91 species and 28 varieties in my country, which are distributed in the northeast and southwest of China. The fruit is rich in antioxidant substances such as anthocyanins, which have health functions such as improving eyesight, anti-oxidation, anti-cancer and delaying brain aging. ". [0003] The preservation of germplasm resources is an important guarantee for lingonberry breeding and maintenance of genetic diversity. Breeding new varieti...

Claims

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Application Information

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IPC IPC(8): A01N3/00A01H4/00
CPCA01H4/005A01N3/00
Inventor 张志东孙海悦刘海广唐雪东李亚东汪立宇
Owner JILIN AGRICULTURAL UNIV
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