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

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

Example Embodiment

[0030] Example 1 Bilberry stem tip droplet vitrification method ultra-low temperature preservation technology and plant regeneration culture method

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

[0032] 1. Seedling training: select tissue culture seedlings of 4 weeks old, and inoculate 0.5 cm stems with apical buds on WPM medium at 4℃ for 3 weeks in the dark;

[0033] 2. Extraction of bilberry stem tip: Under aseptic conditions, peel off the top buds after refining seedlings under a stereo microscope, the stem tip length is 1.5-2.0mm;

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

[0035] 4. Loading: Transfer the pre-cultured stem apex into a loading solution consisting of WPM+2M glycerol+1.0M sucrose, and load it for 30min at room temperature;

[0036] 5. Vitrification treatment: The stem apex after the loading treatment is t...

Example Embodiment

[0041] Example 2 Effect of seedling tempering time and loading time on ultra-low temperature storage of bilberry stem tips

[0042] The operation steps are the same as in Example 1, only changing the cultivation time, setting the cultivation time to 0, 1, 2, 3, 4, 5 weeks, and the loading time to 0, 10, 20, 30, 40 min. The survival rate of the stem tip of the bilberry and the regeneration of the bilberry were counted after 7 days of recovery. The results showed that the cultivation time and loading time had an important influence on the survival rate and regeneration rate of bilberry shoot tips after preservation. The survival rate and regeneration rate of shoot tips show a trend of rising first and then falling with the change of seedling time, such as figure 1 As shown, as the loading time changes, it basically rises first and then falls, such as figure 2 Shown. When the seedling time is 3 weeks, the survival and regeneration effect of the shoot tip is better, and when the lo...

Example Embodiment

[0043] Example 3 The effect of pre-culture time and vitrification time on cryopreservation of bilberry stem tips

[0044] The procedure is the same as in Example 1, except that the pre-culture time is changed, the peeled bilberry stem tips are inoculated into WPM solid medium containing 0.3 M sucrose, and pre-cultured in the tissue culture room for 0-48 hours in the dark, and the vitrification time is set It is 0, 20, 40, 60, 80min. The survival rate of the stem tip of the bilberry and the regeneration of the bilberry were counted after 7 days of recovery. The results showed that the pre-cultivation time and vitrification time had an important influence on the survival rate and regeneration rate of bilberry shoot tips after preservation. The survival rate and regeneration rate of shoot tips basically increased first and then decreased with the change of pre-culture time, such as image 3 As shown, with the change of the vitrification time, it first rises, then falls, and then ri...

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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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