Method for preserving and purifying smallanthus sonchifolius germplasms based on ultra-low temperature technology
A purification method, the technology of yacon, applied in the field of agricultural biology, can solve the problems of difficult control of natural environmental diseases and insect pests, infected virus species, loss of germplasm resources, etc., and achieve the effect of good plant growth, rapid growth and long storage time
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Embodiment 1
[0039] A method for preserving and purifying yacon fruit germplasm based on ultra-low temperature technology, the technical scheme of the method is composed of the following steps:
[0040] S1 obtains yacon clonal propagation line
[0041] In 2013, using the yacon seedlings provided by Hunan Qidong Alpine Yacon R&D Base as the test material, the axillary buds of the yacon seedlings were cut and soaked in 0.1% washing powder water for 15-20 minutes, then rinsed with tap water, and the axillary buds were peeled off. After reaching the growth point, first disinfect the surface with 70% alcohol for 30s, then rinse with sterile water for 2-3 times, then place it in 0.1% mercuric chloride with a few drops of Tween 20 for 10min, and then rinse with sterile water for 4 -5 times, the filter paper absorbs the water, and then inoculates on the MS medium supplemented with KT 2.0 mg / L, IAA 0.5 mg / L, hydrolyzed protein 35 mg / L, and polyvinyl alcohol 0.2 mg / L to induce adventitious buds; Th...
Embodiment 2
[0055] Test the influence of the pre-medium of the present invention on the cryopreservation effect of the yacon stem tip: only change the pre-medium in Step S1 of Example 1 (the remaining 2 pre-mediums are commonly used pre-mediums in cryopreservation), other steps Similar to Example 1, the shoot apex survival rate and regeneration rate were counted, and the results are shown in Table 1 below.
[0056]
[0057] The above results show that using the pre-medium of the present invention, the survival rate and regeneration rate of yacon stem tips are the highest after ultra-low temperature preservation for 3 years; the pre-medium of the present invention reduces nutrients and adds butyrate hydrazide to reduce the yacon stem tips. the growth rate of the plant; the addition of pomegranate seed powder, betaine, salicylic and acid brassinolide can improve the stress resistance of the shoot tip and enhance its tolerance to low temperature; the addition of high concentration of sucro...
Embodiment 3
[0059] The impact of the cryoprotective solution of the present invention on the ultra-low temperature preservation effect of the yacon stem tip: only change the cryoprotective solution in step S4 of Example 1 (the remaining 2 kinds of cryoprotective solutions are commonly used in ultra-low temperature preservation), and other steps are the same as Example 1 is the same, and the plant regeneration rate is counted, and the results are shown in Table 2 below.
[0060]
[0061] The above results show that, using the cryoprotective solution of the present invention, the survival rate and the regeneration rate of the yacon stem tips after ultra-low temperature preservation for 3 years are the highest; the cryoprotective solution of the present invention additionally adds trehalose, L-carnitine, glutathione, helix Algal flour helps to maintain the integrity of cell structure, and water-soluble carbon nanotubes can change the properties of cryoprotective solution and reduce the mec...
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